Methods for Treating Skin Disorders

Abstract

The present invention relates to treating skin disorders in a subject in need thereof by treating a skin disorder comprising administering to a subject a compound of formula (I): The present invention relates to a method of treating a human by administering to the human an effective dose of a compound represented by JPEG2024504410000022.jpg32141.

Description

[Technical field]

[0001] TECHNICAL FIELD TO WHICH THEINVENTION BELONGS The present invention relates to the field of drug therapy, in particular to the treatment of skin disorders with compounds capable of inhibiting the ion channel exchangers ENaC, NHE and NCX. [Background technology]

[0002] background Skin is a semi-permeable barrier that is important in maintaining homeostasis and preventing excess water loss, microbial attack, thermoregulation, and sensation. To maintain these functions, skin needs to tightly regulate its functions, including proliferation, differentiation, and epidermal immune interactions. Ion channel flux is important in regulating these functions, and cations such as sodium and calcium are tightly regulated both extracellularly and intracellularly by ion channel exchangers. Changes in intracellular Na and membrane potential can modulate the activity of the basolateral Na / H and Na / Ca exchangers that affect this crosstalk (Harvey et al., 1995). This means that changes in one of these channels can lead to compensatory mechanisms in the other channels. An example of this is when regulation of all individual conductances does not need to be done in the right direction in a homeostatic sense if a sufficient subset of the remaining conductances is appropriately regulated (O'Leary et al., 2013).

[0003] Although the regulation of these channels is complex, it is widely accepted that in many different electrically excitable cells, voltage-gated calcium channels provide an indirect measure of membrane voltage changes, and the subsequent influx of calcium ions can cause changes in ion channel expression. Pharmacological demonstration of this phenomenon is that treatment with the sodium channel blocker mexiletine results in upregulation of sodium channel transcription and expression, and treatment with the calcium channel blocker verapamil produces effects similar to those seen with sodium channel blockade, suggesting that the sodium channel blockade response is mediated by changes in calcium ion flux (Rosati et al, 2004).

[0004] Another example is that sodium overload can lead to NCX hyperactivity, whereas sodium depletion can promote NCX1-mediated responses and proinflammatory cytokine release from immune cells ( Harvey et al., 1995 ).

[0005] WO9009792 suggests amiloride and some of its derivatives for the treatment of inflammatory skin and eye disorders.

[0006] WO2015168574 suggests the use of epithelial ion channel (ENaC) blockers, such as benzamil, for the treatment of psoriasis.

[0007] WO2020150606 suggests a method for treating an immune dysregulation disease in a subject, the method comprising, for example, administering a therapeutically effective amount of benzamil to the subject. The immune dysregulation disease may be an inflammatory disease, and the inflammatory disease may be a dermatological disorder. Summary of the Invention

[0008] overview The present invention describes the use of certain amiloride derivatives that have a directionality that mimics benzamil and show a complex off-target profile with activity against all three ion channels ENaC, NHE1 and NCX1.Compared with certain ENaC inhibitors that have reduced or no activity against NHE and NCX, such amiloride derivatives have distinct and different effects on skin inflammation.

[0009] The present invention utilizes the combined inhibition of multiple ion channels (ENaC, NHE and NCX) by amiloride derived drugs to allow restoration of abnormal signaling by cytokines involved in skin inflammation where the expression or activity of some of these channels is dysregulated, making these drugs usable as treatments for a variety of skin disorders. Amiloride derivatives useful in the present invention have the formula (I): [ka] [In the formula, R is -C(CH3)CH2C(CH3)3; [ka] Select from and pharma- ceutically acceptable salts thereof.

[0010] According to a first aspect, a skin disorder in a subject in need thereof is treated with an anti-inflammatory drug comprising a compound represented by formula (I): [ka] [In the formula, R is -C(CH3)CH2C(CH3)3; [ka] Select from and pharma- ceutical acceptable salts thereof, comprising administering to a human an effective dose of a compound represented by the formula: A method is provided wherein when the compound is phenamil or benzamil, the skin disorder is not psoriasis.

