Crystalline forms of NK3R antagonists and their preparation methods and uses

Abstract

The present invention discloses crystalline form A of the NK3R antagonist represented by formula (I), as well as its preparation and use. Crystalline form A has characteristic peaks at 10.40±0.20°, 11.79±0.20°, 19.51±0.20°, and 20.81±0.20° in X-ray powder diffraction using Cu-Kα radiation at 2θ angles. Crystalline form A according to the present invention is stable, has low hygroscopicity, is suitable for long-term storage, is suitable for pharmaceutical formulation, and can effectively inhibit NK3R. [Formula 1] JPEG2025528595000025.jpg37169

Description

Detailed Description of the Invention

[0001] [Technical Field] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims priority from a prior application filed with the State Intellectual Property Office of China on October 19, 2022, bearing patent application number 202211282020.3 and entitled "Crystalline Form of NK3R Antagonist, and Methods for Preparation and Use Thereof," and a prior application filed with the State Intellectual Property Office of China on October 11, 2023, bearing patent application number 202311315851.0 and entitled "Crystalline Form of NK3R Antagonist, and Methods for Preparation and Use Thereof," both of which are incorporated herein by reference in their entireties.

[0002] The present invention relates to the field of pharmaceutical compounds, particularly to the crystalline form of NK3R antagonist and its preparation method and use.

[0003] [Background technology] Tachykinin receptors are targets for a family of structurally related peptides, collectively termed "tachykinins," including substance P (SP), neurokinin A (NKA), and neurokinin B (NKB). Tachykinins are synthesized in the central nervous system (CNS) and peripheral tissues, where they exert various biological activities. Currently, three tachykinin receptors are known: neurokinin-1 (NK-1), neurokinin-2 (NK-2), and neurokinin-3 (NK-3) receptors. Tachykinin receptors belong to the rhodopsin-like seven-transmembrane G protein-coupled receptor family. SP has the highest affinity and is thought to be the endogenous ligand for the NK-1 receptor, NKA is the endogenous ligand for the NK-2 receptor, and NKB is the endogenous ligand for the NK-3 receptor. NK-1, NK-2, and NK-3 receptors have already been identified in various species. NK-1 and NK-2 receptors are expressed in a variety of peripheral tissues, and NK-1 receptors are also expressed in the CNS, whereas NK-3 receptors are expressed primarily in the CNS.

[0004] Neurokinin receptors mediate a variety of biological effects stimulated by tachykinins, including the transmission of excitatory neuronal signals (e.g., pain) in the CNS and periphery, the regulation of smooth muscle contractile activity, the modulation of immune and inflammatory responses, the induction of antihypertensive effects by dilation of the peripheral vasculature, and the stimulation of endocrine and exocrine secretions.

[0005] The NK-3 receptor is encoded by the TACR3 gene and is involved in regulating the hypothalamus-pituitary-gonadal axis. TACR3 gene knockout or mutant mice both exhibit abnormal development of reproductive organs, reduced levels of sex hormones, and significantly reduced fertility. TACR3 gene mutations cause abnormal release of gonadotropins in patients, resulting in sexual immaturity and infertility, and a significant portion of familial hypogonadism is caused by TACR3 gene mutations.

[0006] Kisspeptin / neurokinin B / dynorphin (KNDy) neurons are involved in the gonadotropin-releasing hormone (GnRH) signaling pathway, promoting estrogen production through the GnRH neuron-pituitary-genital pathway. This signaling pathway is regulated by a negative feedback mechanism, thereby maintaining hormone levels within a reasonable range. KNDy neurons are also involved in the thermoregulatory signaling pathway, releasing NKB ligands to bind to NK-3 receptors in the median preoptic nucleus, suppressing shivering and vasoconstriction, promoting sweating and vasodilation, and regulating body temperature within a certain range. In menopausal women, the decline in estrogen levels in the body causes a loss of the negative feedback mechanism, resulting in the overactivation of KNDy neurons, which releases large amounts of endogenous NKB ligands that bind to NK-3 receptors in the median preoptic nucleus, resulting in symptoms such as sweating, vasodilation, and hot flashes. Therefore, the development of antagonists against the NK-3 receptor in KNDy neurons and the median preoptic nucleus is expected to have a positive therapeutic effect on hot flash symptoms.

[0007] In the CNS, NK-3 receptors are expressed in regions including the medial prefrontal cortex, hippocampus, thalamus, and amygdala. NK-3 receptors are expressed in dopaminergic neurons. Activation of NK-3 receptors has been shown to regulate the release of dopamine, acetylcholine, and serotonin, indicating the therapeutic utility of NK-3 receptor modulators in the treatment of various diseases, including psychotic disorders, anxiety disorders, depression, schizophrenia, and obesity, pain, or inflammation.

[0008] The development of NK-3R antagonists having novel structures and pharmaceutical solid forms suitable for the formulation of such compounds, for example, solid forms with improved stability, hygroscopicity, and / or efficacy, thereby achieving good results in the formulation and administration stages, is a technical problem that those skilled in the art are constantly working to solve.

[0009] Summary of the Invention In order to solve the above technical problems, the present invention provides a crystalline form A of the compound represented by formula (I),

[0010] [ka]

[0011] The crystalline form A has characteristic peaks in X-ray powder diffraction expressed in 2θ angles at 10.40±0.20°, 11.79±0.20°, 19.51±0.20°, and 20.81±0.20° using Cu-Kα radiation.

[0012] Preferably, the crystalline form A has characteristic peaks in X-ray powder diffraction, expressed in 2θ angles, using Cu-Kα radiation at the following positions: 10.40±0.20°, 11.79±0.20°, 15.92±0.20°, 16.92±0.20°, 19.51±0.20°, 20.81±0.20°, 21.19±0.20°, and 22.91±0.20°.

[0013] In some embodiments, the crystalline form A further has characteristic peaks in X-ray powder diffraction, expressed in 2θ angles, at 3.17±0.20°, 8.50±0.20°, 13.24±0.20°, 15.15±0.20°, 19.03±0.20°, 25.03±0.20°, and / or 26.95±0.20° using Cu-Kα radiation.

[0014] In some embodiments of the present invention, the crystalline form A has an XRPD pattern substantially as shown in FIG.

[0015] In some embodiments of the present invention, the analytical data of the XRPD pattern of crystalline form A is shown in Table 1:

[0016] [Table 1]

[0017] As shown in the figure.

[0018] In some embodiments of the present invention, the analytical data of the XRPD pattern of crystalline form A is shown in Table 2:

[0019] [Table 2]

[0020] As shown in the figure.

[0021] In some embodiments of the present invention, the crystalline form A exhibits a weight loss of 0.244% at 150±3°C.

[0022] In some embodiments of the present invention, the crystalline form A has a TGA pattern substantially as shown in FIG.

[0023] In some embodiments of the present invention, the crystalline form A has an endothermic peak at 272.53±3°C.

[0024] In some embodiments of the present invention, the crystalline form A has a DSC pattern substantially as shown in FIG.

