Novel salts of GLP-1 receptor agonist compounds, process for the preparation thereof and pharmaceutical compositions

Abstract

The present invention relates to a GLP-1 receptor agonist compound having excellent in vivo solubility, stability and bioavailability, a novel salt thereof, a method for preparing the same, and a pharmaceutical composition comprising the same.

Description

Technical Field

[0001] The present invention relates to novel salts of glucagon-like peptide-1 receptor (GLP-1R) agonist compounds showing improved physical properties in terms of stability, solubility and the like and excellent bioavailability, a method for preparing the same and a pharmaceutical composition containing the same. Background Art

[0002] It is well known based on large-scale clinical studies that strict control of blood sugar to normal blood sugar levels in the treatment of diabetes is crucial to prevent various complications caused by diabetes. Candidate compounds that can lower blood sugar by strongly stimulating insulin secretion include a hormone called glucagon like peptide-1 (GLP-1). GLP-1 was first discovered in 1985 as an incretin hormone secreted by L-cells in the ileum and colon. GLP-1 increases insulin secretion by acting on a receptor called glucagon like peptide-1 receptor (GLP-1R). GLP-1 is secreted through stimulation of absorbed nutrients or blood sugar levels. Diabetes treatment using GLP-1 has the advantage that hypoglycemia will not occur because insulin is secreted according to glucose concentration. In addition, this hormone is known to effectively reduce the motility of the upper digestive system and suppress appetite, and proliferate existing β cells in the pancreas.

[0003] Currently, various GLP-1 analogs resistant to the DPP-4 enzyme that destroys GLP-1 in the blood have been developed and are being used as treatments for type 2 diabetes. Since these GLP-1 analogs have a significantly longer half-life than GLP-1, they have the advantage of maintaining a hypoglycemic effect for a long time, but there is a problem of low convenience of drug treatment because these GLP-1 analogs cannot be orally administered and should be used in the form of injections. Therefore, in recent years, some studies have been conducted to find small molecule GLP-1R agonists that can be orally administered and develop agonists as diabetes therapeutic agents.

[0004] In this regard, improved physicochemical properties, such as solubility, stability, non-hygroscopicity and the like, as well as improved bioavailability, and reduced toxicity are desired for small molecule GLP-1R agonists.

[0005] In this context, the present inventors have discovered that the tromethamine salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid as a GLP-1 receptor agonist compound has excellent physicochemical properties, high purity and excellent bioavailability compared to other commonly used pharmaceutically acceptable salt compounds, and have completed the present invention. Summary of the invention [Technical issues]

[0006] The object of the present invention is to provide a novel salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid.

[0007] Specifically, the present invention provides a novel salt, a preparation method thereof and a pharmaceutical composition comprising the same, wherein the novel salt has excellent physicochemical and / or pharmaceutical properties, such as hygroscopicity, thermal stability, solubility and the like, so as to have excellent formulation processing performance and storage stability as well as excellent bioavailability. [Technical solution]

[0008] In a general aspect, the present invention provides novel salts of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid as an active ingredient, methods for preparing the same, and pharmaceutical compositions comprising the same.

[0009] Hereinafter, each detailed description is provided below.

[0010] Novel salts of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid

[0011] For the above purpose, the present invention provides a (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid tromethamine salt.

[0012] The tromethamine salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid can be represented by the following Chemical Formula I:

[0013] [Chemical Formula I]

[0014]

[0015] The novel salts according to the present invention exhibit excellent physicochemical and pharmacokinetic properties in various aspects, such as thermal stability, in vivo solubility, bioavailability and the like.

[0016] The tromethamine salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid of the present invention may have a thermogravimetric analysis (TGA) pattern showing a weight loss of less than 1.0 wt%, 0.9 wt%, 0.8 wt%, 0.7 wt%, 0.6 wt%, 0.5 wt%, 0.4 wt%, 0.3 wt% or 0.2 wt% at 170° C. or less. Specifically, it may show Figure 2 Thermogravimetric analysis (TGA) patterns.

[0017] The tromethamine salt of the present invention (specifically, a crystalline form thereof) is characterized by having an endothermic transition peak at 174 to 204° C. in a differential scanning calorimetry (DSC) chart when the heating rate is 10° C. / min, and preferably, characterized by having an endothermic transition peak at 179 to 199° C., more preferably at 184 to 194° C., and more preferably at 189±2° C.

[0018] In addition, the tromethamine salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid of the present invention can show Figure 2 Differential scanning calorimetry.

[0019] Additionally, the novel tromethamine salts of the present invention are preferably in crystalline form.

[0020] The crystalline form of the tromethamine salt of the present invention may comprise at least three diffraction peaks at 2-theta (2θ) angle values ​​selected from the group consisting of: 4.03±0.2, 8.09±0.2, 10.04±0.2, 15.07±0.2, 15.71±0.2, 17.90±0.2, 19.60±0.2, 22.07±0.2, 24.92±0.2 and 25.39±0.2 in an X-ray powder diffraction (XRPD) pattern.

[0021] More specifically, the crystalline form of the tromethamine salt of the present invention may include diffraction peaks at the following 2-θ (2θ) angles in an X-ray powder diffraction (XRPD) pattern: 4.03±0.2, 8.09±0.2, 10.04±0.2, 15.07±0.2, 15.71±0.2, 17.90±0.2, 19.60±0.2, 22.07±0.2, 24.92±0.2, and 25.39±0.2.

[0022] Further specifically, the crystalline form of the tromethamine salt of the present invention may further comprise in the XRPD pattern any one or more diffraction peaks at 2θ (2θ) angle values ​​selected from the group consisting of: 12.17±0.2, 14.39±0.2, 17.02±0.2, 20.18±0.2 and 21.81±0.2.

[0023] In the present invention, the thiamethoxam salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid having the aforementioned characteristics can be referred to as crystalline form 1.

[0025] In addition, the crystalline form of the tromethamine salt of the present invention may further comprise any one or more diffraction peaks at 2-theta (2θ) angle values ​​selected from the group consisting of 19.80±0.2 and 24.53±0.2 in the XRPD pattern.

[0026] In the present invention, the crystalline form of the tromethamine salt of the present invention may further comprise diffraction peaks at 2-θ (2θ) angle values ​​of 19.80±0.2 and 24.53±0.2 in the XRPD pattern, and the crystalline form may be crystalline form 1A.

[0027] In addition, the tromethamine salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid of the present invention can exhibit Figure 1X-ray powder diffraction spectroscopy pattern.

