Pharmaceutical composition for preventing or treating alopecia areata that acts as a functional antagonist of S1PR1 and S1PR4

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating alopecia areata, which acts as a functional antagonist against S1P receptor subtypes S1PR1 and S1PR4, and more specifically, to a pharmaceutical composition comprising as an active ingredient a sphingolipid compound that acts as a functional antagonist against S1PR1 and S1PR4, and has the effect of preventing or treating alopecia areata without causing side effects of cardiovascular disease.

Description

[Technical field]

[0001] The present invention relates to a composition for preventing or treating alopecia areata that acts as a functional antagonist against S1PR1 and S1PR4. More specifically, the present invention relates to a pharmaceutical composition comprising a sphingolipid compound that acts as a functional antagonist against S1PR1 and S1PR4, does not cause side effects of cardiovascular disease, and has the effect of preventing or treating alopecia areata. [Background technology]

[0002] Alopecia is a condition in which excessive hair loss occurs. It usually refers to hair loss from the head, but can also refer to abnormal hair loss from beards, eyebrows, pubic hair, underarm hair, and other areas.

[0003] Hair is generated from hair follicles, which are located in the dermis layer, which lies beneath the epidermis layer and above the subcutaneous fat. Hair follicles are found on all parts of the body except the lips, palms, and soles of the feet. New hairs are produced from the hair matrix at the bottom of the follicle. Living cells in the hair matrix multiply and push upwards, and as these cells rapidly dry out and die, they are compressed into a dense, hard mass that forms the hair shaft. The hair shaft, made of dead protein, is covered by a delicate layer of thin, plate-like scales (cuticle).

[0004] The hair growth cycle consists of three stages: the anagen phase, during which hair grows most actively, followed by the catagen phase, during which hair begins to degenerate, and the telogen phase, during which hair growth stops. At the end of the telogen phase, the hair falls out and a new hair grows from the hair follicle, and a new cycle begins again. The growth phase of eyebrows and eyelashes lasts about 1-6 months, while the growth phase of scalp hair lasts about 2-6 years, and generally about 50-100 hairs fall out each day as they reach the end of the telogen phase.

[0005] Alopecia areata is a type of hair loss that causes hair to fall out in a circular pattern. In the early stages, it shows one or two circular hair loss spots, but when it gets worse, it can occur in multiple locations at the same time, or the hair loss spots can merge, causing hair loss in the eyebrows and beard in addition to the scalp. Alopecia areata is an autoimmune disease in which the immune system changes and immune cells attack hair follicles, causing inflammation, and is known to be caused by stress and genetic factors. In less than 10% of patients with alopecia areata, other autoimmune diseases such as atopic dermatitis, thyroid disease, and malignant anemia may be present. It occurs in both sexes and at all ages, but is most commonly seen in children and young adults, with a prevalence of about 1-2% of the total population.

[0006] The representative treatment for alopecia areata is steroid preparations. To date, steroid injections, oral steroid drugs, immunosuppressants, etc. have been used as treatments, and although steroid preparations are effective to some extent in the early stages of treatment, they can worsen symptoms during long-term treatment, or induce side effects such as scalp inflammation, scalp depression, high blood pressure, weight gain, heartburn, and gastritis, and tend to recur.

[0007] Currently, the drugs most commonly used to treat hair loss include the FDA-approved 2,4-diamino-6-piperidinopyrimidine-3-oxide (also known as "Minoxidil"; see U.S. Pat. Nos. 4,139,619 and 4,596,812) and finasteride, a specific inhibitor of type II 5α-reductase.

[0008] Minoxidil preparations are drugs that induce hair growth by increasing blood flow through vasodilatory effects and supplying nutrients to hair roots, and are known to be particularly effective in easing hair loss symptoms at the crown of the head, and medicines that use it as an active ingredient are commercially available under the name Rogaine (a trademark of Pharmacia & Upjohn Company). Rogaine is known to reduce hair loss by up to 10% and promote hair growth in men suffering from male pattern baldness, but it has the disadvantages of having to be applied directly to the scalp from the outside and used regularly for a long period of time, and not being very effective at treating hair loss in areas other than the crown of the head.

[0009] A drug using finasteride as an active ingredient is commercially available under the name Propecia (a trademark of Merck & Co., Inc.), which is an oral pill that is known to inhibit the function of type II 5α-reductase and prevent testosterone from being converted to dihydrotestosterone (DHT), thereby inhibiting hair loss. However, this drug also requires continuous and regular administration, and some patients experience side effects such as decreased libido and erectile dysfunction. It can only be used by adult men, and is not effective for alopecia areata.

[0010] Recently, Olumiant (active ingredient baricitinib), an oral JAK inhibitor known as a treatment for rheumatoid arthritis, received FDA approval in June 2022 as the third hair loss drug and the first systemic treatment for alopecia areata. Pfizer also announced efficacy data similar to Olumiant after completing phase 3 trials of PF-06651600 (active ingredient ritlecitinib), a type of JAK inhibitor, and was recently approved by the FDA. However, research results have shown that JAK inhibitors increase the risk of heart attack, stroke, and cancer, and there is a continuing need to develop safe drugs that have excellent efficacy in treating alopecia areata but have low toxicity and side effects.

[0011] Therefore, the inventors have been making efforts to develop a pharmaceutical agent that has excellent therapeutic efficacy for alopecia areata while having less toxicity and side effects, and have confirmed that the compound according to the present invention has a preventive or therapeutic effect on alopecia areata by specifically binding to S1PR1 and S1PR4 receptors and acting as a functional antagonist, thereby completing the present invention. Summary of the Invention [Problem to be solved by the invention]

[0012] An object of the present invention is to provide a pharmaceutical composition containing, as an active ingredient, a compound having pharmacological activity for the prevention or treatment of alopecia areata, without causing side effects of cardiovascular disease.

