Composition for treating alopecia areata and promoting hair growth, comprising zipalertinib or pharmaceutically acceptable salt thereof

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating alopecia areata, comprising zipalertinib or a pharmaceutically acceptable salt thereof. The composition, according to the present invention, has anti-inflammatory and immune cell-suppressing properties by regulating the secretion of inflammatory markers (IL-6, IL-1β, IL-8, and TNF-α) and inducers of Th1 and Tc1 cells (CXCL9, CXCL10, and CXCL11). Accordingly, the composition according to the present invention has a therapeutic effect on alopecia areata, and can be efficiently used for preventing and treating alopecia areata.

Description

Composition for treating hair loss and promoting hair growth comprising zipalertinib or a pharmaceutically acceptable salt thereof

[0001] The present invention relates to a composition for treating hair loss and promoting hair growth, and more preferably, to a composition for treating hair loss and promoting hair growth comprising zipalertinib or a pharmaceutically acceptable salt thereof.

[0002] Hair loss is a highly prevalent condition, caused by both genetic and environmental factors (e.g., environmental factors, dietary habits, and autoimmune diseases). Worldwide, more than 140 million people, or 14% of the world's population, are affected by hair loss. The number of people with hair loss in Korea is estimated to be increasing annually, reaching 10 million. In particular, those in their 20s and 30s, a time when people are most sensitive to their appearance, account for the largest proportion of hair loss patients, at 48.5%.

[0003] Hair loss occurs due to irregular lifestyle habits, stress, male hormones, aging, genetics, and nutritional imbalances. Hair becomes thinner and weaker due to vellus hair loss, the growth phase is shortened, hair falls out in the resting or catagen phase, and hair follicles atrophy, which prevents the supply of nutrients to the hair follicles, causing them to lose their function, and eventually, hair loss progresses.

[0004] Traditional treatments for hair loss include hormone-based preparations based on female hormones. However, due to reports of skin inflammation and side effects from hormone administration, these treatments are now discontinued. Recently, representative hair growth agents in use were initially developed to promote blood circulation, but some patients reported experiencing hair growth side effects. Since then, minoxidil and finasteride have been approved by the U.S. Food and Drug Administration (FDA) as hair growth agents and are now used as hair growth treatments.

[0005] Minoxidil and finasteride, both 5-alpha reductase inhibitors, are widely used as hair loss treatments. However, they have side effects such as depression and sexual dysfunction, require long-term use, and reports of side effects and toxicity, such as hair loss recurring upon discontinuation, are being published. As such, the efficacy of currently available hair loss treatments is limited, long-term use is difficult, and toxicity and side effects are reported, the number of patients seeking surgical options is increasing every year.

[0006] Alopecia areata is a systemic autoimmune disease in which the body attacks its own hair follicles, inhibiting or stopping hair growth. Alopecia areata is the second most common type of hair loss in humans, affecting over 2% of individuals and characterized by non-scarring hair loss. While hair loss itself does not cause physical distress, it can negatively impact body image and self-esteem, leading to significant psychological problems and a diminished quality of life.

[0007] Treatment for alopecia areata includes topical immunotherapy, corticosteroids, and immunosuppressants such as psoralen, minoxidil, dithranol (or anthralin), and cyclosporine, combined with ultraviolet A phototherapy (PUVA). However, these treatments all have potential side effects, ranging from minor dermatitis and folliculitis to major skin cancer, requiring careful monitoring. Therefore, the field needs a treatment that is safe and effective for long-term use.

[0008] Meanwhile, zipalertinib is known as an epidermal growth factor receptor (EGFR) inhibitor, and a phase 3 clinical trial is underway for the first-line treatment of locally advanced or metastatic non-small cell lung cancer positive for EGFR exon 20 insertion mutation.

[0009] Against this backdrop, the inventors of the present invention have completed the present invention by confirming that zipalertinib or a composition comprising it inhibits T cell activity in alopecia and alleviates symptoms. Therefore, the composition according to the present invention is expected to be useful as a pharmaceutical composition for the prevention or treatment of hair loss.

[0010] An object of the present invention is to provide a pharmaceutical composition for preventing or treating hair loss comprising zipalertinib or a pharmaceutically acceptable salt thereof.