[0025] The present invention further provides a method for preparing crystalline form A of the compound represented by formula (I), which includes preparing the compound represented by formula (I) as a raw material by a poor solvent method, a solvent evaporation method, a temperature-lowering precipitation method, a suspension stirring method, a temperature cycle method, a gas-solid permeation method, a high polymer derivatization method, or a pulverization method.

[0026] Preferably, the process for preparing crystalline form A comprises: (a) adding a compound of formula (I) to a solvent to form a suspension, wherein the solvent is selected from an organic solvent, water, or a mixture of an organic solvent and water; (b) stirring the suspension at 25 to 60°C for 8 to 120 hours; (c) after step (b) is completed, centrifuging and drying (e.g., drying for 8 to 16 hours) to obtain crystalline form A.

[0027] In some embodiments of the present invention, the organic solvent may be selected from one or more of methanol, ethanol, isopropanol, acetone, methyl isobutyl ketone, heptane, toluene, m-xylene, dichloromethane, trichloromethane, anisole, methyl tert-butyl ether, dimethylacetamide, N-methylpyrrolidone, dimethyl sulfoxide, 2-methyltetrahydrofuran, 1,4-dioxane, acetonitrile, tetrahydrofuran, ethyl acetate, and isopropyl acetate, and the like.

[0028] The present invention further provides a pharmaceutical composition containing the above crystalline form A.

[0029] According to an embodiment of the present invention, the pharmaceutical composition further contains a pharmaceutically acceptable pharmaceutical excipient, for example, including, but not limited to, one or more of an excipient, a filler, a lubricant, an adhesive, a disintegrant, an inorganic salt, a solvent, a solubilizing agent, a suspending agent, an isotonicity agent, a buffer, a preservative, an antioxidant, a coloring agent, a foaming agent, and a flavoring agent.

[0030] According to an embodiment of the present invention, the pharmaceutical composition may further comprise one or more active ingredients other than the crystalline form A.

[0031] In the pharmaceutical composition, the dosage of the crystalline form A and the other active ingredient(s) is / are therapeutically effective.

[0032] The present invention further provides a use of the above-mentioned crystalline form A or the pharmaceutical composition in preparing a pharmaceutical preparation, wherein the pharmaceutical preparation is an NK-3 receptor antagonist.

[0033] According to an embodiment of the present invention, the pharmaceutical preparation is for preventing and / or treating an NK-3 receptor mediated disease, such as depression, anxiety disorder, psychosis, schizophrenia, psychotic disorder, bipolar disorder, cognitive impairment, Parkinson's disease, Alzheimer's disease, attention deficit hyperactivity disorder (ADHD), pain, convulsions, obesity, inflammatory diseases, vomiting, pre-eclampsia, airway-related diseases, reproductive disorders, contraception and sex hormone-dependent diseases and / or gynecological-related diseases.

[0034] According to an embodiment of the present invention, the sex hormone-dependent diseases include, but are not limited to, benign prostatic hyperplasia (BPH), benign prostatic hyperplasia, metastatic prostate cancer, testicular cancer, breast cancer, ovarian cancer, androgen-dependent acne, male pattern baldness, endometriosis, pubertal abnormalities, uterine fibroids, uterine fibroids, hormone-dependent cancers, hyperandrogenemia, hirsutism, virilization, polycystic ovary syndrome (PCOS), premenstrual dysphoric disorder (PMDD), HAIR-AN syndrome (hyperandrogenemia, insulin resistance, and acanthosis nigricans), ovarian theca cell hyperplasia (HAIR-AN with luteinizing theca cell proliferation in the ovarian stroma), other manifestations of high intraovarian androgen levels (e.g., arrest of follicular maturation, atresia, anovulation, dysmenorrhea, dysfunctional uterine bleeding, infertility), androgen-secreting tumors (virilizing oophoroma or adrenal tumor), menorrhagia, and / or adenomyosis.

[0035] According to embodiments of the present invention, the airway-related disease includes, but is not limited to, chronic obstructive pulmonary disease, asthma, airway hyperresponsiveness, bronchoconstriction and / or cough.

[0036] In some embodiments, the pharmaceutical formulation is for treating and / or preventing menopausal syndrome-related disorders, wherein the menopausal syndrome includes symptoms such as hot flashes, sweating, palpitations, dizziness, and / or obesity.

[0037] According to an embodiment of the present invention, the pharmaceutical formulation may be in the form of a powder, a tablet (e.g., a coated tablet, a sustained- or controlled-release tablet), a troche, a capsule (e.g., a soft capsule or a hard capsule), a granule, a pill, a dispersible powder, a suspension, a solution, an emulsion, an elixir, a syrup, an aerosol, a cream, an ointment, a gel, an injection, a lyophilized powder injection, or a suppository.

[0038] According to an embodiment of the present invention, the pharmaceutical preparation can be administered orally, bucally, sublingually, by inhalation, topically, parenterally via intravenous, subcutaneous, acupuncture or intramuscular injection, or rectally.

[0039] The present invention further provides a method for preventing and / or treating a disease, comprising administering a therapeutically effective amount of said crystalline form A or said pharmaceutical composition to a patient, the disease is an NK-3 receptor mediated disease, preferably depression, anxiety disorder, psychosis, schizophrenia, psychotic disorder, bipolar disorder, cognitive impairment, Parkinson's disease, Alzheimer's disease, attention deficit hyperactivity disorder, pain, convulsions, obesity, inflammatory diseases, vomiting, pre-eclampsia, airway-related diseases, reproductive disorders, contraception and sex hormone-dependent diseases and / or gynecological diseases; Alternatively, the disease is a disease associated with menopausal syndrome, and the menopausal syndrome includes hot flashes, sweating, palpitations, dizziness, and / or obesity symptoms.

[0040] [Beneficial Effects] The crystalline form A according to the present invention has stable properties, low hygroscopicity, is suitable for long-term storage, is suitable for pharmaceutical preparation, and can effectively inhibit NK3R.

[0041] [Definitions and explanations of terms] Unless otherwise specified, the definitions of terms in the specification and claims of this application, including their exemplary definitions, exemplary definitions, preferred definitions, definitions of specific compounds in the examples, etc., may be arbitrarily combined and combined with each other, and such combinations and combinations should be within the scope described in the specification of this application.

[0042] The term "crystalline form" refers to crystalline forms that have the same chemical composition but differ in the spatial arrangement of the molecules and / or ions that form the crystal.

[0043] The compound of formula (I) is the "free base", and "free base crystalline form A" is "crystalline form A of the compound of formula (I)".

[0044] The term "therapeutically effective amount" refers to an amount of crystalline Form A, another active ingredient, or ingredients described in the present invention sufficient to achieve the intended use (including, but not limited to, disease treatment as defined below). The therapeutically effective amount can vary depending on the intended use (in vitro or in vivo), or the subject and disease state to be treated, such as the subject's weight and age, the severity of the disease state, and the mode of administration, and can be readily determined by one skilled in the art. The specific dose will vary depending on the particular active ingredient selected, the dosing regimen followed, whether it is administered in combination with other compounds, the schedule of administration, the tissue to which it is administered, and the physical delivery system involved.