[0013] In another aspect, the present invention aims to provide a method for preventing or treating alopecia areata, comprising administering to a subject in need thereof the composition or an active ingredient therein.

[0014] In yet another aspect, the present invention aims to provide a use of the composition or an active ingredient in the composition for the prevention or treatment of alopecia areata.

[0015] In yet another aspect, the present invention aims to provide a use of the composition or an active ingredient in the composition for the manufacture of a medicament for the prevention or treatment of alopecia areata. [Means for solving the problem]

[0016] In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating alopecia areata, comprising, as an active ingredient, a compound represented by the following chemical formula 1, an optical isomer thereof, or a pharma- ceutical acceptable salt thereof:

[0017] [ka]

[0018] Where: R 1 is hydrogen; R 2 is hydrogen or an acetyl group; X is a single bond, C 2 Alkylene, or C 2 alkenylene; A is a 5-membered heteroarylene ring containing 3 N atoms; B is C 2-11 is a straight or branched chain alkylene; C is a single bond or phenylene; and D is hydrogen, phenyl and C 1-6 It is selected from the group consisting of alkyl.

[0019] R 2 may be hydrogen.

[0020] The X is C 2 It may be an alkylene.

[0021] The B is C 2-11 It may also be a straight chain alkylene.

[0022] The compound represented by Chemical Formula 1 may be any one compound selected from the following compound group: (1) 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol; (2) 2-amino-2-(2-(1-octyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol; (3) 2-amino-2-(2-(1-(4-hexylphenethyl)-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol; (4) 2-amino-2-(1-dodecyl-1H-1,2,3-triazol-4-yl)propane-1,3-diol; (5) (E)-2-amino-2-(1-decyl-1H-1,2,3-triazol-4-yl)vinyl-1,3-diol; (6) 2-amino-2-(2-(1-(8-phenyloctyl)-1H-1,2,3-triazolebutyl-4-yl)ethyl)propane-1,3-diol; (7) N-(2-(1-dodecyl-1H-1,2,3-triazol-4-yl)-1,3-dihydroxypropan-2-yl)acetamide; (8) N-(4-(1-decyl-1H-1,2,3-triazol-4-yl)-1-hydroxy-2-(hydroxymethyl)butan-2-yl)acetamide; and (9) N-(4-(1-(4-hexylphenethyl)-1H-1,2,3-triazol-4-yl)-1-hydroxy-2-(hydroxymethyl)butan-2-yl)acetamide.

[0023] The compound represented by Chemical Formula 1 may be a compound represented by Chemical Formula 2 below (hereinafter referred to as "NXC736").

[0024] [ka]

[0025] The pharmaceutical composition according to the present invention may be a formulation for oral administration or a formulation for parenteral administration.

[0026] Pharmaceutical compositions according to the invention may further comprise one or more other therapeutic agents suitable for the treatment of alopecia areata.

[0027] The pharmaceutical compositions according to the present invention can act as functional antagonists against S1PR1 and S1PR4.

[0028] The pharmaceutical composition according to the present invention is expected to be effective in not causing side effects of cardiovascular diseases.

[0029] The present invention also provides a composition comprising the compound represented by Chemical Formula 1, an optical isomer thereof, or a salt thereof.

[0030] The present invention also provides a method for preventing or treating alopecia areata, comprising administering the composition or an active ingredient in the composition to a subject in need thereof.

[0031] The present invention also provides a use of the composition or an active ingredient in the composition for the prevention or treatment of alopecia areata.

[0032] The present invention also provides a use of the composition or an active ingredient in the composition for the manufacture of a medicament for the prevention or treatment of alopecia areata. Effect of the Invention

[0033] The pharmaceutical composition according to the present invention acts as a functional antagonist against S1PR1 and S1PR4 and has a preventive or therapeutic effect on alopecia areata.

[0034] In particular, the pharmaceutical composition according to the present invention acts as a functional antagonist against S1PR1 and S1PR4 among the S1P receptor subtypes (S1P1, S1P2, S1P3, S1P4 and S1P5), and has a preventive or therapeutic effect on alopecia areata while at the same time having no side effects related to cardiovascular disease. [Brief description of the drawings]

[0035] [Figure 1] FIG. 1 is a graph showing changes in the number of total T cells and T cell subtypes in mice after oral administration of NXC736 (3 mg / kg / day). [Diagram 2] FIG. 2 is a graph showing changes in the number of total T cells and T cell subtypes in rats after oral administration of NXC736 (10 mg / kg / day). [Diagram 3]FIG. 3 shows the results of comparing the alopecia areata lesion area (AA lesion area) between a control group and a group administered NXC736 (30 mg / kg / day) orally in an alopecia areata mouse model C3H / HeJ. [Figure 4] FIG. 4 shows the results of comparing the disease-free ratio between a control group and a group orally administered NXC736 (30 mg / kg / day) in an alopecia areata mouse model C3H / HeJ. [Diagram 5] FIG. 5 shows the results of comparing the effect of oral administration of NXC736 (30 mg / kg / day) to inhibit induction of alopecia areata in a C3H / HeJ mouse model of alopecia areata. [Figure 6] FIG. 6 is a table showing the results of an evaluation test of the effect of NXC736 (1, 3, 10, 30 μM) on HEK293 cells in which the hERG gene was overexpressed. [Figure 7] FIG. 7 is a graph showing the measurement results of heart rate in beagle dogs after oral administration of NXC736 (12.5, 25, 50 mg / kg). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0036] The descriptions disclosed in this specification or application are provided merely for the purpose of illustrating embodiments according to the technical ideas of the present invention, and the embodiments according to the technical ideas of the present invention may be implemented in various forms other than the embodiments disclosed in this specification or application, and it is not to be understood that the technical ideas of the present invention are limited to the embodiments described in this specification or application.