[0011] An object of the present invention is to provide a method for preventing or treating hair loss, comprising administering a therapeutically effective amount of zipalertinib or a pharmaceutically acceptable salt thereof to a subject in need thereof.

[0012] An object of the present invention is to provide a use of zipalertinib of the present invention or a pharmaceutically acceptable salt thereof for preparing a medicament for preventing or treating hair loss.

[0013] An object of the present invention is to provide a composition comprising zipalertinib or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of hair loss.

[0014] The present invention provides a pharmaceutical composition for preventing or treating hair loss comprising zipalertinib or a pharmaceutically acceptable salt thereof.

[0015] In the present invention, zipalertinib has a structure represented by the following chemical formula 1.

[0016] [Chemical Formula 1]

[0017]

[0018] The above-mentioned zipalertinib is known as an EGFR (epidermal growth factor receptor) inhibitor.

[0019] In one embodiment of the present invention, zipalertinib was confirmed to have an inhibitory effect on JAK3. JAK3, a JAK-type tyrosine kinase, can regulate autoimmune responses through the JAK-STAT mechanism, which is attracting attention as a new target for hair loss treatments. This inhibits T cell activation that occurs in hair loss and alleviates hair loss symptoms, suggesting potential therapeutic potential for hair loss.

[0020] The composition according to the present invention has an inhibitory effect on immune response activity and can exhibit excellent therapeutic efficacy against hair loss, particularly alopecia areata.

[0021] Hair loss refers to the phenomenon of hair falling out from the skin or a condition in which hair becomes thicker or thinner, and is a term that can be used interchangeably with alopecia.

[0022] In the present invention, hair includes hair roots and hair follicles of the head, hair and eyelashes, eyebrows, beard, armpits, pubic hair, and all parts of the body with hair roots and hair follicles.

[0023] In the present invention, "hair loss" may be at least one selected from the group consisting of nutritional alopecia, endocrine alopecia, vascular alopecia, middle-aged alopecia, alopecia areata, neurogenic alopecia, alopecia pityriasis, trichotillomania, malignant alopecia, female pattern alopecia, male pattern alopecia, androgenetic alopecia, telogen effluvium, tinea capitis, alopecia totalis, hypotrichosis, hereditary hypotrichosis simplex, drug-induced alopecia universalis, mechanical alopecia, traumatic alopecia, compression alopecia, anagen alopecia, alopecia pityriasis, syphilitic alopecia, seborrheic alopecia, symptomatic alopecia, cicatricial alopecia, and congenital alopecia.

[0024] More specifically, it could be alopecia areata. Alopecia areata is a systemic autoimmune disease in which the body attacks its own hair follicles, inhibiting or stopping hair growth. Alopecia areata is the second most common type of hair loss in humans, affecting over 2% of people and characterized by scarless hair loss.

[0025] Specifically, alopecia areata is characterized by the sudden appearance of clearly bordered bald patches without any separate prodromal symptoms or subjective symptoms. Those affected by alopecia areata typically have one or more circular or patchy bald lesions on the scalp, and the incidence is similar in men and women. In approximately 5% of cases, alopecia universalis develops, which extends to the entire scalp (alopecia totalis) or hair-bearing areas of the body. Less than 10% of alopecia universalis patients recover. It can present in various forms, from partial to universal alopecia.

[0026] The composition according to the present invention has the effect of inhibiting the secretion of any one or more proteins selected from the group consisting of inflammatory markers IL-6, IL-1β, IL-8, and TNF-α.

[0027] In addition, the composition according to the present invention has the effect of inhibiting the expression of any one or more mRNAs selected from the group consisting of inflammatory markers IL-6, IL-1β, IL-8, and TNF-α.

[0028] In addition, the composition according to the present invention has the effect of suppressing the expression of one or more cytokines selected from the group consisting of Th1, Tc1 cell inducers CXCL9, CXCL10, and CXCL11.

