Composition for preventing or treating psoriasis comprising crenolanib or pharmaceutically acceptable salt thereof

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating psoriasis, comprising crenolanib or a pharmaceutically acceptable salt thereof. The composition according to the present invention has an effect of alleviating symptoms of psoriasis, such as erythema, epidermal hardening, and scaling, by regulating the secretion of inflammatory markers (NO, PGE2, TNF-α, IL6), allergic markers (histamine, TNF-α), and inflammatory markers of keratinocytes (RANTES, MDC, TARC). Therefore, the composition according to the present invention can be advantageously used for preventing and treating psoriasis.

Description

Composition for preventing or treating psoriasis comprising crenolanib or a pharmaceutically acceptable salt thereof

[0001] The present invention relates to a composition for preventing or treating psoriasis, and more preferably, to a composition for preventing or treating psoriasis comprising crenolanib or a pharmaceutically acceptable salt thereof.

[0002] Psoriasis is an immune-mediated autoimmune skin disease caused by chronic activation of inflammatory cell infiltration in the skin and dysregulation of epidermal keratinocytes. Typical symptoms include erythema, rash, epidermal hardening, scaling, and itching, and lesions can spread to surrounding areas. Severe cases, in particular, can lead to significant psychological problems and a decline in quality of life.

[0003] While the cause of psoriasis remains unclear, it is known that autoimmune diseases, including psoriasis, can be triggered or exacerbated by abnormal autophagy associated with ULK2. ULK2, a serine / threonine kinase, acts upstream of PIK3C3 to regulate autophagy. Furthermore, ULK1, a member of the same protein family, is known to maintain immune homeostasis in psoriatic skin when inhibited, suggesting its potential as a therapeutic target for psoriasis.

[0004] Immunomodulators such as cyclosporine are used to treat psoriasis. However, long-term use can cause nephrotoxicity and hypertension, requiring careful monitoring. Therefore, the industry needs a treatment that is safe and effective even for long-term use.

[0005] Meanwhile, Crenolanib is a drug currently undergoing clinical trials as a treatment for acute myeloid leukemia (AML) and gastrointestinal stromal tumors (GIST). It inhibits the activity of Class III receptor tyrosine kinases (RTKs), such as FLT3 (FMS-like Tyrosine Kinase 3), PDGFR α (Platelet-Derived Growth Factor Receptor), and PDGFR β. These kinases are known to be closely related to the PI3K-AKT-mTOR signaling pathway, a representative inflammation-inducing signal.

[0006] Against this backdrop, the present inventors have confirmed that crenolanib or a composition comprising the same has an inhibitory effect on immune response and autophagy activity based on its inhibitory activity against the PI3K-AKT-mTOR signal pathway, which is a basic inhibition target, and the newly identified ULK2, and have also confirmed that it has an effect of improving symptoms of psoriasis such as erythema, epidermal sclerosis, and keratinization, thereby completing the present invention. Accordingly, it is expected that the composition according to the present invention can be usefully utilized as a pharmaceutical composition for the prevention or treatment of psoriasis.

[0007] An object of the present invention is to provide a pharmaceutical composition for preventing or treating psoriasis comprising crenolanib or a pharmaceutically acceptable salt thereof.

[0008] An object of the present invention is to provide a method for preventing or treating psoriasis, comprising administering a therapeutically effective amount of crenolanib or a pharmaceutically acceptable salt thereof to a subject in need thereof.

[0009] An object of the present invention is to provide a use of the present invention's crenolanib or a pharmaceutically acceptable salt thereof for preparing a medicament for preventing or treating psoriasis.

[0010] An object of the present invention is to provide a composition comprising crenolanib or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of psoriasis.

[0011] The present invention provides a pharmaceutical composition for preventing or treating psoriasis, comprising crenolanib or a pharmaceutically acceptable salt thereof.

[0012] In the present invention, crenolanib has a structure represented by the following chemical formula 1.

[0013] [Chemical Formula 1]

[0014]

[0015] The above-mentioned crenolanib is an anticancer drug developed to target FLT3 and PDGFRα / β, and is known to have the effect of inhibiting the PI3K-AKT-mTOR signal pathway, a representative inflammation-inducing signal, by inhibiting the activity of RTKs corresponding to Class Ⅲ, such as FLT3, PDGFRα, and PDGFRβ.