[0037] In the present invention, "prevention" refers to any action of administering an active ingredient to an individual to suppress or delay the onset of a target disease.

[0038] In the present invention, "treatment" refers to any action of administering an active ingredient to an individual to improve or benefit the symptoms of a target disease.

[0039] The present invention will be described in detail below.

[0040] The pharmaceutical composition for preventing or treating alopecia areata according to the present invention comprises a compound represented by the following chemical formula 1, an optical isomer thereof, or a pharma- ceutical acceptable salt thereof as an active ingredient.

[0041] [ka]

[0042] Where: R 1 is hydrogen; R 2 is hydrogen or an acetyl group; X is a single bond, C 2 Alkylene, or C 2 alkenylene; A is a 5-membered heteroarylene ring containing 3 N atoms; B is C 2-11 is a straight or branched chain alkylene; C is a single bond or phenylene; and D is hydrogen, phenyl and C 1-6 It is selected from the group consisting of alkyl.

[0043] R 2 may be hydrogen.

[0044] The X is C 2 It may be an alkylene.

[0045] The B is C 2-11 It may also be a straight chain alkylene.

[0046] The compound represented by Chemical Formula 1 may be any one compound selected from the following compound group: (1) 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol; (2) 2-amino-2-(2-(1-octyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol; (3) 2-amino-2-(2-(1-(4-hexylphenethyl)-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol; (4) 2-amino-2-(1-dodecyl-1H-1,2,3-triazol-4-yl)propane-1,3-diol; (5) (E)-2-amino-2-(1-decyl-1H-1,2,3-triazol-4-yl)vinyl-1,3-diol; (6) 2-amino-2-(2-(1-(8-phenyloctyl)-1H-1,2,3-triazolebutyl-4-yl)ethyl)propane-1,3-diol; (7) N-(2-(1-dodecyl-1H-1,2,3-triazol-4-yl)-1,3-dihydroxypropan-2-yl)acetamide; (8) N-(4-(1-decyl-1H-1,2,3-triazol-4-yl)-1-hydroxy-2-(hydroxymethyl)butan-2-yl)acetamide; and (9) N-(4-(1-(4-hexylphenethyl)-1H-1,2,3-triazol-4-yl)-1-hydroxy-2-(hydroxymethyl)butan-2-yl)acetamide.

[0047] The compound represented by Chemical Formula 1 may be 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol (also known as NXC736), which is a compound represented by Chemical Formula 2 below.

[0048] [ka]

[0049] The compound represented by Chemical Formula 1 may be used in the form of a pharma- ceutically acceptable salt. In this case, the salt may be an acid addition salt formed with a pharma- ceutically acceptable free acid.

[0050] In addition, the pharmaceutical composition according to the present invention may contain not only the compound represented by Chemical Formula 1 or a pharma- ceutically acceptable salt thereof as an active ingredient, but also a substance selected from a solvate, optical isomer, hydrate, etc. that can be produced from the compound as an active ingredient.

[0051] In the present invention, the compound represented by Chemical Formula 1, its optical isomer, or a pharma- ceutically acceptable salt thereof may be contained in a pharma- ceutically effective amount, for example, any amount within a range of 0.1 to 99.9% by weight based on the total weight of the pharmaceutical composition.

[0052] The pharmaceutical composition for preventing or treating alopecia areata, which contains the compound represented by Chemical Formula 1, its optical isomer, or a pharma- ceutically acceptable salt thereof as an active ingredient, may be modified into an oral preparation or other types of preparations within the range of exhibiting pharmacological activity.

[0053] Formulations for oral administration may include, but are not limited to, the following forms: troches, lozenges, tablets, aqueous suspensions, oily suspensions, preparative powders, granules, emulsions, hard capsules, soft capsules, syrups, or elixirs.

[0054] In addition, in order to prepare the pharmaceutical composition according to the present invention into a preparation for oral administration, binders such as lactose, sucrose, sorbitol, mannitol, starch, amylopectin, cellulose, gelatin, etc., excipients such as dicalcium phosphate, etc., disintegrants such as corn starch or sweet potato starch, etc., lubricants such as magnesium stearate, calcium stearate, sodium stearyl fumarate, polyethylene glycol wax, etc., sweeteners, flavorings, syrups, etc. may be used. In addition, in the case of capsules, in addition to the above-mentioned substances, liquid carriers such as fatty oils, etc. may be further used.

[0055] A pharmaceutical composition for preventing or treating alopecia areata, comprising the compound represented by Chemical Formula 1, an optical isomer thereof, or a pharma- ceutical acceptable salt thereof as an active ingredient, can be formulated into various dosage forms suitable for parenteral administration and used.

[0056] Preparations for parenteral administration include, but are not limited to, injections, suppositories, powders for respiratory inhalation, aerosols for spraying, ointments, powders for application, oils, creams, and the like.

[0057] In addition, in order to formulate the pharmaceutical composition according to the present invention into a preparation for parenteral administration, a sterile aqueous solution, a non-aqueous solvent, a suspension, an emulsion, a freeze-dried preparation, an external preparation, etc. can be used. Specifically, the non-aqueous solvent and the suspension can be propylene glycol, polyethylene glycol, a vegetable oil such as olive oil, an injectable ester such as ethyl oleate, etc.