[0028] In the present invention, novel salts that have never been used in the prior art are prepared. Specifically, the amine salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid can have excellent thermal stability and stability according to temperature and humidity, thereby stably maintaining the amount unchanged over a long period of time. Specifically, the amine salt has excellent thermal stability and similar properties to high temperatures, showing excellent storage stability. Therefore, the raw materials of the novel salts of the present invention can be obtained with high yield and high purity, and the increase of related substances is significantly lower even when stored for a long time, and thus high purity can be maintained for a long period of time.

[0029] In addition, the novel salts of the present invention can exhibit excellent solubility values ​​under various pH conditions (especially biologically relevant media conditions), thereby having high bioavailability, thereby achieving excellent pharmacological effects, and can be effectively used as novel active ingredients of pharmaceutical compositions capable of treating various indications.

[0030] According to one embodiment of the present invention, when simulated gastric fluid (SGF), fasting state simulated intestinal fluid (FaSSIF) and fed state simulated intestinal fluid (FeSSIF) are prepared and tested for measuring solubility and dissolution under conditions close to the in vivo environment, good solubility is exhibited. Specifically, the solubility of tromethamine salt in FaSSIF is excellent. It is thus found that the novel salts according to the present invention have significantly excellent in vivo solubility and exhibit high bioavailability. In addition, since the novel salts according to the present invention have high in vivo solubility, they exhibit high exposure in the body and maintain high blood concentrations, thereby having a significant effect of excellent bioavailability.

[0031] In addition, since the novel salts of the present invention exhibit high bioavailability when orally administered, they are likely to exhibit excellent therapeutic effects even when taken in small amounts, thereby significantly improving drug compliance in patients.

[0032] Furthermore, the novel salts of the present invention can act quickly and have a thermodynamically stable form, and can be extremely advantageous in terms of processing and storage of pharmaceutical products, are easy to formulate, and furthermore, can maintain the same state even after the preparation of the preparation, so that the uniformity of the preparation amount can be stably maintained for a long period of time, and thus the novel salts can be easily applied to mass production.

[0033] In the present invention, the thiamethoxam salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (crystalline form 1) can be transformed into crystalline form 1A after exposure to atmospheric humidity under specific relative humidity conditions, specifically at 20°C and 55% RH or higher.

[0034] However, crystalline Form 1A can be restored to crystalline Form 1 after desorption by changing the relative humidity conditions to 20°C, below 55% RH, preferably 20°C, 50% RH or lower, more preferably 20°C, 45% RH or lower.

[0035] The tromethamine salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (crystalline form 1) according to the present invention is a non-stoichiometric hydrate, and when exposed to moisture in the atmosphere over a wide range of up to 90% RH, only lattice expansion and contraction due to water molecules occur, and there is no collapse of the lattice structure or crystal phase transition during adsorption and desorption. Therefore, even if the crystalline form of tromethamine according to the present invention adsorbs moisture when exposed to a wide range of humidity, it can still return to the original crystalline form, i.e., crystalline form 1, upon desorption.

[0036] Preparation method of tromethamine salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazine-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid

[0037] In another general aspect, the present invention provides a method for preparing a tromethamine salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid represented by the following Chemical Formula I:

[0038] [Chemical Formula I]

[0039]

[0040] Specifically, the preparation method of the present invention can be divided into the following steps:

[0041] (1) mixing a compound represented by the following chemical formula II with a single organic solvent or a mixed organic solvent, and reacting with tromethamine represented by the following chemical formula III;

[0042] [Chemical Formula II]

[0043]

[0044] [Chemical Formula III]

[0045]

[0046] (2) heating the reaction solution obtained in step (1) and stirring while warming;

[0047] (3) cooling and stirring the reaction solution obtained in step (2); and

[0048] (4) Filter and dry the product of step (3).

[0049] In the preparation method of the present invention, the compound represented by the following chemical formula II is (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid, which can be the free base prepared according to Comparative Example 1 of the present invention.

[0050] In the preparation method of the present invention, the organic solvent used for the reaction in step (1) may be a single organic solvent or a mixture of organic solvents. The organic solvent may be one or a combination of two or more selected from the group consisting of methanol, ethanol, isopropanol, acetonitrile, n-propanol, acetone, methyl ethyl ketone, methyl acetate, ethyl acetate, tetrahydrofuran and 2-methyltetrahydrofuran. Preferably, the organic solvent may be one or a combination of two or more selected from the group consisting of methanol, ethanol, isopropanol, acetonitrile, acetone and ethyl acetate.

[0051] The mixing ratio of the mixed solvent may be 1:1 to 1:20 by volume.

[0052] In the present invention, tromethamine represented by chemical formula III is called tris(hydroxymethyl)aminomethane, and is also called Tris. Therefore, as used herein, Tris refers to tromethamine.

[0053] The tromethamine in step (1) is preferably used in an amount of 0.6 to 1.5 equivalents, more preferably in an amount of 0.7 to 1.3 equivalents, and further more preferably in an amount of 0.9 to 1.1 equivalents, based on 1.0 equivalent of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid.

[0054] In the preparation method of the present invention, the reaction in step (1) may be performed at a temperature of 20 to 80°C, preferably 40 to 70°C, and more preferably 55 to 60°C.

[0055] Step (2) may be a step of precipitating a solid precipitate, and step (3) may be a step of separating the solid precipitate produced in step (2), wherein the solid precipitate may be separated by a process commonly performed in the relevant field (such as a principle separation method and the like).

[0056] In the method for preparing the novel salts of the present invention, the reaction temperature and time can be adjusted according to the type and amount of the sample and solvent used in the reaction, but are not limited to the above ranges.

[0057] Pharmaceutical composition

[0058] The present invention provides a pharmaceutical composition for preventing or treating metabolic diseases, comprising a tromethamine salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid represented by the following chemical formula I:

[0059] [Chemical Formula I]

[0060]

[0061] The novel tromethamine salt according to the present invention may have excellent stability, thermal stability and pH stability, and may exhibit excellent in vivo solubility under biologically relevant medium conditions close to the in vivo environment, thereby exhibiting excellent pharmacological effects.