[0016] In one embodiment of the present invention, it was confirmed that crenolanib has an inhibitory effect on ULK2. ULK2 is a serine / threonine kinase known to act upstream of PIK3C3 to regulate autophagy. Abnormal autophagy may be associated with the onset or worsening of symptoms of autoimmune diseases, including psoriasis.

[0017] That is, the composition according to the present invention has the effect of suppressing the PI3K-AKT-mTOR signaling pathway, which is a signal that induces inflammation. Furthermore, the composition according to the present invention has the effect of suppressing the secretion of ULK2, an autophagy regulatory protein. This indicates that the composition according to the present invention has an inhibitory effect on immune response and autophagy activity, and may exhibit therapeutic effects on autoimmune diseases, including psoriasis.

[0018] "Psoriasis" in the present invention is an immune-mediated autoimmune skin disease caused by chronic activation of inflammatory cell infiltration in the skin and dysregulation of epidermal keratinocytes. Representative symptoms include erythema, rash, epidermal hardening, scaling, and itching, and lesions may spread to surrounding areas.

[0019] Specifically, psoriasis includes plaque psoriasis, pustular psoriasis, intertrigo psoriasis, guttate psoriasis, and erythrodermic psoriasis, depending on the lesion type. Plaque psoriasis is the most common form of the disease, and is characterized by well-defined, reddish plaques with silvery scales, usually on the extensor surfaces of the knees and elbows.

[0020] The composition according to the present invention is effective in improving symptoms of psoriasis such as erythema, epidermal hardening, and keratinization.

[0021] According to one embodiment of the present invention, the composition according to the present invention significantly reduced symptoms of psoriasis, such as erythema, epidermal hardening, and keratinization, as observed by visual inspection. In other words, the composition according to the present invention is effective in improving the symptoms of psoriasis and treating psoriasis.

[0022] In addition, the composition according to the present invention has an effect of inhibiting the secretion of any one or more inflammatory markers selected from the group consisting of NO, PGE2, TNF-α, and IL6.

[0023] In addition, the composition according to the present invention has the effect of inhibiting the secretion of any one or more selected from the group consisting of histamine and tumor necrosis factor-α, which are allergic markers. In addition, the composition of the present invention has the effect of inhibiting the activity of mast cells (human mast cell lines), which are known to be the main cause of allergic reactions and the cause of worsening symptoms in psoriasis, by inhibiting the secretion of histamine or TNF-α, which are allergic markers.

[0024] Additionally, the composition according to the present invention has the effect of inhibiting the secretion of at least one inflammatory marker selected from the group consisting of RANTES, MDC, and TARC of keratinocytes. This indicates that the composition according to the present invention has anti-inflammatory, anti-allergic, and anti-dermatitis properties, and is effective in the treatment of psoriasis.

[0025] The term "prevention" as used herein refers to any action that suppresses or delays the onset of psoriasis by administering a composition. In the present invention, "treatment" refers to any action that improves or beneficially alters the symptoms of the disease by administering a composition.

[0026] In the present invention, “pharmaceutically acceptable salt” means a salt commonly used in the pharmaceutical industry, and for example, inorganic ionic salts manufactured with calcium, potassium, sodium, and magnesium, etc.; inorganic acid salts manufactured with hydrochloric acid, nitric acid, phosphoric acid, hydrobromic acid, iodic acid, perchloric acid, and sulfuric acid, etc.; organic acid salts manufactured with acetic acid, trifluoroacetic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, and hydroiodic acid, etc.; sulfonic acid salts manufactured with methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, and naphthalenesulfonic acid, etc. Amino acid salts made from glycine, arginine, lysine, etc.; and amine salts made from trimethylamine, triethylamine, ammonia, pyridine, picoline, etc.; however, the types of salts meant in the present invention are not limited by these listed salts.

[0027] In addition, the crenolanib of the present invention includes not only pharmaceutically acceptable salts but also all salts, hydrates and solvates that can be prepared by conventional methods.

[0028] The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier, and may be formulated in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods.

[0029] The pharmaceutically acceptable carriers include, but are not limited to, those commonly used in the art, such as lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil. In addition, the pharmaceutical composition of the present invention may include, but is not limited to, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants, and other pharmaceutically acceptable additives.

