6-((5,6-Diphenyl-1,2,4-triazin-3-yl)(isopropyl)amino)-N-(methylsulfonyl)hexanamide Polymorph B, its uses and preparation method

Abstract

The present invention discloses polymorph B of compound I (6-((5,6-diphenyl-1,2,4-triazin-3-yl)(isopropyl)amino)-N-(methylsulfonyl)hexanamide), its pharmaceutical composition, use and preparation method. The polymorph B of compound I described in the present invention has diffraction peaks in its X-ray powder diffraction pattern at least at the following diffraction angles 2θ: 3.36 ± 0.2°, 10.07 ± 0.2°, 13.90 ± 0.2°, 16.51 ± 0.2°, 20.20 ± 0.2°, 21.10 ± 0.2°. The polymorph B of compound I of the present invention has the advantages of low hygroscopicity and good stability, which is of great significance for drug development.

Description

[0001] Cross - reference to related applications

[0002] This application claims the priority of Chinese Patent Application CN202410169645.1 with an application date of February 6, 2024, and this application incorporates the entire text of the above - mentioned Chinese patent application by reference. Technical field

[0003] The present invention belongs to the field of pharmacy, and particularly relates to the crystal forms of the compound 6 - ((5,6 - diphenyl - 1,2,4 - triazin - 3 - yl)(isopropyl)amino)-N-(methylsulfonyl)hexanamide, its pharmaceutical compositions, uses, and preparation methods. Background art

[0004] Pulmonary arterial hypertension (PAH) is a rare and incurable pulmonary vascular disease that can gradually lead to right - heart failure and ultimately death. The characteristics of PAH are pulmonary microvascular remodeling, which leads to a progressive increase in pulmonary vascular resistance (PVR) and thus right - heart failure, making it a progressive and fatal disease. 75% of PAH patients die within 5 years after diagnosis, and the average survival period after the onset of symptoms is 1.9 years. Therefore, it is also known as the "malignant tumor in the field of cardio - pulmonary vessels".

[0005] Currently, global treatment methods for PAH include conventional treatment and targeted treatment. Conventional treatment often only improves symptoms and cannot effectively prevent the progression of the disease. In terms of targeted treatment, there are three pathways for PAH targeted drugs, namely the nitric oxide pathway, the endothelin pathway, and the prostacyclin pathway (PGI2). PGI2 is an important vascular endothelial relaxing factor, which causes pulmonary vascular smooth muscle relaxation and inhibits smooth muscle growth by stimulating the production of cyclic adenosine monophosphate (cAMP). The lack of PGI2 can cause pulmonary arterial hypertension. Therefore, PGI2 - type drugs are currently the most active method for treating PAH. PGI2 - type drugs include PGI2 analogs and PGI2 receptor agonists. PGI2 analogs have a natural PGI2 skeleton in their structure, have a fast metabolism rate in the body, a very short biological half - life, and require high - frequency administration or intravenous infusion, resulting in poor patient compliance. In addition, their target selectivity is poor, and it is difficult to separate the therapeutic effect from other effects, and adverse reactions are likely to occur.

[0006] Compound I, with the chemical name 6-((5,6-diphenyl-1,2,4-triazin-3-yl)(isopropyl)amino)-N-(methylsulfonyl)hexanamide, and its structural formula shown in Formula I, is a PGI2 receptor agonist with a novel structure and good drug-likeness. Compound I has strong target selectivity, and its agonist activity for the prostacyclin IP receptor is more than 1000 times that of the other 7 prostacyclin receptor targets. It mainly activates the IP receptor, promotes the generation of cAMP in pulmonary artery smooth muscle cells, and then inhibits the abnormal contraction of the pulmonary artery, inhibits the proliferation of pulmonary artery smooth muscle cells, and reduces pulmonary artery pressure, achieving the effect of treating pulmonary hypertension. Compared with the marketed drugs of the same kind, it has higher efficacy and safety.

[0007]

[0008] Currently, the crystal form of Compound I has not been publicly reported. This invention focuses on the crystal form research of Compound I during the drug development process, and provides a drug crystal form with good stability, as well as its pharmaceutical composition, uses, and preparation method. Summary of the Invention

[0009] Through a large number of exploratory studies, it was found that Compound I exists in different crystalline forms. A large number of studies on the crystal forms of Compound I were carried out to determine and prepare the crystalline forms that meet the pharmaceutical requirements. Based on these studies, this invention provides Crystal Form B of Compound I, which has no hygroscopicity and good storage stability, and is suitable for formulation development. This invention also provides a pharmaceutical composition and uses of Crystal Form B of Compound I, and provides a preparation method of Crystal Form B of Compound I, whose preparation process conditions are mild and suitable for large-scale production.