[0062] The term metabolic disease as used herein includes, for example, diabetes (T1D and / or T2DM, such as prediabetes), idiopathic T1D (type 1b), latent autoimmune diabetes of adults (LADA), early onset T2DM (EOD), young-onset diabetes mellitus (YOAD), maturity-onset diabetes of the young (MODY), malnutrition-related diabetes, gestational diabetes, hyperglycemia, insulin resistance, hepatic insulin resistance, impaired glucose tolerance, diabetic neuropathy, diabetic nephropathy, renal disease (e.g., acute renal failure, tubular dysfunction), disorders, pro-inflammatory changes in the proximal tubules), diabetic retinopathy, adipocyte dysfunction, visceral fat accumulation, sleep apnea, obesity (e.g., hypothalamic obesity and monogenic obesity) and related comorbidities (e.g., osteoarthritis and urinary incontinence), eating disorders (e.g., bulimia nervosa, anorexia nervosa, and obesity syndromes such as Prader-Willi syndrome and Bardet-Biedl syndrome), syndrome), weight gain due to use of other medications (e.g., use of steroids and antipsychotics), excessive sugar intake, dyslipidemia (including hyperlipidemia, hypertriglyceridemia, increased total cholesterol, high LDL cholesterol, and low HDL cholesterol), hyperinsulinemia, NAFLD (including related diseases such as steatosis, NASH, fibrosis, cirrhosis, and hepatocellular carcinoma), cardiovascular disease, atherosclerosis (including coronary artery disease), peripheral vascular disease, hypertension, endothelial dysfunction, abnormal vascular compliance, congestive heart failure, myocardial infarction (e.g., necrosis and apoptosis), stroke, hemorrhagic stroke, ischemic stroke, traumatic brain injury, pulmonary hypertension, restenosis after angioplasty, intermittent claudication, postprandial lipidemia, metabolic acidosis, ketosis, arthritis, osteoporosis, Parkinson's disease disease), left ventricular hypertrophy, peripheral arterial disease, vision loss, cataracts, glomerulosclerosis, chronic renal failure, metabolic syndrome, syndrome X, premenstrual syndrome, angina, thrombosis, atherosclerosis, transient ischemic attack, vascular restenosis, abnormal glucose metabolism, symptoms of impaired fasting glucose, hyperuricemia, gout, erectile dysfunction, skin and connective tissue disorders, psoriasis, foot ulcers, ulcerative colitis, hyper-apo B-lipoproteinemia, Alzheimer's disease, schizophrenia, cognitive impairment, inflammatory bowel disease, short bowel syndrome, Crohn's disease, colitis, irritable bowel syndrome, polycystic ovary syndrome, and addiction (e.g., alcohol and / or drug abuse).

[0063] Specifically, the metabolic disease may be at least any one selected from the group consisting of diabetes, idiopathic type 1 diabetes, latent autoimmune diabetes in adults (LADA), early onset type 2 diabetes (EOD), young-onset diabetes (YOAD), maturity-onset diabetes of the young (MODY), malnutrition-related diabetes, gestational diabetes, hyperglycemia, insulin resistance, hepatic insulin resistance, impaired glucose tolerance, diabetic neuropathy, diabetic nephropathy, kidney disease, diabetic retinopathy, visceral fat accumulation, sleep apnea, obesity, eating disorders, dyslipidemia, hyperinsulinemia, non-alcoholic fatty liver disease (NAFLD), cardiovascular disease, atherosclerosis, peripheral vascular disease, hypertension, congestive heart failure, myocardial infarction. thrombosis, stroke, hemorrhagic stroke, ischemic stroke, traumatic brain injury, pulmonary hypertension, restenosis after angioplasty, intermittent claudication, postprandial lipidosis, metabolic acidosis, ketosis, arthritis, osteoporosis, Parkinson's disease, left ventricular hypertrophy, peripheral arterial disease, vision loss, cataracts, glomerulosclerosis, chronic renal failure, metabolic syndrome, syndrome X, premenstrual syndrome, angina pectoris, thrombosis, transient ischemic attack, vascular restenosis, abnormal glucose metabolism, symptoms of abnormal fasting glucose, hyperuricemia, gout, erectile dysfunction, psoriasis, foot ulcers, ulcerative colitis, hyperlipoproteinemia, Alzheimer's disease, schizophrenia, cognitive impairment, inflammatory bowel disease, short bowel syndrome, Crohn's disease, colitis, irritable bowel disease, and polycystic ovary syndrome.

[0064] Specifically, non-alcoholic fatty liver disease (NAFLD) may be, for example, at least any one selected from the group consisting of steatosis, non-alcoholic steatohepatitis (NASH), fibrosis, cirrhosis, and hepatocellular carcinoma. The exemplary non-alcoholic fatty liver disease described above includes a class of metabolic diseases significantly associated with various medical diseases other than the class of alcoholic liver disease.

[0065] In one aspect, the present invention provides a pharmaceutical composition comprising: a tromethamine salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid represented by the following chemical formula I, and a pharmaceutically acceptable carrier:

[0066] [Chemical Formula I]

[0067]

[0068] As used herein, the term "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible.

[0069] The compositions according to the invention may be in various forms. The compositions according to the invention may be in the form of, for example, liquid, semisolid and solid dosage forms, such as liquid solutions (e.g., injectable and injectable solutions), dispersions or suspensions, tablets, pills, powders, liposomes and suppositories. The form depends on the intended route of administration and its therapeutic purpose.

[0070] Typical compositions are in the form of injectable and infusible solutions.One mode of administration is parenteral mode (eg, intravenous, subcutaneous, intraperitoneal, intramuscular mode).

[0071] Oral administration of solid preparations can be achieved, for example, based on hard or soft capsules, pills, cachets, lozenges or tablets, each of which contains a predetermined amount of one or more compounds according to the present invention. In some embodiments, oral administration can be achieved based on powder or granule forms.

[0072] In still other embodiments, oral administration can be achieved in a liquid dosage form. Liquid dosage forms for oral administration include, for example, pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs containing inert diluents commonly used in the art (eg, water).

[0073] Other carrier materials and modes of administration known in the pharmaceutical art may be used. The pharmaceutical composition according to the present invention may be prepared by any well-known pharmaceutical techniques, such as effective formulation and administration procedures.

[0074] These preparations can be prepared by conventional methods for formulation in this art or methods disclosed in the literature [see Remington's Pharmaceutical Science (latest version), Mack Publishing Company, Easton PA], and can be formulated into various preparations depending on each disease or component.

[0075] The composition of the present invention can be administered orally or parenterally (e.g., intravenously, subcutaneously, intraperitoneally or topically) according to the desired method, and the dosage varies depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate and severity of the disease. The daily dosage of the novel salts of the present invention is about 0.01 to 500 mg / kg, preferably 0.1 to 100 mg / kg, and can be divided and administered once or several times a day.