[0011] The skin disorder may involve dysregulation of CXCL1, TNFα and / or IL1α (e.g., the presence of abnormal levels of CXCL1, TNFα and / or IL1α) and / or may involve the pathogenesis of, and / or may involve the pathology of, acne, atopic dermatitis, seborrheic dermatitis, nummular dermatitis, perioral dermatitis, rosacea, vitiligo, hidradenitis suppurativa, lupus, morphea, scleroderma, skin ulcers, inflammatory seborrheic keratosis, nevi, fibrous papules, pityriasis rubra pilaris, pemphigus vulgaris and pemphigus vegetans, bullous pemphigoid, pemphigus including IgA pemphigus, follicular Hyperkeratosis, lamellar ichthyosis, epidermolytic ichthyosis, Netherton syndrome, congenital ichthyosiform erythroderma, cutaneous vasculitis, Behçet's disease, cutaneous graft-versus-host disease, SAPHO (synovitis, acne, pustulosis, hyperostosis and osteitis) syndrome, sarcoidosis, panniculitis, Stevens-Johnson syndrome, toxic epidermal necrolysis, neutrophilic dermatosis, pyoderma gangrenosum, Sweet's syndrome, subcorneal pustulosis, acute generalized exanthematous pustulosis (AGEP), dermatitis herpetiformis, dermatomyositis, cryoporin-associated periodic fever syndrome periodic syndromes), familial Mediterranean fever, xeroderma pigmentosum, pruritus, pain, ultraviolet light-induced skin damage, irritant contact dermatitis, Still's disease, lichen planus, eczema, wounds, lesions, ulcers, alopecia, and inflammatory skin neoplasms including actinic keratosis, squamous cell carcinoma in situ, squamous cell carcinoma, basal cell carcinoma, extramammary Paget's disease, dermatofibrosarcoma protuberans, atypical fibroxanthomas, sebaceous gland carcinoma; and Merkel cell carcinoma.

[0012] The compounds represented by formula (I) may be administered locally or systemically.

[0013] The compounds may be administered by injection or orally.

[0014] The subject to be treated can be a mammalian subject, such as a human or an animal.

[0015] When the compound is in the form of a pharma- ceutically acceptable salt, the salt may be selected from lactate, acetate, and phosphate salts.

[0016] In some embodiments, the compound is administered in an amount effective to achieve a serum concentration in the subject of 5-50 ng / mL.

[0017] According to a second aspect, there is provided a compound of formula (I): [ka] [In the formula, R is -C(CH3)CH2C(CH3)3; [ka] Select from and pharma- ceutically acceptable salts thereof.

[0018] According to a third aspect, in the manufacture of a pharmaceutical composition for use in the above method of treatment, a compound of formula (I): [ka] [In the formula, R is -C(CH3)CH2C(CH3)3; [ka] Select from The present invention provides the use of a compound represented by the formula: [Brief description of the drawings]

[0019] [Figure 1a] FIG. 1a shows that ENaC and NCX1 are upregulated and NHE is downregulated in patients with acne compared to non-acne controls. [Figure 1b] FIG. 1b shows that CXCL1 and IL1α are upregulated in patients with acne. [Figure 2a] FIG. 2a shows that NCX1 is upregulated and ENA C is downregulated in patients with atopic dermatitis compared to controls without atopic dermatitis. [Figure 2b] FIG. 2b shows that CXCL1 and TNFα are upregulated in patients with eczema. [Diagram 3] FIG. 3 shows that benzamil reduces the expression of CXCL1 in stimulated keratinocytes. [Figure 4] FIG. 4 shows that benzamil reduces the expression of IL1α in stimulated keratinocytes. [Diagram 5] FIG. 5 shows that benzamil reduces the expression of TNFα in stimulated keratinocytes. [Figure 6] FIG. 6 shows that benzamil does not inhibit proliferation of keratinocytes under low levels of inflammation. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0020] Detailed Description Benzamil, an amiloride derivative, has been proposed for use in the treatment of psoriasis (WO2015168574, A1) and cystic fibrosis (Hirsh et al., 2004). Benzamil is a more potent ENaC inhibitor than amiloride in vitro, but the in vivo potency of benzamil and phenamil specifically against ENaC has previously been shown to be comparable to that of amiloride (Hirsh et al., 2004). Thus, if ENaC is the specific target underlying the effect on keratinocyte proliferation, higher doses of amiloride would be expected to confer partial or similar effects.