[0010] To achieve the purpose of this invention, the following technical solutions are adopted:

[0011] One object of this invention is to provide Crystal Form B of Compound I, whose X-ray powder diffraction pattern expressed in 2θ angles has diffraction peaks at 3.36±0.2°, 10.07±0.2°, 13.90±0.2°, 16.51±0.2°, 20.20±0.2°, 21.10±0.2°.

[0012] In some embodiments, Crystal Form B of Compound I, whose X-ray powder diffraction pattern expressed in 2θ angles has characteristic peaks at 3.36±0.2°, 6.72±0.2°, 10.07±0.2°, 13.90±0.2°, 16.51±0.2°, 17.67±0.2°, 19.50±0.2°, 20.20±0.2°, 21.10±0.2°.

[0013] In some embodiments, Crystal Form B of Compound I, whose X-ray powder diffraction pattern expressed in 2θ angles is asFigure 1 as shown

[0014] In some embodiments, for polymorph B of Compound I, when characterized by TGA / DSC, its TGA graph can determine that polymorph B does not contain water of crystallization or solvates.

[0015] In some embodiments, for polymorph B of Compound I, when characterized by TGA / DSC, its DSC graph shows that the melting point (extrapolated onset temperature) of polymorph B is 140.5 ± 2 °C.

[0016] In some embodiments, for polymorph B of Compound I, its TGA / DSC graph is as Figure 2 as shown

[0017] The second object of the present invention is to provide a method for preparing polymorph B of Compound I, which comprises the following steps: heating and dissolving Compound I in a solvent, cooling to 50 °C to 60 °C, keeping warm until solids precipitate, then continuing to crystallize, separating, and drying to obtain polymorph B.

[0018] In some embodiments, the solvent is one or a mixed solvent of alcohols, ethers, esters, alkanes, ketones, acetonitrile, and water.

[0019] In some embodiments, the solvent is one or a mixed solvent of methanol, ethanol, isopropanol, acetonitrile, tetrahydrofuran, ethyl acetate, n - hexane, acetone, and water.

[0020] In certain preferred embodiments, the solvent is one or a mixed solvent of isopropanol, ethanol, water, tetrahydrofuran, and n - hexane.

[0021] In certain preferred embodiments, the solvent is isopropanol.

[0022] In some embodiments, the mass - volume ratio (g / mL) of Compound I to the solvent is 1:3.5 to 15.

[0023] In some embodiments, the heating and dissolving can be heating and dissolving in a single solvent or a mixed solvent, or first heating and dissolving in one solvent and then adding another solvent or two or more solvents.

[0024] In some embodiments, the heating condition is heating under reflux.

[0025] In some embodiments, when cooling to 50 °C to 60 °C, there are various cooling methods. It can be naturally and slowly cooling by turning off the heating, or slowly cooling by adding other solvents under heating or non - heating conditions, as long as the temperature range of the cooling can be ensured, and various cooling methods can be selected.

[0026] In some embodiments, the temperature is reduced to 50°C to 60°C, and the more preferred temperature reduction range is to reduce the temperature to 55°C to 60°C.

[0027] In some embodiments, the mixture is kept warm until a solid precipitates. The crystallization method can be static crystallization or crystallization under stirring, such as paddle stirring, suspension stirring, etc. Stirring crystallization is preferred as it can achieve crystallization faster than static crystallization.

[0028] In some embodiments, the mixture is kept warm until a solid precipitates, and the holding temperature fluctuates by 1°C to 2°C based on the previously reduced temperature.

[0029] In some embodiments, after the mixture is kept warm until a solid precipitates, further crystallization is continued. There are various crystallization methods, and conventional crystallization methods can all be adopted. For example, it can be natural cooling crystallization, followed by further cooling to 0°C to 10°C and holding for crystallization.

[0030] Furthermore, the natural cooling crystallization is carried out at room temperature.

[0031] Furthermore, for the natural cooling crystallization, the crystallization method can be static crystallization or crystallization under stirring, such as paddle stirring, suspension stirring, etc. Stirring crystallization is preferred as it can achieve crystallization faster than static crystallization.

[0032] Furthermore, for the natural cooling crystallization, the crystallization time is 1 to 15 h, preferably 1 to 2 h.

[0033] Furthermore, for the further cooling to 0°C to 10°C, the cooling is carried out in a cold water bath, an ice bath, or an ice-salt bath. As long as the temperature range of the cooling can be ensured, various cooling conditions can be selected.