[0076] The pharmaceutical composition of the present invention may further contain at least one active ingredient exhibiting the same or similar medical effects in addition to the novel salts.

[0077] Therapeutic use and method for treating metabolic diseases and use thereof in the manufacture of a medicament for treatment

[0078] The present invention provides a pharmaceutical composition for preventing or treating metabolic diseases, comprising: a tromethamine salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid represented by the following chemical formula I:

[0079] [Chemical Formula I]

[0080]

[0081] The present invention provides a method for treating a metabolic disease, comprising: administering to a subject in need thereof a therapeutically effective amount of the tromethamine salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid as described above.

[0082] The language "a subject in need thereof" refers to a mammal, including a human, which includes mammals such as humans, monkeys, cows, horses, dogs, cats, rabbits, rats and mice.

[0083] As used herein, the term "therapeutically effective amount" refers to the amount of novel salts or pharmaceutical compositions containing them that can effectively prevent or treat metabolic diseases, and for example, any amount of pharmaceutical compositions containing the above salts can be included as the amount of novel salts administered to the subject to be treated to prevent the occurrence or recurrence of metabolic diseases, relieve symptoms, inhibit direct or indirect pathological results, prevent cancer metastasis, reduce the rate of progression, or relieve or temporarily improve symptoms or improve prognosis. In other words, the therapeutically effective amount can be interpreted as covering all doses that improve or cure the symptoms of metabolic diseases through pharmaceutical compositions.

[0084] The method for preventing or treating metabolic diseases of the present invention not only includes treating the disease itself before the onset of symptoms, but also includes suppressing or avoiding its symptoms by administering the above-mentioned salts or pharmaceutical compositions containing them. In the management of the disease, the preventive or therapeutic dose of a specific active ingredient will vary depending on the nature and severity of the disease or condition and the route of administration of the active ingredient. The dose and frequency of administration will vary depending on the age, weight and response of the individual patient. A person of ordinary skill in the art can naturally consider these factors and easily select a suitable dosing regimen. In addition, the method for treating metabolic diseases using the pharmaceutical composition of the present invention may further include administering a therapeutically effective amount of an additional active agent suitable for treating the disease together with the above-mentioned salt, wherein the additional active agent may exhibit a synergistic or auxiliary effect with the above-mentioned salt as an active ingredient according to the present invention.

[0085] The present invention also provides the use of the tromethamine salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid for the manufacture of a medicament for treating metabolic diseases. The above salts used to prepare the medicament can be mixed with acceptable adjuvants, diluents, carriers and the like, and can be prepared in the form of a combined preparation with other active agents to have a synergistic effect of the active ingredients.

[0086] The substances mentioned in the uses, compositions and treatment methods of the present invention are equally applicable, as long as they do not conflict with each other. [Beneficial effects of the present invention]

[0087] The novel salts according to the present invention may have excellent stability, thereby increasing the stability of the formulation, and may have improved solubility (particularly, in vivo solubility) and bioavailability, and be effectively used as active ingredients of pharmaceutical compositions. BRIEF DESCRIPTION OF THE DRAWINGS

[0088] The following drawings attached to this specification illustrate preferred embodiments of the present invention and are used to further understand the technical concept of the present invention and the above-mentioned contents of the present invention, and therefore the present invention should not be considered to be limited to the contents described in these drawings.

[0089] Figure 1 This is an X-ray powder diffraction analysis (XRPD) pattern of the tromethamine salt compound (crystalline form 1) represented by Chemical Formula I prepared in Example 3 of the present invention.

[0090] Figure 2 The thermogravimetric analysis (TGA) pattern analysis diagram and the differential scanning calorimetry (DSC) pattern diagram of the tromethamine salt compound (crystalline form 1) represented by the chemical formula I prepared in Example 3 of the present invention.

[0091] Figure 3 This is a thermogravimetric analysis (TGA) pattern analysis diagram of the (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid free base prepared in Comparative Example 1.

[0092] Figure 4 This is a differential scanning calorimetry (DSC) pattern analysis chart of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid free base prepared in Comparative Example 1.

[0093] Figure 5 This is the X-ray powder diffraction analysis (XRPD) pattern of the L-arginine salt compound of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid prepared in Comparative Example 2.

[0094] Figure 6 This is a thermogravimetric analysis (TGA) pattern analysis diagram of the L-arginine salt compound of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid prepared in Comparative Example 2.

[0095] Figure 7 This is a differential scanning calorimetry (DSC) pattern analysis chart of the L-arginine salt compound of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid prepared in Comparative Example 2.

[0096] Figure 8 This is the X-ray powder diffraction analysis (XRPD) pattern of the meglumine salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid prepared in Comparative Example 3.

[0097] Fig. 9This is a thermogravimetric analysis (TGA) pattern analysis diagram of the meglumine salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid prepared in Comparative Example 3.

[0098] Fig.10 This is a differential scanning calorimetry (DSC) pattern analysis chart of the meglumine salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid prepared in Comparative Example 3.

[0099] Fig.11 This is the XRPD pattern of the potassium salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid prepared in Comparative Example 4.

[0100] Fig.12 This is the TGA pattern analysis chart of the potassium salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid prepared in Comparative Example 4.

[0101] Fig.13 This is the DSC pattern analysis chart of the potassium salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid prepared in Comparative Example 4.

[0102] Fig.14 It is a graph showing the results of dynamic vapor sorption (DVS) measurement of the tromethamine salt compound (crystalline form 1) represented by Chemical Formula I prepared in Example 3 of the present invention.

[0103] Fig.15 It is a graph showing the DVS measurement results of the L-arginine salt compound of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid prepared in Comparative Example 2.

[0104] Fig.16A graph showing the DVS measurement results of the meglumine salt compound of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid prepared in Comparative Example 3.

[0105] Fig.17 A graph showing the DVS measurement results of the potassium salt compound of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid prepared in Comparative Example 4.

[0106] Fig.18 An X-ray powder diffraction (XRPD) graph of the tromethamine salt compound (crystalline form 1) represented by Chemical Formula I and crystalline form 1A prepared in Example 3 of the present invention, analyzed at 20 °C under conditions of 55% RH or higher. Detailed Description

[0107] Hereinafter, examples and the like will be described in detail to assist in understanding the present invention. However, these examples of the present invention can be modified in various other forms, and the scope of the present invention should not be considered limited to the following examples. These examples of the present invention are provided to more fully explain the present invention to those of ordinary skill in the art.