[0030] When the pharmaceutical composition of the present invention is formulated as an oral solid preparation, it includes tablets, pills, powders, granules, capsules, etc., and such solid preparations may include at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc., and include, but are not limited to, lubricants such as magnesium stearate and talc.

[0031] When the pharmaceutical composition of the present invention is formulated as an oral liquid, it includes a suspension, a solution, an emulsion, a syrup, etc., and includes, but is not limited to, a diluent such as water or liquid paraffin, a wetting agent, a sweetener, a fragrance, a preservative, etc.

[0032] When the pharmaceutical composition of the present invention is formulated for parenteral use, it includes a sterile aqueous solution, a non-aqueous solvent, a suspension, an emulsion, a lyophilized preparation, and a suppository. Non-aqueous solvents and suspensions include, but are not limited to, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a base for a suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin, and the like can be used, but are not limited to these.

[0033] The composition may be administered in single or multiple doses in a pharmaceutically effective amount. The term “pharmaceutically effective amount” as used herein means an amount sufficient to prevent or treat a disease at a reasonable benefit-risk ratio applicable to medical prevention or treatment, and the effective dosage level may be determined according to factors including the severity of the disease, the activity of the drug, the patient’s age, weight, health, sex, the patient’s sensitivity to the drug, the time of administration of the composition of the present invention used, the route of administration and the excretion rate, the treatment period, drugs used in combination or concurrently with the composition of the present invention used, and other factors well known in the medical field. For example, crenolanib or a pharmaceutically acceptable salt thereof may be administered at 0.0001 to 100 mg / kg per day, and the administration may be administered once a day or in several divided doses.

[0034] The pharmaceutical composition of the present invention can be administered to mammals such as rats, mice, livestock, and humans via various routes, including, but not limited to, oral administration, intrathecal, intra-auricular, intraperitoneal, intravenous, intramuscular, subcutaneous, intrauterine, sublingual, or intracerebrovascular injection. In addition, it can be applied directly to the affected area as needed.

[0035] The pharmaceutical composition of the present invention may contain 0.01 to 95 wt%, preferably 1 to 80 wt%, of crenolanib or a pharmaceutically acceptable salt thereof based on the total weight of the composition.

[0036] The pharmaceutical composition of the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents. Furthermore, the pharmaceutical composition of the present invention can be administered singly or in multiple doses. Taking all of the above factors into consideration, it is important to administer the amount that achieves maximum efficacy with the minimum amount possible without causing side effects, a determination readily made by those skilled in the art.

[0037] The term "subject" of the present invention encompasses an animal or human whose symptoms can be improved by administration of a pharmaceutical composition according to the present invention. By administering a therapeutic composition according to the present invention to a subject, psoriasis can be effectively prevented and treated.

[0038] The term "administration" in the present invention refers to introducing a given substance into a human or animal by any suitable method. The therapeutic composition according to the present invention may be administered orally or parenterally via any conventional route, as long as it can reach the target tissue. Furthermore, the therapeutic composition according to the present invention may be administered by any device capable of transporting the active ingredient to target cells.

[0039] Another aspect of the present invention provides a method for preventing or treating psoriasis, comprising administering a therapeutically effective amount of crenolanib or a pharmaceutically acceptable salt thereof to a subject in need thereof.

[0040] The term "subject" of the present invention refers to any animal that exhibits or may exhibit psoriasis, and typically refers to an animal that may exhibit a beneficial effect from treatment with the present invention's crenolanib or a pharmaceutically acceptable salt thereof, but includes, without limitation, any subject that has psoriasis or is likely to have such a disease. As described above, psoriasis can be effectively prevented or treated by administering the pharmaceutical composition of the present invention to a subject.

[0041] Another aspect of the present invention provides the use of crenolanib or a pharmaceutically acceptable salt thereof of the present invention for preparing a medicament for preventing or treating psoriasis.

[0042] Another aspect of the present invention provides the use of crenolanib or a pharmaceutically acceptable salt thereof for the prevention or treatment of psoriasis.

[0043] Another aspect of the present invention provides a composition comprising crenolanib or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of psoriasis.

[0044] The above psoriasis may be at least one selected from the group consisting of plaque psoriasis, pustular psoriasis, intertrigo psoriasis, guttate psoriasis, and erythrodermic psoriasis.