[0029] The above Th1 and Tc1 cells are subsets of helper T (Th) and cytotoxic T (Tc) lymphocytes, which recognize tumor antigens and secrete cytokines to promote or suppress tumor growth. For example, Th1 cells mainly produce interferon (IFN)-γ to enhance cell-mediated immunity against tumors, and Tc1 cells secrete IFN-γ and can destroy tumor cells through perforin- or Fas-mediated pathways (Arch Immunol Ther Exp (Warsz). 2014 Dec;62(6):449-58.).

[0030] Meanwhile, CXCL9, CXCL10, and CXCL11 are cytokines that play a key role in the differentiation, migration, and activation of Th1 cells. CXCL9, CXCL10, and CXCL11 are ligands for the CXCR3 receptor protein, and are cytokines whose expression is induced by IFN. The signaling axis consisting of IFN-CXCL9, CXCL10, CXCL11-CXCR3 is known as an important target in the fields of immune response and immuno-oncology.

[0031] In particular, it is known that CXCL9 and CXCL10 cytokines are increased in the blood of patients with alopecia areata, and it is known that the progression of alopecia areata can be suppressed by inhibiting the signaling leading to CXCL9, CXCL10, and CXCL11-CXCR3.

[0032] Through this, it can be seen that the composition according to the present invention has anti-inflammatory and immune cell suppression properties and is effective in treating hair loss.

[0033] The term "prevention" as used herein refers to any action that inhibits or delays the onset of hair loss through the administration of a composition. In the present invention, "treatment" refers to any action that improves or beneficially alters the symptoms of a disease through the administration of a composition.

[0034] 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.

[0035] In addition, the zipalertinib of the present invention includes not only pharmaceutically acceptable salts but also all salts, hydrates, and solvates that can be prepared by conventional methods.

[0036] 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.

[0037] 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.

[0038] 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.

[0039] 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.

[0040] 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.

[0041] 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, zipalertinib 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.

[0042] 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.

[0043] The pharmaceutical composition of the present invention may contain 0.01 to 95 wt%, preferably 1 to 80 wt%, of zipalertinib or a pharmaceutically acceptable salt thereof based on the total weight of the composition.

[0044] 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.

[0045] The term "subject" of the present invention includes 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, hair loss can be effectively prevented and treated.

[0046] 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.

[0047] Another aspect of the present invention provides a method for preventing or treating hair loss, comprising administering to a subject in need thereof a therapeutically effective amount of zipalertinib or a pharmaceutically acceptable salt thereof.

[0048] The term "subject" in the present invention refers to any animal that exhibits or may exhibit hair loss, and typically refers to an animal that may exhibit a beneficial effect from treatment with zipalertinib or a pharmaceutically acceptable salt thereof of the present invention, but includes, without limitation, any subject that exhibits hair loss or is likely to exhibit such a disease. As described above, hair loss can be effectively prevented or treated by administering the pharmaceutical composition of the present invention to a subject.

[0049] Another aspect of the present invention provides a use of zipalertinib or a pharmaceutically acceptable salt thereof of the present invention for preparing a medicament for preventing or treating hair loss.

[0050] Another aspect of the present invention provides a use of zipalertinib or a pharmaceutically acceptable salt thereof of the present invention for the prevention or treatment of hair loss.

[0051] Another aspect of the present invention provides a composition comprising zipalertinib or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of hair loss.

[0052] The above hair loss may be at least one selected from the group consisting of nutritional alopecia, endocrine alopecia, vascular alopecia, middle-aged alopecia, alopecia areata, neurogenic alopecia, alopecia pityriasis, trichotillomania, malignant alopecia, female pattern alopecia, male pattern alopecia, androgenetic alopecia, telogen effluvium, tinea capitis, alopecia totalis, hypotrichosis, hereditary hypotrichosis simplex, drug-induced alopecia universalis, mechanical alopecia, traumatic alopecia, compression alopecia, anagen alopecia, alopecia pityriasis, syphilitic alopecia, seborrheic alopecia, symptomatic alopecia, cicatricial alopecia, and congenital alopecia.

[0053] The above zipalertinib or a pharmaceutically acceptable salt thereof may a) inhibit the activity of JAK3, b) inhibit the secretion and mRNA expression of any one or more proteins selected from the group consisting of IL-6, IL-1β, IL-8, and TNF-α, and / or c) inhibit the expression of any one or more cytokines selected from the group consisting of CXCL9, CXCL10, and CXCL11.