[0058] When the pharmaceutical composition according to the present invention is formulated into an injection solution, it can be mixed with a stabilizer or buffer in water to prepare a solution or suspension, which can be formulated for unit administration in an ampoule or vial. When the pharmaceutical composition according to the present invention is formulated into an aerosol, it can be mixed with an additive such as a propellant so that the water-dispersed concentrate or wet powder can be dispersed. When the pharmaceutical composition according to the present invention is formulated into an ointment, cream, etc., it can be formulated using animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, etc. as a carrier.

[0059] Pharmaceutical compositions according to the invention may further comprise one or more other therapeutic agents suitable for the treatment of alopecia areata.

[0060] One or more other therapeutic agents suitable for treating alopecia areata can be administered together or separately.When administered separately, they can be administered simultaneously or sequentially in any order.The dosage and administration time of the compound of formula 1 or its pharmacologic acceptable salt and other therapeutic agents can be selected to achieve the desired combined therapeutic effect.

[0061] The pharmaceutical composition according to the present invention may further comprise a pharma- ceutically acceptable carrier. The term "pharma-ceutically acceptable" means a compound that does not stimulate an organism when administered and does not inhibit the biological activity and properties of the compound to be administered, which is commonly used in the pharmaceutical field.

[0062] The type of carrier is not particularly limited, and any carrier commonly used in the art can be used.Non-limiting examples of carriers include saline, sterilized water, Ringer's solution, buffered saline, albumin injection solution, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, maltodextrin, glycerol, ethanol, etc.These may be used alone or in combination of two or more.

[0063] If necessary, the pharmaceutical composition according to the present invention may be used by adding other pharma- ceutically acceptable additives such as excipients, diluents, antioxidants, buffers, bacteriostatic agents, etc. If necessary, the pharmaceutical composition according to the present invention may be used by additionally adding fillers, extenders, wetting agents, disintegrants, dispersants, surfactants, binders, lubricants, etc.

[0064] The pharmacologically effective amount and the dosage of the pharmaceutical composition according to the present invention to the human body can be varied depending on the formulation method, administration method, administration time and / or administration route of the pharmaceutical composition. The dosage can be varied depending on various factors including the type and degree of the reaction to be achieved by administration of the pharmaceutical composition, the type, age, weight, general health condition, symptoms and degree of disease, sex, diet, excretion, other components of the composition such as drugs used simultaneously or at different times in the individual, and similar factors widely known in the medical field. In addition, the administration route and administration method of the pharmaceutical composition according to the present invention may be independent of each other and are not particularly limited in the method. As long as the pharmaceutical composition according to the present invention is administered and the active ingredient can reach the desired site, any administration route and administration method can be followed.

[0065] The pharmaceutical composition according to the present invention can be administered orally or parenterally. For example, parenteral administration can be intravenous, intraperitoneal, intramuscular, transdermal or subcutaneous. In addition, the pharmaceutical composition according to the present invention can be applied to the diseased area, sprayed or inhaled, but is not limited thereto. In addition, the pharmaceutical composition according to the present invention can be used for the treatment of alopecia areata in a subject in need thereof. In this case, the type of subject is not particularly limited, but may be a mammal, preferably a human.

[0066] In another aspect, the present invention provides a composition comprising a compound represented by Chemical Formula 1, or an optical isomer thereof, or a salt thereof.

[0067] In still another aspect, the present invention provides a method for preventing or treating alopecia areata, comprising administering the compound represented by Chemical Formula 1, or an optical isomer thereof, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition comprising the same to an individual in need thereof. The individual may be a human or a non-human animal.

[0068] In yet another aspect, the present invention provides a use of the compound represented by Chemical Formula 1, or an optical isomer thereof, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition containing the same, for the prevention or treatment of alopecia areata.

[0069] In yet another aspect, the present invention provides a use of the compound represented by Chemical Formula 1, or an optical isomer thereof, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition containing the same, for the manufacture of a medicament for the prevention or treatment of alopecia areata.

[0070] S1PR1 receptor is mainly present in lymphocytes, and is mainly responsible for the role of regulating lymphocytes' release from immune cells.S1PR4 receptor is also present in lymphocytes, and not only acts on immune cell trafficking, but also acts on the regulation of T cells, and is involved in inflammatory reaction.Therefore, the compound of the present invention not only acts as a functional antagonist on S1PR1 receptor to inhibit lymphocyte release, but also acts on S1PR4 to show effective anti-inflammatory effect, and has the effect of preventing or treating alopecia areata.

[0071] Specifically, the compound of the present invention has the effect of preventing or treating alopecia areata by acting as a functional antagonist against the receptors through a mechanism of specifically binding to S1PR1 and S1PR4, and then influxing them into cells and eliminating them.In addition, the pharmaceutical composition according to the present invention can act as a functional antagonist against S1PR1 and S1PR4, and not cause side effects of cardiovascular disease.

[0072] The compound represented by Chemical Formula 1, its optical isomer, or a pharma- ceutically acceptable salt thereof acts as a functional antagonist of S1PR1 and S1PR4, and can prevent or treat alopecia areata without causing side effects of cardiovascular disease.

[0073] The present invention will be described in more detail with reference to the following examples. It will be obvious to those skilled in the art that these examples are merely for the purpose of illustrating the present invention and should not be construed as limiting the scope of the present invention.

[0074] Example: Preparation of the compound represented by formula 1 The compound represented by Chemical Formula 1 of the present invention can be prepared by the method described in Korean Patent Publication No. 10-2017-0087813, which is described in the following examples. However, the method for preparing the compound represented by Chemical Formula 1 of the present invention is not limited thereto, and the compound can be prepared by a method modified within the scope of modification by a person skilled in the art.

