Novel salts of 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol and pharmaceutical compositions containing same

Abstract

The present invention relates to novel salts of 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol and pharmaceutical compositions containing same. The novel salt of 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol according to the present invention exhibits excellent effects in terms of low hygroscopicity, stability of standard stock solution, photostability, oxidation stability, pH stability, solubility, etc., as compared with hydrochloride and other salts of 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol. The pharmaceutical composition of the present invention containing the novel salt as an active ingredient is effective in treating multiple sclerosis, ischemic storke, focal segmental glomerulosclerosis (FSGS), inflammatory bowel disease, interstitial fibrosis and tubular atrophy. The present invention can be useful in the prevention or treatment of alopecia areata (IFTA) or alopecia areata (AA).

Description

[Technical field]

[0001] The present invention relates to a novel salt of 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol and a pharmaceutical composition containing the same for a novel use. More specifically, the present invention relates to a fumarate or hemifumarate salt of 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol and a pharmaceutical composition containing the same for a novel use. [Background technology]

[0002] 2-Amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol compound (hereinafter referred to as compound 1) selectively binds to each receptor of S1P subtype and can be useful for preventing or treating multiple sclerosis or cerebral ischemia.

[0003] However, Compound 1 is highly hygroscopic and physically unstable after exposure to high humidity conditions, and no new salt has been found that is more stable than the free base or hydrochloride of Compound 1. Therefore, there is a need to develop a new salt of the compound that overcomes the above-mentioned shortcomings of the free base and the hydrochloride, has excellent solid-state properties in the natural state, has reduced hygroscopicity, maintains a constant purity even during long-term storage, and is easy to formulate. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Korean Patent Publication No. 10-2017-0087813 (July 31, 2017) Summary of the Invention [Problem to be solved by the invention]

[0005] The object of the present invention is to provide a novel salt of 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol (compound 1) which has reduced hygroscopicity in the natural state, maintains a constant purity even during long-term storage, and has improved stability, is easy to develop as a drug, is amenable to mass production, and is obtained in a high-purity, homogeneous quality; a method for producing the same; and a pharmaceutical composition containing the same as an active ingredient and having a novel use. [Means for solving the problem]

[0006] In order to solve the above problems, the present invention provides a hemifumarate salt (Compound 1a, NXC736) of 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol (Compound 1), which is a compound represented by the following chemical formula 1, and a pharmaceutical composition containing the same as an active ingredient.

[0007] [ka]

[0008] In one embodiment of the invention, the hemifumarate salt is in crystalline form.

[0009] In one embodiment of the present invention, compound 1a exhibits an X-ray powder diffraction spectroscopy (XRPD) pattern with major diffraction peaks at 2θ values ​​of 6.3±0.2°, 9.4±0.2°, 18.5±0.2°, 19.0±0.2°, 19.9±0.2°, 20.7±0.2°, 25.5±0.2°, 28.7±0.2°, and 29.0±0.2°.

[0010] In one embodiment of the present invention, compound 1a exhibits a melting endotherm onset temperature of about 82° C. (82.42° C.) with an onset melting point in differential scanning calorimetry (DSC).

[0011] In one embodiment of the present invention, compound 1a exhibits a water adsorption rate of less than 2% at 90% RH in DVS.

[0012] In one embodiment of the present invention, compound 1a maintains its crystalline form and purity of 99% or more even after 2 weeks under high humidity conditions (40°C / 75±5% RH or 60°C / 75±5% RH).

[0013] In one embodiment of the present invention, compound 1a exhibits a purity of 98% or more after 2 weeks in a phosphate buffer having a pH of 6.8 at 40±2° C.

[0014] In one embodiment of the present invention, compound 1a may exist as a crystalline composition.

[0015] In one embodiment of the present invention, there is provided a pharmaceutical composition comprising compound 1a as an active ingredient, which may further comprise one or more pharma- ceutically acceptable carriers, diluents and / or excipients.

[0016] The present invention also provides a fumarate salt (Compound 1b) of 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol (Compound 1), which is a compound represented by the following chemical formula 1, and a pharmaceutical composition containing the same as an active ingredient.

[0017] [ka]

[0018] In one embodiment of the invention, the Fumarate Salt is in a crystalline form.

[0019] In one embodiment of the present invention, compound 1a exhibits an X-ray powder diffraction spectroscopy (XRPD) pattern with major diffraction peaks at 2θ values ​​of 13.9±0.2°, 18.9±0.2°, 19.4±0.2°, 22.1±0.2°, and 22.5±0.2°.

[0020] In one embodiment of the present invention, compound 1a exhibits a moisture adsorption rate of more than 3% and less than 5% under conditions of 90% RH in DVS.

[0021] In one embodiment of the present invention, the pharmaceutical composition of the present invention comprising a novel salt of compound 1 according to the present invention, for example, a hemifumarate or fumarate salt of compound 1, as an active ingredient, can be useful for preventing or treating multiple sclerosis, ischemic storke, focal segmental glomerulosclerosis (FSGS), inflammatory bowel disease, interstitial fibrosis and tubular atrophy (IFTA), or alopecia areata (AA).

[0022] In one embodiment of the invention, the inflammatory bowel disease may be ulcerative colitis (UC) or Crohn's disease (CD).

[0023] Among the above-mentioned medicinal uses, the pharmacological effects on multiple sclerosis and cerebral ischemia are described in Korean Patent Registration No. 10-1830244, and the pharmacological effects on other medicinal uses will be described later.

[0024] Those skilled in the art of organic chemistry will appreciate that many organic compounds can form complexes with solvents in which they are reacted or from which they are precipitated or crystallized. Such complexes are known as "solvates." For example, complexes with water are known as "hydrates." Pharmaceutically acceptable solvates of the compounds of the present invention are within the scope of the present invention.

[0025] The novel salt of compound 1 according to the present invention exhibits excellent effects in terms of low hygroscopicity, stability of standard stock solution, photostability, oxidation stability, pH stability, solubility, etc., compared with the hydrochloride salt and other salts of compound 1, and can be stably maintained for a long period of time without change in content or increase in impurities.

[0026] The novel salt of compound 1 according to the present invention can be obtained from high purity raw materials, and has the advantage of being able to maintain high purity for a long period of time due to the extremely low increase in softening materials even during long-term storage, and exhibiting low hygroscopicity in the range of relative humidity. Therefore, it is very advantageous in processing and storing pharmaceuticals, and is easy to formulate, and even after preparations containing it are produced, the same state is maintained, and the uniformity of the formulation content can be stably maintained for a long period of time, making it easy to apply to mass production.

[0027] The novel salt of compound 1 of the present invention was orally administered to mice and rats, and was confirmed to have an immunosuppressive effect. It also showed excellent anti-fibrotic effects in a UUO (Unilateral Urinary Obstruction) renal fibrosis model and an adenine-induced renal fibrosis model. Therefore, it can be usefully used as an active ingredient of a pharmaceutical composition that can prevent or treat multiple sclerosis, cerebral ischemia, focal segmental glomerulosclerosis, inflammatory bowel disease, interstitial fibrosis, tubular atrophy, alopecia areata, etc.