[0021] Loss of ENaC, NCX1 or NHE individually does not alter intracellular ion channel signaling to the same extent as inhibition of multiple channels (Rosati et al., 2004). It has been demonstrated that differences in expression of NCX1 (Staiano et al., 2009), NHE1 and ENaC correlate with activation, and they may complement each other.

[0022] The inventors discovered that there is a distinct off-target profile of benzamil compared with amiloride or specific sodium channel inhibitors that allows for the therapeutic efficacy of benzamil, inhibiting pro-inflammatory cytokine production in skin cells that underlies inflammation in inflammatory skin diseases through combined inhibition of ENaC, NCX1 and NHE.

[0023] Thus, the present invention relates to a method of treatment using a compound sharing the property of combined inhibition of ENaC, NCX1 and NHE by benzamil for the treatment of conditions caused or complicated by proinflammatory cytokine production in skin cells, wherein when the compound is phenamil or benzamil, the skin disorder is not psoriasis. In some embodiments, when the compound is benzamil, the skin disorder is not atopic dermatitis, alopecia areata, bullous pemphigus, chronic eczema, dermatomyositis, erythema nodosum, epidermolysis bullosa, hydradenitis suppurativa, lichen planus, pemphigus vulgaris, pyoderma gangrenosum, scleroderma, or vitiligo.

[0024] The results of the experiments disclosed herein demonstrate that a compound with a combined inhibitory effect on all three ion channels, ENaC, NCX1 and NHE, exemplified by benzamil, is sufficient and necessary to prevent pro-inflammatory signaling through the inflammatory cytokines CXCL1, TNFα and IL1α.

[0025] Many amiloride derivatives having a combined inhibitory effect on all three ion channels ENaC, NCX1 and NHE are known in the art, including those represented by formula (I): [ka] [In the formula, R is -C(CH3)CH2C(CH3)3; [ka] Select from Examples of compounds include those represented by the formula: (Kleyman et al., 1988).

[0026] The particular amiloride derivatives disclosed above (also referred to herein as "compounds of the invention") have a similar profile of targeting all three ion channels ENaC, NHE and NCX1 and therefore have therapeutic potential in the treatment of skin disorders combining inflammation and abnormal keratinocyte proliferation, as described herein.

[0027] The proinflammatory cytokines CXCL1, TNFα and / or IL1a have been demonstrated to underlie the development of skin disorders in which aberrant epithelial signaling leads to an inflammatory immune response. These conditions include acne (Li, 2006), perioral dermatitis (Farley, SM, et al., 2006), rosacea (Li, N., et al., 2014), vitiligo (Jimbo, H., et al., 2020), hidradenitis suppurativa (Gupta and Skinner, 2004), lupus (Norman, R., et al., 2006), Morphea (Fett, N., 2013), scleroderma, skin ulcers, inflammatory seborrheic keratosis, nevi, fibrous papules, pityriasis pilaris (Muller, H., et al., 2008), pemphigus vulgaris and proliferative pemphigus (Tavakolpour, S., et al., 2020), bullous pemphigoid (Tabatabaei-Panah, PS, et al., 2019), pemphigus including IgA pemphigus (Howell, SM, et al., 2005), follicular keratosis (Mayuzumi, N., et al., 2005), and foliar ichthyosis (Malik, K., et al., 2019), epidermolytic ichthyosis (Malik, K., et al., 2019), Netherton syndrome (Malik, K., et al., 2019), congenital ichthyosiform erythroderma (Malik, K., et al., 2019), and cutaneous vasculitis (Carbone, F. and F.Montecucco, 2015), Behçet's disease (van der Houwen and van Laar, 2020), graft-versus-host disease (Gupta and Skinner, 2004), SAPHO (synovitis, acne, pustulosis, hyperostosis, and osteitis) syndrome (Gupta and Skinner, 2004), sarcoidosis (Gupta and Skinner, 2004), panniculitis (Gupta and Skinner, 2004), Stevens-Johnson syndrome (Wang, C. W., et al., 2018), toxic epidermal necrolysis (Gupta and Skinner, 2004), neutrophilic dermatosis (Ahn, C. et al., 2018), pyoderma gangrenosum (Ahn, C. et al., 2018), Sweet syndrome (Gupta and Skinner, 2004), subcorneal pustulosis (Gupta and Skinner, 2004), acute generalized exanthematous pustulosis (AGEP) (Feldmeyer, L., et al., 2016), dermatitis herpetiformis (Amerio, P., et al., 2000), dermatomyositis (Alexis and Strober, 2005), cryopyrin-associated periodic syndromes (Luo, X. Y., et al., 2020), familial Mediterranean fever (Samuels and Ozen, 2006), xeroderma pigmentosum (Kunisada, M., et al., 2017), pruritus (Deftu, A. F., et al., 2018), pain (Deftu, A. F., et al., 2018), skin damage caused by ultraviolet rays (Fukunaga, A., et al., 2021), irritant contact dermatitis (Funch, A. B., et al., 2021), Still's disease (Kadavath, S. and P. Efthimiou, 2015), lichen planus (Wu, T., et al., 2013), eczema (Farley, S. M., et al., 2006), wound (Zheng, R., et al., 2019), lesion, ulcer, alopecia (Loh, S. H., et al., 2018), as well as cutaneous neoplasms with inflammation including actinic keratosis (Maru, GB, et al., 2014), squamous cell carcinoma in situ (Maru, GB, et al., 2014), squamous cell carcinoma, basal cell carcinoma (Maru, GB, et al., 2014), extramammary Paget's disease, dermatofibrosarcoma protuberans, atypical fibroxanthoma, sebaceous carcinoma; and Merkel cell carcinoma.