[0045] The above crenolanib or a pharmaceutically acceptable salt thereof may a) inhibit the PI3K-AKT-mTOR signaling pathway, b) inhibit the activity of ULK2, c) inhibit the secretion of any one or more selected from the group consisting of NO, PGE2, TNF-α, and IL6, d) inhibit the secretion of Histamine, TNF-α, or all of them, and / or e) inhibit the secretion of any one or more selected from the group consisting of RANTES, MDC, and TARC.

[0046] The present invention provides a food composition for preventing or improving psoriasis, comprising crenolanib or a food-related acceptable salt thereof.

[0047] The term “improvement” of the present invention means any action by which psoriasis is improved or beneficially changed by administration of the composition of the present invention.

[0048] In the food composition of the present invention, the crenolanib and its salt are preferably included in an amount of 0.00001 to 0.01 wt% relative to the food composition. If the amount is less than 0.00001 wt%, the effect is minimal, and if the amount exceeds 0.01 wt%, the increase in effect relative to the amount used is minimal, making it uneconomical.

[0049] Food-grade acceptable salt may be applied in the same manner as pharmaceutically acceptable salt.

[0050] When the food composition of the present invention is used as a food additive, it can be added as is or used together with other foods or food ingredients, and can be used appropriately according to a conventional method.

[0051] In the present invention, the food composition includes a health functional food.

[0052] The above “health functional food” refers to food manufactured and processed using raw materials or ingredients with functionality useful to the human body as defined in Act No. 6727 on Health Functional Foods, and “functionality” refers to consumption for the purpose of obtaining a useful effect for health purposes such as regulating nutrients for the structure and function of the human body or physiological effects.

[0053] The food composition and health functional food of the present invention may include additional ingredients. For example, they may include biotin, folate, pantothenic acid, vitamins A, C, D, E, B1, B2, B6, B12, niacin, etc. In addition, they may include minerals such as chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), zinc (Zn), iron (Fe), and calcium (Ca). In addition, they may include amino acids such as cysteine, valine, lysine, and tryptophan. In addition, food additives such as preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), coloring agents (tar color, etc.), coloring agents (sodium nitrite, sodium nitrite, etc.), bleaching agents (sodium sulfite), bactericides (bleaching powder and high-purity bleaching powder, sodium hypochlorite, etc.), leavening agents (alum, D-potassium hydrogen tartrate, etc.), reinforcing agents, emulsifiers, thickeners (pasting agents), film-forming agents, antioxidants (butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), etc.), seasonings (MSG, monosodium glutamate, etc.), sweeteners (dulcin, cyclamate, saccharin, sodium, etc.), flavorings (vanillin, lactones, etc.), gum bases, antifoaming agents, solvents, and improvers can be added. The above additives can be selected depending on the type of food and used in an appropriate amount.

[0054] The composition according to the present invention has anti-inflammatory, anti-allergic, and anti-dermatitis properties through the regulation of the secretion of inflammatory markers (NO, PGE2, TNF-α, IL6), allergic markers (histamine, TNF-α), and inflammatory markers of keratinocytes (RANTES, MDC, TARC). In addition, it is effective in improving symptoms of psoriasis such as erythema, epidermal hardening, and keratinization. Therefore, the composition according to the present invention can be usefully used for the prevention and treatment of psoriasis.

[0055] Figure 1 is a graph showing the inhibitory efficacy of crenolanib against ULK2.

[0056] Figure 2 is a graph showing the effect of crenolanib on improving the psoriasis area and severity index (PASI) in a psoriasis-induced mouse model.

[0057] Figure 3 is an image of psoriasis lesion tissue observed by H&E staining in a psoriasis-induced mouse model.

[0058] Figure 4 is a graph showing the effect of crenolanib on improving epidermal thickness in a psoriasis-induced mouse model.

[0059] Figure 5 is an image of skin tissue stained and observed using CD8+ antibody in a psoriasis lesion tissue of a psoriasis-induced mouse model.

[0060] Figure 6 is a graph showing the CD8+ cell infiltration inhibitory effect of crenolanib in a psoriasis-induced mouse model.

[0061] Figure 7 is an image of skin tissue stained and observed using IL-17+ antibody in a psoriasis-induced mouse model.

[0062] Figure 8 is a graph showing the inhibitory effect of crenolanib on IL-17+ cell infiltration in a psoriasis-induced mouse model.

[0063] Figure 9 is an image visually observing the spleen transformation inhibitory effect of crenolanib in a psoriasis-induced mouse model.