[0028] In one embodiment of the present invention, the novel salt of Compound 1 contained in the above-mentioned pharmaceutical composition may be in a crystalline form, more specifically, a crystalline form exhibiting an X-ray powder diffraction spectroscopy (XRPD) pattern with major diffraction peaks at 2θ values ​​of 6.3±0.2°, 9.4±0.2°, 18.5±0.2°, 19.0±0.2°, 19.9±0.2°, 20.7±0.2°, 25.5±0.2°, 28.7±0.2°, and 29.0±0.2°.

[0029] In one embodiment of the present invention, the pharmaceutical composition of the present invention may further include one or more pharma- ceutically acceptable carriers in addition to the novel salt of Compound 1, which is an active ingredient, for administration. The pharma-ceutically acceptable carriers may be saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, or a mixture of one or more of these components, and other conventional additives such as antioxidants, buffers, bacteriostatic agents, etc. may be added as necessary. In addition, the composition of the present invention may be formulated into injection forms such as aqueous solutions, suspensions, emulsions, pills, capsules, granules, or tablets by additionally adding diluents, dispersants, surfactants, binders, and lubricants. Thus, the composition of the present invention may be a patch, liquid, pill, capsule, granule, tablet, suppository, etc. These preparations may be prepared by conventional methods used in the art for formulation or by methods disclosed in Remington's Pharmaceutical Science (latest edition), Mack Publishing Company, Easton PA, and may be formulated into various preparations according to each disease or ingredient.

[0030] In one embodiment of the present invention, the pharmaceutical composition of the present invention can be administered orally or parenterally, for example, intravenously, subcutaneously, intraperitoneally or topically, depending on the desired method, and the dosage range varies depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and severity of the disease.

[0031] In one embodiment of the present invention, there is provided a method for treating multiple sclerosis, cerebral ischemia, focal segmental glomerulosclerosis, inflammatory bowel disease, interstitial fibrosis and renal tubular atrophy or alopecia areata, comprising administering to an individual a pharma- ceutical effective amount of a novel salt of compound 1 according to the present invention.

[0032] In one embodiment of the present invention, the novel salt of Compound 1 according to the present invention used in the method may be in a crystalline form, more specifically, in a crystalline form exhibiting an X-ray powder diffraction spectroscopy (XRPD) pattern with major diffraction peaks at 2θ values ​​of 6.3±0.2°, 9.4±0.2°, 18.5±0.2°, 19.0±0.2°, 19.9±0.2°, 20.7±0.2°, 25.5±0.2°, 28.7±0.2°, and 29.0±0.2°.

[0033] In one embodiment of the present invention there is provided the use of the novel salts of compound 1 according to the invention in the manufacture of a medicament for the treatment of multiple sclerosis, cerebral ischemia, focal segmental glomerulosclerosis, inflammatory bowel disease, interstitial fibrosis and renal tubular atrophy or alopecia areata.

[0034] In one embodiment of the present invention, the novel salt of compound 1 according to the present invention for the above-mentioned use may be in a crystalline form, more specifically, in a crystalline form exhibiting an X-ray powder diffraction spectroscopy (XRPD) pattern with major diffraction peaks at 2θ values ​​of 6.3±0.2°, 9.4±0.2°, 18.5±0.2°, 19.0±0.2°, 19.9±0.2°, 20.7±0.2°, 25.5±0.2°, 28.7±0.2°, and 29.0±0.2°. Effect of the Invention

[0035] The novel salt of 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol (compound 1) according to the present invention exhibits superior effects in terms of hygroscopicity, stability of standard stock solution, photostability, oxidation stability, pH stability, solubility, etc., as compared with the hydrochloride salt and other salts of compound 1, and the pharmaceutical composition of the present invention containing the novel salt as an active ingredient can be usefully used for the prevention or treatment of multiple sclerosis, cerebral ischemia, focal segmental glomerulosclerosis, inflammatory bowel disease, interstitial fibrosis, and tubular atrophy or alopecia areata. [Brief description of the drawings]

[0036] [Figure 1] 1 shows DSC data of compound 1a. [Diagram 2] 1 shows DVS data for compound 1a. [Diagram 3] XRPD data of compound 1a. [Figure 4] 1 is a graph showing changes in the number of total T cells and T cell subtypes in mice after oral administration of Compound 1a (3 mg / kg / day). [Diagram 5] 1 is a graph showing changes in the number of total T cells and T cell subtypes in rats after oral administration of Compound 1a (10 mg / kg / day). [Figure 6] 1 is a graph showing the effect of compound 1a (5, 10 mg / kg / day) on suppressing inflammatory bowel disease in a mouse model of DSS-induced colitis. [Figure 7] 1 is a table showing the results of an evaluation test of the effect of compound 1a (1, 3, 10, 30 μM) on HEK293 cells in which the hERG gene was overexpressed. [Figure 8] 1 is a graph showing the results of measuring the heart rate in beagle dogs after oral administration of compound 1a (12.5, 25, 50 mg / kg). [Figure 9] 1 is a graph showing the effect of compound 1a (10, 30, 90 mg / kg / day) on renal fibrosis in a UUO mouse model. [Figure 10] 1 is a graph showing the effect of compound 1a (10, 30, 90 mg / kg / day) on renal fibrosis in an adenine-induced CKD mouse model. [Figure 11] 1 shows photographs of the free base of Compound 1, Compound 1a, and the hydrochloride of Compound 1, each of which was placed under conditions of 60° C. / 75% RH, and the changes in properties were observed initially and after 2 weeks. [Figure 12] 1 is a photograph showing the degree of corrosivity of a sample after storing the free base of Compound 1, Compound 1a, and the hydrochloride of Compound 1 together with the sample at 60° C. for two weeks. [Figure 13] Compound 1a was orally administered at 30 mg / kg for 12 weeks to 10-16 week-old C3H / HeJ mice in which alopecia areata had been induced, and the AA lesion area was then measured. [Figure 14] The results are shown in Table 1. Compound 1a was orally administered at 30 mg / kg for 12 weeks to 10-16 week-old C3H / HeJ mice in which alopecia areata had been induced, and the disease-free ratio was then measured. [Figure 15] Photographs comparing the induction of alopecia areata after administration of compound 1a to 10-16 week old C3H / HeJ mice in which alopecia areata had been induced. [Figure 16] 1 shows DSC data of compound 1b. [Figure 17] 1 shows DVS data of compound 1b. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0037] General Definitions and Terminology Unless specifically defined otherwise, ratios (including percentages) or parts are calculated herein on a weight basis.