[0045] The term "patient" refers to any animal, including a mammal, preferably a mouse, rat, other rodent, rabbit, dog, cat, pig, cow, sheep, horse or primate, and most preferably a human.

[0046] The term "plurality" refers to two or more, for example, two or more than two.

[0047] The term "150±3°C" refers to 147-153°C, i.e., it may be 147°C, 148°C, 149°C, 150°C, 151°C, 152°C, 153°C, or any value between both of the aforementioned.

[0048] The term "272.53±3°C" refers to 269.53 to 275.53°C, i.e., may be 269.53°C, 270.00°C, 271.00°C, 272.00°C, 275.53°C, or any value between both of the aforementioned.

[0049] BRIEF DESCRIPTION OF THE DRAWINGS [Figure 1] XRPD pattern of crystalline form A of the compound represented by formula (I).

[0050] [Figure 2] TGA pattern of crystalline form A of the compound of formula (I).

[0051] [Figure 3] DSC pattern of crystalline form A of the compound of formula (I).

[0052] [Figure 4] DVS diagram of crystalline form A of the compound of formula (I).

[0053] [Figure 5] Comparison of XRPD patterns of crystalline form A of the compound of formula (I) before and after DVS test.

[0054] [Figure 6] Comparison of XRPD patterns of crystalline form A of the compound of formula (I) before and after stability evaluation.

[0055] [Mode for Carrying Out the Invention] The following provides a more detailed description of the technical solutions of the present invention through specific examples. It should be understood that the following examples are merely for illustrative purposes and are not intended to limit the scope of protection of the present invention. Any technology realized based on the above content of the present invention is included within the intended scope of protection of the present invention.

[0056] Unless otherwise specified, all raw materials and reagents used in the following examples are commercially available or can be prepared by known methods.

[0057] The following are the equipment, parameters, characterization and test methods used in the examples: (1) X-ray powder diffraction (XRPD) method Model number: PANalytical X-ray powder diffraction analyzer X'Pert3; X-rays: Cu, kα, Kα1(Å): 1.540598, Kα2(Å): 1.544426, Kα2 / Kα1 intensity ratio: 0.50, X-ray tube settings: 45kV, 40mA, Divergence slit: fixed 1 / 8°, Scan mode: Continuous, Scan range (°2 Theta): 3~40, Scan time per step (s): 46.7, Scan step size (°2Theta): 0.0263, Test time (min): 5

[0058] (2) Thermal Gravimetric Analyzer (TGA) method Device model number: Discovery 5500, Sample pans: open or closed aluminum pans, Starting temperature: room temperature (lower than 35°C), End temperature: 300°C or weight < 80% (w / w), stop the next step. (The weight loss of the compound is 20% (w / w) or less.) Heating rate: 10℃ / min Nitrogen gas flow rate: stable at 10 mL / min, sample chamber 25 mL / min, Sample size: approximately 2 to 10 mg.

[0059] (3) Differential Scanning Calorimetry (DSC) Method Device model number: TA Discovery2500 or Q2000, Sample pan: Tzero pan and Tzero sealing cover, with a pinhole of 0.7 mm diameter. Temperature range: 30 to 300°C or before decomposition Heating rate: 10℃ / min Nitrogen gas flow rate: 50 mL / min Sample size: approximately 0.5 to 2 mg.

[0060] (4) Dynamic Water Sorption (DVS) method DVS curves are acquired with DVS Intrinsic via SMS (Surface Measurement Systems). Relative humidity at 25°C is calibrated based on the deliquescence points of LiCl, Mg(NO3)2, and KCl. Temperature: 25℃, Sample amount: 10-20mg, Protective gas and flow rate: N2, 200mL / min, dm / dt: 0.002% / min, Minimum dm / dt equilibrium time: 10min, Maximum equilibration time: 180min, RH range: 0%RH~95%RH~0%RH, RH gradient: 10%RH (0%RH-90%RH&90%RH-0%RH), 5%RH (90%RH-95%RH&95%RH-90%RH).

[0061] (5) Ultra-high performance liquid chromatography (UPLC) method Waters H-Class ultra-high performance liquid chromatography was used to collect solubility and purity data in the project. The specific equipment and test parameters are shown in Table 3:

[0062] [Table 3]

[0063] Patent application PCT / CN2022 / 087947 describes the compound of formula (I), the entire contents of which are incorporated herein by reference.

[0064] [Table 4]

[0065] Preparation Example Compound of formula (I) The structures of the compounds of the present invention are confirmed by nuclear magnetic resonance (NMR) and / or liquid chromatography-mass spectrometry (LC-MS). NMR chemical shifts (δ) are given in parts per million (ppm). NMR measurements were performed using a Bruker AVANCE-400 nuclear magnetometer, with deuterated dimethyl sulfoxide (DMSO-d), deuterated methanol (CD3OD), and deuterated chloroform (CDCl3) as solvents, and tetramethylsilane (TMS) as the internal standard.

[0066] Liquid chromatography-mass spectrometry (LC-MS) was performed using an Agilent 1200 Infinity Series mass spectrometer. HPLC was performed using an Agilent 1200DAD high-pressure liquid chromatograph (Sunfire 18°C, 150 × 4.6 mm chromatography column) and a Waters 2695-2996 high-pressure liquid chromatograph (Gimini 18°C, 150 × 4.6 mm chromatography column).

[0067] Thin-layer chromatography silica gel plates are Yantai Yellow Sea HSGF254 or Qingdao GF254 silica gel plates, with TLC using 0.15mm to 0.20mm specifications, and thin-layer chromatography product separation and purification using 0.4mm to 0.5mm specifications. Column chromatography generally uses Yantai Yellow Sea silica gel 200-300 mesh silica gel as the carrier.

[0068] Unless otherwise specified, all reactions of the present invention are carried out under a dry nitrogen or argon atmosphere with continuous magnetic stirring, the solvents are dry solvents, and the reaction temperatures are in degrees Celsius.

[0069] 1.1 Synthesis of Intermediate 001 Preparation of (4-fluorophenyl)(8-methyl-3-(3-methyl-1,2,4-thiadiazol-5-yl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)methanone (001)

[0070] [ka]

[0071] Step 1, Preparation of 2-(chloromethyl)-3-methylpyrazine 2,3-Dimethylpyrazine 001a (10 g, 92.47 mmol) was added to carbon tetrachloride (250 mL), followed by N-chlorosuccinimide (14.83 g, 110.96 mmol) and benzoyl peroxide (224 mg, 9.25 mmol). The reaction mixture was heated at 80 °C for 16 h under nitrogen gas protection. After the reaction was confirmed by LCMS, the solvent was spun dry, and water (150 mL) and dichloromethane (3 × 100 mL) were added. The mixture was extracted and the resulting solution was separated. The extract was then washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, and filtered under reduced pressure. The filtrate was concentrated under reduced pressure and purified using a silica gel column (petroleum ether / ethyl acetate = 10:1) to give the title product, 2-(chloromethyl)-3-methylpyrazine 001b (3.20 g, colorless oil) in 22% yield.