[0075] Example 1: Preparation of 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol [ka]

[0076] Step 1: Preparation of tert-butyl (2,2-dimethyl-5-((triisopropylsilyl)buta-1,3-diyn-1-yl)-1,3-dioxan-5-yl)carbamate [ka]

[0077] N 2 Under gaseous conditions, CuCl (10 mg, 0.10 mmol), NH 2 OH·HCl (1.02g, 14.67mmol), n-BuNH 2 To a solution of tert-butyl 5-ethynyl-2,2-dimethyl-1,3-dioxan-5-ylcarbamate (1.25 g, 4.89 mmol) dissolved in methanol (30 mL) was added, followed by dropwise addition of 2-bromo-1-triisopropylsilylacetylene (2.56 g, 9.78 mmol) dissolved in methanol (20 mL). The reaction mixture was stirred well at room temperature for 2 hours, after which the reaction was quenched with water and concentrated. The concentrate was diluted with ethyl acetate, washed with brine, and the organic layer was then washed with MgSO 4The mixture was dried at 40° C., filtered under vacuum, and concentrated, and purified by flash column chromatography (hexane / EtOAc, 7:1) to give the target compound as a white solid (1.85 g, 4.25 mmol, 87%).

[0078] Step 2: Preparation of tert-butyl (5-((1-decyl-1H-1,2,3-triazol-4-yl)ethynyl)-2,2-dimethyl-1,3-dioxan-5-yl)carbamate [ka]

[0079] To a solution of the compound (334 mg, 1.20 mmol) prepared in step 1 in anhydrous DMF (12 mL), 1-azidodecane (339 mg, 1.86 mmol), CuI (114 mg, 0.60 mmol), DIPEA (N,N-Diisopropylethylamine) (0.63 mL, 3.6 mmol) and AgF (182 mg, 1.44 mmol) were added. The reaction mixture was stirred well at room temperature for 12 hours, and then the reaction was diluted with saturated NH 4 After quenching with Cl, the mixture was extracted twice with ethyl acetate, and the organic extracts were washed with saturated NH 4 Cl and brine. This was then washed with MgSO 4 After drying at 40° C., the mixture was concentrated under reduced pressure and the concentrate was purified by flash column chromatography (hexane / EtOAc, 6:1) to produce the target compound as a white solid (439 mg, 0.95 mmol, 79%).

[0080] Step 3: tert-Butyl (5-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)-2,2-dimethyl-1,3-dioxan-5-yl)carbamate [ka]

[0081] The compound prepared in step 2 (149 mg, 0.32 mmol) was dissolved in methanol (30 mL) and 10% Pd / C (45 mg, 30 wt % of the compound prepared in step 2) was added. The flask was evacuated and then H 2 After filling with gas and stirring at room temperature for 1 h, the mixture was filtered through Celite and concentrated, and the precipitate was purified by flash column chromatography (hexane / EtOAc, 7:1 or 2:1) to produce the target compound as a colorless oil (110 mg, 0.24 mmol, 74%).

[0082] Step 4: Preparation of 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol [ka]

[0083] The compound prepared in step 3 (65 mg, 0.14 mmol) was dissolved in CH 2 Cl 2 To a solution of 1.40 mL of TFA was added (1.40 mL). The reaction was stirred for 12 h, and the solvent and reagents were removed under reduced pressure. The concentrate was then purified by flash column chromatography (CH 2 Cl 2 / MeOH / NH 4 OH, 100:10:1) to produce the desired compound as a white solid (36 mg, 0.11 mmol, 78%). 1 H NMR (400 MHz, CDCl 3 )δ0.86(t, J=6.8Hz, 3H), 1.23-1.29(m, 14H), 1.85(t, J=6.8Hz, 2H), 1.92(t, J=7.7Hz, 2H), 2.47(b rs, 4H), 2.78(t, J=7.8Hz, 2H), 3.57(dd, J=11.2, 26.9Hz, 4H), 4.27(t, J=7.3Hz, 2H), 7.31(s, 1H); 13 C NMR (75 MHz, CDCl 3)δ14.03, 19.2, 22.57, 26.50(2C), 28.95(2C), 29.03(2C), 30.19, 31.70, 50.42, 60.05, 63.09(2C), 121.34, 146.82;IR(CHCl 3 )υHRMS(FAB)calcd for C 17 H 35 N 4 O 2 ([M+H] + )327.2760, found 327.2762.

[0084] Example 2: Preparation of 2-amino-2-(2-(1-octyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol [ka]

[0085] The target compound was prepared by carrying out the same reaction process as in Example 1, except that 1-azidooctane was used instead of 1-azidodecane. 1 H NMR (400 MHz, CDCl 3 )δ0.86(t, J=6.8Hz, 3H), 1.24-1.29(m, 10H), 1.84-1.91(m, 4H), 2.48(brs, 4H), 2 .77(t, J=7.4Hz, 2H), 3.54(q, J=12.2Hz, 4H), 4.28(t, J=7.2Hz, 2H), 7.28(s, 1H).

[0086] Example 3: Preparation of 2-amino-2-(2-(1-(4-hexylphenethyl)-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol [ka]

[0087] The target compound was prepared by carrying out the same reaction process as in Example 1, except that 1-(2-azidoethyl)-4-hexylbenzene was used instead of 1-azidodecane.

[0088] Example 4: Preparation of 2-amino-2-(1-dodecyl-1H-1,2,3-triazol-4-yl)propane-1,3-diol [ka]

[0089] The target compound was prepared by the same procedure as in Example 1, except that bromotriisopropylsilane was used instead of 2-bromo-1-triisopropylsilylacetylene used in step 1 of Example 1, and 1-azidododecane was used instead of 1-azidododecane in step 2. 1 H NMR (300MHz, MeOD) δ0.89(t, J=6.7Hz, 3H), 1.28-1.32(m, 18H), 1.90-1.92(m, 2H), 3.89-3.98(m, 4H), 4.41(t, J=7.1Hz, 2H), 8.07(s, 1H).