[0038] As used herein, the term "comprise" or its cognate terms "containing," "including," or "having" is open-ended and does not exclude elements, steps or ingredients not otherwise stated.

[0039] Use of the term "about" includes and describes a value or parameter by itself. For example, "about x" includes and describes "x" by itself. In some embodiments, the term "about" when used in connection with a measurement or to define a value, unit, constant, or range of values, refers to a deviation of ±1% to ±10%, unless otherwise stated. For example, in some embodiments, "about 82°C" includes 82°C to 83°C.

[0040] The term "pharmaceutical composition" refers to an active ingredient, optionally combined with one or more pharma- ceutically acceptable ingredients (such as, without limitation, carriers and / or excipients), exemplified by the compound of formula I or a hemifumarate salt thereof, one or more crystalline forms according to the present invention, or one or more crystalline compositions according to the present invention.

[0041] The terms "administration" or "administering" and the like refer to methods that allow for the delivery of a compound or composition to a desired site of biological action, including, but not limited to, oral, parenteral (including intravenous, subcutaneous, intraperitoneal, intramuscular, intravascular injection or infusion), topical, rectal administration, and the like.

[0042] For a pharmaceutical or pharmacologically active agent, the term "effective amount" refers to an amount of the drug or agent sufficient to achieve a desired effect while not being toxic. In this application, an "effective amount" of an active agent in a composition in the context of an oral dosage form refers to the amount required to achieve a desired effect together with other active agents in the composition. An effective amount can be determined on an individual basis and will vary depending on the recipient as well as the age and general conditions of the particular active agent. In a particular case, the effective amount can be determined by one skilled in the art through routine testing.

[0043] The terms "active ingredient," "therapeutic agent," "active substance," or "active agent" refer to a chemical entity useful for effectively treating or preventing a target disorder, disease, or condition. As used herein, such terms can refer to, for example, a compound of Formula I or a hemifumarate salt thereof, one or more crystalline forms according to the present invention, or one or more crystalline compositions according to the present invention.

[0044] The term "low hygroscopicity" as used in the present invention means the property of a material not absorbing or adsorbing moisture from the air, and is a concept that is contrasted with hygroscopicity, which attracts and retains water molecules from the material's surrounding environment.

[0045] The term "stability" used in the present invention refers to the degree to which the salt of the compound represented by Chemical Formula 1 of the present invention or a composition containing the same is chemically or physically stable under specific conditions and for a certain period of time. The purpose of the stability test is to provide evidence of how the quality of the salt of the present invention changes over time under the influence of various environmental factors such as temperature, humidity, and light, and subsequently to set the usage period (or expiration period) and recommended storage conditions of the finished drug when manufacturing a drug using the same.

[0046] The Korean Food and Drug Safety Agency's notification on the stability testing standards for pharmaceuticals and other products stipulates "severe tests" to confirm the decomposition process and decomposition products of pharmaceuticals and other products under severe conditions, and states that the test conditions for light, temperature, and humidity should be set taking into consideration the characteristics of the specimen. If the specimen is a raw material, test conditions for the aqueous solution state should be included, and the effects of oxidation and photolysis on the raw pharmaceutical material and sensitivity to hydrolysis depending on the pH range should be evaluated.

[0047] Accordingly, the "stability of standard stock solution" among the stability test criteria of the present invention was determined by dissolving the salt of the present invention in a standard stock solution, storing it under specific temperature and humidity conditions for a specific time and period, and then checking the purity and measuring the presence or absence of softening substances (impurities). In addition, the "photostability", "oxidative stability" and "stability according to pH" of the salt of the present invention were evaluated according to specific conditions specified in the following examples.

[0048] In X-ray powder diffraction (XRPD or XRD) spectra, the diffraction pattern obtained from a crystalline compound is generally characteristic of a particular crystal form, where the relative intensities of bands (especially at low angles) vary due to orientation effects resulting from differences in crystallization conditions, particle size, and other measurement conditions. Thus, the relative intensities of the diffraction peaks are not characteristic of a given crystal form. When determining whether a crystal form is the same as one known in the art, it is more important to note the relative positions of the peaks rather than their relative intensities. In addition, there may be some errors in the peak positions for any given crystal form, which is also well known in the crystallography field. For example, the positions of the peaks may shift during sample analysis due to changes in temperature, sample movement, or instrument calibration; the measurement error of the 2θ values ​​may sometimes be about ±0.2°, typically about ±0.1°. Thus, such errors must be taken into account when determining crystal structures. If a crystalline form according to the invention is described substantially as shown in a drawing, the term "substantially" is also intended to include such differences in diffraction peaks.

[0049] In XRPD patterns, the peak positions are generally expressed as 2θ or the crystal surface distance d, and the simple conversion between d and θ is d=λ / 2sinθ, where d is the crystal surface distance, λ is the wavelength of the incident X-ray, and θ is the diffraction angle. In the same crystal form of the same compound, the peak positions of the XRPD pattern are generally similar, and the relative intensity error may be large. It should also be noted that in identifying mixtures, some diffraction lines may disappear due to factors such as content reduction, and therefore it is not necessary to rely on the total bands observed in high purity samples, and one band is characteristic for a given crystal.

[0050] Differential scanning calorimetry (DSC) is used to measure the transition temperatures at which crystals absorb and release heat due to changes in their crystal structure or when the crystal melts. The error in thermal transition temperatures and melting points is typically within about 5°C, and generally within about 3°C, for the same crystalline form of the same compound in successive analyses. When a compound is described as having a given DSC peak or melting point, this means ±5°C of the DSC peak or melting point. A complementary method is provided by DSC to distinguish different crystalline forms. Different crystalline forms can be distinguished by their different transition temperatures. The DSC peak or melting point of a mixture can vary over a wide range. The melting point also relates to the rate of heating due to decomposition during melting of the substance.

[0051] Dynamic Vapor Sorption (DVS) is a method that has long been used to investigate the interaction between active pharmaceutical ingredients (APIs) and pharmaceutical products and moisture. To analyze the moisture absorption rate of a sample at a specific temperature, a DVS moisture sorption device (hygroscopicity analyzer) is used to expose the sample to a desired relative humidity until the weight change stabilizes.

[0052] The method for producing the salt of the present invention and the process for selecting the salt therethrough are as follows. First, it was confirmed that the free base of Compound 1 has a slight basicity (pKa=8.12) in the natural state, and therefore, various methods were used to select acids that can be appropriately matched among the acidic counter ions. Specifically, acids that are appropriately matched with the pKa of the free base of Compound 1 were calculated by computer, and it was confirmed that these acids mainly have a pKa of around 2 to 5.

[0053] Therefore, various experiments were conducted with the above range of acids and various solvents to obtain a total of 22 crystalline solids, and the crystallinity of each of these was confirmed and DSC and DVS analyses were performed. As a result, it was confirmed that the hemifumarate or fumarate salt was the most excellent salt in terms of crystallinity, melting point, hygroscopicity, etc.

