Salts of dipeptide compounds, and methods for preparing and using the same

Abstract

The present invention relates to novel salts of Compound (II) and their crystals, as well as methods for preparing said salts and their crystals. The present invention further provides pharmaceutical compositions comprising the hydrochloride salt of Compound (II), and the use of said salts in the preparation of medicaments for the prevention and treatment of thrombin-mediated and / or thrombin-related diseases. TIFF2026504578000018.tif41170

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This invention claims priority to Chinese Application No. 202310121070.1, filed on February 16, 2023, the entire contents of which are incorporated herein by reference.

[0002] The present invention relates to the field of pharmaceutical chemical industry, and specifically to salts of dipeptide compounds, as well as methods for preparing and using the same. [Background technology]

[0003] US Patent US17289379 discloses a compound of formula II (abbreviated as Compound II), which is used to mediate the activity of tryptase serine protease, and is used as an anticoagulant and as a drug that modulates or inhibits the serine protease activity of tryptase, thereby enabling the treatment of thromboembolic diseases and other related cardiovascular diseases. [ka]

[0004] Blood possesses both a coagulation system and an anticoagulation system (fibrinolytic system). Under physiological conditions, coagulation factors in blood are continuously and limitedly activated, generating thrombin and forming minute amounts of fibrin. This fibrin deposits in the endothelium of the cardiovascular system after formation, but activation of the fibrinolytic system rapidly dissolves the small amount of deposited fibrin. Therefore, the coagulation system and anticoagulation system (fibrinolytic system) ensure both the blood's potential coagulability and its fluidity. When a factor that induces clot formation appears, the dynamic balance described above is disrupted, triggering a coagulation reaction. The coagulation reaction primarily consists of a primary coagulation process and a secondary coagulation process. The secondary coagulation process involves a series of biochemical reactions (the blood coagulation cascade). This cascade includes the activation of inactive enzymes (or proenzymes)—blood coagulation factors—to serine proteases (from factor X to factor Xa). Serine proteases can subsequently activate subsequent coagulation factors (factor Xa activates factor II to form factor IIa), ultimately converting the soluble plasma protein fibrinogen into an insoluble plasma protein. Fibrin-thrombin, an extracellular insulin-like serine protease, plays a key role in the blood coagulation cascade and is the final enzyme in this process. Inhibition of thrombin activity can block clot formation. After oral administration, compound II is degraded in the body by carboxylesterase-1 / carboxylesterase-2 (CES-1 / CES-2) via two intermediate states to the active metabolite compound V. Compound V exerts its anticoagulant effect by selectively inhibiting thrombin. [ka]

[0005] However, excessive inhibition of thrombin activity can lead to bleeding risks, so the amount of anticoagulant used must be precisely controlled in clinical settings. Summary of the Invention

[0006] According to one aspect, the present invention provides a crystalline form of the hydrochloride salt of the compound of formula II. [ka]

[0007] In some embodiments of the present invention, the stoichiometric ratio of the compound of Formula II to HCl is 1:1 or 1:2.

[0008] According to another aspect, the present invention provides crystalline form A of the monohydrochloride salt of compound of formula II, having diffraction peaks at 6.7±0.2°, 16.2±0.2°, 21.8±0.2°, and 22.9±0.2° 2θ in an X-ray diffraction spectrum measured using Cu-Kα radiation.

[0009] Typically, it has diffraction peaks at 2θ of 6.7±0.2°, 16.2±0.2°, 16.5±0.2°, 20.7±0.2°, 21.8±0.2°, and 22.9±0.2°.

[0010] More typically, it has diffraction peaks at 2θ of 6.7±0.2°, 7.8±0.2°, 16.2±0.2°, 16.5±0.2°, 19.4±0.2°, 20.7±0.2°, 21.8±0.2°, 22.9±0.2°, 23.8±0.2°, and 27.5±0.2°.

[0011] In some embodiments of the present invention, crystalline Form A of the monohydrochloride salt of the compound of formula II has an X-ray diffraction spectrum substantially the same as the X-ray diffraction spectrum shown in FIG. 1, measured using Cu-Kα radiation.

[0012] In some embodiments of the present invention, crystalline Form A of the monohydrochloride salt of the compound of Formula II has a DSC spectrum substantially the same as the DSC spectrum shown in FIG. 2 or a TGA spectrum substantially the same as the TGA spectrum shown in FIG. 3.