[0072] MS m / z (ESI): 143.2[M+1] + . 1 H NMR (400 MHz, CDCl3) δ 8.45 (d, J = 2.0 Hz, 1H), 8.38 (d, J = 2.0 Hz, 1H), 4.71 (s, 2H), 2.69 (s, 3H).

[0073] Step 2, Preparation of 2-((3-methylpyrazin-2-yl)methyl)isoindoline-1,3-dione 2-(Chloromethyl)-3-methylpyrazine 001b (3.20 g, 22.44 mmol) was added to N,N-dimethylformamide solvent, and potassium phthalimide (6.23 g, 33.66 mmol) was added. The reaction mixture was reacted at 110° C. for 8 hours under nitrogen gas protection. After the reaction was complete by LCMS, the solvent was spin-dried, and water and ethyl acetate were added. The mixture was extracted and the liquid was separated. Then, it was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, and suction-filtered. The filtrate was concentrated under reduced pressure and purified on a silica gel column with petroleum ether / ethyl acetate=1:1 to obtain the title product 2-((3-methylpyrazin-2-yl)methyl)isoindoline-1,3-dione 001c as a pale yellow solid (3.10 g, 95% yield).

[0074] MS m / z (ESI): 254.2[M+1]+. 1 H NMR (400 MHz, CDCl3) δ 8.33 (d, J = 2.4 Hz, 1H), 8.24 (d, J = 2.4 Hz, 1H), 7.90 (dd, J = 5.6, 3.2 Hz, 2H), 7.75 (dd, J = 5.6, 3.2 Hz, 2H), 5.02 (s, 2H), 2.70 (s, 3H).

[0075] Step 3. Preparation of 2-(aminomethyl)-3-methylpyrazine 2-((3-Methylpyrazin-2-yl)methyl)isoindoline-1,3-dione 001c (2.00 g, 7.90 mmol) was added to ethanol (50 mL) and hydrazine hydrate (3.95 g, 79 mmol) was added. The reaction mixture was reacted at 80 °C for 6 hours under nitrogen gas protection. After the reaction was confirmed to be complete by LCMS, the solvent was spin-dried, water (150 mL) was added, and the mixture was extracted with dichloromethane / methanol (1 / 1, v / v, 50 mL). The liquid was separated, the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and suction filtered. The filtrate was concentrated under reduced pressure to give crude product 2-(aminomethyl)-3-methylpyrazine 001d (300 mg, yellow oil) in 28% yield.

[0076] MS m / z (ESI): 124.3[M+1] + . 1 H NMR (400 MHz, CDCl3) δ 8.36 (s, 1H), 8.33 (d, J = 2.4 Hz, 1H), 4.02 (s, 2H), 2.54 (s, 3H).

[0077] Step 4. Preparation of 3-methyl-N-((3-methylpyrazin-2-yl)methyl)-1,2,4-thiadiazole-5-carboxamide 3-Methyl-1,2,4-thiadiazole-5-carboxylic acid (280 mg, 1.94 mmol) was added to dichloromethane (10 mL), followed by 0.5 mL of oxalyl chloride (0.5 mL) and N,N-dimethylformamide (0.1 mL). The reaction mixture was allowed to react at 25°C for 0.5 hours. After completion of the reaction was monitored by LCMS, the solvent was spun down to give the crude product, 3-methyl-1,2,4-thiadiazole-5-carbonyl chloride.

[0078] 2-(Aminomethyl)-3-methylpyrazine 001d (200 mg, 1.62 mmol) and triethylamine (246 mg, 2.43 mmol) were added to dichloromethane (10 mL), and the crude product 3-methyl-1,2,4-thiadiazole-5-carbonyl chloride dissolved in dichloromethane (5 mL) was slowly added dropwise thereto. The reaction mixture was reacted at 25°C for 0.5 hours. After monitoring the complete reaction by LCMS, water (30 mL), dichloromethane (3 x 20 mL) were added and extracted, the liquid was separated, then washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, suction filtered, the filtrate was concentrated under reduced pressure and purified by silica gel column (petroleum ether / ethyl acetate=1:1) to give the title product 3-methyl-N-((3-methylpyrazin-2-yl)methyl)-1,2,4-thiadiazole-5-carboxamide 001e (170 mg, yellow solid), the yield is 40%.

[0079] MS m / z (ESI): 250.2[M+1] + . 1 H NMR (400 MHz, CDCl3) δ 8.59 (s, 1H), 8.46 (s, 2H), 4.78 (d, J = 4.8 Hz, 2H), 2.76 (s, 3H), 2.65 (s, 3H).

[0080] Step 5. Preparation of 3-methyl-5-(8-methylimidazo[1,5-a]pyrazin-3-yl)-1,2,4-thiadiazole 3-Methyl-N-((3-methylpyrazin-2-yl)methyl)-1,2,4-thiadiazole-5-carboxamide 001e (400 mg, 1.60 mmol) was added to acetonitrile (10 mL), followed by phosphoryl chloride (0.74 g, 4.80 mmol) and N,N-dimethylformamide (0.2 mL). The reaction mixture was reacted at 85°C for 48 hours under nitrogen gas protection. After completion of the reaction was monitored by LCMS, the solvent was spun off. Saturated sodium bicarbonate solution (50 mL), ethyl acetate (3 × 20 mL) was added, extracted, the liquid was separated, then washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, suction filtered, the filtrate was concentrated under reduced pressure and purified by silica gel column (petroleum ether / ethyl acetate = 1:1) to obtain the title product 3-methyl-5-(8-methylimidazo[1,5-a]pyrazin-3-yl)-1,2,4-thiadiazole 001f (250 mg, yellow solid), the yield was 60%.

[0081] MS m / z (ESI): 232.2[M+1] + . 1 H NMR (400 MHz, CDCl3) δ 9.58 (d, J = 3.2 Hz, 1H), 8.51 (s, 1H), 7.81 (s, 1H), 3.22 (s, 3H), 2.83 (s, 3H).

[0082] Step 6. Preparation of 7-(4-methoxybenzyl)-8-methyl-3-(3-methyl-1,2,4-thiadiazol-5-yl)imidazo[1,5-a]pyrazin-7-ium 3-Methyl-5-(8-methylimidazo[1,5-a]pyrazin-3-yl)-1,2,4-thiadiazole 001f (1.0 g, 4.3 mmol) was added to acetonitrile (6 mL), followed by potassium iodide (357 mg, 2.15 mmol) and 1-(chloromethyl)-4-methoxybenzene (1.30 g, 8.60 mmol). The reaction mixture was reacted at 8 °C for 16 hours under nitrogen gas protection. After completion of the reaction was monitored by LCMS, the solvent was spun down to afford the crude title product, 7-(4-methoxybenzyl)-8-methyl-3-(3-methyl-1,2,4-thiadiazol-5-yl)imidazo[1,5-a]pyrazin-7-ium 001g (600 mg, yellow solid), in 34% yield.