[0034] Furthermore, after cooling to 0°C to 10°C, crystallization is carried out while holding the temperature. The crystallization method can be static crystallization or crystallization under stirring, such as paddle stirring, suspension stirring, etc. Stirring crystallization is preferred as it can achieve crystallization faster than static crystallization.

[0035] Furthermore, after cooling to 0°C to 10°C, crystallization is carried out while holding the temperature, and the crystallization time is 1 to 15 h, preferably 1 to 2 h.

[0036] In some embodiments, after the mixture is kept warm until a solid precipitates, further crystallization is continued. The crystallization method can also be to turn on the heating, raise the temperature to 55°C to 60°C, hold the temperature while stirring, then carry out natural cooling crystallization, and further cool to 0°C to 10°C and hold for crystallization.

[0037] Furthermore, for the holding and stirring, the stirring time is 20 min to 3 h, preferably 30 min to 1 h.

[0038] Further, the natural cooling and crystallization is achieved by turning off the heating and slowly cooling naturally in a hot oil bath.

[0039] Further, for the natural cooling and crystallization, the crystallization method can be static crystallization or crystallization under stirring, such as paddle stirring, suspension stirring, etc. Stirring crystallization is preferred as it can achieve crystallization faster than static crystallization.

[0040] Further, for the natural cooling and crystallization, the crystallization time is 5 - 24 h, preferably 5 - 8 h.

[0041] Further, for the continued cooling to 0°C - 10°C, the cooling condition is to carry out the cooling in a cold water bath, an ice bath or an ice - salt bath. As long as the temperature range of the cooling can be ensured, various cooling conditions can be selected.

[0042] Further, for the heat - preservation crystallization, the crystallization method can be static crystallization or crystallization under stirring, such as paddle stirring, suspension stirring, etc. Stirring crystallization is preferred as it can achieve crystallization faster than static crystallization.

[0043] Further, for the heat - preservation crystallization, the crystallization time is 1 - 20 h, preferably 1 - 2 h.

[0044] In some embodiments, after heat - preservation until solid precipitation occurs and then continuing crystallization, the crystallization method can also be natural cooling crystallization.

[0045] Further, for the natural cooling crystallization, it can be achieved by removing the oil bath and cooling naturally at room temperature, or by natural cooling in a hot oil bath.

[0046] Further, for the natural cooling crystallization, the crystallization method can be static crystallization or crystallization under stirring, such as paddle stirring, suspension stirring, etc. Stirring crystallization is preferred as it can achieve crystallization faster than static crystallization.

[0047] Further, for the natural cooling crystallization, the crystallization time is 5 - 24 h, preferably 10 - 20 h.

[0048] In some embodiments, crystal seeds of Compound I can be further added.

[0049] In some embodiments, the addition amount of the crystal seeds is 0.1% - 5% of the mass of Compound I.

[0050] In some embodiments, after the heating and dissolution, during the process of cooling to 50°C - 60°C or during the heat - preservation process after cooling to 50°C - 60°C, crystal seeds of Compound I can be added to further accelerate the precipitation of Crystal Form B.

[0051] Further, the seed crystal can be various crystal forms of Compound I, such as the seed crystal of Crystal Form A, the seed crystal of Crystal Form B, etc. The Crystal Form A of Compound I is described in the patent "6-((5,6-diphenyl-1,2,4-triazin-3-yl)(isopropyl)amino)-N-(methylsulfonyl)hexanamide Crystal Form A and Its Pharmaceutical Composition, Use and Preparation Method" filed by the present applicant on the same filing date.

[0052] In some embodiments, the separation can be a conventional method, such as centrifugation or filtration.

[0053] In some embodiments, the drying is a conventional drying method, such as vacuum drying.

[0054] In some embodiments, the drying needs to be dried to a constant weight.

[0055] The third object of the present invention is to provide a pharmaceutical composition, which comprises a therapeutically effective dose of Compound I Crystal Form B and a pharmaceutically acceptable excipient.

[0056] Further, the pharmaceutical composition can be formulated into various dosage forms for easy administration. For example, oral preparations (such as tablets, capsules, solutions or suspensions); injectable preparations (such as injectable solutions or suspensions, or injectable dry powders that can be used immediately after adding a drug solvent before injection).

[0057] The fourth object of the present invention is to provide the use of a therapeutically effective dose of Compound I Crystal Form B or the pharmaceutical composition in the preparation of a drug for preventing and / or treating a disease or disorder.