[0054] Sphingosine-1-phosphate (S1P) receptors 1-5 constitute a family of G protein-coupled receptors with seven transmembrane domains. These receptors, designated S1P1 through S1P5, are activated through binding to sphingosine-1-phosphate generated by sphingosine kinase-catalyzed phosphorylation of sphingosine. S1P1, S1P4, and S1P5 receptors activate Gi but not Gq, whereas S1P2 and S1P3 receptors activate both Gi and Gq. S1P3 receptors respond to nootropics by increasing intracellular calcium, whereas S1P1 receptors do not.

[0055] The compound 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol (hereinafter referred to as compound 1) selectively binds to the receptors of each S1P subtype and acts as a functional antagonist to reduce lymphocytes in the peripheral bloodstream.

[0056] Compound 1 is useful for the treatment of multiple sclerosis and cerebral ischemia, see Korean Patent Publication No. 10-2017-0087813, which is incorporated herein by reference.

[0057] Meanwhile, inflammatory bowel disease is a chronic disease in which abnormal inflammation occurs repeatedly inside the intestines. It occurs due to an immune response to the intestinal flora, and typical examples are ulcerative colitis and Crohn's disease. It mainly occurs in Westerners, but as Asians' dietary habits gradually become more Westernized, the number of patients in Korea is also increasing. Ulcerative colitis was a disease with a high incidence in Europe and North America, where meat is the main diet, but recently, due to Westernized eating habits, the incidence has been increasing in East Asian countries, including Korea. Crohn's disease is a chronic inflammatory bowel disease that can occur anywhere in the entire digestive tract, from the mouth to the anus, and there is still no drug treatment that can completely cure ulcerative colitis.

[0058] In addition, there were approximately 120,700 organ transplants worldwide in 2015, an increase of 5.8% from the previous year, of which approximately 60% were kidney transplants. Interstitial fibrosis and tubular atrophy (IFTA) occurs in approximately 25% of patients one year after kidney transplantation. Histopathological changes in the transplanted kidney cause the interstitial space to expand and replace normal tissue, resulting in tubular atrophy and interstitial fibrosis, which leads to a decrease in the filtrate filtration rate and an increase in proteinuria. The incidence rate is 66% in the fifth year after transplantation and 90% in the tenth year after transplantation. In the early stages, these symptoms are mainly caused by immunological causes, but in the later stages, they are mainly caused by the toxicity of calcineurin inhibitors, which are immunosuppressants.

[0059] The mechanism by which calcineurin inhibitors, which are primarily used in post-transplant immunosuppressant maintenance therapy, induce IFN-γ is unclear, but they are known to provide a cause for fibrosis by inducing vasoconstriction, oxidative stress, TGF-β, etc.

[0060] Currently, calcineurin inhibitors, as well as antimetabolic agents and combined steroid therapy are used to treat chronic transplant rejection, but there is no clearly established treatment for IFTA. We hope that the development of a drug that simultaneously exhibits immunosuppressive and anti-fibrotic effects will improve the quality of life of post-transplant patients.

[0061] Focal segmental glomerular sclerosis (FSGS) is a filtration disorder and proteinuria-inducing disease caused by structural damage (e.g., sclerosis) to the glomerular epithelial cells, which are the filtration system of the kidney. It is the most common cause of end-stage renal failure. There are more than 300,000 patients worldwide, including approximately 80,000 patients in the United States, accounting for one-third of adult nephritic syndrome. Currently, steroids, immunosuppressants, and angiotensin receptor blockers are used, but they induce serious complications such as renal failure and anemia, and there is a need to develop drugs that have improved side effects and efficacy compared to conventional FSGS treatments.

[0062] Alopecia areata is a disease in which hair falls out in a circular pattern. It may occur in one or two places, or in severe cases, in many places at the same time. The bald spots may merge and cause hair loss in the eyebrows and beard in addition to the scalp. When the entire scalp is lost, it is called alopecia frontalis, and when hair is lost not only on the scalp but also on the whole body, it is called alopecia universalis. When it occurs in a band along the outer edge of the sides and back of the head, it is called serpentine alopecia. Alopecia areata is an autoimmune disease in which immune cells attack hair follicles and cause inflammation due to changes in the immune system, and is thought to be caused by stress and genetic factors. Less than 10% of patients with alopecia areata may have other autoimmune diseases such as atopic dermatitis, thyroid disease, and pernicious anemia. It occurs in both men and women and in all age groups, but is most common among children and young adults, with a prevalence of about 1-2% of the total population.

[0063] The most common treatment for alopecia areata is steroids. Steroid injections, oral steroids, and immunosuppressants are recognized and used as treatments, but after long-term treatment, although some initial effects are seen, the condition worsens or there are frequent side effects such as scalp inflammation, scalp depression, high blood pressure, weight gain, stomach irritation, and gastritis, and the condition tends to recur. Olumiant (active ingredient baricitinib), an oral JAK inhibitor currently known globally as a treatment for rheumatoid arthritis, was approved by the US Food and Drug Administration in June 2022 as the first systemic treatment for alopecia areata. Following this, Pfizer, the world's number one pharmaceutical company, also announced efficacy data similar to Olumiant after completing phase 3 trials of PF-06651600 (active ingredient ritlecitinib), a type of JAK inhibitor, and in particular, no major adverse cardiac events (MACE) or infectious side effects, which are of concern when administered long-term with JAK inhibitors, were observed, and based on these results, they applied for marketing approval from the FDA. Therefore, there is a continuing need for development of a drug that can be used to treat alopecia areata, has excellent efficacy, and is safe in terms of side effects and toxicity.

[0064] As a result, the present inventors have been making efforts to develop an independent compound that selectively acts on S1P subtype receptors among sphingolipid compounds, and have confirmed that the compound according to the present invention specifically binds to S1PR1 and S1PR4 receptors and acts as a functional inhibitor (antagonist), thereby not causing side effects of cardiovascular diseases due to non-selectivity for S1PR subtypes, and at the same time has preventive and therapeutic effects for focal segmental glomerulosclerosis, interstitial fibrosis and tubular atrophy, preventive or therapeutic effects for inflammatory bowel disease, and preventive or therapeutic effects for alopecia areata, thereby completing the present invention. EXAMPLES

[0065] The present invention will be described in more detail with reference to the following examples. However, the following examples are provided to more specifically explain the present invention, and the scope of the present invention is not limited to the following examples. The following examples may be appropriately modified or altered by those skilled in the art within the scope of the present invention.