[0083] MS m / z (ESI): 352.2[M+1] + ..

[0084] Step 7. Preparation of 5-(7-(4-methoxybenzyl)-8-methyl-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazin-3-yl)-3-methyl-1,2,4-thiadiazole 7-(4-Methoxybenzyl)-8-methyl-3-(3-methyl-1,2,4-thiadiazol-5-yl)imidazo[1,5-a]pyrazin-7-ium 001g (600mg, 1.7mmol) was added to ethanol (10mL) solvent, and acetic acid (0.1mL) and sodium cyanoborohydride (320mg, 5.1mmol) were added successively. The reaction solution was heated under nitrogen gas protection until 0°C. ℃The mixture was reacted with 50 mL of HCl for 0.5 hours. After the reaction was complete by LCMS, the solvent was spun off. Water (50 mL) and dichloromethane (3 × 20 mL) were added, followed by extraction. The liquid was separated, washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, and filtered under reduced pressure. The filtrate was concentrated under reduced pressure and purified using a silica gel column (petroleum ether / ethyl acetate = 1:1) to obtain the title product 5-(7-(4-methoxybenzyl)-8-methyl-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazin-3-yl)-3-methyl-1,2,4-thiadiazole 001h (300 mg, yellow solid), with a yield of 24%.

[0085] MS m / z (ESI): 356.2[M+1] + ..

[0086] Step 8. Preparation of 3-methyl-5-(8-methyl-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazin-3-yl)-1,2,4-thiadiazole 5-(7-(4-Methoxybenzyl)-8-methyl-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazin-3-yl)-3-methyl-1,2,4-thiadiazole 001h (200 mg, 0.56 mmol) was added to trifluoroacetic acid (3 mL). Under nitrogen gas protection, the reaction mixture was reacted at 100°C for 16 hours. After completion of the reaction was monitored by LCMS, the reaction mixture was cooled to room temperature. The solvent was spin-dried to obtain the crude product, which was purified using a reverse-phase column (acetonitrile / water = 1:10) to obtain the title product, 3-methyl-5-(8-methyl-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazin-3-yl)-1,2,4-thiadiazole 001i (120 mg, white solid), in a 72% yield.

[0087] MS m / z (ESI): 236.2[M+1] + . HNMR: 1 H NMR (400 MHz, DMSO- d6) δ 9.47 (s, 1H), 7.32 (s, 1H), 4.95- 4.88 (m, 1H), 4.70 (q, J = 6.4 Hz, 1H), 4.45-4.35 (m, 1H), 3.86-3.79 (m, 1H), 3.60-3.51 (m, 1H), 2.66 (s, 3H), 1.63 (d, J = 6.8 Hz, 3H).

[0088] Step 9, Preparation of (4-fluorophenyl)(8-methyl-3-(3-methyl-1,2,4-thiadiazol-5-yl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)methanone 3-Methyl-5-(8-methyl-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazin-3-yl)-1,2,4-thiadiazole 001i (100 mg, 0.42 mmol) was dissolved in dichloromethane (4 mL), and triethylamine (64 mg, 0.63 mmol) and p-fluorobenzoyl chloride (80 mg, 0.50 mmol) were added sequentially. The reaction mixture was incubated at 25 °C for 2 hours. After the reaction was completed, water (20 mL) and dichloromethane (2 × 20 mL) were added, followed by extraction. The liquid was separated, washed with 20 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated. The resulting residue was purified using a reverse-phase chromatography column (acetonitrile / water = 1:1) to give 001 (10.20 mg, white solid) in 25% yield.

[0089] MS m / z (ESI): 358.0 [M+1] + . 1H NMR (400 MHz, CDCl3) δ 7.47 (dd, J = 8.6, 5.3 Hz, 2H), 7.16 (t, J = 8.6 Hz, 2H), 7.07 (s, 1H), 5.71 (br s, 1H), 5.06 (dd, J = 13.8, 2.4 Hz, 1H), 4.43 - 4.35(m, 1H), 4.24 - 4.17(m, 1H), 3.54 (t, J = 12.7 Hz, 1H), 2.68 (s, 3H), 1.61 (d, J = 6.8 Hz, 3H).

[0090] 1.2 Resolution of Intermediate 001 Preparation of (R)-(4-fluorophenyl)(8-methyl-3-(3-methyl-1,2,4-thiadiazol-5-yl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)methanone (002) and (S)-(4-fluorophenyl)(8-methyl-3-(3-methyl-1,2,4-thiadiazol-5-yl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)methanone (003)

[0091] [ka]

[0092] (4-Fluorophenyl)(8-methyl-3-(3-methyl-1,2,4-thiadiazol-5-yl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)methanone 001 (100 mg) was resolved by perp-SFC (CO₂ / MeOH (0.2N H₄OH)) and (R)-(4-fluoro ... (S)-(4-fluorophenyl)(8-methyl-3-(3-methyl-1,2,4-thiadiazol-5-yl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)methanone (002) (39.20 mg, white solid) and (S)-(4-fluorophenyl)(8-methyl-3-(3-methyl-1,2,4-thiadiazol-5-yl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)methanone (003) (39.20 mg, white solid).

[0093] The data corresponding to intermediate 002 are as follows: t R =3.62min HNMR: 1 H NMR (400 MHz, CDCl3) δ 7.47 (dd, J = 8.6, 5.6 Hz, 2H), 7.16 (t, J = 8.6 Hz, 2H), 7.06 (s, 1H), 5.93-5.52 (m, 1H), 5.06 (dd, J = 13.8, 2.4 Hz, 1H), 4.54-4.11 (m, 2H), 3.54 (t, J = 12.4 Hz, 1H), 2.68 (s, 3H), 1.61 (d, J = 6.8 Hz, 3H).

[0094] The data corresponding to intermediate 003 are as follows: t R =1.82min MS m / z (ESI): 358.0 [M+1] + . HNMR: 1 H NMR (400 MHz, CDCl3) δ 7.51-7.43 (m, 2H), 7.16 (t, J = 8.6 Hz, 2H), 7.06 (s, 1H), 5.91-5.47 (m, 1H), 5.05 (dd, J = 13.8, 2.4 Hz, 1H), 4.52-4.06 (m, 2H), 3.54 (t, J = 12.6 Hz, 1H), 2.67 (s, 3H), 1.61 (d, J = 6.8 Hz, 3H).

[0095] SFC division conditions: Chromatography column: Daicel CHIRALPAK OZ-H 250mm*20mm ID, 5μm Mobile phase: CO2 / MeOH(0.2%NH4·OH)=70 / 30 Flow rate: 50g / min.