[0108] <XRPD, DVS, and NMR Analysis Conditions>

[0109] 1) XRPD (X-ray powder diffractometer)

[0110] An X-ray powder diffraction (XRPD) pattern was measured using a DYY884-AERIS-300 manufactured by PANalytical, and analyzed at a step size of approximately 0.02° in the range of approximately 2° to 45° in the 2θ angle. The usage conditions are set as shown in Table 1 below.

[0111] [Table 1]

[0113] 2) DVS (Dynamic vapor sorption)

[0114] The moisture adsorption / desorption profile of a sample with an amount of approximately 5 to 25 mg was tested using a DVS Intrinsic 1 manufactured by Surface Measurement System according to the parameters in Table 2 below under a 0% to 90% relative humidity (RH) cycle at 25 °C.

[0115] [Table 2]

[0116] 3) 1 H-NMR (Nuclear Magnetic Resonance)

[0117] About 3 mg of compound was weighed into an NMR tube, and 0.5 mL of deuterated dimethyl sulfoxide was added to completely dissolve the sample. The tube was placed in a rotor and placed in the open position of the autosampler and scanned with a BRUKER AVANCE III (400 MHz).

[0118] <Example 1> Preparation of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid tromethamine salt

[0119] To 500 mg of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid free base prepared according to Comparative Example 1, 5 mL of acetone was added and mixed at room temperature. 1.1 equivalents of tromethamine was added to the mixture and reacted under reflux stirring for 1 hour. The reaction solution was cooled to room temperature and stirred for 12 hours. The obtained solid was filtered, washed with acetone, and dried to obtain 0.4 g of tromethamine salt (yield: 65.6%).

[0120] The results of NMR analysis of the tromethamine salt prepared according to Example 1 are as follows.

[0121] 1H NMR (400MHz, DMSO-d6): δ8.99 (dd, Ja=2.0Hz, Jb=0.4Hz, 1H), 8.30 (dd, Ja=8.0Hz, Jb=2.0Hz, 1H), 8.19 (d, J=0.8Hz, 1H), 7 .82(dd,Ja=8.4Hz,Jb=1.2Hz,1H),7.58-7.54(m,2H),7.51(t,Ja=16.0Hz,Jb=8.0Hz,1H),6.33(d,J=8.0Hz,1H),6.20(d, J=7.6Hz,1H),5.42(s,2H),5.09-5.07(m,1H),4.78-4.72(m,1H),4.63-4.59(m,1H),4.51-4.46(m,1H),4.40-4.35(m,1H) ), 3.94 (d, J = 13.2Hz, 1H), 3.77 (d, J = 13.6Hz, 1H), 3.40 (s, 6H), 3.33-3.32 (m, 4H), 2.73-2.65 (m, 1H), 2.48-2.41 (m, 5H).

[0122] <Example 2> Preparation of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid amine triol salt

[0123] To 16 g of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid free base prepared according to Comparative Example 1, 3 mL of methanol and 15 mL of ethanol were added and mixed at room temperature. 1.05 equivalents of tromethamine were added to the mixture, and reacted by stirring at 70° C. for 1 hour. The reaction solution was cooled to room temperature and stirred for 12 hours. The obtained solid was filtered, washed with ethanol, and dried to obtain 17.64 g of tromethamine salt (yield: 90.0%).

[0124] <Example 3> Preparation of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid tromethamine salt

[0125] To 3.4 kg of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid free base prepared according to Comparative Example 1, 17 L of methanol and 34 L of ethyl acetate were added and mixed at room temperature. 1.05 equivalents of tromethamine were added to the mixture, and the reaction was carried out by stirring at 55 to 60° C. for 1 hour. The reaction solution was cooled to room temperature and stirred for 12 hours. The obtained solid was filtered, washed with ethyl acetate, and dried to obtain 3.84 kg of tromethamine salt (yield: 92.31%).

[0126] As a result of measuring XRPD of the tromethamine salt prepared in Example 3, it was confirmed that Figure 1 and the 2θ diffraction patterns shown in Table 3, which confirm that the tromethamine salt prepared in Example 3 has the same crystalline form as the tromethamine salts of Examples 1 and 2. In addition, the above results only show one crystalline form.

[0127] [Table 3]

[0128] <Comparative Example 1> Preparation of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid free base

[0129] (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid free base can be prepared in the same manner as disclosed in Korean Patent Laid-Open No. 10-2021-0059653 and the specific method thereof is as follows.

[0130] Step (1): Synthesis of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid methyl ester

[0131] 6-(((6-(piperazin-1-yl)pyridin-2-yl)oxy)methyl)nicotinonitrile (233 mg), (S)-methyl 2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate (232 mg) and potassium carbonate (436 mg) were placed in a round-bottom flask, dissolved in acetonitrile (10 mL) and stirred at 60° C. for one day. After the completion of the reaction was confirmed by TLC, the solution was diluted with ethyl acetate, and the organic layer was washed with a saturated aqueous solution of NaHCO 3 , a saturated aqueous solution of NH 4 Cl and brine. Subsequently, the organic layer was dried over anhydrous magnesium sulfate and filtered under reduced pressure to obtain a filtrate. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to give the target compound (S)-methyl 2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate (390 mg, yield 89%) as a clear syrup.

[0132] Step (2): Synthesis of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid

[0133] The compound (387mg) obtained in step (1) is placed in a round-bottom flask, dissolved in acetonitrile (10mL), and stirred. 1.0M TBD aqueous solution (1.4mL) is added dropwise while stirring. Purified water (0.6mL) is added to the mixture and stirred at 60 ° C for one day. After confirming the completion of the reaction by TLC, the mixture is neutralized to pH 7 with 1N HCl aqueous solution, extracted with 10% DCM / MeOH solution, dried over anhydrous magnesium sulfate, and filtered under reduced pressure to obtain a filtrate. The filtrate is concentrated under reduced pressure. The residue is purified by silica gel column chromatography (DCM / MeOH) to obtain the final compound (225mg, yield 60%) in the form of a light green solid.

[0134] <Comparative Example 2> Preparation of L-arginine salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid

[0135] To a 2 mL vial containing 20 mg of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid free base was added 1 mL of acetonitrile and the resulting mixture was mixed at room temperature for 5 seconds. 1.1 equivalents of L-arginine were added to the mixture and heating and cooling cycles were performed using a ThermoMixer C (Eppendorf). After the temperature cycle, the suspension was centrifuged to obtain a precipitate, and the resulting precipitate was dried under vacuum at room temperature to obtain L-arginine acid salt. XRPD analysis of the L-arginine acid salt was performed and the results are shown in Figure 5 middle.