[0028] The present amiloride derivatives activate and / or alter the immune response, thereby affecting the carcinomas listed above.

[0029] Skin disorders in which CXCL1 is dysregulated include acne, xeroderma pigmentosum, pruritus, pain, irritant contact dermatitis, lichen planus, and atopic dermatitis.

[0030] Skin disorders in which TNFα is dysregulated include pityriasis rubra pilaris, pemphigus vulgaris and pemphigus vegetans, bullous pemphigoid, pemphigus including IgA pemphigus, follicular keratosis, lamellar ichthyosis, epidermolytic ichthyosis, Netherton syndrome, congenital ichthyosa-like erythroderma, cutaneous vasculitis, Behçet's disease, cutaneous graft-versus-host disease, SAPHO (synovitis, acne, pustulosis, hyperostosis and osteitis) syndrome, sarcoidosis, panniculitis, Stevens-Johnson syndrome, toxic epidermal necrolysis, neutrophilic dermatosis, pyoderma gangrenosum, Sweet's syndrome, subcorneal pustulosis, AGEP, dermatitis herpetiformis, scleroderma, morphea, dermatomyositis, cryoporin-associated periodic fever syndrome, and erythroderma. These conditions include cutaneous dermatosis, familial Mediterranean fever, ultraviolet light-induced dermatosis, Still's disease, hidradenitis suppurativa, alopecia, vitiligo, basal cell carcinoma, actinic keratosis, and squamous cell carcinoma including squamous cell carcinoma in situ (Bowen's disease).

[0031] Skin disorders in which IL1α is dysregulated include bullous pemphigoid, follicular keratosis, cutaneous vasculitis, cryoporin-associated periodic syndrome, familial Mediterranean fever, UV-induced skin damage, Still's disease, lichen planus, wounds, seborrheic dermatitis, and alopecia.

[0032] A composition comprising an effective dose of benzamil and a compound or salt having the same combined inhibitory effect on all of ENaC, NCX1 and NHE, and optionally combined with additional therapeutic agents, can be provided to an individual suffering from any of the above-listed conditions. This administration can be preferably oral or topical, etc. In some embodiments, topical administration is preferred. The dosage and periodicity of administration are selected to provide therapeutic efficacy.