[0066] Example 1: Preparation of the hemifumarate salt of 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol (compound 1) 98mg of fumaric acid (0.55 times the amount) and 500mg of compound 1-free base were added to 25mL of MIBK (Methyl Isobutyl Ketone) (23.9mg / mL) and mixed at room temperature for 5 minutes while stirring. The suspension was subjected to the following cooling crystallization process. First, the suspension was heated at 40℃ for 10 hours and then stirred at 5℃ for 12 hours. When the reaction was completed, a thick white suspension was formed. The solid material in the suspension was separated by centrifugal separation to separate the precipitate, and dried at room temperature for 16 hours under vacuum (-700mmHg) to obtain 525.5mg (89.2%) of the final product. The prepared hemifumarate salt exhibits an X-ray powder diffraction spectroscopy (XRPD) pattern with major diffraction peaks at 2θ values ​​of 6.3±0.2°, 9.4±0.2°, 18.5±0.2°, 19.0±0.2°, 19.9±0.2°, 20.7±0.2°, 25.5±0.2°, 28.7±0.2°, and 29.0±0.2°. 1 H-NMR (DMSO-d6): δ0.8(3H,t), 1.2(15H,m), 1.75(4H,m), 2.65(2H,m), 3.4(4H,t), 4.25(2H,t), 6.4(1H,m), 7.8(1H,s) The above 1 In the H-NMR data, the proton number (integrated numerical standard) of the >CH proton constituting fumaric acid appearing at δ 6.4 ppm was calculated to be half the proton number of the corresponding free base, and the ratio of the salt formation components of the final product was confirmed to be 1:0.5.

[0067] Example 2: Preparation of the fumarate salt of 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol (NXC736) 196 mg of fumaric acid (1.1 times equivalent amount) and 500 mg of NXC736-free base were added to 25 mL of acetone and mixed with stirring at room temperature for 5 minutes. The suspension was subjected to the same cooling crystallization process as in Example 1 and dried under vacuum (-700 mmHg) to obtain 562 mg of the final product. The final product, the fumarate salt, exhibits an X-ray powder diffraction spectroscopy (XRPD) pattern with major diffraction peaks at 2θ values ​​of 13.9±0.2°, 18.9±0.2°, 19.4±0.2°, 22.1±0.2°, and 22.5±0.2°.

[0068] Comparative Examples 1 to 6: Preparation of other salts of Compound 1 Salts of Compound 1 were prepared by mixing various acids and the free base of Compound 1. The acids and solvents used are shown in Table 1.

[0069] Experimental example 1: Hygroscopicity test The salts prepared in Comparative Examples 1 to 6 and the novel salt of the present invention were characterized by the DVS method, and the results are shown in Table 1 below.

[0070] [Table 1]

[0071] According to the DVS analysis of the hydrochloride salt of Compound 1 (Comparative Example 1), the compound exhibited a water adsorption rate of 49.5% at 90% RH, indicating deliquescence in the natural state. At the same time, the salts of Comparative Examples 2 to 6 also showed relatively high hygroscopicity.

[0072] Meanwhile, compared with the above-mentioned Comparative Examples 1 to 6, Example 1 was confirmed to have very excellent low moisture absorption, with a moisture adsorption rate of less than 2% under a condition of 90% RH according to the DVS evaluation results.

[0073] In addition, the DVS evaluation results of Example 2, which was repeated several times, showed that the water vapor adsorption rate was more than 3% and less than 5% under a condition of 90% RH, and thus the material was evaluated to have excellent moisture absorption properties.

[0074] Experimental Example 2: Solubility study of compound 1a The solubility of compound 1a prepared in Example 1 was evaluated in several different media, including hydrochloric acid buffer at pH 1.2, citrate buffer at pH 3.0, acetate buffer at pH 4.7, phosphate buffer at pH 6.8, simulated gastric fluid, simulated intestinal fluid in fasted state, simulated intestinal fluid in fed state, and water. Approximately 2.17 mg of sample (compound 1-free base basis) per media was taken into a 2 mL clear glass vial and 0.5 mL of media was added (salt composition considered as compound 1: relative ion=1:0.5; target concentration: 2 mg / mL). The mixture was stirred at 250 RPM for 24 hours at 25°C using a magnetic stirrer. The pH and appearance of the mixture were then recorded, and the mixture was filtered using a MultiScreen® HTS-HV 96-well plate (manufacturer: Merck; 0.45 μm hydrophilic, low protein binding Durapore® Membrane). Solubility was determined using HPLC and a calibration curve was generated by injecting known concentrations of compound 1a. Filtered samples were injected (diluted appropriately if necessary) and the concentration of compound 1a (mg / mL) in the filtrate was calculated from the calibration curve (taking into account the salt factor). The analytical results are as follows:

[0075] [Table 2]

[0076] The solubility of compound 1a was 2 mg / mL or more in all aqueous media mentioned above except for phosphate buffer (pH 6.8), and virtually all samples were completely dissolved, whereas undissolved suspensions were observed in some samples of the hydrochloride of compound 1 under the same conditions.

[0077] Experimental Example 3: Stability Study The standard stock solution, pH, salinity, light, and oxidation stability of compound 1a were confirmed under various conditions as follows: Experimental Example 3-1: Stability study of standard stock solution - 40℃ / 75±5%RH The stability of the crystalline compound was investigated under the conditions of 40℃ / 75±5%RH. The physical stability was confirmed through comparative XRPD analysis before and after the test, and the chemical stability was confirmed by HPLC. The experimental conditions and test results used in the stability test for compound 1a are as follows.

[0078] Stability conditions and test methods: 40℃ / 75±5%RH: Vial with the lid open and standing upright The required amount (~1 mg) of solid material was dissolved in 1 mL of diluent (1:1 methanol / water) and the prepared solution was analyzed by HPLC.

[0079] [Table 3]

[0080] [Table 4]

[0081] In Table 3, the hydrochloride of compound 1 changed into a liquid state that was considered physically unstable after one week under high humidity conditions (40°C / 75±5%RH), and some samples showed deliquescence. However, according to Table 4, in the case of compound 1a, the crystalline form was maintained even after two weeks under high humidity conditions (40°C / 75±5%RH), and the purity was maintained at 99% or more, indicating that the stability of the standard stock solution is very good.

[0082] Experimental Example 3-2: Stability study of standard stock solution - 60℃ / 75±5%RH The stability of the crystalline compound was investigated under the conditions of 60°C / 75±5%RH, and the purity was confirmed by HPLC. The experimental conditions and test results used in the stability test for compound 1a are as follows:

[0083] Stability conditions and test methods: 60℃ / 75±5%RH: Vial with the lid open and standing upright The required amount (~1 mg) of solid material was dissolved in 1 mL of diluent (1:1 methanol / water) and the prepared solution was analyzed by HPLC.

[0084] [Table 5]

[0085] In Table 5, compound 1a exhibited very low generation of softening substances and changes in content after 1 and 2 weeks at high humidity (60°C / 75±5% RH) conditions compared to compound 1 free base, and the purity was maintained at 98% or more, indicating that the stability of the standard stock solution was relatively excellent.