[0096] 1.3 Synthesis of compounds of formula (I)

[0097] [ka]

[0098] Step 1: Preparation of (R)-(1-bromo-8-methyl-3-(3-methyl-1,2,4-thiadiazol-5-yl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)(4-fluorophenyl)methanone (004) (R)-(4-Fluorophenyl)(8-methyl-3-(3-methyl-1,2,4-thiadiazol-5-yl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)methanone 002 (50 mg, 0.013 mmol) was dissolved in dichloromethane (10 mL) and N-bromosuccinimide (25 mg, 0.013 mmol) was added. The mixture was stirred at room temperature for 1 hour. The reaction mixture was extracted with dichloromethane (10 mL), washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and filtered under suction. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by reverse-phase column chromatography (40% acetonitrile / water) to give (R)-(1-bromo-8-methyl-3-(3-methyl-1,2,4-thiadiazol-5-yl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)(4-fluorophenyl)methanone 004 (16 mg, white solid), yield 26%.

[0099] MS m / z (ESI): 436.20 [M+1] + .

[0100] 1 H NMR (400 MHz, CDCl3) δ 7.49-7.45 (m, 2H), 7.19-7.17 (m, 2H), 6.02- 5.81 (m, 1H), 5.13- 4.92 (m, 2H), 4.25- 4.13 (m, 1H), 3.68- 3.49 (m, 1H), 2.68 (s, 3H), 1.64 (d, J = 6.4 Hz, 3H). HPLC: 254nm (99.76%), 214nm (99.53%).

[0101] Step 2: Preparation of (R)-1-(7-(4-fluorobenzoyl)-8-methyl-3-(3-methyl-1,2,4-thiadiazol-5-yl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazin-1-yl)-pyrrolidin-2-one (R)-(1-Bromo-8-methyl-3-(3-methyl-1,2,4-thiadiazol-5-yl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)(4-fluorophenyl)methanone 004 (40 mg, 0.09 mmol) was dissolved in 1,4-dioxane (2 mL), and pyrrolidin-2-one (100 mg, 0.19 mmol), potassium carbonate (38 mg, 0.28 mmol), cuprous iodide (1 mg, 0.005 mmol), 2-dicyclohexylphosphine-2',6'-diisopropoxy-1,1'-biphenyl (18 mg, 0.04 mmol), and (1R,2R)-N1,N2-dimethylcyclohexane-1,2-diamine (3 mg, 0.02 mmol) were added sequentially. The reaction mixture was heated to 120°C under nitrogen gas protection and stirred for 16 hours. After completion of the reaction, the mixture was quenched with water, extracted with ethyl acetate (3 x 10 mL), and the combined organic phases were washed with saturated brine (20 mL). The combined organic phases were dried over anhydrous sodium sulfate and concentrated. The resulting residue was subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 50 / 50) to obtain a crude product. The crude product was separated and purified using a reverse phase column (mobile phase: acetonitrile / water = 52 / 48) to obtain (R)-1-(7-(4-fluorobenzoyl)-8-methyl-3-(3-methyl-1,2,4-thiadiazol-5-yl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazin-1-yl)-pyrrolidin-2-one (6.53 mg), i.e., the compound represented by Formula (I), in a 15% yield.

[0102] MS m / z (ESI): 441.1 [M+1] + .

[0103] HPLC: 90.94% (214 nm), 97.04% (254 nm). 1 H NMR (400 MHz, CDCl3) δ 7.56 - 7.45 (m, 2H), 7.21 - 7.12 (m, 2H), 5.99 (s, 1H), 5.11 (d, J = 12.8 Hz, 1H), 5.01 - 4.72 (m, 1H), 4.28 - 4.12 (m, 2H), 3.64 (s, 1H), 3.43 (s, 1H), 2.69 (s, 3H), 2.52 (s, 2H), 2.30 - 2.12 (m, 2H), 1.36 (s, 3H).

[0104] Example 1: Preparation of crystalline form A of the compound of formula (I) Antisolvent addition test A total of 13 anti-solvent addition experiments were conducted using different solvents. Approximately 15 mg of the starting sample of the compound of Formula (I) was weighed and placed in a 20 mL vial. After dissolving in a certain amount of solvent, the corresponding anti-solvent was gradually added to the clear solution, and the solution was stirred dropwise (500 rpm) until a solid precipitated. If no solid precipitated after adding 5 mL of anti-solvent, the experiment was stopped. The clear solution was transferred to 5°C / -20°C and stirred to induce solid precipitation. If the solution was still clear, it was transferred to room temperature and evaporated. The solid was collected and subjected to XRPD testing. The results are shown in Table 4. The anti-solvent addition experiment yielded free base crystalline form A, with no other crystalline forms observed.

[0105] [Table 5]

[0106] *: No solid was obtained by adding a poor solvent, and the mixture remained transparent even after being transferred to 5°C / -20°C and stirred, and then transferred to room temperature for evaporation.

[0107] #: No solid was obtained by adding antisolvent, but after transferring to 5°C and stirring, a solid precipitated.

[0108] Slow evaporation test A total of five slow evaporation experiments were conducted using different solvents. Approximately 15 mg of the starting sample of the compound of Formula (I) was weighed and placed in a 3 mL vial. An appropriate amount of the corresponding solvent was added to dissolve the sample. The solution was filtered (using a 0.45 μm PTFE filter membrane) to obtain a clear solution. The solution was then sealed with a sealing film at room temperature, punctured with 4-5 small holes, and allowed to evaporate naturally. The resulting solid was collected and subjected to XRPD testing. The test results are shown in Table 5. The slow evaporation experiment yielded free base crystalline form A, with no other crystalline forms observed.

[0109] [Table 6]

[0110] Slow temperature drop test A total of eight slow cooling experiments were conducted using different solvents. Approximately 15 mg of a starting sample of the compound of Formula (I) was weighed and placed in a 3 mL vial. The corresponding solvent was added and stirred at 50°C for 2 hours. The supernatant was then removed and filtered. The resulting filtrate was cooled from 50°C to 5°C at a rate of 0.1°C / min and maintained at 5°C. If the sample was a clear solution, it was transferred to -20°C to induce solid precipitation. If it was still clear, it was transferred to room temperature and evaporated. The precipitated solid was collected and subjected to XRPD testing. The test results are shown in Table 6. The free base crystalline form A was obtained in the slow cooling experiment, with no other crystalline forms observed.

[0111] [Table 7]

[0112] *: No solid was obtained by slowly cooling the solution, and it remained a transparent solution even when transferred to -20°C. It was then transferred to room temperature and evaporated.

[0113] #: No solid was obtained by slowly cooling the temperature, but after transferring to -20°C, a solid precipitated.

[0114] Suspension stirring (room temperature / 50℃) test A total of 38 suspension stirring tests were conducted using different solvents at different temperatures. Approximately 15 mg of the starting sample of the compound of Formula (I) was weighed and placed in an HPLC vial, and 1.0 mL of each of the solvents listed in Table 7 was added. The resulting suspension was stirred at the corresponding temperature for a certain period of time, then centrifuged, and the solid was collected and subjected to XRPD testing. If the sample was a clear solution, it was transferred to -20°C and stirred to induce solid precipitation. If it was still clear, it was transferred to room temperature and evaporated. The precipitated solid was collected and subjected to XRPD testing. The test results are shown in Table 7. The suspension stirring test yielded free base crystalline form A, with no other crystalline forms observed.