[0013] According to another aspect, the present invention provides crystalline form B of the monohydrochloride salt of compound of formula II, having diffraction peaks at 6.6±0.2°, 15.2±0.2°, 21.2±0.2°, and 21.6±0.2° 2θ in an X-ray diffraction (XRD) spectrum measured using Cu-Kα radiation. Typically, form B has diffraction peaks at 6.6±0.2°, 15.2±0.2°, 20.3±0.2°, 21.2±0.2°, 21.6±0.2°, and 23.7±0.2° 2θ. More typically, it has diffraction peaks at 2θ of 6.6±0.2°, 9.2±0.2°, 15.2±0.2°, 17.6±0.2°, 20.3±0.2°, 21.2±0.2°, 21.6±0.2°, 22.4±0.2°, 23.7±0.2°, and 25.7±0.2°.

[0014] In some embodiments of the present invention, crystalline form B of the monohydrochloride salt of compound of formula II has an X-ray diffraction spectrum substantially the same as the X-ray diffraction spectrum shown in FIG. 4, measured using Cu-Kα radiation.

[0015] In some embodiments of the present invention, crystalline Form B of the monohydrochloride salt of the compound of Formula II has a DSC spectrum substantially the same as the DSC spectrum displayed in FIG. 5 or a TGA spectrum substantially the same as the TGA spectrum displayed in FIG. 6.

[0016] The present invention further provides the dihydrochloride salt of the compound of formula II.

[0017] According to another aspect, the present invention provides crystalline Form I of the dihydrochloride salt of compound of Formula II, having diffraction peaks at 6.0±0.2°, 17.1±0.2°, 19.9±0.2°, and 23.3±0.2° 2θ in an X-ray diffraction (XRD) spectrum measured using Cu-Kα radiation. Typically, Form I has diffraction peaks at 6.0±0.2°, 17.1±0.2°, 19.9±0.2°, 23.3±0.2°, 25.4±0.2°, and 25.7±0.2° 2θ. More typically, it has diffraction peaks at 2θ of 6.0±0.2°, 13.1±0.2°, 17.1±0.2°, 17.5±0.2°, 19.9±0.2°, 20.2±0.2°, 22.0±0.2°, 23.3±0.2°, 25.4±0.2°, and 25.7±0.2°.

[0018] In some embodiments of the present invention, crystalline Form I of the dihydrochloride salt of compound of Formula II has an X-ray diffraction spectrum substantially the same as the X-ray diffraction spectrum shown for Form I in FIG. 7, measured using Cu-Kα radiation.

[0019] In some embodiments of the present invention, crystalline Form I of the dihydrochloride salt of the compound of Formula II has a DSC spectrum substantially the same as the DSC spectrum shown in FIG. 8 or a TGA spectrum substantially the same as the TGA spectrum shown in FIG. 9.

[0020] According to another aspect, the present invention further provides crystalline Form II of the dihydrochloride salt of compound of Formula II, having diffraction peaks at 6.0±0.2°, 17.1±0.2°, 20.0±0.2°, and 22.8±0.2° 2θ in an X-ray diffraction (XRD) spectrum measured using Cu-Kα radiation. Typically, Form II has diffraction peaks at 6.0±0.2°, 13.4±0.2°, 17.1±0.2°, 20.0±0.2°, 21.2±0.2°, and 22.8±0.2° 2θ. More typically, it has diffraction peaks at 6.0±0.2°, 9.9±0.2°, 13.4±0.2°, 17.1±0.2°, 20.0±0.2°, 20.5±0.2°, 21.2±0.2°, 22.8±0.2°, 26.9±0.2°, and 30.2±0.2° 2θ.

[0021] In some embodiments of the present invention, crystalline Form I of the dihydrochloride salt of the compound of Formula II is Form I has an X-ray diffraction spectrum measured using Cu-Kα radiation that is substantially the same as the X-ray diffraction spectrum shown for crystalline Form II in FIG.

[0022] In some embodiments of the present invention, crystalline Form II of the dihydrochloride salt of the compound of Formula II has a DSC spectrum substantially the same as the DSC spectrum shown in FIG. 10 or a TGA spectrum substantially the same as the TGA spectrum shown in FIG. 11.