[0086] Figure 11 shows photographs of the initial and two-week changes in properties of the free base of Compound 1, Compound 1a, and the hydrochloride of Compound 1 after storing them under conditions of 60°C / 75±5%RH, and it can be seen that the hydrochloride absorbed moisture and turned into a liquid. In addition, in Figure 12, the free base of Compound 1, Compound 1a, and the hydrochloride of Compound 1 were stored together with the sample for two weeks under conditions of 60°C, and the corrosiveness of the sample was checked. As a result, it was confirmed that corrosion occurred in the case of the hydrochloride, indicating that moisture absorption occurred due to the hydrochloride.

[0087] Experimental Example 3-3: pH and Salinity Stability The pH and salt stability tests of compound 1-hemifumarate were carried out under the condition of an initial target concentration of 2 mg / mL. The pH and salt stability conditions and evaluation results used in the evaluation of compound 1-hemifumarate are as follows:

[0088] Stability conditions and test methods: 40±2℃: Vial with the lid closed and standing upright Storage container for stability: 2 mL screw-neck clear glass vial (manufacturer: La-Pha-Pack) Target concentration: ~2mg / mL (sample is added to 1mL of media, then vortex mixed for 30 seconds) 500 μL of the test mixture was dissolved in 500 μL of diluent (1:1 methanol / water), and the prepared solution was analyzed by HPLC without filtration.

[0089] [Table 6]

[0090] [Table 7]

[0091] As seen in Table 7, compound 1a exhibited a purity of more than 98% after 2 weeks in a phosphate buffer solution of pH 6.8 at 40±2° C., and showed a tendency to maintain almost the same purity despite various pH changes and the addition of salt, and thus exhibited excellent pH stability and salt stability overall.

[0092] However, looking at Table 6, which shows the data for the hydrochloride of Compound 1 under the same conditions, it was confirmed that the purity was less than 98% after two weeks in phosphate buffer solution of pH 6.8 at 40±2°C, and that the purity was still less than 98% even one week after the addition of salt.

[0093] Therefore, it was confirmed that compound 1a has relatively superior pH stability and salt stability compared to the hydrochloride salt of compound 1.

[0094] Experimental Example 3-3: Photostability and oxidative stability For the photostability test, 10 mg of compound 1a was exposed to ultraviolet and visible light at room temperature for 1.2 million lux hours. For the oxidation stability test, 30% H 2 O 2 0.2 mL of the above and 1 mg of compound 1a were placed in a 2 mL vial and stored at 25° C. for up to 24 hours. The light and oxidation stability conditions used in the evaluation of compound 1-fumarate and the evaluation results are as follows.

[0095] [Table 8]

[0096] [Table 9]

[0097] As seen in Table 9, compound 1a was confirmed to have excellent photostability since there was no significant change in purity and physical properties even when exposed to ultraviolet light and visible light at 1.2 million lux hours, and decomposition was observed after 24 hours under oxidative stress conditions. However, as seen in Table 8, compound 1 hydrochloride was observed to have some change in purity when exposed to ultraviolet light and visible light at 1.2 million lux hours, and showed a decomposition rate of about 8% or more after 20 hours under oxidative stress conditions. Therefore, compound 1a was confirmed to have relatively excellent photostability and oxidative stability compared to compound 1 hydrochloride.

[0098] In conclusion, the hemifumarate salt of 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol (Compound 1) according to the present invention exhibits excellent effects in terms of low hygroscopicity, stability of standard stock solution, light stability, oxidation stability, pH stability, solubility, etc., as compared with the hydrochloride salt and other salts of Compound 1, and has been confirmed to have the most excellent physical properties when judged based on solid-state properties (crystallinity, melting point, low hygroscopicity).

[0099] In addition, compound 1a according to the present invention was confirmed to be a salt that satisfies all of the following minimum conditions for mass production and storage for clinical use: 1) low hygroscopicity requirement of less than 2% water adsorption rate under 90% RH in DVS; and 2) stability requirement of standard stock solution that must be stable for 2 weeks or more under high humidity conditions (e.g., 40°C / 75% RH). Also, compound 1a according to the present invention was confirmed to be a salt that satisfies all of the following minimum conditions for smooth clinical progress: 1) low hygroscopicity requirement of less than 2% water adsorption rate under 90% RH in DVS; and 2) stability requirement of standard stock solution that must be stable for 2 weeks or more under high humidity conditions (e.g., 40°C / 75% RH). Also, solubility requirement of 2 mg / mL or more in all aqueous media, which is the minimum condition for smooth clinical progress.

[0100] Also, it was confirmed that compound 1b according to the present invention is a salt exhibiting similar physical and chemical properties to compound 1a overall, except for a slight difference in water adsorption rate.

[0101] Experimental Example 4: Confirmation of the inhibitory effect of compound 1a on mouse immune cells FTY720 (Fingolimod) is a multiple sclerosis treatment that works by reversibly capturing a portion of lymphocytes in lymph nodes or bone marrow, isolating them in secondary lymphatic organs and inhibiting their entry into the central nervous system, or by reducing the number of lymphocytes circulating in the bloodstream, thereby reducing the number of activated lymphocytes that reach the brain.

[0102] To confirm whether compound 1, which acts on S1P receptors, also has an effect on lymphocyte reduction, the effect of compound 1 on immune cells was evaluated. Compound 1 was orally administered to mice at 3 mg / kg / day, and lymphocyte changes in the peripheral blood (PB) and bone marrow (BM) were classified into T cells, CD4 T cells, and CD8 T cells, and FACS analysis was performed at 0, 4, 8, 12, 24, and 36 hours after administration of the test substance. The cell number was measured and expressed as a percentage (% of control) based on the 0 time.

[0103] Referring to Figure 4, it was confirmed that administration of 3 mg / kg of compound 1a reduced T cells, CD4 T cells, and CD8 T cells in the peripheral blood flow (PB) from 4 hours to 40.6% / 41.6%, 16.4% / 17.6%, and 5.9% / 9.3% levels at 0 hours. The reduction was maintained until 12 hours, and then recovered to 88.9% at 36 hours. Dose-dependency was observed in T cells 24 hours after administration, and a dose-dependent reduction effect was observed in CD4 T cells and CD8 T cells from 4 hours to 24 hours after administration.

[0104] In the bone marrow (BM), after administration of 3 mg / kg of compound 1a, the distribution of T cells was confirmed at 135% and 120% 4 hours later, and at 131% and 118% 24 hours later. This is considered to be a phenomenon caused by the receptor internalization caused by compound 1, which inhibits the release of T cells.

[0105] After administration of 3mg / kg of compound 1a, the cell number of T cells began to decrease from 4 hours, and it was confirmed that the inhibitory effect was maintained to a certain extent for up to 24 hours before recovery. In conclusion, it was confirmed that the administration of compound 1a reduced the number of lymphocytes circulating in the bloodstream, demonstrating an immunosuppressive effect.