[0058] In certain preferred embodiments, the disease or disorder is related to PGI2 receptor agonistic effect.

[0059] In certain preferred embodiments, the disease or disorder is selected from: pulmonary hypertension, cardiovascular and cerebrovascular diseases related to platelet aggregation, and diabetic nephropathy, etc.

[0060] The fifth object of the present invention is to provide the use of a therapeutically effective dose of Compound I Crystal Form B or the pharmaceutical composition in the preparation of a PGI2 receptor agonist drug.

[0061] Term Definition and Explanation

[0062] As used herein, the term "room temperature" or "RT" refers to an ambient temperature of 20 to 25 °C (68 - 77 °F).

[0063] On the basis of conforming to common general knowledge in the art, the above preferred conditions can be combined arbitrarily to obtain various preferred examples of the present invention.

[0064] The positive and progressive effects of the present invention are as follows: The compound I polymorph B of the present invention has low hygroscopicity, a higher melting point, more prominent thermal stability, high purity, is not prone to generating impurities, exhibits extremely strong drug stability and polymorph stability in forced degradation tests, and is suitable for formulation development. At the same time, its preparation process conditions are mild and suitable for large-scale production. Therefore, polymorph B has obvious advantages in aspects such as drug formation and facilitating industrial production, and is of great significance for drug development. BRIEF DESCRIPTION OF THE DRAWINGS

[0065] Figure 1 It represents the XRPD spectrum of compound I polymorph B prepared in Example 1;

[0066] Figure 2 It represents the TGA / DSC spectrum of compound I polymorph B prepared in Example 1;

[0067] Figure 3 It represents that of compound I polymorph B prepared in Example 1 1 1H-NMR spectrum;

[0068] Figure 4 It represents the IR spectrum of compound I polymorph B prepared in Example 1;

[0069] Figure 5 It represents the superimposed XRPD diagram of compound I polymorph B prepared in Example 1 at high temperature, high humidity, and under light for 30 days and day 0;

[0070] Figure 6 It represents the average plasma drug-time curve of compound I in the first group of animals after intravenous administration in the bioavailability test of Experimental Example 7;

[0071] Figure 7 It represents the average plasma drug-time curve of compound I in the second group of animals after oral administration in the bioavailability test of Experimental Example 7. DETAILED DESCRIPTION OF THE INVENTION

[0072] The present invention will be further described in detail below through specific embodiments, but it is only used to help understand the present invention, enabling those skilled in the art to implement or use the present invention, and does not constitute any limitation to the present invention.

[0073] Example 1

[0074] 10 g of the raw material drug of compound I was heated under reflux and dissolved in 120 mL of isopropanol. The heating of the oil bath was turned off, and the temperature was slowly decreased naturally to 58 °C. It was stirred at 58 ± 1 °C until solids precipitated. Subsequently, it was cooled and stirred naturally at room temperature for 2 h for crystal precipitation, and then further cooled to 10 °C in a cold water bath and stirred for 1.5 h for crystal precipitation. It was filtered and vacuum dried to constant weight to obtain a sample of compound I polymorph B, with a yield of 93.5%.

[0075] Example 2

[0076] 10 g of the raw material drug of Compound I was dissolved by heating under reflux in 80 mL of absolute ethanol. The heating of the oil bath was turned off, and the temperature was slowly decreased naturally to 52 °C. It was kept at 52 ± 1 °C with stirring until solid precipitated. Subsequently, it was cooled naturally with stirring at room temperature for 1.5 h for crystallization. Then it was cooled to 5 °C in an ice bath and kept with stirring for crystallization for 2.0 h. It was filtered and dried in vacuo to constant weight to obtain a sample of crystalline form B of Compound I, with a yield of 85.3%. The X-ray powder diffraction pattern of the sample obtained by this method was consistent with that of the sample of crystalline form B of Compound I obtained in Example 1.

[0077] Example 3

[0078] 10 g of the raw material drug of Compound I was dissolved by heating under reflux in 50 mL of methanol. The heating of the oil bath was turned off, and the temperature was slowly decreased naturally to 50 °C. 0.1 g of the seed crystal of crystalline form B was added. It was kept at 50 ± 1 °C with stirring until solid precipitated. Subsequently, heating was started and the temperature was raised to 58 ± 2 °C and kept with stirring for 30 min. The heating of the oil bath was turned off, and it was cooled and stirred for crystallization naturally and slowly in a hot oil bath for 5 h. Then it was further cooled to 0 °C in an ice-salt bath and kept with stirring for crystallization for 1 h. It was filtered and dried in vacuo to constant weight to obtain a sample of crystalline form B of Compound I, with a yield of 77.4%. The X-ray powder diffraction pattern of the sample obtained by this method was consistent with that of the sample of crystalline form B of Compound I obtained in Example 1.