[0023] The present invention further provides crystalline Form III of the dihydrochloride salt of the compound of Formula II, which has diffraction peaks at 6.0±0.2°, 17.2±0.2°, 20.1±0.2°, and 22.0±0.2° 2θ in an X-ray diffraction (XRD) spectrum measured using Cu-Kα radiation. Typically, Form III has diffraction peaks at 6.0±0.2°, 13.5±0.2°, 17.2±0.2°, 20.1±0.2°, 21.2±0.2°, and 22.0±0.2° 2θ. More typically, it has diffraction peaks at 2θ of 6.0±0.2°, 9.3±0.2°, 13.5±0.2°, 17.2±0.2°, 18.7±0.2°, 20.1±0.2°, 22.0±0.2°, 21.2±0.2°, 24.0±0.2°, and 26.4±0.2°.

[0024] In some embodiments of the present invention, crystalline Form III of the dihydrochloride salt of compound of Formula II has an X-ray diffraction spectrum substantially the same as the X-ray diffraction spectrum shown for Form III in Figure 7, measured using Cu-Kα radiation.

[0025] In some embodiments of the present invention, crystalline Form III of the dihydrochloride salt of the compound of Formula II has a DSC spectrum substantially the same as the DSC spectrum displayed in FIG. 12 or a TGA spectrum substantially the same as the TGA spectrum displayed in FIG. 13.

[0026] The present invention provides Step (a) dissolving the free base of compound of formula II in a mixture of methyl ethyl ketone and n-heptane and stirring to obtain a clear solution; (b) slowly adding a solution of HCl in ethyl acetate to the solution, stirring, filtering and drying; The present invention further provides a process for preparing crystalline Form A of the monohydrochloride salt of compound of formula II, comprising:

[0027] In some embodiments of the present invention, in the method for preparing crystalline form A, in step (a), the volume ratio of methyl ethyl ketone / n-heptane may be 1:1. In step (b), the acid-base charging ratio of HCl to the free base of the compound of Formula II is 1:1 to 2:1, for example, 1.3:1, 1.4:1, 1.5:1, 1.6:1, 1.7:1, 1.8:1, 1.9:1, or 2:1, preferably 1.1:1 to 1.5:1.

[0028] In some embodiments of the present invention, the method for preparing crystalline form A comprises dissolving the free base of the compound of formula II at room temperature in step (a), and stirring at room temperature for 24 hours in step (b).

[0029] According to another aspect, the present invention provides a method for manufacturing a method of manufacturing a semiconductor device comprising: (a) dissolving the free base of compound of formula II in ethyl acetate and stirring to obtain a clear solution; (b) slowly adding a solution of HCl in ethyl acetate to the solution, stirring, filtering and drying; The present invention provides a process for preparing crystalline form B of the monohydrochloride salt of compound of formula II, comprising:

[0030] In some embodiments of the present invention, in the process for preparing crystalline form B, in step (b), In this case, the acid-base charging ratio of HCl to the free base of the compound represented by formula II is 1:1 to 2:1, for example, 1.1:1, 1.3:1, 1.4:1, 1.5:1, 1.6:1, 1.7:1, 1.8:1, 1.9:1, 2:1, preferably 1.1:1 to 1.5:1.

[0031] In some embodiments of the present invention, the method for preparing crystalline form B comprises dissolving the free base of the compound of formula II at room temperature in step (a), and stirring at room temperature for 24 hours in step (b).

[0032] According to another aspect, the present invention provides a method for manufacturing a method of manufacturing a semiconductor device comprising: (a) dissolving the free base of compound of formula II in ethyl acetate and stirring to obtain a clear solution; (b) slowly adding a solution of HCl in ethyl acetate to the solution, stirring, filtering and drying; The present invention provides a process for preparing crystalline Form I of the dihydrochloride salt of compound of formula II, comprising:

[0033] In some embodiments of the present invention, in the method for preparing Crystalline Form I, in step (b), the acid-base charge ratio of HCl to the free base of the compound of Formula II is 2:1 to 5:1, for example, 2.5:1, 3:1, 3.5:1, 4:1, 4.2:1, preferably 2.5:1 to 4:1.

[0034] In some embodiments of the present invention, the method for preparing crystalline Form I comprises dissolving the free base of the compound of Formula II at room temperature in step (a), and stirring at low temperature for 5 hours in step (b).