[0033] Thus, the present invention relates to and uses of pharma-ceutical compositions comprising a therapeutically effective amount of at least one compound according to the present invention, optionally in the form of a pharma-ceutical acceptable salt, formulated together with one or more pharma-ceutical acceptable excipients, optionally combined with one or more additional agents for the treatment of related skin disorders. The active ingredient and excipient(s) may be formulated into compositions and dosage forms according to methods known in the art. The pharmaceutical compositions of the present invention may be formulated for administration in solid, liquid, or semi-liquid form, including those adapted for oral administration (e.g., tablets, capsules, powders, granules, pastes for application to the tongue, aqueous or non-aqueous solutions or suspensions, drenches, or syrups); or topical application (e.g., as lotions, creams, ointments, sprays, patches, microneedle arrays, etc. applied to the skin). The medicaments, pharmaceutical compositions, or therapeutic combinations according to the present invention may be in any form suitable for application to humans and / or animals, preferably humans, including infants, children, and adults, and may be manufactured by standard procedures known to those skilled in the art.The medicament, (pharmaceutical) composition or therapeutic combination can be manufactured by standard procedures known to those skilled in the art, e.g., "Pharmaceutics: The Science of Dosage Forms", 2nd Edition, Aulton, ME (ED. Churchill Livingstone, Edinburgh (2002); "Encyclopedia of Pharmaceutical Technology", 2nd Edition, Swarbrick, J. and Boylan JC (eds.), Marcel Dekker, Inc. New York (2002); "Modern Pharmaceutics", 4th Edition, Banker GS and Rhodes CT (eds.) Marcel Dekker, Inc. New York 2002, "The Theory and Practice of Industrial Pharmacy", Lachman L., Lieberman H. And Kanig J. (Eds.), Lea & Febiger, Philadelphia (1986).

[0034] The effective dose of benzamil and the compound or salt with similar combined inhibitory effect of ENaC, NCX1 and NHE may include "therapeutically effective dose or amount" or "prophylactically effective dose or amount". "Therapeutically effective dose / amount" means an amount effective at the dosage and time period necessary to achieve the desired therapeutic result. The therapeutically effective amount may vary depending on factors such as the individual's condition, age, sex, and weight, and the ability to elicit a desired response in the individual. The therapeutically effective dose / amount is also an amount in which any toxic or harmful effects are outweighed by the therapeutically beneficial effects. "Prophylactically effective dose / amount" means an amount effective at the dosage and time period necessary to achieve the desired prophylactic result. Generally, the prophylactically effective amount will be less than the therapeutically effective amount, since the prophylactic dose is used in subjects before or at an early stage of the disease.

[0035] In animal studies by the inventors, a serum concentration of about 20 nanograms / mL (62.5 nM) has been found to be therapeutically effective. Thus, in one embodiment, the compounds of the invention are provided to a subject at a dose sufficient to achieve a serum concentration of about 20 ng / mL, for example, 5-50 ng / mL. Currently, it is estimated that a serum concentration of 5-50 ng / mL of the compounds of the invention corresponds to a dose of about 0.1-20 mg administered to a human subject. By way of comparison, oral administration of the sodium channel inhibitor amiloride to humans requires 10 mg for a serum concentration of 20.6 ng / mL in adults (Jones et al., 1997). The dose administered depends on the route of administration, the age and weight of the subject, and the bioavailability of the active ingredient administered, which may be affected by the dosage form used, as known in the art (Adajare, 2020). Methods for assessing the bioavailability of topical drugs are known in the art, for example, (Herkenne et al., 2008).

[0036] Dosage regimen can be adjusted to provide the optimum desired response (e.g., therapeutic or prophylactic response).For example, a single dose can be administered, several divided doses can be administered over time, or the dose can be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation.This dose can be administered to the subject when the symptoms of skin disease appear or before the onset of symptoms.

[0037] It should be noted that dosage values ​​may vary depending on the type and severity of the condition to be alleviated.Furthermore, it should be understood that for any particular subject, specific dosage regimens must be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the composition, and that dosage ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed compositions.

[0038] As used herein, the term "dose" refers to the amount of a substance administered to a subject, e.g., milligrams (mg). In one embodiment, the dose is a fixed dose, e.g., not dependent on the body weight of the subject to whom the substance is administered. In another embodiment, the dose is not a fixed dose, e.g., dependent on the body weight of the subject to whom the substance is administered, or, in the case of topical therapy, the dose can be related to the surface area to be treated (e.g., 1 m of skin). 2 (dose per serving).