[0090] Example 5: Preparation of (E)-2-amino-2-(1-decyl-1H-1,2,3-triazol-4-yl)vinyl-1,3-diol [ka]

[0091] The temperature of the compound prepared in step 2 of Example 1 dissolved in THF (5 mL) was lowered to -78°C, and then a THF solution (0.3 mmol) of LAH (lithium aluminum hydride) was slowly added dropwise. After stirring at 0°C for 3 hours, the reaction was quenched with water and then dissolved in CH 2 Cl 2 The organic layer was washed with brine and extracted twice with MgSO 4The concentrate was purified by flash column chromatography (CH 2 Cl 2 / MeOH, 20:1) to give the target compound as a white solid. The reaction was then carried out in the same manner as in step 4 of Example 1 to give the target compound (24 mg, 0.074 mmol, 74%).

[0092] Example 6: Preparation of 2-amino-2-(2-(1-(8-phenyloctyl)-1H-1,2,3-triazolbutyl-4-yl)ethyl)propane-1,3-diol [ka]

[0093] The target compound was prepared by the same procedure as in Example 1, except that 8-azidooctylbenzene was used instead of 1-azidodecane used in step 2 of Example 1.

[0094] Example 7: Preparation of N-(2-(1-dodecyl-1H-1,2,3-triazol-4-yl)-1,3-dihydroxypropan-2-yl)acetamide [ka]

[0095] The compound prepared in Example 4 (50 mg, 1 eq, 0.15 mmol) was dissolved in acetic anhydride (0.2 mL), MeOH (1 mL), saturated NaHCO 3 Aqueous solution (1 mL) was added and stirred for 30 min, then concentrated and the reaction was quenched with aqueous ammonium chloride solution and then added to CH 2 Cl 2 Extract twice with MgSO 4 The mixture was dried at 40° C. and concentrated to give the desired compound in 98% yield (54 mg, 98%) without any additional purification steps. 1H NMR (300MHz, MeOD) δ0.89(t, J=6.7Hz, 3H), 1.28-1.32(m, 18H), 1.86-1.96(m, 2H), 2.00(s, 3H), 4.02(q, J=11.2Hz, 4H), 4.35(t, J=7.2Hz, 2H), 7.84(s, 1H).

[0096] Example 8: Preparation of N-(4-(1-decyl-1H-1,2,3-triazol-4-yl)-1-hydroxy-2-(hydroxymethyl)butan-2-yl)acetamide [ka]

[0097] The compound prepared in Example 1 was used as a starting material and the acetylation reaction was carried out in the same manner as in Example 7 to prepare the target compound. 1 H NMR (300MHz, MeOD) δ0.86(t, J=6.8Hz, 3H), 1.23-1.32(m, 14H), 1.86-1.92(m, 2H), 1.94(s, 3H), 1 .99-2.02(m, 2H), 2.47(m, 2H), 3.62(dd, J=11.2, 26.9Hz, 4H), 4.31(t, J=7.3Hz, 2H), 7.31(s, 1H).

[0098] Example 9: Preparation of N-(4-(1-(4-hexylphenethyl)-1H-1,2,3-triazol-4-yl)-1-hydroxy-2-(hydroxymethyl)butan-2-yl)acetamide [ka]

[0099] The compound prepared in Example 3 was used as a starting material and the acetylation reaction was carried out in the same manner as in Example 7 to prepare the target compound.

[0100] Experimental Example 1: Confirmation of the inhibitory effect of NXC736 on mouse immune cells FTY720 (Fingolimod) is a multiple sclerosis treatment with a mechanism of action that involves reversibly capturing a portion of lymphocytes from lymph nodes or bone marrow, isolating them in secondary lymphoid organs and preventing them from entering the central nervous system, or reducing the number of lymphocytes circulating in the bloodstream, thereby reducing the number of activated lymphocytes that reach the brain.

[0101] To confirm whether NXC736, which acts on the S1P receptor, also has an effect on lymphocyte reduction, we evaluated the effect of NXC736 on immune cells. After a single oral administration of 3 mg / kg / day of NXC736 to C57BL / 6 mice (DBL, control group: 4 mice, test group: 18 mice), changes in lymphocytes in the peripheral blood (PB) and bone marrow (BM) were measured by classifying them into T cells (using anti-mouse CD3ε-PE / cyanine7 antibody), CD4 T cells (using anti-mouse CD4-FITC antibody), and CD8 T cells (using anti-mouse CD8a-PerCP antibody). Specifically, after administration of the test substance, measurements were performed using a flow cytometer (FC500, Beckman Coulter) at 0, 4, 8, 12, 24, and 36 hours, and analysis was performed using FlowJoTM V10 (Flowjo, LLC). The numbers of T cells, CD4 T cells, and CD8 T cells were expressed as a percentage (% of control) based on the number at 0 hour.

[0102] Referring to Figure 1, it was confirmed that administration of 3 mg / kg of NXC736 in the peripheral blood (PB) reduced T cells, CD4 T cells, and CD8 T cells to levels of 40.6-41.6%, 16.4-17.6%, and 5.9-9.3%, respectively, after 4 hours. In the case of T cells, the reduction was maintained for up to 12 hours, after which it gradually recovered, recovering to 88.9% at 36 hours.

[0103] In bone marrow (BM), administration of 3 mg / kg of NXC736 increased T cell counts to 120-135% after 4 hours and 118-131% after 24 hours. This is believed to be due to receptor internalization caused by NXC736, which inhibits T cell release.