[0064] Hereinafter, the present invention will be described in more detail through examples. However, these examples are intended to illustrate the present invention and the scope of the present invention is not limited to these examples.

[0065] <Example>

[0066] Example 1. Confirmation of the inhibitory effect of crenolanib on ULK2.

[0067] A kinase-tagged T7 phage strain was prepared from an E. coli host derived from the BL21 strain. E. coli was cultured, infected with the T7 phage, and incubated with shaking at 32°C until lysis. After incubation, the lysate was centrifuged, and the supernatant was separated. Streptavidin-coated magnetic particles were reacted with the separated supernatant for 30 minutes. Unbound and nonspecifically bound ligands were then removed from the particles using locking buffer (SeaBlock (Pierce), 1% BSA, 0.05% Tween 20, 1 mM DTT). The sample was reacted by adding 1x binding buffer (20% SeaBlock, 0.17x PBS, 0.05% Tween 20, 6 mM DTT), and a stock (x111) solution was prepared. The sample was diluted to a final concentration of 0.9% DMSO in 0.02 mL and washed with wash buffer (1x PBS, 0.05% Tween 20). After resuspending in elution buffer (1x PBS, 0.05% Tween 20, 0.5 μM nonbiotinylated affinity ligand), the reaction was performed at room temperature for 30 minutes and measured by qPCR.

[0068] As a result, as shown in Fig. 1, it was confirmed that crenolanib had excellent inhibitory efficacy against ULK2.

[0069] Example 2. Cell-based efficacy evaluation of crenolanib

[0070] Example 2-1. Confirmation of the anti-inflammatory activity of crenolanib

[0071] Mouse macrophage cell line (RAW 264.7) cells were cultured in DMEM (Dulbecco's Modified Eagle's Medium) containing 10% FBS (Fetal Bovine Serum) and 1% P&S (Penicillin-Streptomycin). Cultured RAW264.7 cells were seeded at 4 × 10 to check cytotoxicity. 3 After seeding cells / well (96-well plate), overnight incubation was performed. Crenolanib was treated at various concentrations (from 50 μM) and reacted for 24 hours (final 100 μL / well). 10 μL / well of Cell Counting Kit-8 (CCK-8, Dojindo, Japan) solution was added and reacted for 4 hours. The absorbance was measured at 450 nm. The measured value was compared with the control well to calculate cell viability%. Among them, the highest concentration showing cell viability of 95% or more was selected as the highest non-toxic concentration (Max. Conc.) and anti-inflammatory experiments were performed.

[0072] To confirm the anti-inflammatory activity, RAW 264.7 cells were seeded in 48-well plates and incubated overnight, and then cultured for 24 hours (final 500 μL / well) under the conditions of creolanib treatment and (±)-LPS treatment (1 μg / mL). For the quantification of NO, PGE2, TNF-α, and IL-6, the culture medium was collected and NO was quantified using Griess reagent (Promega), and PGE2, TNF-α, and IL-6 were quantified using each ELISA kit. The inhibition rate of inflammatory mediators and cytokine production induced by LPS treatment was calculated and shown in Table 1.

[0073] As a result, as shown in Table 1 below, the composition of Example 1 was found to inhibit the secretion of NO (31%), PGE2 (23%), TNF-α (39%), and IL6 (51%) by 23% to 51% at the non-toxic maximum concentration of 100 nM. This confirmed that crenolanib has anti-inflammatory activity.

[0074] NoCodeMaximum Non-Atoxic Concentration (Max Conc.)SecondaryNOPGE2TNF-αIL-6IC 50 % inh. at Max Conc.IC 50 % inh. at Max Conc.IC 50 % inh. at Max Conc.IC 50 % inh. at Max Conc.1 Example 1 (Crenolanib) 100 nM - 30.71 - 22.71 - 39.37 62.56 nM 50.69

[0075] Example 2-2. Confirmation of anti-allergic activity using mast cell lines

[0076] Human mast cell line (HMC-1.2) cells were cultured in IMDM (Iscove's Modified Dulbecco's Medium) containing 10% FBS (Fetal Bovine Serum) and 1% P&S (Penicillin-Streptomycin). Cultured HMC-1.2 cells were maintained at a density of 2X10 4 After seeding cells / well (96-well plate), overnight incubation was performed. Crenolanib was treated at various concentrations (from 50 μM) and reacted for 6 hours (final 100 μL / well). 10 μL / well of CCK-8 solution was added and reacted for 4 hours. The absorbance was measured at 450 nm. The measured value was compared with the control well to calculate cell viability %. Among them, the highest concentration showing cell viability of 95% or more was selected as the highest non-toxic concentration (Max. Comc.) and an anti-allergy experiment was conducted.