[0115] [Table 8] JPEG0007818738000013.jpg176169

[0116] *: After suspending and stirring at room temperature or 50°C, the solution became transparent, and after being transferred to -20°C and stirred, the solution remained transparent, and was then transferred to room temperature and evaporated.

[0117] Temperature Cycle Test A total of 12 temperature cycling experiments were conducted using different solvents. Approximately 15 mg of the starting sample of the compound of Formula (I) was weighed and placed in an HPLC bottle. An appropriate amount of solvent was added, and the suspension was placed in a biochemical incubator at 50°C for a temperature cycling experiment. (Procedure: The temperature was kept constant at 50°C for 2 hours, then cooled to 5°C at a rate of 0.1°C / min, and then kept constant at 5°C for 2 hours, and then heated to 50°C at a rate of 0.1°C / min. A total of two cycles were performed. Finally, all samples were kept at 5°C.) After the temperature cycling procedure was completed, the resulting solid was characterized by XRPD. If the sample was a clear solution, it was transferred to -20°C and stirred to induce solid precipitation. If it was still clear, it was transferred to room temperature and evaporated. The test results are shown in Table 8. The free base crystalline form A was obtained in the temperature cycling experiment, and no other crystalline forms were observed.

[0118] [Table 9]

[0119] *: After the temperature was increased and decreased in cycles and stirring, the solution became clear, and after being transferred to -20°C and stirred, it remained a clear solution, and was then transferred to room temperature and evaporated.

[0120] Gas-solid permeation test A total of 13 gas-solid permeation tests were conducted in different solvents. Approximately 15 mg of the starting sample of the compound of formula (I) was weighed and placed in a 3 mL vial. A 20 mL vial was taken and the corresponding solvent was added thereto. The 3 mL vial was placed in an open state in another 20 mL vial, sealed, and left at room temperature for one week. The solid was then collected and subjected to XRPD testing. When the solid was completely dissolved, it was transferred to room temperature and evaporated. The test results are shown in Table 9. The free base crystalline form A was obtained in the gas-solid permeation test, and no other crystalline forms were observed.

[0121] [Table 10]

[0122] *: One week after gas-solid permeation, the solution became clear and was transferred to room temperature for evaporation.

[0123] High polymer induction test A total of six high-polymer derivatization experiments were conducted using different solvents. Approximately 15 mg of the starting sample of the compound of formula (I) was weighed and placed in a 3 mL vial. After dissolving in a certain amount of solvent, a clear solution was obtained. If the solution did not become clear even after adding 1.5 mL of solvent, it was filtered (using a 0.45 μm PTFE filter membrane) to obtain a clear solution. Approximately 2 mg of the corresponding polymer was added to the corresponding 3 mL vial. The vial was sealed with a sealing film at room temperature, punctured with four small holes, and then placed in a fume hood for natural evaporation. The solid was collected and subjected to XRPD testing. The results are shown in Table 10. The high-polymer derivatization experiment yielded free base crystalline form A, with no other crystalline forms observed.

[0124] [Table 11]

[0125] Mixed High Polymer A: Polyvinylpyrrolidone, polyvinyl alcohol, polyvinyl chloride, polyvinyl acetate, hydroxypropyl methylcellulose and methylcellulose (mixed by equal weight), Mixed High Polymer B: Polycaprolactone, polyethylene glycol, polymethyl methacrylate, sodium alginate and hydroxyethyl cellulose (mixed in equal amounts by mass).

[0126] Crushing test A total of five grinding tests were conducted using different solvents. Approximately 15 mg of the starting sample of the compound of formula (I) was taken and placed in a mortar. No corresponding solvent was added to the mortar, or an appropriate amount was added, and the solid was manually ground (for about 5 minutes). After grinding, the obtained solid sample was subjected to XRPD testing. The test results are shown in Table 11, and all grinding tests yielded free base crystalline form A.

[0127] [Table 12]

[0128] Example 2 Characterization of Crystalline Form A of Compound of Formula (I) The XRPD pattern of the free base crystalline form A prepared in Example 1 is shown in FIG. 1, and its analytical data are shown in Table 12.

[0129] [Table 13] JPEG0007818738000019.jpg229169

[0130] The free base crystalline form A had a TGA pattern as shown in Figure 2 and showed a weight loss of 0.244% at 150°C.

[0131] The DSC pattern of the free base crystalline form A is shown in Figure 3, with the endothermic peak temperature being 272.53°C.

[0132] The hygroscopicity of free base crystalline form A was evaluated by DVS testing at a constant temperature of 25°C. The DVS results are shown in Figure 4, which indicate that the water adsorption of free base crystalline form A at 25°C / 80% RH was approximately 0.15%, indicating that free base crystalline form A has almost no hygroscopicity. The XRPD results before and after the DVS test are shown in Figure 5, which indicate that the crystalline form of free base crystalline form A did not change before and after the DVS test.

[0133] The solid-state stability of free base crystalline form A was evaluated. Appropriate amounts of sample were weighed and left open for 1 day at 60°C, and then left open for 1 week and 4 weeks at 25°C / 60%RH and 40°C / 75%RH. After leaving, the samples were subjected to XRPD and HPLC characterization to detect changes in crystalline form and chemical purity. The test results are summarized in Table 13, and the XRPD comparison diagram is shown in Figure 6. The results showed that the purity of free base crystalline form A did not change significantly (<0.2%, area%) and the crystalline form did not change at all after leaving open for 1 day at 60°C, and then left open for 1 week and 4 weeks at 25°C / 60%RH and 40°C / 75%RH, respectively, indicating that free base crystalline form A has good physicochemical stability under the current evaluation conditions.

[0134] [Table 14]

[0135] Example 3 Biological evaluation of compounds of formula (I) Determination of the activity of compounds of formula (I) against human NK-3 receptor The method is for measuring the agonistic effect of the compound represented by formula (I) on the activity of human NK-3 receptor protein expressed in human NK-3R / HEK293 stably transfected cell line.

[0136] 1. Test materials and instruments 1.1 Culture medium F12 (Gibco, Cat#11765-047), FBS (Corning, Cat#35-076-CV), Geneticin (Invitrogen, Cat# 10131), Penicillin / Streptomycin (Invitrogen, Cat# 15140).

[0137] 1.2 Reagents Fluo-4 Direct (Invitrogen, Cat# F10471), HBSS (Gibco, Cat#14025076), HEPES (Gibco, Cat#15630080), Bonine Serum Albumin(Sgima,Cat#B2064-100G).

[0138] 1.3 Equipment Consumables 384 well Poly-D-Lysine protein coating plate (Greiner, Cat#781946), FLIPR (Molecular Devices), Vi-cell XR Cell Viability Analyzer (Beckman Coulter), Incubator (Thermo).