[0104] After administration of 3mg / kg of NXC736, the cell number of T cells in the peripheral blood (PB) steadily decreased for up to 12 hours, and the inhibitory effect was maintained to some extent until 24 hours, at which point recovery was observed. In conclusion, it was confirmed that administration of NXC736 reduces the number of lymphocytes circulating in the bloodstream, resulting in an immunosuppressive effect.

[0105] Experimental Example 2: Confirmation of the inhibitory effect of NXC736 on rat immune cells To evaluate the effect of NXC736 on rat immune cells, NXC736 was orally administered once at 10 mg / kg / day to SD rats (Daehan Biolink (DBL), control group 3 rats, test group 15 rats), and then T cells, CD4 T cells, and CD8 T cells in the peripheral blood flow (PB) and bone marrow (BM) were classified and measured using the same antibodies as in Experimental Example 1. Specifically, after administration of the test substance, measurements were performed using a flow cytometer (FC500, Beckman Coulter) at 0, 4, 8, 12, 24, 36, and 48 hours, and analysis was performed using FlowJoTM V10 (Flowjo, LLC) to measure the numbers of T cells, CD4 T cells, and CD8 T cells, and the results were expressed as a percentage (% of control) based on 0 time.

[0106] Referring to Figure 2, after administration of NXC736, a rapid decrease in T cells was observed in the peripheral blood (PB) for up to 4 hours. The immunosuppressive effect was observed to be maintained for up to 24 hours with administration of 10 mg / kg of NXC736. Conversely, no change in lymphocyte count or a tendency for it to increase was observed in the bone marrow (BM).

[0107] Through this study, it was confirmed that NXC736, after oral administration, exhibits an immunosuppressive effect by decreasing the number of lymphocytes in the peripheral blood (PB). In addition, in the bone marrow (BM), it was confirmed that the number of lymphocytes remained unchanged or tended to increase. It is believed that this effect is related to the mechanism by which NXC736 acts as a functional antagonist on the S1P receptors present in lymph nodes and bone marrow (BM), suppressing lymphocyte release.

[0108] Experimental Example 3: Confirmation of efficacy in suppressing alopecia areata using a C3H / HeJ alopecia areata mouse model The efficacy of NXC736 against alopecia areata was evaluated using a mouse model of alopecia areata, C3H / HeJ (Jackson Labs, Theron Bio). C3H / HeJ mice are experimental animals in which alopecia areata naturally progresses with age. Lymph node cells obtained from C3H / HeJ mice over 20 weeks old in which alopecia areata was naturally induced were expanded and injected into 10-16 week old C3H / HeJ mice in which alopecia areata was not induced to induce alopecia areata. The conditions of the animal breeding room were maintained at a temperature of 23±3℃, relative humidity of 50±10%, and night and day were maintained at 12-hour intervals. The experimental animals were kept in an isolation breeding box (ThreeShine) and were given experimental animal feed and water ad libitum.

[0109] Mice were divided into two groups, a control group (treated with sterile distilled water) and an experimental group treated with NXC736, with five C3H / HeJ mice randomly allocated to each group. Lymph node cells were expanded and injected into 10-16 week-old C3H / HeJ mice in which alopecia areata had not been induced. From that point on, 30mg / kg of NXC736 was orally administered to the NXC736 treatment group and the same volume of sterile distilled water to the control group once a day at the same time for 12 weeks, and the AA lesion area and disease-free ratio were measured at 2-week intervals.

[0110] The area of ​​the alopecia areata lesion was measured using the Image J program (Version 1.44p) after taking a photograph.

[0111] Referring to FIG. 3, the area of ​​alopecia areata lesions (AA lesion area) was about 80% in the control group and about 10% in the NXC736 administration group, and it was confirmed that the area of ​​alopecia areata lesions (AA lesion area) was significantly smaller in the NXC736 administration group than in the control group.

[0112] Disease-free rates were defined as the percentage of drug-treated mice that showed no alopecia symptoms at the designated time points.

[0113] Referring to Figure 4, the disease-free ratio results show that it took 56 days for 100% alopecia areata to be observed in the control group, whereas it took 84 days in the NXC736-administered group, demonstrating that administration of NXC736 delayed the progression of alopecia areata.

[0114] In addition, referring to FIG. 5, the effect of inhibiting induction of alopecia areata by oral administration of NXC736 was compared with that of the control group on the 84th day after drug administration. As a result, it was confirmed by visual observation that the progression of alopecia areata was significantly delayed when NXC736 was administered.

[0115] For reference, no significant difference in body weight was observed between the control group and the NXC736-treated group.

[0116] JPEG0007607376000018.jpg48169

[0117] On the day of the experiment, hERG channel currents were measured, and cells that were measured at more than 500 pA and less than 3000 pA at the -50 mV repolarization step were selected. Then, cells were re-adjusted to the external bath solution (137 mM NaCl, 4 mM KCl, 10 mM HEPES, 1 mM MgCl 2 , 10mM D-glucose, 1.8mM CaCl 2After treatment for 9 minutes with various concentrations of the test substances dissolved in 100 mM NaCl (pH 7.4), the channel current was recorded using the Notocord program (Notocord System, France).

[0118] As a negative control, recordings were made using only the external bath solution, and as a positive control, a random cell was treated with 0.1 μM positive control substance (E-4031 n-Hydrate, FUJIFILM Wako Pure Chemical Corporation, Japan) after washing, and the suitability of the cell line was confirmed by recording and confirming that hERG channel currents were suppressed.

[0119] Referring to Figure 6, the suppression rates (%) of hERG channel currents of NCX736 (B-E groups) at concentrations of 1, 3, 10 and 30μM were 9.94±1.96, 13.98±4.48, 35.03±12.09 and 82.62±7.31% (n=3), respectively, and the compensated suppression rate (%) of hERG channel currents of the negative control group (A group) was 0±4.12 (n=3). That is, it was confirmed that NCX736 (D and E groups) at concentrations of 10 and 30μM showed a statistically significant difference compared to the negative control group (A group).