[0054] The present invention provides a food composition for preventing or improving hair loss, comprising zipalertinib or a food-wise acceptable salt thereof.

[0055] The term “improvement” of the present invention means any act in which hair loss is improved or beneficially changed by administration of the composition of the present invention.

[0056] In the food composition of the present invention, the zipalertinib 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.

[0057] Food-grade acceptable salt may be applied in the same manner as pharmaceutically acceptable salt.

[0058] 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.

[0059] In the present invention, the food composition includes a health functional food.

[0060] 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.

[0061] 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.

[0062] The present invention provides a cosmetic composition for preventing or improving hair loss, comprising zipalertinib or a pharmaceutically acceptable salt thereof.

[0063] According to a specific embodiment of the present invention, a composition comprising zipalertinib or a pharmaceutically acceptable salt thereof promotes hair production and growth, thereby exhibiting an excellent effect in preventing or improving hair loss.

[0064] The cosmetic composition of the present invention can be formulated in the form of, for example, a solution, a sol-gel, a suspension, an emulsion, a paste, a microcapsule, an oil, a wax, or an aerosol, but is not limited thereto. Specific formulations of the cosmetic composition include, for example, a hair tonic, a hair conditioner, a hair lotion, a hair nourishing lotion, a hair shampoo, a hair rinse, a hair treatment, a hair cream, a hair nourishing cream, a hair moisture cream, a hair massage cream, a hair wax, a hair aerosol, a hair pack, a hair nourishing pack, a hair soap, a hair cleansing foam, a hair oil, a hair drying agent, a hair preservative, a hair dye, a hair waving agent, a hair bleaching agent, a hair gel, a hair glaze, a hair dressing agent, a hair lacquer, a hair moisturizer, a hair mousse, a hair spray, a patch, an ointment, a sealant, a liniment, a paste, a cataplasma agent, and a powder, but are not limited thereto.

[0065] The cosmetic composition of the present invention may additionally include a cosmetically acceptable carrier. The type of cosmetically acceptable carrier that can be used in the present invention is not specifically suggested, and any carrier commonly used in this technical field may be used. The carrier may be used alone or in combination of two or more types.

[0066] The cosmetic composition of the present invention may further include an adjuvant commonly used in cosmetic compositions, as long as the effects of the present invention are not impaired. The types of adjuvants that can be used in the present invention are not particularly limited, and any adjuvant commonly used in this technical field may be used. Examples of such adjuvants include, but are not limited to, gelling agents, activators, preservatives, antioxidants, stabilizers, solubilizers, vitamins, solvents, fragrances, fillers, blocking agents, pigments, deodorants, and dyes. The adjuvants may be used alone or in combination of two or more.

[0067] The cosmetic composition of the present invention can be used by methods such as directly applying or spraying it on the hair or scalp.

[0068] The present invention provides a pharmaceutical composition for preventing or improving hair loss, comprising zipalertinib or a pharmaceutically acceptable salt thereof.

[0069] The pharmaceutical composition of the present invention can be effectively used for preventing or improving hair loss.

[0070] Specific formulations of the above-mentioned pharmaceutical composition include, but are not limited to, disinfectants, bath agents, wet tissues, ointments, creams, lotions, soaps, shampoos, rinses, essences, sprays, and patches.

[0071] The quasi-drug composition of the present invention may further include adjuvants commonly used in cosmetic compositions, as long as the effects of the present invention are not impaired. The types of adjuvants that can be used in the present invention are not particularly limited, and any adjuvant commonly used in this technical field may be used. The adjuvants may include, for example, conventional adjuvants such as gelling agents, activators, preservatives, fillers, antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances.

[0072] The present invention provides a composition for promoting hair growth comprising zipalertinib or a pharmaceutically acceptable salt thereof.

[0073] In the present invention, “hair growth” means the production and growth of hair, and is a broad concept including the meaning of the terms wool or hair growth used in this field.

[0074] The composition of the present invention can be effectively used for promoting hair growth by significantly promoting hair production and growth.