[0077] To confirm the anti-allergic activity, HMC-1.2 cells were seeded in 48-well plates and incubated overnight. Then, they were cultured for 6 hours (final 500 μL / well) under the conditions of creolanib treatment and PMA and A23187 treatment (20 nM + 1 μM each, PA). To quantify the released mediators, the culture medium was collected and histamine and TNF-α were quantified using each ELISA kit. The inhibition rate of mediator production induced by PA treatment was calculated and shown in Table 2.

[0078] As a result, as shown in Table 2 below, the composition of Example 1 was found to inhibit the secretion of histamine and TNF-α by 48% and 22%, respectively, at the non-toxic maximum concentration of 50 nM. This confirmed that crenolanib has anti-allergic activity.

[0079] NoCodeNon-toxic Highest Concentration (Max Conc.)Secondary HistamineTNF-αIC 50 % inh. at Max Conc.IC 50 % inh. at Max Conc.1 Example 1 (Crenolanib) 50 nM - 47.56 - 21.91

[0080] Example 2-3. Confirmation of anti-dermatitis activity using keratinocyte cell lines

[0081] Human keratinocyte cell line (HaCaT) cells were cultured in DMEM (Dulbecco's Modified Eagle's Medium) containing 10% FBS (Fetal Bovine Serum) and 1% P&S (Penicillin-Streptomycin). Cultured HaCaT cells were maintained at a density of 3X10 3After seeding cells / well (96-well plate), overnight incubation was performed. Crenolanib was treated at various concentrations (from 50 μM) and reacted for 24 hours (final 100 μL / well). 10 μL / well of CCK-8 solution was added and reacted for 4 hours. The absorbance was measured at 450 nm. The measured value was compared with the control well to calculate cell viability %. Among them, the highest concentration showing cell viability of 95% or more was selected as the highest non-toxic concentration (Max. Conc.) and anti-dermatitis experiment was performed.

[0082] To confirm the anti-dermatitis efficacy, HaCaT cells were seeded in 6-well plates and incubated overnight. Then, they were cultured for 24 hours (final 1000 μL / well) under conditions of creolanib treatment and TNF-α and IFN-γ treatment (each 10 ng / mL, TI). To quantify the released chemokines, the culture medium was collected and RANTES, TARC, and MDC were quantified using each ELISA kit. The inhibition rate of chemokine production induced by TI treatment was calculated and is shown in Table 3.

[0083] As a result, as shown in Table 3 below, crenolanib was found to inhibit the secretion of RANTES, MDC, and TARC, which are inflammatory markers of keratinocytes, by 52%, 46%, and 53%, respectively, at the non-toxic maximum concentration of 2 μM. This confirmed that crenolanib possesses anti-dermatitis activity.

[0084] NoCodeMaximum Non-Toxic Concentration (Max Conc.)2ndRANTESMDCTARCIC 50 % inh. at Max Conc.IC 50 % inh. at Max Conc.IC 50% inh. at Max Conc.1Example 1 (Crenolanib)2 μM1.90 μM52.04-46.101.64 μM52.85

[0085] Experimental Example 1. Establishment of a psoriasis-induced mouse model.

[0086] To conduct animal experiments using creolanib, a mouse model of psoriasis induced by imiquimod (IMQ) was established.

[0087] The experimental groups for animal experiments were organized as follows.

[0088]

[0089] The experimental groups for the above animal experiments were prepared as follows. First, to induce psoriasis, the hair on the back of the mice was removed with depilatory cream the day before the experiment began, and then 62.5 mg of Aldara cream was applied daily from the start day (Day 0) to the end day to induce psoriasis.

[0090] Here, for the crenolanib PO group, a mixed solution containing 2 mg crenolanib in 1 mL of excipient (10% DMSO + 70% Corn Oil + 20% Saline) was orally administered once daily at 200 μL (20 mpk) from the start to the end of the experiment. In addition, for the crenolanib TA group, a mixed solution containing 4 mg crenolanib in 1 mL of excipient (10% DMSO + 70% Corn Oil + 20% Saline) was appropriately applied to the affected area at 100 μL (20 mpk) once daily from the start to the end of the experiment. The test substance was completely dissolved by adding a solvent, vortexing for 1 to 2 minutes, water bath sonication for 12 minutes, and vortexing and sonication three times before use.