[0136] <Comparative Example 3> Preparation of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid meglumine salt

[0137] To a 2 mL vial containing 20 mg of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid free base was added 1 mL of acetonitrile and the resulting mixture was mixed at room temperature for 5 seconds. 1.1 equivalents of meglumine was added to the mixture and heating and cooling cycles were performed using a ThermoMixer C (Eppendorf). After the temperature cycle, the suspension was centrifuged to obtain a precipitate, and the resulting precipitate was dried under vacuum at room temperature to obtain the meglumine salt. The meglumine salt was subjected to XRPD analysis and the results are shown in Figure 8 middle.

[0138] <Comparative Example 4> Preparation of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid potassium salt

[0139] To a 2 mL vial containing 20 mg of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid free base was added 1 mL of acetonitrile and the resulting mixture was mixed at room temperature for 5 seconds. 1.1 equivalents of potassium hydroxide (KOH) was added to the mixture and heating and cooling cycles were performed using a ThermoMixer C (Eppendorf). After the temperature cycle, the suspension was centrifuged to obtain a precipitate, and the resulting precipitate was dried under vacuum at room temperature to obtain the potassium salt. The potassium salt was subjected to XRPD analysis and the results are shown in Fig.11 middle.

[0140] <Experimental Example 1> Thermogravimetric (TGA) analysis

[0141] Thermogravimetric (TGA) analysis was performed on the tromethamine salt prepared in Example 3, the free base prepared in Comparative Example 1, the L-arginine acid salt prepared in Comparative Example 2, the meglumine salt prepared in Comparative Example 3, and the potassium salt prepared in Comparative Example 4. Thermogravimetric analysis was performed using TGAQ50 manufactured by TA Instruments at a heating rate of 10°C / min from 20°C to 350°C under nitrogen conditions. The weight loss of each salt at a temperature of about 170°C or lower was measured, and the results are shown in Table 1. Figure 2 (tromethamine salt), Figure 3 (free base), Figure 6 (L-arginine salt), Fig. 9 (meglumine salt) and 12 (potassium salt), and are summarized in Table 4 below.

[0142] [Table 4]

[0143] As can be seen from the above results, the free base of Comparative Example 1 and the other salts of Comparative Examples 2 to 4 showed weight loss caused by containing water or an organic solvent, but the tromethamine salt according to the present invention showed very little weight loss. Therefore, it can be confirmed that the tromethamine salt according to the present invention has excellent physicochemical properties without containing water or an organic solvent.

[0144] <Experimental Example 2> Differential Scanning Calorimetry (DSC) Analysis

[0145] The tromethamine salt prepared in Example 3, the free base prepared in Comparative Example 1, and the other salts prepared in Comparative Examples 2 to 4 were subjected to differential scanning calorimetry (DSC) analysis. Specifically, using a DSC analyzer, DSC was measured in an airtight pan under nitrogen purge at a scanning rate of 10°C / min in the range of 30°C to 300°C. Therefore, the DSC results of the tromethamine salt prepared in Example 3 are shown in Figure 2The DSC results of the free base of Comparative Example 1 are shown in Figure 4 The DSC results of the other salts obtained in Comparative Examples 2 to 4 are shown in Figure 7 , 10 and 13, and are summarized in Table 5 below.

[0146] [Table 5]

[0147] As can be confirmed in Table 5, (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid free base has the disadvantage of poor thermodynamic stability due to its low melting point. On the other hand, it can be confirmed that the tromethamine salt of the present invention has a significantly higher melting point than the other salts and free bases obtained in Comparative Examples 2 to 4, thereby exhibiting excellent thermodynamic stability.

[0148] <Experimental Example 3> Hygroscopicity test

[0149] (1) 25°C and 90% RH conditions

[0150] The hygroscopicity of the tromethamine salt prepared in Example 3, the L-arginine salt prepared in Comparative Example 2, the meglumine salt prepared in Comparative Example 3, and the potassium salt prepared in Comparative Example 4 was measured. The results are shown in Figures 14 to 17 The results according to the percentage of weight increase of moisture adsorption are shown in the following Table 6.

[0151] [Table 6]

[0152] Figures 14 to 17 As can be confirmed in Table 6, the tromethamine salt showed a weight change of about 1.44% when the hygroscopicity was measured by DVS at 25°C and 90% humidity, and exhibited significantly lower hygroscopicity than the L-arginine salt, the meglumine salt and the potassium salt. Therefore, it was confirmed that the novel tromethamine salt of the present invention had excellent storage stability.

[0153] (2) 25°C and 80% RH conditions

[0154] Under the hygroscopicity test conditions of the European Pharmacopoeia (EP), the tromethamine salt prepared in Example 3 and the free base prepared in Comparative Example 1 were stored at a temperature of 25° C. and a relative humidity of 80% for 24 hours, and then the mass change of the samples due to hygroscopicity was measured.

[0155] The results are shown in Figure 7 middle.

[0156] [Table 7]

[0157] As can be confirmed in Table 7, the tromethamine salt of the present invention has a moisture absorption coefficient of 0.22% and exhibits significantly superior non-hygroscopicity compared to the free base of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid.

[0158] Therefore, it was confirmed that the tromethamine salt of the present invention has significantly superior storage stability compared to the free base and is suitable for formulation.

[0159] <Experimental Example 4> Evaluation of solubility in bio-related media

[0160] The solubility of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid free base and tromethamine salt in water and biologically relevant media, namely simulated gastric fluid (SGF), fasted state simulated intestinal fluid (FaSSIF), and fed state simulated intestinal fluid (FeSSIF), was demonstrated.

[0161] Buffers for biorelevant media are prepared as follows.

[0162] Water: Laboratory Milli-Q purified water.

[0163] SGF (Simulated Gastric Fluid): Use 2.0 g of sodium chloride and 7 mL of hydrochloric acid dissolved in 1000 mL of water.

[0164] FaSSIF (Fasted State Simulated Intestinal Fluid): A commercial product purchased from Bio-Relevant and manufactured according to the instructions was used.

[0165] FeSSIF (Fed State Simulated Intestinal Fluid): A commercial product purchased from Bio-Relevant and manufactured according to the instructions was used.