[0079] Example 4

[0080] 10 g of the raw material drug of Compound I was dissolved by heating under reflux in 50 mL of absolute ethanol and 50 mL of isopropanol. The heating of the oil bath was turned off, and the temperature was slowly decreased naturally to 53 °C. It was kept at 53 ± 1 °C with stirring until solid precipitated. Subsequently, it was cooled naturally at room temperature and allowed to stand for crystallization for 13 h. Then it was cooled to 3 °C in an ice bath and kept with stirring for crystallization for 1 h. It was filtered and dried in vacuo to constant weight to obtain a sample of crystalline form B of Compound I, with a yield of 88.7%. The X-ray powder diffraction pattern of the sample obtained by this method was consistent with that of the sample of crystalline form B of Compound I obtained in Example 1.

[0081] Example 5

[0082] Dissolve 10 g of the raw material of Compound I in 40 mL of ethyl acetate by heating under reflux. Turn off the heating of the oil bath, and slowly cool it naturally to 50 °C. Keep it at 50 ± 1 °C and stir until solid precipitates. Then turn on the heating, raise the temperature to 55 ± 2 °C and keep stirring for 1 h. Turn off the heating, and let it cool slowly and stand for crystal precipitation in the hot oil bath for 20 h. Then cool it to 2 °C in an ice bath, keep it at this temperature and stand for crystal precipitation for 18 h. Filter and dry it under vacuum until constant weight to obtain the sample of Compound I polymorph B, with a yield of 68.8%. The X-ray powder diffraction pattern of the sample obtained by this method is consistent with that of the sample of Compound I polymorph B obtained in Example 1.

[0083] Example 6

[0084] Dissolve 10 g of the raw material of Compound I in 90 mL of 95% ethanol by heating under reflux. Turn off the heating of the oil bath, and slowly cool it naturally to 55 °C. Keep it at 55 ± 1 °C and stir until solid precipitates. Then let it cool naturally at room temperature for crystal precipitation and stir for 1 h. Then cool it to 5 °C in an ice bath, keep it at this temperature and stir for crystal precipitation for 2.0 h. Filter and dry it under vacuum until constant weight to obtain the sample of Compound I polymorph B, with a yield of 86.2%. The X-ray powder diffraction pattern of the sample obtained by this method is consistent with that of the sample of Compound I polymorph B obtained in Example 1.

[0085] Example 7

[0086] Dissolve 10 g of the raw material of Compound I in a mixed solvent of 60 mL of acetonitrile and 40 mL of water by heating under reflux. Turn off the heating of the oil bath, and slowly cool it naturally to 51 °C. Keep it at 51 ± 1 °C and stir until solid precipitates. Then let it cool naturally at room temperature for crystal precipitation and stir for 1.5 h. Then cool it to 3 °C in an ice bath, keep it at this temperature and stir for crystal precipitation for 1 h. Filter and dry it under vacuum until constant weight to obtain the sample of Compound I polymorph B, with a yield of 76.1%. The X-ray powder diffraction pattern of the sample obtained by this method is consistent with that of the sample of Compound I polymorph B obtained in Example 1.

[0087] Example 8

[0088] Dissolve 10 g of the raw material of Compound I in a mixed solvent of 20 mL of acetone and 15 mL of water by heating under reflux. Turn off the oil bath heating, and slowly cool it naturally to 55 °C. Keep it at 55 ± 1 °C and stir until solid precipitates. Then let it cool naturally at room temperature and stir for crystal precipitation for 1 h. Then cool it to 3 °C in an ice bath, keep it at this temperature and stand for crystal precipitation for 15 h. Filter and dry it under vacuum until constant weight to obtain the sample of Compound I polymorph B, with a yield of 80.3%. The X-ray powder diffraction pattern of the sample obtained by this method is consistent with that of the sample of Compound I polymorph B obtained in Example 1.

[0089] Example 9

[0090] 10 g of the API of Compound I was dissolved by heating under reflux in 30 mL of tetrahydrofuran. 120 mL of n-hexane was slowly added, and the temperature was lowered to 60 °C. Subsequently, it was stirred at 60 ± 2 °C until a solid precipitated. The oil bath was removed, and it was allowed to cool naturally and stirred at room temperature for crystallization for 16 h. It was filtered and dried under vacuum to a constant weight to obtain a sample of polymorph B of Compound I, with a yield of 89.3%. The X-ray powder diffraction pattern of the sample obtained by this method was consistent with the pattern of the sample of polymorph B of Compound I obtained in Example 1.