[0106] Experimental Example 5: Confirmation of the inhibitory effect of compound 1a on rat immune cells To evaluate the effect of compound 1a on rat immune cells, compound 1a was orally administered to SD rats at 10 mg / kg / day, and then classified into T cells, CD4 T cells, and CD8 T cells in the peripheral blood (PB) and bone marrow (BM), and FACS analysis was performed at 0, 4, 8, 12, 24, 36, and 48 hours after administration of the test substance. The cell number was measured and expressed as a percentage (% of control) based on the 0 time, and the number of T cells, CD4 T cells, and CD8 T cells was measured and expressed as a % of control based on the 0 time.

[0107] Referring to Figure 5, it was confirmed that T cells were reduced in the peripheral blood (PB) from 4 hours after administration of compound 1a. The inhibitory effect was dose-dependent (70%, 80%, 87%), and it was confirmed that the inhibitory effect was maintained up to 24 hours by administration of 110 mg / kg of the compound. Conversely, no change or a tendency to increase was confirmed in the bone marrow (BM).

[0108] Through this study, it was confirmed that compound 1a, after oral administration, exhibits an immunosuppressive effect by decreasing lymphocyte counts in the peripheral blood (PB). It was also confirmed that there was no change or a tendency for lymphocyte counts to increase in the bone marrow (BM). This effect is believed to be related to the mechanism by which compound 1 acts as a functional antagonist on S1P receptors present in lymph nodes, bone marrow (BM), etc., suppressing lymphocyte release.

[0109] Experimental Example 6: Confirmation of the inhibitory effect on inflammatory bowel disease using a mouse DSS-induced model The efficacy of compound 1a against inflammatory bowel disease was evaluated using a dextran sulfate sodium (DSS)-induced colitis model. In the mouse model, inflammatory bowel disease was induced in mice by administering 2.5% DSS to the drinking water. The treatment groups consisted of a test group administered with compound 1a, a positive control group administered with ozanimod or MTX (methotrexate), a vehicle group, and a normal control group, each group having n=10. Compound 1, which is a compound of the present invention, was orally administered once a day for 7 days at doses of 5 and 10 mg / kg to the model in which inflammatory bowel disease was induced, and ozanimod, a positive control substance, was orally administered once a day at 5 mg / kg, and MTX (methotrexate), a positive control substance, was administered by intraperitoneal injection at 3 mg / kg three times a week.

[0110] Figure 6 shows the disease activity index (DAI) and the area under the curve (AUC) of the disease activity index in mice with DSS-induced colitis. The DAI indicates the total score of weight loss compared to the initial weight of the mouse, stool consistency, and the degree of intestinal bleeding. The calculation criteria for DAI are shown in Table 10 below, and DAI was evaluated on days 0, 1, 2, 3, 4, 5, 6, 7, and 8.

[0111] [Table 10]

[0112] Referring to FIG. 6, a statistically significant reduction effect was observed in all groups administered the positive control substances MTX at 3 mg / kg, Ozanimod at 5 mg / kg, Compound 1 at 5 mg / kg, and Compound 1 at 10 mg / kg compared to the negative control group (vehicle).

[0113] In particular, when compound 1a was administered at 5 mg / kg and 10 mg / kg, it was confirmed that the clinical indicators of weight loss, stool concentration, and intestinal bleeding level were equal to or greater than those of ozanimod, a conventional targeted treatment for ulcerative colitis.

[0114] The evaluation results were subjected to statistical analysis between the negative control group and the test group or between two test groups, and the p-value was shown to be significant at 0.05 or less in the Student's t-test or Mann Whitney U test to compare the groups. This confirmed that compound 1a is a substance that has an inhibitory effect on inflammatory bowel disease.

[0115] Experimental Example 7: Evaluation of the effect of compound 1a on cardiovascular function 1) Evaluation of the effect of compound 1a on HEK293 cells overexpressing the hERG gene Compound 1a was administered to HEK-293 cells stably expressing hERG potassium ion channels by introducing hERG (human Ether-a-go-go Related Gene) to evaluate the effect on hERG channel currents. Compound 1a was set at concentrations of 1, 3, 10, and 30 μM. On the day of each experiment, compound 1a was administered to a random cell after recording of hERG channel currents for the negative control substance or each concentration of test substance.

[0116] Referring to Figure 7, the suppression rates (%) of hERG channel currents of compound 1 (B to E groups) at concentrations of 1, 3, 10 and 30 μM were 9.94 ± 1.96, 13.98 ± 4.48, 35.03 ± 12.09 and 82.62 ± 7.31% (n = 3), respectively, and the 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 30 μM showed a statistically significant difference compared to the negative control group (A group).

[0117] Under the same conditions, E-4031 was treated at a concentration of 0.1 μM, which was used in the positive control group (F group). The compensated suppression rate (%) of hERG channel currents was a high value of 92.64 ± 1.66% (n = 5). Based on these results, this test method is considered to be suitable for evaluating the effect of compound 1 on hERG channel currents.

[0118] In the evaluation of the effect on hERG channel currents using HEK-293 cells transfected with the hERG gene under the above conditions, the test substance Compound 1a was treated up to a maximum concentration of 30 μM, and the compensated suppression rate (%) of the hERG channel currents was 82.62 ± 7.31%, and K +The ion channel inhibitory potential (IC50) was calculated to be 12.94 μM (Hill coefficient: 1.527). Therefore, it was confirmed that compound 1a is a substance that does not inhibit hERG potassium channel activity and does not induce cardiac abnormalities.

[0119] 2) Confirmation of effects on cardiovascular function using beagle dogs Compound 1a was orally administered to four unanesthetized and unrestrained male beagle dogs with remote transmitters inserted. Thereafter, the heart rates of the four beagle dogs were measured to evaluate the effect of compound 1 on the cardiovascular system, and the occurrence of any abnormal symptoms was confirmed with the naked eye.

[0120] In this experiment, four beagle dogs were administered with the experimental substance in four stages. In the first administration stage (0 mg / kg of compound 1), 0.5% MC aqueous solution was administered as a control substance, in the second administration stage, 12.5 mg / kg of compound 1a was administered, in the third administration stage, 25 mg / kg of compound 1a was administered, and in the fourth administration stage, 50 mg / kg of compound 1a was administered. The first to fourth administrations were performed at weekly intervals. In addition, the heart rates of the beagle dogs were measured 0 hours before administration of the experimental substance, 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.

[0121] Referring to FIG. 8, it was confirmed that administration of Compound 1 at doses of 12.5, 25, and 50 mg / kg did not cause any change in heart rate.

[0122] Furthermore, as a result of observing the general symptoms in beagle dogs with the naked eye, no abnormal symptoms were observed following administration of compound 1a at doses of 12.5, 25, and 50 mg / kg.