[0166] For the experiment, about 2 mg of compound (2 mg in free base form) was weighed into each glass bottle, and then 1 mL of medium was added (final concentration was 2 mg / mL). The samples were then stirred continuously at 5000 rpm in a magnetic stirrer at room temperature for 24 hours. The pH was then measured and the concentration was determined using 96-well plate (Merck; 0.45 μm hydrophilic low protein binding The mixture was filtered through a 4% paraformaldehyde (1% paraformaldehyde) membrane. The solubility was analyzed using HPLC. The experimental results are shown in Table 8.

[0167] [Table 8]

[0168] As can be confirmed in Table 8, the tromethamine salt exhibits excellent solubility in all SGF, FaSSIF and FeSSIF, which have pH adjusted to be similar to the stomach, pre-meal small intestine and post-meal small intestine in vivo, but particularly exhibits excellent solubility under the conditions of fasting state simulated intestinal fluid (FaSSIF) of the small intestine simulating the occurrence of most drug absorption. This effect shows that the tromethamine salt can exhibit high solubility regardless of whether a meal is taken or not, and has a higher absorption rate than the free base, thereby having excellent bioavailability and similar characteristics.

[0169] <Experimental Example 5> Drug stability evaluation

[0170] The free base and tromethamine salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid were weighed into individual vials and stored under pressure conditions to study stability.

[0171] The pressure conditions are as follows:

[0172] -High temperature open chamber conditions: 80℃;

[0173] Samples were analyzed by XRPD and HPLC at the beginning of storage (day 0) and week 1. The HPLC results are shown in Table 9.

[0174] [Table 9]

[0175] As can be seen in Table 9, the tromethamine salt according to the present invention has a purity change rate of less than 1% under harsh conditions, showing significantly superior stability compared to the free base. In addition, the tromethamine salt according to the present invention was confirmed to be an extremely stable material under harsh conditions, which did not show any change in the XRPD pattern and had no change in appearance.

[0176] <Experimental Example 6> Analysis of crystal forms under various relative humidity conditions (reversible adsorption / desorption).

[0177] XRPD analysis of the crystalline form 1 of the tromethamine salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid was performed at 20° C. under conditions of different relative humidity from 15% RH to 90% RH, and the change of the crystalline form according to the relative humidity was confirmed. The results are shown in Fig.18, Table 10 and Table 11. Table 10 shows the crystalline form when the relative humidity changes from 15% RH to 60% RH, and Table 11 shows the crystalline form during desorption from 50% RH to 15% RH.

[0178] [Table 10]

[0179] [Table 11] Relative humidity (%RH, 20℃) 50 40 30 15 Crystalline form Intermediate response between Form 1 and Form 1A Form 1 Form 1 Form 1

[0180] Fig.18 It can be confirmed that the crystalline form 1A has the 2θ diffraction pattern (2θ±0.2°) shown in Table 3 above, and further includes diffraction peaks at 2-θ (2θ) angles of 19.8019±0.2 and 24.5314±0.2.

[0181] As can be confirmed in Tables 10 and 11, the tromethamine salt of the present invention maintains the form of crystalline Form 1 under the conditions of 15% RH or more and less than 55% RH at 20°C.

[0182] However, Form 1A exhibited an XRPD peak change at 55% RH or higher at 20°C (forming Form 1A), but returned to the form of crystalline Form 1 when desorbed at 15% RH or higher and below 55% RH at 20°C.

[0183] The above results show that the crystalline form of the tromethamine salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid is a non-stoichiometric hydrate, and the lattice expansion and contraction only occur during the adsorption and desorption of water, and there is no collapse of the lattice structure.

Claims

1. A (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid tromethamine salt represented by the following chemical formula I: [Chemical Formula I] 2. The tromethamine salt according to claim 1, wherein the tromethamine salt has a thermogravimetric analysis (TGA) pattern showing a weight loss of less than 0.2 wt% at 170°C or less.

3. The tromethamine salt according to claim 1, wherein the tromethamine salt has an endothermic transition peak at 174 to 204°C in a differential scanning calorimetry (DSC) chart.

4. The tromethamine salt according to claim 1, wherein the tromethamine salt has an endothermic transition peak at 189±2°C in a differential scanning calorimetry (DSC) chart.

5. The tromethamine salt according to claim 1, wherein the tromethamine salt is in crystalline form.

6. The tromethamine salt of claim 5, wherein the crystalline form comprises at least three diffraction peaks in an X-ray powder diffraction (XRPD) pattern at 2-theta (2θ) angle values ​​selected from the group consisting of: 4.03±0.2, 8.09±0.2, 10.04±0.2, 15.07±0.2, 15.71±0.2, 17.90±0.2, 19.60±0.2, 22.07±0.2, 24.92±0.2, and 25.39±0.

2.

7. The tromethamine salt of claim 6, wherein the crystalline form comprises diffraction peaks at the following 2-theta (2θ) angles in the XRPD pattern: 4.03±0.2, 8.09±0.2, 10.04±0.2, 15.07±0.2, 15.71±0.2, 17.90±0.2, 19.60±0.2, 22.07±0.2, 24.92±0.2, and 25.39±0.

2.

8. The tromethamine salt of claim 6, wherein the crystalline form further comprises in the XRPD pattern any one or more diffraction peaks at 2-theta (2θ) angle values ​​selected from the group consisting of: 12.17±0.2, 14.39±0.2, 17.02±0.2, 20.18±0.2, and 21.81±0.

2.

9. A pharmaceutical composition comprising the tromethamine salt according to any one of claims 1 to 8; and a pharmaceutically acceptable carrier.

10. A method for preparing a tromethamine salt of (S)-2-((4-(6-((5-cyanopyridin-2-yl)methoxy)pyridin-2-yl)piperazin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid represented by the following chemical formula I, comprising: (1) mixing a compound represented by the following chemical formula II with a single organic solvent or a mixed organic solvent, and reacting with tromethamine represented by the following chemical formula III; (2) heating the reaction solution obtained in step (1) and stirring while warming; (3) cooling and stirring the reaction solution obtained in step (2); and (4) filtering and drying the product of step (3): [Chemical Formula I] [Chemical Formula II] [Chemical Formula III] 11. The preparation method according to claim 10, wherein the organic solvent in step (1) is one or a combination of two or more selected from the group consisting of: methanol, ethanol, isopropanol, acetonitrile, acetone and ethyl acetate.

12. A pharmaceutical composition for preventing or treating metabolic diseases, comprising the tromethamine salt according to any one of claims 1 to 8.