[0091] Experimental Example 1: Determination of the diffraction angle (2θ) of the X-ray powder diffraction peaks

[0092] Regarding the sample of polymorph B of Compound I prepared in Example 1, its X-ray powder diffraction (XRPD) spectrum is shown in Figure 1 , and the values of the diffraction angle (2θ) of its X-ray powder diffraction peaks are shown in Table 1.

[0093] It should be noted that the physical property data of each crystal recorded in Experimental Example 1 were determined under the following conditions.

[0094] Detection instrument: Rigaku SmartLab SE fully automatic multifunctional X-ray diffractometer from Japan.

[0095] Operating conditions: X-ray tube anode: anticathode: copper; tube voltage: 40 kV; tube current: 30 mA; scanning mode: one-dimensional scanning; scanning rate: 10 ° / min; scanning axis: θ / 2θ; scanning range: 3 - 35 °; step interval: 0.01 °.

[0096] Detection results: Polymorph B showed characteristic peaks in the X-ray powder diffraction at diffraction angles 2θ of 3.36 ± 0.2 °, 6.72 ± 0.2 °, 10.07 ± 0.2 °, 13.90 ± 0.2 °, 16.51 ± 0.2 °, 17.67 ± 0.2 °, 19.50 ± 0.2 °, 20.20 ± 0.2 °, 21.10 ± 0.2 °.

[0097] Table 1 Values of the diffraction angle (2θ) of the X-ray powder diffraction peaks of the sample of polymorph B of Compound I prepared in Example 1

[0098]

[0099] Experimental Example 2: Thermogravimetric-differential scanning calorimetry analysis

[0100] Regarding the sample of polymorph B of Compound I prepared in Example 1, its thermogravimetric-differential scanning calorimetry (TGA / DSC) spectrum is shown in Figure 2 .

[0101] Detection instrument model: Synchronous thermal analyzer STA449F3

[0102] Test conditions: Temperature: 25 °C, Humidity: 35% RH

[0103] Crucible: DSC / TG pan Al2O3

[0104] Atmosphere: AIR(80 / 20)-- / NITROGEN / 50 / NITROGEN / 20

[0105] Test results: The TGA graph can determine that crystal form B does not contain crystal water or solvates; the DSC graph can confirm that the melting point (extrapolated onset temperature) of crystal form B is 140.5 ± 2 °C.

[0106] Experimental Example 3: Nuclear Magnetic Resonance Spectrum ( 1 H-NMR)

[0107] Regarding the sample of Compound I crystal form B prepared in Example 1, its nuclear magnetic resonance ( 1 H-NMR) spectrum is shown in Figure 3 .

[0108] Measurement conditions: Collected on a Bruker 500 MHz nuclear magnetic resonance spectrometer, with CDCl3 as the solvent.

[0109] Test results: 1 1H-NMR: δ: 10.181 (br, 1H), 7.493~7.439 (m, 4H), 7.392~7.378 (m, 1H), 7.317~7.261 (m, 5H), 5.072 (m, 1H), 3.608 (m, 2H), 3.230 (s, 3H), 2.376 (m, 2H), 1.729 (m, 4H), 1.460 (m, 2H), 1.303~1.289 (m, 6H), ppm.

[0110] Experimental Example 4: Infrared Spectrum (IR)

[0111] Regarding the sample of Compound I crystal form B prepared in Example 1, its infrared (IR) spectrum is shown in Figure 4 .

[0112] Instrument model: Nicolet.

[0113] Detection method: Take an appropriate amount of this product (about 1 - 2 mg), take an appropriate amount of dried potassium bromide after grinding, place it in an agate mortar, mix and grind, take an appropriate amount, press into a tablet, and measure the infrared spectrum.

[0114] Test results: The infrared spectrum of the sample is at 3105 ± 5 cm -1 、2952 ± 5 cm -1 、1684 ± 5 cm -1 、1532 ± 5 cm -1, 1473 ± 5 cm -1 , 1457 ± 5 cm -1 , 1441 ± 5 cm -1 , 1370 ± 5 cm -1 , 1150 ± 5 cm -1 , 1137 ± 5 cm -1 , 697 ± 5 cm -1 , 688 ± 5 cm -1 There are characteristic peaks at...