[0035] According to another aspect, the present invention provides a process for preparing crystalline Form II of the dihydrochloride salt of compound of Formula II, comprising suspending and slurrying crystalline Form I of the dihydrochloride salt of compound of Formula II in a ketone solvent, filtering and drying.

[0036] For the crystalline Form II of the compound of Formula II, the ketone solvent is one or more of acetone, butanone, and methyl ethyl ketone.

[0037] In some embodiments of the present invention, in the method for preparing crystalline Form II, the slurrying is carried out at room temperature to 50° C. for 24 hours.

[0038] According to another aspect, the present invention provides a process for preparing crystalline Form III of the dihydrochloride salt of compound of Formula II, comprising suspending and slurrying crystalline Form I of the dihydrochloride salt of compound of Formula II in methyl t-butyl ether, filtering and drying.

[0039] In some embodiments of the present invention, in the method for preparing crystalline Form III, the slurrying is carried out at room temperature to 50° C. for 24 hours.

[0040] The present invention further provides a pharmaceutical composition comprising a crystalline form of the hydrochloride salt of the compound of Formula II or the monohydrochloride and / or dihydrochloride salts of the compound of Formula II and one or more pharmaceutically acceptable inactive excipients.

[0041] The present invention further provides a pharmaceutical composition comprising 95% or more (e.g., 95%, 96%, 97%, 98%, 99%, or any value or range therebetween) of the dihydrochloride salt of the compound of Formula II.

[0042] The present invention further provides a crystalline composition comprising 80% or more (e.g., 80%, 90%, 95%, or any value or range therebetween) crystalline Form I of the hydrochloride salt of the compound of Formula II.

[0043] In some embodiments of the present invention, the pharmaceutical compositions of the present invention are formulated for oral administration.

[0044] According to another aspect, the present invention provides the use of a crystalline form of the hydrochloride salt of compound of formula II, the monohydrochloride salt, the dihydrochloride salt of compound of formula II, a pharmaceutical composition, or a crystalline composition as defined above in the preparation of a medicament for the prevention and treatment of diseases, wherein said diseases are thrombin-mediated diseases and / or thrombin-related diseases.

[0045] According to another aspect, the present invention provides the use of a crystalline form of the hydrochloride salt of compound of formula II, the monohydrochloride salt, the dihydrochloride salt of compound of formula II, a pharmaceutical composition, or a crystalline composition as defined above in the preparation of a medicament for treating a disease, said disease being a venous thrombotic disease and / or an arterial thrombotic disease. [Brief explanation of the drawings]

[0046] [Figure 1]FIG. 1 shows the XRD spectrum of crystalline form A of the monohydrochloride salt of compound of formula II. [Figure 2] FIG. 2 shows the DSC spectrum of crystalline form A of the monohydrochloride salt of compound of formula II. [Figure 3] FIG. 3 shows the TGA spectrum of crystalline form A of the monohydrochloride salt of compound of formula II. [Figure 4] FIG. 4 shows the XRD spectrum of crystalline form B of the monohydrochloride salt of compound of formula II. [Figure 5] FIG. 5 shows the DSC spectrum of crystalline form B of the monohydrochloride salt of compound of formula II. [Figure 6] FIG. 6 shows the TGA spectrum of crystalline form B of the monohydrochloride salt of compound of formula II. [Figure 7] FIG. 7 shows the XRD spectra of Form I, Form II, and Form III of the dihydrochloride salt of the compound of Formula II. [Figure 8] FIG. 8 shows the DSC spectrum of crystalline Form I of the dihydrochloride salt of compound of Formula II. [Figure 9] FIG. 9 shows the TGA spectrum of crystalline Form I of the dihydrochloride salt of compound of Formula II. [Figure 10] FIG. 10 shows the DSC spectrum of crystalline Form II of the dihydrochloride salt of the compound of Formula II. [Figure 11] FIG. 11 shows the TGA spectrum of crystalline Form II of the dihydrochloride salt of compound of Formula II. [Figure 12] FIG. 12 shows the DSC spectrum of crystalline Form III of the dihydrochloride salt of compound of Formula II. [Figure 13] FIG. 13 shows the TGA spectrum of crystalline Form III of the dihydrochloride salt of compound of Formula II. [Figure 14] FIG. 14 shows the XRD spectrum of crystalline Form I of the dihydrochloride salt of compound of Formula II. [Figure 15] FIG. 15 shows the XRD spectrum of crystalline Form II of the dihydrochloride salt of compound of Formula II. [Figure 16]FIG. 16 shows the XRD spectrum of crystalline Form III of the dihydrochloride salt of compound of Formula II. DETAILED DESCRIPTION OF THE INVENTION

[0047] In the following description, specific details are included to provide a comprehensive understanding of the disclosed embodiments, however, one skilled in the art will recognize that the embodiments may be practiced using other methods, components, materials, etc., without one or more of these specific details.