13. The pharmaceutical composition according to claim 12, wherein the metabolic disease is at least one selected from the group consisting of diabetes, idiopathic type 1 diabetes, latent autoimmune diabetes in adults (LADA), early onset type 2 diabetes (EOD), young-onset diabetes (YOAD), maturity-onset diabetes of the young (MODY), malnutrition-related diabetes, gestational diabetes, hyperglycemia, insulin resistance, hepatic insulin resistance, impaired glucose tolerance, diabetic neuropathy, diabetic nephropathy, nephropathy, diabetic retinopathy, visceral fat accumulation, sleep apnea, obesity, eating disorders, dyslipidemia, hyperinsulinemia, non-alcoholic fatty liver disease (NAFLD), cardiovascular disease, atherosclerosis, peripheral vascular disease, hypertension, congestion. heart failure, myocardial infarction, stroke, hemorrhagic stroke, ischemic stroke, traumatic brain injury, pulmonary hypertension, restenosis after angioplasty, intermittent claudication, postprandial lipidosis, metabolic acidosis, ketosis, arthritis, osteoporosis, Parkinson's disease, left ventricular hypertrophy, peripheral arterial disease, vision loss, cataracts, glomerulosclerosis, chronic renal failure, metabolic syndrome, syndrome X, premenstrual syndrome, angina pectoris, thrombosis, transient ischemic attack, vascular restenosis, abnormal glucose metabolism, symptoms of impaired fasting glucose, hyperuricemia, gout, erectile dysfunction, psoriasis, foot ulcers, ulcerative colitis, hyperlipoproteinemia, Alzheimer's disease, schizophrenia, cognitive impairment, inflammatory bowel disease, short bowel syndrome, Crohn's disease, colitis, irritable bowel disease, and polycystic ovary syndrome. 14 . The pharmaceutical composition according to claim 13 , wherein the non-alcoholic fatty liver disease (NAFLD) is at least any one selected from the group consisting of steatosis, non-alcoholic steatohepatitis (NASH), fibrosis, cirrhosis, and hepatocellular carcinoma.

15. A glucagon-like peptide-1 receptor (GLP-1R) agonist comprising the tromethamine salt according to any one of claims 1 to 8.

16. A method for treating a metabolic disease, comprising: A therapeutically effective amount of the tromethamine salt according to any one of claims 1 to 8 is administered to a subject in need thereof.

17. The method according to claim 16, wherein the metabolic disease is at least any one selected from the group consisting of: Diabetes, idiopathic type 1 diabetes, latent autoimmune diabetes in adults (LADA), early onset type 2 diabetes (EOD), young onset diabetes (YOAD), maturity onset diabetes of the young (MODY), malnutrition-related diabetes, gestational diabetes, hyperglycemia, insulin resistance, hepatic insulin resistance, impaired glucose tolerance, diabetic neuropathy, diabetic nephropathy, kidney disease, diabetic retinopathy, visceral fat accumulation, sleep apnea, obesity, eating disorders, dyslipidemia, hyperinsulinemia, non-alcoholic fatty liver disease (NAFLD), cardiovascular disease, atherosclerosis, peripheral vascular disease, hypertension, congestive heart failure, myocardial infarction, stroke, hemorrhagic stroke, ischemic stroke Stroke, traumatic brain injury, pulmonary hypertension, restenosis after angioplasty, intermittent claudication, postprandial lipidosis, metabolic acidosis, ketosis, arthritis, osteoporosis, Parkinson's disease, left ventricular hypertrophy, peripheral arterial disease, vision loss, cataracts, glomerulosclerosis, chronic renal failure, metabolic syndrome, syndrome X, premenstrual syndrome, angina pectoris, thrombosis, transient ischemic attack, vascular restenosis, abnormal glucose metabolism, symptoms of impaired fasting glucose, hyperuricemia, gout, erectile dysfunction, psoriasis, foot ulcers, ulcerative colitis, hyperlipoproteinemia, Alzheimer's disease, schizophrenia, cognitive impairment, inflammatory bowel disease, short bowel syndrome, Crohn's disease, colitis, irritable bowel disease, and polycystic ovary syndrome. 18 . The method according to claim 17 , wherein the non-alcoholic fatty liver disease (NAFLD) is at least any one selected from the group consisting of steatosis, non-alcoholic steatohepatitis (NASH), fibrosis, cirrhosis, and hepatocellular carcinoma.

19. Use of a tromethamine salt according to any one of claims 1 to 8 for the manufacture of a medicament for the treatment of metabolic diseases.

20. The use according to claim 19, wherein the metabolic disease is at least any one selected from the group consisting of: Diabetes, idiopathic type 1 diabetes, latent autoimmune diabetes in adults (LADA), early onset type 2 diabetes (EOD), young onset diabetes (YOAD), maturity onset diabetes of the young (MODY), malnutrition-related diabetes, gestational diabetes, hyperglycemia, insulin resistance, hepatic insulin resistance, impaired glucose tolerance, diabetic neuropathy, diabetic nephropathy, kidney disease, diabetic retinopathy, visceral fat accumulation, sleep apnea, obesity, eating disorders, dyslipidemia, hyperinsulinemia, non-alcoholic fatty liver disease (NAFLD), cardiovascular disease, atherosclerosis, peripheral vascular disease, hypertension, congestive heart failure, myocardial infarction, stroke, hemorrhagic stroke, ischemic stroke Stroke, traumatic brain injury, pulmonary hypertension, restenosis after angioplasty, intermittent claudication, postprandial lipidosis, metabolic acidosis, ketosis, arthritis, osteoporosis, Parkinson's disease, left ventricular hypertrophy, peripheral arterial disease, vision loss, cataracts, glomerulosclerosis, chronic renal failure, metabolic syndrome, syndrome X, premenstrual syndrome, angina pectoris, thrombosis, transient ischemic attack, vascular restenosis, abnormal glucose metabolism, symptoms of impaired fasting glucose, hyperuricemia, gout, erectile dysfunction, psoriasis, foot ulcers, ulcerative colitis, hyperlipoproteinemia, Alzheimer's disease, schizophrenia, cognitive impairment, inflammatory bowel disease, short bowel syndrome, Crohn's disease, colitis, irritable bowel disease, and polycystic ovary syndrome.

21. The use according to claim 20, wherein the non-alcoholic fatty liver disease (NAFLD) is at least any one selected from the group consisting of steatosis, non-alcoholic steatohepatitis (NASH), fibrosis, cirrhosis, and hepatocellular carcinoma.

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