[0091] Example 3. Confirmation of the effect of crenolanib on improving the psoriasis area and severity index (PASI) in a psoriasis-induced mouse model.

[0092] In order to confirm the degree of progression of psoriasis and the treatment efficacy, the degree of skin erythema, hardening, and keratinization was measured once a day from day 0 to the end of the experiment according to the analysis criteria below, and each measurement value was finally added up and expressed as a PASI score (psoriasis severity index, maximum 12 points) and is shown in Figure 2.

[0093] Comparison of symptoms between each group revealed that erythema, induration, and keratosis symptoms all increased in the Vehicle PO and Vehicle TA groups compared to the normal control group, and the PASI index also increased accordingly. The Crenolanib PO group, which was orally administered a composition containing Crenolanib, showed similar symptoms to the Vehicle PO group.

[0094] On the other hand, in the case of the Crenolanib TA group, in which a composition including Crenolanib was applied to the skin, erythema, hardening, and keratin symptoms were significantly reduced compared to the Vehicle TA group, and accordingly, the PASI index was also lower than that of the Vehicle TA group.

[0095] That is, in visual examination, it was observed that the symptoms of psoriasis, such as erythema, induration, and keratinization, were improved in the crenolanib TA group compared to the vehicle TA group, confirming that crenolanib is effective in treating psoriasis caused by IMQ.

[0096] Example 4. Confirmation of the effect of crenolanib on improving epidermal thickness in a psoriasis-induced mouse model.

[0097] To confirm the therapeutic efficacy of psoriasis, the degree of epidermal improvement was assessed. Skin tissue from each experimental group was stained with hematoxylin and eosin (H&E), observed under a 100x magnification microscope, and epidermal thickness was measured. These results are shown in Figures 3 and 4, respectively.

[0098] As a result, the epidermal thickness increased in Vehicle TA compared to the normal control group, and the epidermal thickness significantly decreased in Crenolanib TA compared to Vehicle TA.

[0099] That is, crenolanib TA was observed to improve the epidermal thickness of psoriasis lesions compared to Vehicle TA, confirming that crenolanib has therapeutic efficacy in treating IMQ-induced psoriasis.

[0100] Example 5. Confirmation of the CD8+ cell infiltration inhibition effect of crenolanib in a psoriasis-induced mouse model.

[0101] To confirm the therapeutic efficacy of psoriasis, the degree of CD8+ antibody penetration was determined. For immunochemical analysis of skin tissue, skin tissue was stained with a specific CD8+ antibody and photographed under a 400x magnification microscope. Cells expressing various immune cells and cytokines were counted using ImageJ software (Wayne Rasband, NIH, USA). In particular, the average optical density (AOD) of areas expressing keratin 16, known as a marker of epidermal hyperproliferation, was analyzed. The stained tissue image is shown in Figure 5, and the results of the image analysis are shown in Figure 6.

[0102] As a result of observing the tissues of each group, the degree of CD8+ cell infiltration increased in the Vehicle TA group compared to the normal control group, and the degree of CD8+ cell infiltration in the Crenolanib TA group significantly decreased compared to the Vehicle TA group.

[0103] That is, it was found that the crenolanib TA group had the effect of inhibiting CD8+ cell infiltration in psoriasis lesions compared to the Vehicle TA group, thereby confirming that crenolanib has therapeutic efficacy in treating IMQ-induced psoriasis.

[0104] Example 6. Confirmation of the inhibitory effect of crenolanib on IL-17+ cell infiltration in a psoriasis-induced mouse model.

[0105] To confirm the therapeutic efficacy of psoriasis, the degree of penetration of IL-17+ antibodies was determined. For immunochemical analysis of skin tissue, skin tissue was stained using a specific IL-17+ antibody and photographed under a microscope at 400x magnification. Cells expressing each immune cell and cytokine were counted using ImageJ software (Wayne Rasband, NIH, USA). In particular, the average optical density (AOD) of the area expressing keratin 16, known as a marker of epidermal hyperproliferation, was analyzed. The stained image is shown in Figure 7, and the image analysis results are shown in Figure 8.