[0115] Experimental Example 5: Hygroscopicity

[0116] Regarding the sample of Compound I polymorph B prepared in Example 1, its hygroscopicity was determined with reference to General Chapter 9103 of the Fourth Volume of the Chinese Pharmacopoeia (2020 Edition). The specific test method is as follows:

[0117] Take a dry stoppered glass weighing bottle (outer diameter 50 mm, height 15 mm), place it in a suitable constant temperature and humidity dryer at 25°C ± 1°C one day before the test, and accurately weigh its weight (m1).

[0118] Take an appropriate amount of the test sample, spread it evenly in the above-mentioned weighing bottle, and the thickness of the test sample is generally about 1 mm, and accurately weigh its weight (m2).

[0119] Open the weighing bottle and place it together with the bottle cap under the above constant temperature and humidity conditions for 24 h. Cover the weighing bottle cap and accurately weigh its weight (m3).

[0120]

[0121] Test results: As shown in Table 2, the hygroscopic weight gain of the 3 batches of samples is less than 0.2%, indicating that the polymorph B obtained in the present invention has no hygroscopicity.

[0122] Table 2 Summary of Hygroscopicity Results

[0123] Batch number Batch 1 Batch 2 Batch 3 Percentage of weight gain 0.02％ 0.02％ 0.03％

[0124] Experimental Example 6: Stability Test

[0125] Regarding the sample of Compound I polymorph B prepared in Example 1, a stability study was carried out. The sample was placed at high temperature (60°C), high humidity (RH 90 ± 5%), and light (4500 Lx ± 500 Lx) for 30 days respectively, and then the XRPD spectra of the sample were measured respectively. By comparing with the XRPD spectrum measured on the 0th day, the polymorph change was determined. At the same time, high performance liquid chromatography was used to determine its purity to investigate the change of related substances.

[0126] The stability test results of polymorph B samples at 0 day and after 30 days under high temperature, high humidity and light are shown in Table 3. The superimposed XRPD patterns of polymorph B samples at the end of 30 days of high temperature, high humidity and light and on the 0th day are shown in Figure 5 .

[0127] Figure 5 The results showed that the XRPD patterns of polymorph B detected after 30 days of placement under the conditions of high temperature (60 °C), high humidity (RH90±5%), and light (4500Lx±500Lx) were compared with the XRPD patterns detected on the 0th day, and the crystal form remained unchanged, indicating that polymorph B is a stable crystal form.

[0128] According to the data in Table 3, there were no significant changes in the related substances of polymorph B samples after 30 days of high temperature, high humidity and light. The purity of polymorph B was very high and stable. The results of the stability study showed that this crystal form was suitable for formulation development.

[0129] Table 3 Stability test results of polymorph B samples under high temperature, high humidity and light for 30 days

[0130] Experimental conditions and time Crystal form Purity Day 0 Crystal form B 99.94％ High temperature (60°C) for 30 days Crystal form B 99.95％ High humidity (RH90±5%) for 30 days Crystal form B 99.94％ Light (4500Lx±500Lx) for 30 days Crystal form B 99.93％

[0131] Experimental Example 7: Bioavailability test

[0132] For the bioavailability test of polymorph B samples of compound I prepared in Example 1, the polymorph B samples of compound I were respectively prepared into clear solutions and suspensions by conventional methods.

[0133] In this study, 12 Beagle dogs (6 males and 6 females) were randomly divided into 2 groups, with 3 male and 3 female animals in each group. The animals in the first group were not fasted and were given 0.5 mg / kg of compound I (clear solution) by single intravenous injection, and blood samples were collected until 24 h after administration. The specific sampling time points were before administration, 0.083, 0.25, 0.5, 1, 2, 3, 4, 5, 6, 8, 12, and 24 h after administration; the animals in the second group were fasted and then given 0.5 mg / kg of compound I (suspension) by single oral gavage, and blood samples were collected until 24 h after administration. The specific sampling time points were before administration, 0.25, 0.5, 1, 2, 3, 4, 5, 6, 8, 12, and 24 h after administration; the validated HPLC-UV analysis method was used for the uniformity and concentration analysis of the administered preparations. The validated liquid chromatography-tandem mass spectrometry (LC-MS / MS) analysis method was used to detect the concentration of compound I in plasma samples. Through the blood drug concentration data, Phoenix 7.0 software was used to calculate its bioavailability by the non-compartmental model.

[0134] The average plasma concentration-time curves of Compound I in the first group of animals after intravenous administration and in the second group of animals after oral administration in the bioavailability test are shown in Figure 6 and Figure 7 .

[0135] The results showed that the bioavailability of polymorph B was 67.48%, which was average. However, since the bioavailability was between 60% and 90%, it could meet the basic requirements for in vivo absorption and drug efficacy.