[0075] The hair growth promoting composition of the present invention can be used as a pharmaceutical, food, cosmetic or quasi-drug composition.

[0076] The hair growth promoting composition of the present invention can be administered orally or parenterally, depending on the intended purpose. In the present invention, parenteral administration includes, for example, intraperitoneal administration, rectal administration, subcutaneous administration, intravenous administration, intramuscular administration, thoracic administration, and transdermal administration. It also includes, but is not limited to, applying the composition to the scalp or hair, spraying it, or inhaling it. Preferably, the composition is applied to the scalp or hair or sprayed it.

[0077] The composition of the present invention may additionally include a hair growth promoting auxiliary ingredient. Hair growth promoting auxiliary ingredients include, but are not limited to, 5-alpha-reductase inhibitors, hair root and hair follicle cell activators, hair follicle cell blood flow enhancing ingredients, bactericidal ingredients, anti-dandruff agents, keratin softeners, refreshing agents, and moisturizers.

[0078] The composition according to the present invention has anti-inflammatory and immune cell suppression properties through regulation of the secretion of inflammatory markers (IL-6, IL-1β, IL-8, and TNF-α) and Th1 and Tc1 cell inducers (CXCL9, CXCL10, CXCL11). Therefore, the composition according to the present invention has therapeutic efficacy for hair loss and can be usefully used for the prevention and treatment of hair loss.

[0079] Figure 1 is a graph showing the inhibitory efficacy of zipalertinib against JAK3.

[0080] Figure 2 is a graph confirming the cytotoxicity of zipalertinib against outer root sheath cells.

[0081] Figure 3 is a graph showing the inhibitory effect of zipalertinib on IL-6, IL-1β, IL-8, and TNF-α protein secretion.

[0082] Figure 4 is a graph showing the inhibitory effect of zipalertinib on the expression of IL-6, IL-1β, IL-8, and TNF-α mRNA.

[0083] Figure 5 is a graph showing the inhibitory effect of zipalertinib on the expression of cytokines CXCL9, CXCL10, and CXCL11.

[0084] 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.

[0085] <Example>

[0086] Example 1. Confirmation of the inhibitory effect of zipalertinib on JAK3.

[0087] 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.

[0088] As a result, as shown in Fig. 1, it was confirmed that zipalertinib has excellent inhibitory efficacy against JAK3.

[0089] Example 2. Evaluation of cytotoxicity of zipalertinib against myoblast cells

[0090] Outer root sheath cells (ORS) were cultured primarily from donated human scalp. Afterwards, an immortalized cell line (SV-ORS) was established using SV40Tag, and experiments were conducted using this line. Cells were cultured using KBM (10% FBS, 1% P&S). Keratinocyte-SFM was used as the basal medium and cultured in a CO2 incubator (37°C, 5% CO2), and the medium was changed every 2–3 days. The experimental substance, zipalertinib, was dissolved in dimethyl sulfoxide (DMSO) and diluted to an appropriate concentration in the cell medium for use. DMSO was used as the control.

[0091] Cell viability when SV-ORS was treated with zipalertinib at each concentration (0, 0.1, 0.5, 1, 5, and 10 μM) is shown in Figure 2.

[0092] As a result, the cell viability was found to be over 90% when treated with zipalertinib at a concentration of 1 μM or less, and thus, it was experimentally confirmed that zipalertinib at a concentration of 1 μM or less did not have significant cytotoxicity.

[0093] Example 3. Inhibitory effect of zipalertinib on IL-6, IL-1β, IL-8, and TNF-α protein secretion

[0094] To induce an inflammatory response resembling alopecia areata, SV-ORS cells were treated with IFNγ (5 ng / ml) and double-stranded RNA, poly(I:C) (10 μg / ml). To evaluate the anti-inflammatory effect related to alopecia areata, outer root sheath cells were seeded in 12-well plates, pretreated with zipalertinib, and then treated with IFNγ and poly(I:C) and cultured for 24 hours. An enzyme-linked immunosorbent assay (ELISA) was performed using the cell culture medium, and the secretion of cytokines IL-6, IL-1β, IL-8, and TNF-α, which are involved in the development and aggravation of alopecia areata, was confirmed, and the results are shown in Fig. 3.