[0039] Exemplary doses for use in treating adult humans, e.g., flat doses, can include about 0.01 mg, about 0.05 mg, about 0.1 mg, about 0.5 mg, about 1 mg, about 5 mg, about 10 mg, about 20 mg, about 50 mg, about 100 mg, about 500 mg, or more.

[0040] Exemplary doses, such as those for topical use to treat adults with the methods of the invention, include those for a surface area of ​​1 m 2 Approximately 0.01 mg per 1 m surface area 2 Approximately 0.05 mg per 1 m surface area 2 Approximately 0.1 mg per 1 m surface area 2 Approximately 0.5 mg per 1 m surface area 2 Approximately 1 mg per square meter, surface area 1 m 2 Approximately 5 mg per square meter, surface area 1 m 2 Approximately 10 mg per square meter, surface area 1 m 2 Approximately 20 mg per square meter, surface area 1 m 2 Approximately 50 mg per square meter, surface area 1 m 2 Approximately 100 mg per square meter, surface area 1 m 2 Approximately 500 mg or more per serving.

[0041] Ranges intermediate to the above-listed ranges are also contemplated, for example ranges having any one of these values ​​as an upper or lower limit, such as about 0.01 mg to about 100 mg, about 1 mg to about 10 mg, etc. are also contemplated as part of the invention.

[0042] The administration of the composition can comprise repeated cycles of administering the composition to the subject.The periodicity of administration of the compound can be about once a week, once every two weeks, about once every 3 weeks, about once every 4 weeks, about once every 5 weeks, about once every 6 weeks, about once every 7 weeks, about once every 8 weeks, about once every 9 weeks, about once every 10 weeks, about once every 11 weeks, about once every 12 weeks, about once every 13 weeks, about once every 14 weeks, about once every 15 weeks, about once every 16 weeks, about once every 17 weeks, about once every 18 weeks, about once every 19 weeks, about once every 20 weeks, about once every 21 weeks, about once every 22 weeks, about once every 23 weeks. The periodicity may be, for example, once every 24 weeks, once every 5-10 days, once every 10-20 days, once every 10-50 days, once every 10-100 days, once every 10-200 days, once every 25-35 days, once every 20-50 days, once every 20-100 days, once every 20-200 days, once every 30-50 days, once every 30-90 days, once every 30-100 days, once every 30-200 days, once every 50-150 days, once every 50-200 days, once every 60-180 days, or once every 80-100 days. Periodicities intermediate to the above-listed times are also contemplated by the present invention. Ranges intermediate to the above-listed ranges are also contemplated by the present invention. For example, ranges having any one of these values ​​as an upper or lower limit, such as about 110 days to about 170 days, about 160 days to about 220 days, etc., are also intended to be part of the invention.

[0043] "Periodic period" refers to the time during which repeated administration cycles are performed.For example, the periodic period of administration of the substance can be up to about 4 weeks, up to about 8 weeks, up to about 12 weeks, up to about 16 weeks or more, up to about 20 weeks, up to about 24 weeks, up to about 28 weeks, up to about 32 weeks or more, during which the periodicity of administration is about once per week.For example, the periodic period can be about 6 weeks, during which the periodicity of administration is about once per 4 weeks, for example, the substance is administered at week 0 and week 4.

[0044] The compounds of the present invention can be provided in the form of pharma- ceutically acceptable salts.As used herein, "pharma- ceutically acceptable salts" refers to derivatives of the disclosed compounds, in which the parent compound is modified by making its acid salt or base salt.Examples of pharma- ceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids.Pharmaceutically acceptable salts include, for example, the conventional non-toxic salts or quaternary ammonium salts of the parent compound formed from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, and the like; and salts prepared from organic acids such as acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfanilic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, isethionic acid, and the like. Presently preferred pharma- ceutical acceptable salts are prepared from lactic acid, acetic acid, and phosphoric acid, which have been found to have improved properties with respect to at least enhanced stability, increased aqueous solubility, and / or reduced polymorphism (as described in co-pending patent application EP21199548.5). Pharmaceutically acceptable salts of the compounds of the present invention can be synthesized from parent compounds containing a basic or acidic moiety by conventional chemical methods. In general, such salts can be prepared by reacting the free acid or free base form of these compounds with a stoichiometric amount of an appropriate base or acid in water or an organic solvent, or in a mixture of the two; generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. A list of suitable salts can be found in Remington: The Science And Practice Of Pharmacy (Adajare, 2020).