[0136] The polymorph B samples of Compound I prepared in Examples 2-9 had comparable properties to the polymorph B sample of Compound I prepared in Example 1, including hygroscopicity, stability, and bioavailability.

[0137] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, or improvements made within the spirit and principle of the present invention should be included within the protection scope of the present invention.

Claims

1. Polymorph B of a compound of formula I, characterized in that, The free base crystalline form B of the compound of formula I, whose X-ray powder diffraction pattern expressed in 2θ angles has diffraction peaks at 3.36±0.2°, 10.07±0.2°, 13.90±0.2°, 16.51±0.2°, 20.20±0.2°, 21.10±0.2°. 。 2. The crystalline form B of the compound of formula I as claimed in claim 1, characterized in that, The free base crystalline form B of the compound of formula I, whose X-ray powder diffraction pattern expressed in 2θ angles has diffraction peaks at 3.36±0.2°, 6.72±0.2°, 10.07±0.2°, 13.90±0.2°, 16.51±0.2°, 17.67±0.2°, 19.50±0.2°, 20.20±0.2°, 21.10±0.2°.

3. The crystalline form B of the compound of formula I as claimed in claim 2, characterized in that, The free base crystalline form B of the compound of formula I, whose X-ray powder diffraction pattern expressed in 2θ angles has the pattern shown in Figure 1.

4. The crystalline form B of the compound of formula I as claimed in claim 1, characterized in that, The free base crystalline form B of the compound of formula I, whose melting point measured by differential scanning calorimetry is 138°C - 143°C.

5. The crystalline form B of the compound of formula I as claimed in claim 1, characterized in that, The free base crystalline form B of the compound of formula I, whose thermogravimetric-differential scanning calorimetry spectrum has the pattern shown in Figure 2.

6. The preparation method of polymorph B according to any one of claims 1-5, characterized in that, Comprising: Heating and dissolving compound I in a solvent, cooling to 50°C - 60°C, maintaining the temperature until a solid precipitates, then continuing to crystallize, separating, and drying to obtain crystalline form B; the solvent is one or more mixed solvents of methanol, ethanol, isopropanol, and water, or ethyl acetate, or a mixed solvent of acetonitrile and water, or a mixed solvent of acetone and water, or a mixed solvent of tetrahydrofuran and n-hexane.

7. The preparation method of polymorph B according to claim 6, wherein, The solvent is one or a mixed solvent of isopropanol, ethanol, and water, or a mixed solvent of tetrahydrofuran and n-hexane.

8. The preparation method of polymorph B according to claim 7, characterized in that, The solvent is isopropanol.

9. The preparation method of crystalline form B according to claim 6, wherein The mass-volume ratio of the compound of formula I to the solvent is 1 g : 3.5 mL - 15 mL.

10. The preparation method of crystalline form B according to claim 6, wherein, The heating and dissolving is: heating and dissolving in a single solvent or a mixed solvent, or first heating and dissolving in one solvent and then adding one or two or more other solvents.

11. The preparation method of polymorph B according to claim 6, characterized in that, The temperature range of cooling to 50°C - 60°C is cooling to 55°C - 60°C.

12. The preparation method of crystalline form B according to claim 6, wherein, The step of maintaining the temperature until a solid precipitates and then continuing to crystallize is: naturally cooling to crystallize, then further cooling to 0°C - 10°C and maintaining the temperature to crystallize; or turning on the heating, raising the temperature to 55°C - 60°C, maintaining the temperature and stirring, naturally cooling to crystallize, then further cooling to 0°C - 10°C and maintaining the temperature to crystallize; or naturally cooling to crystallize.

13. The preparation method of crystalline form B according to claim 6, characterized in that, During the preparation process, crystal seeds of compound I can be further added.

14. A pharmaceutical composition, characterized in that, Comprising a pharmaceutically acceptable excipient and a therapeutically effective dose of the crystalline form B of the compound of formula I according to any one of claims 1 - 5.

15. Use of a therapeutically effective dose of the crystalline form B of the compound of formula I according to any one of claims 1 - 5 or the pharmaceutical composition according to claim 14 in the preparation of a drug for preventing and / or treating pulmonary hypertension, cardiovascular and cerebrovascular diseases related to platelet aggregation, or diabetic nephropathy.

16. Use of a therapeutically effective dose of the crystalline form B of the compound of formula I according to any one of claims 1 - 5 or the pharmaceutical composition according to claim 14 in the preparation of a PGI2 receptor agonist drug.

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