[0095] When SV-ORS cells were treated with IFNγ, IL-6 and IL-1β protein secretion increased. When zipalertinib (0.5 or 1 μM) was additionally treated, IL-6 and IL-1β protein secretion was significantly reduced compared to the IFNγ-only treatment group.

[0096] In addition, when SV-ORS cells were treated with IFNγ, IL-8 and TNF-α protein secretion decreased. When zipalertinib (0.5 or 1 μM) was additionally treated, IL-8 and TNF-α protein secretion was confirmed to be significantly reduced compared to the IFNγ-only treatment group.

[0097] That is, it was confirmed that zipalertinib has an inhibitory effect on the secretion of inflammatory markers IL-6, IL-1β, IL-8, and TNF-α proteins, thereby confirming that zipalertinib has anti-inflammatory efficacy.

[0098] Example 4. Inhibitory effect of zipalertinib on IL-6, IL-1β, IL-8, and TNF-α mRNA expression

[0099] After 2 hours of pretreatment with zipalertinib, IFNγ and poly(I:C) were treated for 1 hour, and RNA was extracted. Using this, qPCR was performed, and the mRNA expression of cytokines IL-6, IL-1β, IL-8, and TNF-α, which are involved in the development and aggravation of alopecia areata, was confirmed, and the results are shown in Figure 4.

[0100] When SV-ORS cells were treated with IFNγ, the expression of IL-6, IL-1β, IL-8, and TNF-α mRNA increased. When baricitinib (0.1 or 0.2 μM) or zipaltertinib (0.5 or 1 μM) was additionally treated, the expression of IL-6, IL-1β, IL-8, and TNF-α mRNA was significantly reduced compared to the IFNγ-treated group alone. In particular, zipaltertinib was confirmed to have a superior inhibitory effect on TNF-α compared to baricitinib.

[0101] That is, it was confirmed that zipalertinib has an inhibitory effect on the expression of inflammatory markers IL-6, IL-1β, IL-8, and TNF-α mRNA, thereby confirming that zipalertinib has anti-inflammatory efficacy.

[0102] Example 5. Inhibitory effect of zipalertinib on the expression of cytokines CXCL9, CXCL10, and CXCL11.

[0103] After 2 hours of pretreatment with zipalertinib, IFNγ and poly(I:C) were treated for 1 hour, and RNA was extracted. Using this, qPCR was performed, and the expression of cytokines CXCL9, CXCL10, and CXCL10, which are involved in the development and exacerbation of alopecia areata, was confirmed, and the results are shown in Figure 5.

[0104] When SV-ORS cells were treated with IFNγ, the expression of the cytokines CXCL9, CXCL10, and CXCL11 increased. When baricitinib (0.1 or 0.2 μM) or zipalertinib (0.25, 0.5, or 1 μM) was additionally treated, the expression of the cytokines CXCL9, CXCL10, and CXCL11 was significantly reduced compared to the IFNγ-only treatment group.

[0105] That is, it was confirmed that zipalertinib has an inhibitory effect on the expression of cytokines CXCL9, CXCL10, and CXCL11, thereby confirming that zipalertinib is effective in treating alopecia areata.

[0106] In summary, the present invention confirmed that zipalertinib has an inhibitory effect on JAK3. In addition, it was confirmed that it has anti-inflammatory effects by suppressing the secretion of IL-6, IL-1β, IL-8, and TNF-α proteins in SV-ORS and suppressing the expression of IL-6, IL-1β, IL-8, and TNF-α mRNA. In addition, it was confirmed that it has therapeutic efficacy for alopecia areata by suppressing the expression of cytokines CXCL9, CXCL10, and CXCL11. Through this, it was confirmed that zipalertinib has therapeutic efficacy for hair loss.

Claims

1. A pharmaceutical composition for preventing or treating hair loss, comprising zipalertinib or a pharmaceutically acceptable salt thereof.

2. A pharmaceutical composition according to claim 1, wherein the composition inhibits the activity of JAK3.

3. A pharmaceutical composition according to claim 1, wherein the composition inhibits the secretion and mRNA expression of at least one protein selected from the group consisting of IL-6, IL-1β, IL-8, and TNF-α.