[0045] All references cited herein are incorporated by reference in their entirety. The following examples are included to illustrate the invention and should not be construed as limiting the scope of the invention, as defined in the appended claims. EXAMPLES

[0046] experiment In silico experiments have found that in conditions involving skin inflammation, as exemplified in skin disorders involving abnormal cytokine signaling, rather than individual ion channels, some of the ion channels ENaC, NCX1 and NHE are dysregulated (up-regulated or down-regulated). Figure 1a shows that ENaC and NCX1 are up-regulated and NHE is down-regulated in those with acne compared to those without acne. Figure 1b shows that the cytokines CXCL1 and IL1α are up-regulated in patients with acne compared to those without acne.

[0047] Figure 2a shows that NCX1 is upregulated and ENA C is downregulated in patients with atopic dermatitis compared to controls without atopic dermatitis, and Figure 2b shows that the cytokines CXCL1 and TNFα are upregulated in patients with atopic dermatitis.

[0048] Primary keratinocytes were stimulated with TNFα (5 ng / ml) for 24 h and the expression of proinflammatory cytokines involved in inflammatory skin diseases was analyzed using benzamil as an inhibitor and compared with inhibitors with different affinities (inhibitors with affinity for ENaC / NCX1 / NHE1 (amiloride), inhibitors with affinity for ENaC but not for NCX1 and NHE1 (camostat mesylate), inhibitors with affinity for NCX1 but not for ENaC or NHE1 (KB-R7943) or inhibitors with affinity for NHE but not for ENaC or NCX1 (zoniporide)).

[0049] Primary human keratinocytes were thawed according to the manufacturer's instructions, washed with growth medium (GM), and viability assessed with Tryphan blue. Cells were incubated at 37°C and 5% CO2. After confluence, 1.5 × 10 5 A stock cell solution of 100 cells / ml was prepared in GM. 200 μL of cell suspension was added to each well of each 96-well plate, for a total of 3 × 10 4 cells / well. Cells were incubated overnight at 37°C, 5% CO2 to allow attachment. After overnight incubation, cell culture medium was removed from each well and 100 μL of fresh GM was added to each well. Appropriate concentrations of inhibitor compounds, vehicle and TNFα were prepared in a final volume of 3 ml GM. Concentrations of non-benzamil inhibitors were chosen to be 0.1-, 1- and 10-fold the IC50 of each primary target. The concentrations of TNFα were chosen to reproduce low levels of proinflammatory cytokine stimulation: 0.1, 1 and 10μm for amiloride (IC50 of ENaC 100nM), 5nm, 50nm and 500nM for camostat mesylate (IC50 of ENaC 50nM), 0.01uM, 0.1uM and 1uM for KB-R7943 (IC50 of NCX 5-10uM), 1nM, 10nM and 100nM for zoniporide (IC50 of NHE 14nM).

[0050] 100 μl of each treatment was added to the appropriate well. Samples were incubated for 24 hours. After 20 hours of TNFα stimulation, 20 μL of Alamar Blue was added to each well. Cells were incubated at 37°C for 4 hours. After 24 hours of TNFα stimulation, the fluorescence of each well was read at 544 / 590 nm for cell viability. Supernatants were collected and stored at -80°C for cytokine analysis. The inflammatory cytokines TNFα, CXCL1 and IL1a in cell culture supernatants were analyzed by Luiminex. Data was exported to Excel and processed using Graphpad Prism 9.1.

[0051] result Benzamil, unlike amiloride, camostat mesylate (extracellular ENaC inhibitors with different off-target profiles), KB-R7943 (NCX1 inhibitor) or zoniporide (NHE inhibitor), dose-dependently reduces the expression of CXCL1 (Figure 3), IL1a (Figure 4) and TNFα (Figure 5). Concentrations above 1 μM proved effective when ENaC, NCX1 and NHE1 were predicted to be targets, but were not sufficient as they completely inhibited ENaC but poorly or not NCX1 or NHE1.