[0106] As a result of observing the tissues of each group, the degree of IL-17+ cell infiltration increased in the Vehicle TA group compared to the normal control group, and the degree of IL-17+ cell infiltration in the Crenolanib TA group significantly decreased compared to the Vehicle TA group.

[0107] That is, it was found that the crenolanib TA group had the effect of inhibiting IL-17+ cell infiltration in psoriasis lesions compared to the vehicle TA group, thereby confirming that crenolanib has therapeutic efficacy in treating psoriasis caused by IMQ.

[0108] Example 7. Confirmation of the inhibitory effect of crenolanib on spleen transformation in a psoriasis-induced mouse model.

[0109] After the above animal test was completed, the toxicity of the drug was confirmed by removing the spleen from the mouse when it was sacrificed and measuring the degree of deformation such as size and weight, and the image is shown in Figure 9.

[0110] Comparison of the spleens of each group showed that the spleen of the Vehicle TA group was larger and longer and deformed compared to the normal control group, while the spleen of the Crenolanib TA group was relatively less deformed.

[0111] That is, it was observed that the crenolanib TA group did not induce splenomegaly compared to the negative control group, the Vehicle TA group, confirming that crenolanib has the efficacy of suppressing spleen transformation caused by IMQ.

Claims

1. A pharmaceutical composition for preventing or treating psoriasis comprising crenolanib or a pharmaceutically acceptable salt thereof.

2. A pharmaceutical composition according to claim 1, wherein the composition inhibits the PI3K-AKT-mTOR signal pathway.

3. A pharmaceutical composition according to claim 1, wherein the composition inhibits the activity of ULK2.

4. A pharmaceutical composition according to claim 1, wherein the composition inhibits the secretion of at least one agent selected from the group consisting of NO, PGE2, TNF-α, and IL6.

5. A pharmaceutical composition according to claim 1, wherein the composition inhibits the secretion of at least one agent selected from the group consisting of Histamine and TNF-α.

6. A pharmaceutical composition according to claim 1, wherein the composition inhibits secretion of at least one compound selected from the group consisting of RANTES, MDC, and TARC.

7. A pharmaceutical composition in claim 1, wherein the psoriasis is at least one selected from the group consisting of plaque psoriasis, pustular psoriasis, intertrigo psoriasis, droplet-shaped psoriasis, and erythrodermic psoriasis.

8. Use of creolanib or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the prevention or treatment of psoriasis.

9. The use according to claim 8, wherein the psoriasis is at least one selected from the group consisting of plaque psoriasis, pustular psoriasis, intertrigo psoriasis, droplet-shaped psoriasis, and erythrodermic psoriasis.

10. In paragraph 8, the crenolanib or a pharmaceutically acceptable salt thereof a) Inhibiting the PI3K-AKT-mTOR signaling pathway, or b) inhibiting the activity of ULK2, or c) inhibiting the secretion of any one or more selected from the group consisting of NO, PGE2, TNF-α and IL6; d) inhibiting the secretion of histamine, TNF-α or both, and / or e) Use for inhibiting the secretion of any one or more selected from the group consisting of RANTES, MDC and TARC.

11. A method for preventing or treating psoriasis, comprising administering a therapeutically effective amount of creolanib or a pharmaceutically acceptable salt thereof to a subject in need thereof.

12. A method in claim 11, wherein the psoriasis is at least one selected from the group consisting of plaque psoriasis, pustular psoriasis, intertrigo psoriasis, droplet-shaped psoriasis, and erythrodermic psoriasis.

13. In clause 11, the crenolanib or a pharmaceutically acceptable salt thereof a) Inhibiting the PI3K-AKT-mTOR signaling pathway, or b) inhibiting the activity of ULK2, or c) inhibiting the secretion of any one or more selected from the group consisting of NO, PGE2, TNF-α and IL6; d) inhibiting the secretion of histamine, TNF-α or both, and / or e) A method for inhibiting the secretion of any one or more selected from the group consisting of RANTES, MDC and TARC.

14. Use of creolanib or a pharmaceutically acceptable salt thereof for the prevention or treatment of psoriasis.

15. A composition comprising crenolanib or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of psoriasis.

16. A food composition for preventing or improving psoriasis comprising creolanib or a food-chemically acceptable salt thereof.

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