4. A pharmaceutical composition according to claim 1, wherein the composition inhibits the expression of at least one cytokine selected from the group consisting of CXCL9, CXCL10, and CXCL11.

5. A pharmaceutical composition according to claim 1, wherein the hair loss is at least one selected from the group consisting of nutritional alopecia, endocrine alopecia, vascular alopecia, middle-aged alopecia, nodular alopecia, alopecia areata, neurogenic alopecia, alopecia pectina, trichotillomania, malignant alopecia, female pattern alopecia, male pattern alopecia, androgenetic alopecia, telogen alopecia, tinea capitis, alopecia totalis, hypotrichosis, hereditary hypotrichosis simplex, drug-induced alopecia universalis, mechanical alopecia, traumatic alopecia, compression alopecia, anagen alopecia, alopecia pectina, syphilitic alopecia, seborrheic alopecia, symptomatic alopecia, cicatricial alopecia, and congenital alopecia.

6. A pharmaceutical composition according to claim 5, wherein the hair loss is circular hair loss.

7. A pharmaceutical composition for preventing or improving hair loss comprising zipalertinib or a pharmaceutically acceptable salt thereof.

8. A cosmetic composition for preventing or improving hair loss comprising zipalertinib or a pharmaceutically acceptable salt thereof.

9. Use of zipalertinib or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for preventing or treating alopecia.

10. The use of claim 9, wherein the hair loss is at least one selected from the group consisting of nutritional alopecia, endocrine alopecia, vascular alopecia, middle-aged alopecia, nodular alopecia, alopecia areata, neurogenic alopecia, alopecia pectina, trichotillomania, malignant alopecia, female pattern alopecia, male pattern alopecia, androgenetic alopecia, telogen alopecia, tinea capitis, alopecia totalis, hypotrichosis, hereditary hypotrichosis simplex, drug-induced alopecia universalis, mechanical alopecia, traumatic alopecia, compression alopecia, anagen alopecia, alopecia pectina, syphilitic alopecia, seborrheic alopecia, symptomatic alopecia, cicatricial alopecia, and congenital alopecia.

11. In paragraph 9, the zipalertinib or a pharmaceutically acceptable salt thereof a) Inhibit the activity of JAK3, or b) inhibiting the secretion and mRNA expression of any one or more proteins selected from the group consisting of IL-6, IL-1β, IL-8 and TNF-α, and / or c) A use for inhibiting the expression of any one or more cytokines selected from the group consisting of CXCL9, CXCL10 and CXCL11.

12. A method for preventing or treating hair loss, comprising administering a therapeutically effective amount of zipalertinib or a pharmaceutically acceptable salt thereof to a subject in need thereof.

13. The method of claim 12, wherein the hair loss is at least one selected from the group consisting of nutritional alopecia, endocrine alopecia, vascular alopecia, middle-aged alopecia, alopecia areata, neurogenic alopecia, alopecia pectinata, trichotillomania, malignant alopecia, female pattern alopecia, male pattern alopecia, androgenetic alopecia, telogen effluvium, tinea capitis, alopecia totalis, hypotrichosis, hereditary hypotrichosis simplex, drug-induced alopecia universalis, mechanical alopecia, traumatic alopecia, compression alopecia, anagen alopecia, alopecia pectinata, syphilitic alopecia, seborrheic alopecia, symptomatic alopecia, cicatricial alopecia, and congenital alopecia.

14. In paragraph 12, the zipalertinib or a pharmaceutically acceptable salt thereof a) Inhibit the activity of JAK3, or b) inhibiting the secretion and mRNA expression of any one or more proteins selected from the group consisting of IL-6, IL-1β, IL-8 and TNF-α, and / or c) A method for inhibiting the expression of any one or more cytokines selected from the group consisting of CXCL9, CXCL10 and CXCL11.

15. Use of zipalertinib or a pharmaceutically acceptable salt thereof for the prevention or treatment of alopecia.

16. A composition comprising zipalertinib or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of alopecia.

17. A food composition for preventing or improving hair loss comprising zipalertinib or a food-chemically acceptable salt thereof.

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