[0123] As described above, based on the results of the evaluation of the inhibitory efficacy against inflammatory bowel disease using a mouse DSS-induced model and the results of the heart rate and the macroscopic observation of abnormal symptoms in beagle dogs administered with compound 1, it was confirmed that the pharmaceutical composition containing the compound represented by formula 1 according to the present invention, its optical isomer or its pharma- ceutical acceptable salt as an active ingredient has a preventive or therapeutic effect against inflammatory bowel disease by acting as a functional inhibitor against S1PR1 and S1PR4. It was also confirmed that the pharmaceutical composition according to the present invention has a preventive or therapeutic effect against inflammatory bowel disease by acting as a functional inhibitor 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.

[0124] Experimental Example 8: Confirmation of efficacy in inhibiting fibrosis using mouse kidney fibrosis model The anti-fibrotic efficacy of compound 1a in renal disease was evaluated using a unilateral ureteral obstruction (UUO) model, a renal fibrosis disease model. After inducing renal fibrosis by ligating one ureter of a mouse, compound 1a was orally administered at 10, 30, and 90 mg / kg / day, and the positive control substance telmisartan at 30 mg / kg / day, once a day for two weeks.

[0125] Referring to FIG. 9, it was confirmed that the fibrosis area (Sirius red positive area (%)) and renal hydroxyproline were significantly increased in the vehicle group compared to the negative control group (sham), and renal fibrosis was induced. Telmisartan, used as a positive control, statistically significantly decreased renal hydroxyproline (p<0.0001). In all groups administered with compound 1a at 10, 30, and 90 mg / kg / day, renal hydroxyproline was statistically significantly decreased (p<0.05, p<0.01, P<0.001), and in particular, it was confirmed that renal hydroxyproline was statistically significantly decreased (p<0.05) when compound 1a was administered at 90 mg / kg / day. As a result, it was confirmed that compound 1 is a substance with a fibrosis inhibitory effect, and it was confirmed that compound 1 exhibited a dose-dependent renal fibrosis inhibitory effect in a mouse renal fibrosis model induced by UUO.

[0126] Experimental Example 9: Confirmation of anti-fibrotic efficacy using adenine-induced renal failure mouse model The anti-fibrotic effect of compound 1a was evaluated using a model of chronic kidney disease induced by adenine. Chronic kidney disease was induced in mice by orally administering 0.5% adenine (in 0.5% CMC) for 4 weeks, and compound 1a was orally administered at 10, 30, and 90 mg / kg / day and the positive control substance Telmisartan at 10 mg / kg / day for 3 weeks. Serum creatinine, serum BUN, and renal hydroxyproline were measured and histopathology was performed. In addition, the tubular-interstitial index value was found based on the measurement results.

[0127] Biochemical analysis showed that in the adenine-induced CKD model, the vehicle group had significantly higher serum creatinine levels of 1.15±0.25 (mg / dl) and serum BUN levels of 202.3±32.1 (mg / dl) than the control group. Compound 1a at 90mg / kg / day reduced serum creatinine levels by 0.90±0.12mg / dl compared to the vehicle group, reduced serum BUN levels by 157.4±32.3 (mg / dl) compared to the vehicle group, and the group administered compound 1a at 30mg / kg / day had a statistically significant reduction in renal hydroxyproline levels compared to the control group. Referring to Figure 10, it was confirmed that the tubular-interstitial index showed a statistically significant reduction in renal fibrosis in the group orally administered 90 mg / kg / day of compound 1a and the group administered telmisartan. In conclusion, the anti-fibrotic effect of compound 1 was confirmed in the adenine-induced chronic renal fibrosis model.

[0128] Experimental Example 10: Confirmation of efficacy in inhibiting alopecia areata using a mouse model of alopecia areata The efficacy of compound 1a against alopecia areata was evaluated using a spontaneous alopecia areata model (C3H / HeJ). The C3H / HeJ mice used in the study are 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. Compound 1a was orally administered at 30 mg / kg for 12 weeks, and the disease free ratio and AA lesion area were measured.

[0129] No significant difference in body weight was observed between the compound 1a administration group and the control group, and alopecia areata was observed later in the compound 1a administration group (100% alopecia areata observation date: control group: 56 days, compound 1a administration group: 84 days). In addition, the compound 1a administration group showed a statistically significant decrease in the number of sites where alopecia areata was observed compared to the control group (AA lesion area on the 84th day - control group: about 80%, compound 1a administration group: about 10%). For reference, the list of abbreviations used herein is as follows:

[0130] [Table 11] TIFF0007604719000014.tif177152

Claims

1. The hemifumarate salt of 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol is represented by the following formula 1a: 【Chemistry 1】

2. The hemifumarate salt of claim 1, wherein the salt is in crystalline form.

3. 3. The hemifumarate salt of claim 2, exhibiting an X-ray powder diffraction spectroscopy (XRPD) pattern comprising 2θ values ​​of 6.3±0.2°, 9.4±0.2°, 18.5±0.2°, 19.0±0.2°, 19.9±0.2°, 20.7±0.2°, 25.5±0.2°, 28.7±0.2°, and 29.0±0.2°.

4. 3. The hemifumarate salt according to claim 1 or 2, which has a moisture adsorption rate of less than 2% under conditions of 90% RH in DVS.

5. 3. The hemifumarate salt according to claim 1 or 2, which has a purity of 99% or more even after being placed under high humidity (40° C. / 75±5% RH) conditions for two weeks.

6. 3. The hemifumarate salt according to claim 1 or 2, having a purity of 98% or more after storage in a phosphate buffer solution of pH 6.8 at 40±2° C. for two weeks.

7. 3. The hemifumarate salt according to claim 1 or 2, having a melting endotherm onset temperature of 82°C in differential scanning calorimetry (DSC).

8. A pharmaceutical composition for preventing or treating multiple sclerosis or ischemic stalk, comprising the hemifumarate salt of 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol according to claim 1 as an active ingredient.

9. A pharmaceutical composition for preventing or treating focal segmental glomerulosclerosis (FSGS), comprising the hemifumarate salt of 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol according to claim 1 as an active ingredient.

10. A pharmaceutical composition for preventing or treating inflammatory bowel disease, comprising the hemifumarate salt of 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol according to claim 1 as an active ingredient.

11. The pharmaceutical composition of claim 10, wherein the inflammatory bowel disease is ulcerative colitis (UC) or Crohn's disease (CD).

12. A pharmaceutical composition for preventing or treating interstitial fibrosis and tubular atrophy (IFTA), comprising the hemifumarate salt of 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol according to claim 1 as an active ingredient.

13. A pharmaceutical composition for preventing or treating alopecia areata (AA), comprising the hemifumarate salt of 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol according to claim 1 as an active ingredient.

14. 14. A pharmaceutical composition according to any one of claims 8 to 13, further comprising one or more pharma- ceutically acceptable carriers, diluents and / or excipients.

15. 14. The pharmaceutical composition according to any one of claims 8 to 13, wherein the pharmaceutical composition does not cause cardiovascular side effects.

Images