[0120] In particular, the test substance NXC736 was treated at a concentration of up to 30 μM, resulting in a compensated suppression rate (%) of hERG channel currents of 82.62 ± 7.31%, and K + The ion channel inhibitory potency (IC50) was calculated to be 12.94 μM (Hill coefficient: 1.527).

[0121] In addition, when E-4031 was treated at a concentration of 0.1 μM as a positive control group (F group) under the same conditions, the compensated suppression rate (%) of hERG channel currents was as high as 92.64 ± 1.66% (n = 5).

[0122] These results confirm that this test method is suitable for evaluating the effects of NCX736 on hERG channel currents, and that NCX736 is a substance that does not inhibit hERG potassium channel activity and does not induce cardiac abnormalities.

[0123] 2) Confirmation of effects on the cardiovascular system in beagle dogs NXC736 was orally administered to four unanesthetized and unrestrained male beagle dogs (ORIENTBIO Inc., Republic of Korea) with remote transmitters inserted.The heart rates of the four beagle dogs were then measured to evaluate the effects of NXC736 on the cardiovascular system, and the occurrence of any abnormal symptoms was visually confirmed.

[0124] In this experiment, four beagle dogs were administered a 0.5% MC aqueous solution as a control substance, and their heart rates were measured. One week later, 12.5 mg / kg of NXC736 was administered and their heart rates were measured. One week later, 25 mg / kg of NXC736 was administered and their heart rates were measured. One week later, 50 mg / kg of NXC736 was administered and their heart rates were measured.

[0125] Specifically, the heart rates of the beagle dogs were measured 0 hours before administration of the experimental substance, and 0.5 hours, 1 hour, 2 hours, 3 hours, 4 hours, 6 hours, 8 hours, 12 hours, and 24 hours after administration of the experimental substance.

[0126] Referring to FIG. 7, it was confirmed that no change in heart rate was observed when NXC736 was administered at doses of 12.5, 25, or 50 mg / kg. Additionally, visual observation of the general symptoms in beagle dogs revealed no abnormal symptoms following administration of NXC736 at doses of 12.5, 25, or 50 mg / kg.

[0127] As described above, according to the evaluation results of the efficacy of suppressing alopecia areata using C3H / HeJ alopecia areata mouse model and the results of confirming the presence or absence of abnormal symptoms by heart rate and visual observation of beagle dogs administered with NXC736, the pharmaceutical composition comprising the compound represented by formula 1 according to the present invention, its optical isomer or its pharma- ceutically acceptable salt as an active ingredient, can be confirmed to have a preventive or therapeutic effect on alopecia areata by acting as a functional antagonist against S1PR1 and S1PR4.In addition, it can be confirmed that the pharmaceutical composition according to the present invention has a preventive or therapeutic effect on alopecia areata by acting as a functional antagonist against S1PR1 and S1PR4 among the subtypes of S1P receptors (S1P1, S1P2, S1P3, S1P4 and S1P5), and at the same time has an effect of not causing side effects of cardiovascular disease.

Claims

1. A pharmaceutical composition for preventing or treating alopecia areata, comprising as an active ingredient a compound represented by the following chemical formula 1, an optical isomer thereof, or a pharma- ceutical acceptable salt thereof: 【Chemistry 1】 Where: R １ is hydrogen; R ２ is hydrogen or an acetyl group; X is a single bond, C ２ Alkylene, or C ２ alkenylene; A is a 5-membered heteroarylene ring containing 3 N atoms; B is C ２－１１ is a straight or branched chain alkylene; C is a single bond or phenylene; and D is hydrogen, phenyl and C １－６ selected from the group consisting of alkyl, A pharmaceutical composition for preventing or treating alopecia areata, wherein the compound represented by Chemical Formula 1 is any one compound selected from the following compound group: (1) 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol; (2) 2-amino-2-(2-(1-octyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol; (3) 2-amino-2-(2-(1-(4-hexylphenethyl)-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol; (4) 2-amino-2-(1-dodecyl-1H-1,2,3-triazol-4-yl)propane-1,3-diol; (5) (E)-2-amino-2-(1-decyl-1H-1,2,3-triazol-4-yl)vinyl-1,3-diol; (6) 2-amino-2-(2-(1-(8-phenyloctyl)-1H-1,2,3-triazolebutyl-4-yl)ethyl)propane-1,3-diol; (7) N-(2-(1-dodecyl-1H-1,2,3-triazol-4-yl)-1,3-dihydroxypropan-2-yl)acetamide; (8) N-(4-(1-decyl-1H-1,2,3-triazol-4-yl)-1-hydroxy-2-(hydroxymethyl)butan-2-yl)acetamide; and (9) N-(4-(1-(4-hexylphenethyl)-1H-1,2,3-triazol-4-yl)-1-hydroxy-2-(hydroxymethyl)butan-2-yl)acetamide.

2. The pharmaceutical composition for preventing or treating alopecia areata according to claim 1, wherein the compound represented by Chemical Formula 1 is a compound represented by Chemical Formula 2: 【Chemistry 2】

3. The pharmaceutical composition for preventing or treating alopecia areata according to claim 1 or 2, characterized in that the pharmaceutical composition is a formulation for oral administration or a formulation for parenteral administration.

4. The pharmaceutical composition for preventing or treating alopecia areata according to claim 1 or 2, characterized in that the pharmaceutical composition acts as a functional antagonist against S1PR1 and S1PR4.

5. The pharmaceutical composition for preventing or treating alopecia areata according to claim 4, characterized in that the pharmaceutical composition does not cause side effects such as cardiac dysfunction.

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