[0139] 2. Experimental steps 2.1 Human NK-3R / HEK293 stable transfected cell line was seeded into a 384-well cell culture plate at a seeding density of 12,000 cells / well / 25 μL and cultured overnight at 37°C and 5% CO2. 2.2 Thaw 20X Component A to room temperature, dilute it to 2X working concentration with Assay Buffer, and let it stand at room temperature. 2.3 Equilibrate the cell culture plate at room temperature for 10 minutes, remove the medium, add 20 μL of Assay Buffer and 20 μL of 2X Component A, centrifuge at 200 g for 3 to 5 seconds at room temperature, and incubate at 37°C for 2 hours. 2.4 The compounds were diluted 3-fold in the 384 PP_DMSO plate with DMSO, and each compound was transferred to the working plate at 240 nl / well using the Echo 550. 40 μl of Assay Buffer was added to the working plate, and the mixture was shaken at 200 g at room temperature for 1 minute. After that, the mixture was mixed evenly on a shaker at 2500 rpm for 20 minutes, and then the mixture was left to stand at 200 g at room temperature for 1 minute. 2.5 Prepare 2.5 nM Neurokinin B TFA (6X) in Assay Buffer, take 50 μL, add to the 3657 plate, and wait. 2.6 Remove the cell culture plate and let it stand at room temperature for 10 minutes. Add 10 μL of the compound diluted in step 2.4 to the corresponding wells and let it stand at 25°C for 30 minutes. 2.7 Using the FLIPR Tetra, 10 μL of the compound diluted in step 2.5 was added to the corresponding experimental wells and data was collected.

[0140] From the above experiments, the antagonistic activity of the compound represented by formula (I) against the human NK-3 receptor was measured, and an inhibition curve of the compound represented by formula (I) was obtained. The concentration of the compound corresponding to the reference agonist that inhibited 50% (IC 50 ) and confirm the specific IC 50 The values ​​are shown in Table 14.

[0141] [Table 15]

[0142] Testing Conclusion: Based on these data, the compounds of formula (I) are potent NK-3 receptor antagonists.

[0143] Although the embodiments of the present invention have been described above, the present invention is not limited to the above embodiments. Any modifications, equivalent replacements, improvements, etc. made without departing from the spirit and principles of the present invention should be included within the scope of the claims of the present invention. [Brief explanation of the drawings]

[0144] [Figure 1] 1 is an XRPD pattern of crystalline form A of the compound of formula (I). [Figure 2] 1 is a TGA pattern of crystalline form A of the compound of formula (I). [Figure 3] 1 is a DSC pattern of crystalline form A of the compound of formula (I). [Figure 4] FIG. 1 is a DVS diagram of crystalline form A of the compound of formula (I). [Figure 5] 1 is a comparison of XRPD patterns of crystalline form A of the compound of formula (I) before and after DVS testing. [Figure 6] 1 is a comparison of XRPD patterns of crystalline form A of the compound of formula (I) before and after stability evaluation.

Claims

1. A crystal of crystalline form A of the compound of formula (I), 【Chemistry 1】 Here, the crystal of crystalline form A is a crystal of a compound having characteristic peaks at 10.40±0.20°, 11.79±0.20°, 19.51±0.20°, and 20.81±0.20° in X-ray powder diffraction expressed at 2θ angles using Cu-Kα radiation.

2. 2. The crystal of crystalline form A according to claim 1, wherein the crystal exhibits characteristic peaks at 10.40±0.20°, 11.79±0.20°, 15.92±0.20°, 16.92±0.20°, 19.51±0.20°, 20.81±0.20°, 21.19±0.20°, and 22.91±0.20° in X-ray powder diffraction analysis using Cu-Kα radiation.

3. The crystal of crystalline form A according to claim 1, wherein the X-ray powder diffraction measured at 2θ angles using Cu-Kα radiation has characteristic peaks at 3.17±0.20°, 8.50±0.20°, 13.24±0.20°, 15.15±0.20°, 19.03±0.20°, 25.03±0.20°, and / or 26.95±0.20°.

4. The crystalline form A of claim 1 has the analytical data of the XRPD pattern shown in Table 1: Table 1

5. 5. A method for preparing a crystal of crystalline form A of the compound of formula (I) according to any one of claims 1 to 4, comprising preparing the crystal of crystalline form A of the compound of formula (I) as a raw material by a poor solvent method, a solvent evaporation method, a temperature-lowering precipitation method, a suspension stirring method, a temperature cycle method, a gas-solid permeation method, a high polymer derivatization method, or a pulverization method.

6. The method for preparing the crystals of crystalline form A includes the steps of: (a) adding a compound of formula (I) to a solvent to form a suspension, wherein the solvent is selected from an organic solvent, water, or a mixture of an organic solvent and water; (b) stirring the suspension at 25-60°C for 8-120 hours; and (c) after step (b) is completed, centrifuging and drying to obtain the crystals of crystalline form A.

7. The method of claim 6, wherein the organic solvent is selected from one or more of methanol, ethanol, isopropanol, acetone, methyl isobutyl ketone, heptane, toluene, m-xylene, dichloromethane, trichloromethane, anisole, methyl tert-butyl ether, dimethylacetamide, N-methylpyrrolidone, dimethyl sulfoxide, 2-methyltetrahydrofuran, 1,4-dioxane, acetonitrile, tetrahydrofuran, ethyl acetate, and isopropyl acetate.

8. A pharmaceutical composition comprising the crystal of crystalline form A according to any one of claims 1 to 4.

9. 9. The pharmaceutical composition of claim 8, further comprising a pharmaceutically acceptable pharmaceutical excipient.

10. The pharmaceutical composition further comprises one or more active ingredients other than the crystals of crystalline form A, 9. The pharmaceutical composition according to claim 8, wherein the dosage of the crystal of crystalline form A and the dosage of one or more other active ingredients other than the crystal of crystalline form A are each therapeutically effective amounts.

11. Use of a crystal of crystalline form A according to any one of claims 1 to 4 or a pharmaceutical composition containing the crystal of crystalline form A according to any one of claims 1 to 4 in the preparation of a pharmaceutical formulation.

12. The pharmaceutical preparation is for preventing and / or treating an NK-3 receptor-mediated disease. Or the use according to claim 11, wherein the pharmaceutical preparation is for treating and / or preventing diseases associated with menopausal syndrome, the menopausal syndrome including symptoms of hot flashes, sweating, palpitations, dizziness and / or obesity.

13. The use according to claim 12, wherein the pharmaceutical preparation is for the prevention and / or treatment of depression, anxiety disorders, psychosis, schizophrenia, psychotic disorders, bipolar disorders, cognitive disorders, Parkinson's disease, Alzheimer's disease, attention deficit hyperactivity disorder, pain, convulsions, obesity, inflammatory diseases, vomiting, pre-eclampsia, airway-related diseases, reproductive disorders, contraception and sex hormone-dependent diseases and / or gynecological-related diseases.

14. The use of claim 11, wherein the pharmaceutical preparation is an NK-3 receptor antagonist.

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