[0052] Benzamil did not suppress proliferation under these conditions (Figure 6), indicating that the reduced expression of inflammatory cytokines was not due to a toxic effect. These results demonstrate that the combined inhibitory effect of benzamil on all three of ENaC, NCX1, and NHE (but not on either one) is sufficient and necessary to prevent proinflammatory signaling. CXCL1, TNFα, and IL1α have been shown to be important in the development of skin inflammation (see, e.g., Figures 1 and 2).

[0053] References JPEG2024504410000012.jpg229168JPEG2024504410000013.jpg233167JPEG2024504410000014.jpg232167JPEG2024504410000015.jpg215168

Claims

1. 2. A compound of formula (I): 【Chemistry 1】 [In the formula, R is 【Chemistry 2】 Selected from or a pharma- ceutical composition comprising a compound represented by the formula: When the compound is phenamil or benzamil, the skin disorder is not psoriasis. Pharmaceutical compositions.

2. The pharmaceutical composition of claim 1, wherein the skin disorder is selected from the group in which CXCL1, TNFα and / or IL1α are dysregulated.

3. The skin disorder may be selected from the group consisting of acne, atopic dermatitis, seborrheic dermatitis, nummular dermatitis, perioral dermatitis, rosacea, vitiligo, hidradenitis suppurativa, lupus, morphea, scleroderma, skin ulcers, inflammatory seborrheic keratosis, nevi, fibrous papules, pityriasis rubra pilaris, pemphigus vulgaris and pemphigus vegetans, bullous pemphigoid, pemphigus including IgA pemphigus, follicular keratosis, ichthyosis lamellar, epidermolytic ichthyosis, Netherton syndrome, congenital ichthyosa-like erythroderma, cutaneous vasculitis, Behcet's disease, cutaneous graft versus host disease, SAPHO (synovitis, acne, pustulosis, hyperostosis and osteitis) syndrome, sarcoidosis, panniculitis, Stevens-Johnson syndrome, toxic 3. The pharmaceutical composition of claim 1 or 2, wherein the skin disease is selected from the group consisting of epidermal necrolysis, neutrophilic dermatosis, pyoderma gangrenosum, Sweet's syndrome, subcorneal pustulosis, acute generalized exanthematous pustulosis (AGEP), dermatitis herpetiformis, dermatomyositis, cryoporin-associated periodic fever syndrome, familial Mediterranean fever, xeroderma pigmentosum, pruritus, pain, UV-induced skin damage, irritant contact dermatitis, Still's disease, lichen planus, eczema, wounds, lesions, ulcers, alopecia, and inflammatory skin neoplasms including actinic keratosis, squamous cell carcinoma in situ, squamous cell carcinoma, basal cell carcinoma, extramammary Paget's disease, dermatofibrosarcoma protuberans, atypical fibroxanthomas, sebaceous gland carcinoma; and Merkel cell carcinoma.

4. 3. The pharmaceutical composition of claim 1 or 2, wherein when the compound is benzamil, the skin disorder is not atopic dermatitis, alopecia areata, bullous pemphigus, chronic eczema, dermatomyositis, erythema nodosum, epidermolysis bullosa, hydradenitis suppurativa, lichen planus, pemphigus vulgaris, pyoderma gangrenosum, scleroderma, or vitiligo.

5. The pharmaceutical composition of claim 1 or 2, wherein the skin disorder is acne.

6. The pharmaceutical composition of claim 5 , wherein the compound is benzamil.

7. 3. The pharmaceutical composition according to claim 1 or 2, wherein the skin disorder is pityriasis rubra pilaris.

8. The pharmaceutical composition of claim 7, wherein the compound is benzamil.

9. The pharmaceutical composition of claim 1 or 2, wherein the compound represented by formula (I) is administered locally or systemically.

10. The pharmaceutical composition according to claim 1 or 2, wherein the subject is a mammalian subject, such as a human or an animal.

11. The method of claim 1 or 2, wherein a pharma- ceutically acceptable salt of the compound is administered to the subject.

12. 3. The pharmaceutical composition of claim 1 or 2, wherein the compound is administered in an amount effective to achieve a serum concentration of 5-50 ng / mL in the subject.

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