[0048] As used herein, the phrase "in some embodiments" means that at least one embodiment includes the associated particular element, structure, or feature described in that embodiment. Thus, the appearances of the phrase "in some embodiments" in different places throughout the specification do not necessarily all refer to the same embodiment. Furthermore, the particular elements, structures, or features may be combined in any suitable manner in one or more embodiments.

[0049] In one aspect, provided herein are hydrochloride salts of the compound of Formula II, including monohydrochloride and dihydrochloride salts. Specifically, the monohydrochloride salt of the compound of Formula II has a stoichiometric ratio of the compound of Formula II:HCl of 1:1, and can exist in forms such as anhydrous crystalline forms, hydrate forms, and solvates. The dihydrochloride salt of the compound of Formula II has a stoichiometric ratio of the compound of Formula II:HCl of 1:2, and can exist in forms such as anhydrous crystalline forms, hydrate forms, and solvates.

[0050] As used herein, the hydrochloride salt of the compound of Formula II is a crystalline salt. Compared with the free base, the hydrochloride salt of the compound of Formula II has improved pharmaceutical properties and excellent storage characteristics (e.g., stability), and is easily formulated into pharmaceutical compositions such as tablets and capsules. For example, the free base of the compound of Formula II is an oily substance, while the monohydrochloride or dihydrochloride salt of the compound of Formula II is a crystalline compound. While the free base of the compound of Formula II is insoluble in water, the monohydrochloride or dihydrochloride salt of the compound of Formula II has excellent aqueous solubility and intrinsic dissolution rate, and improved bioavailability, making it particularly suitable for pharmaceutical applications. Furthermore, after conversion of the free form to the monohydrochloride or dihydrochloride form, any process impurities that were difficult to remove from the free form are reduced to surprisingly low levels, and the monohydrochloride or dihydrochloride salt is easier to separate than the free base, so the formation of the salt may provide a purification method.

[0051] Any suitable method known in the art can be used to separate the crystallized hydrochloride salt. Suitably, the monohydrochloride or dihydrochloride salt of the compound of Formula II obtained is separated and recovered by filtration. Preferably, the recovered monohydrochloride or dihydrochloride salt of the compound of Formula II obtained is dried under vacuum.

[0052] The various crystalline forms of the hydrochloride salt described herein are in substantially pure form.

[0053] As used herein, the term "substantially pure" means at least 95% purity, preferably 98% purity, where 95% purity means that no more than 5% of other forms of the compound of Formula II (e.g., other crystalline forms, amorphous forms, etc.) are present, and 98% purity means that no more than 2% of other forms of the compound of Formula II (e.g., other crystalline forms, amorphous forms, etc.) are present.

[0054] In another aspect, the present invention provides crystalline Form A of the monohydrochloride salt of the compound of formula II.

[0055] In some embodiments of the present invention, crystalline Form A of the monohydrochloride salt of the compound of Formula II is In the X-ray diffraction (XRD) spectrum measured using Cu-Kα radiation, it has diffraction peaks at 2θ of 6.7±0.2°, 16.2±0.2°, 21.8±0.2°, and 22.9±0.2°. Typically, it has diffraction peaks at 2θ of 6.7±0.2°, 16.2±0.2°, 16.5±0.2°, 20.7±0.2°, 21.8±0.2°, and 22.9±0.2°. More typically, it has diffraction peaks at 2θ of 6.7±0.2°, 7.8±0.2°, 16.2±0.2°, 16.5±0.2°, 19.4±0.2°, 20.7±0.2°, 21.8±0.2°, 22.9±0.2°, 23.8±0.2°, and 27.5±0.2°.

[0056] In some embodiments of the present invention, crystalline Form A of the monohydrochloride salt of the compound of formula II has an X-ray diffraction spectrum substantially the same as the X-ray diffraction spectrum shown in FIG. 1, measured using Cu-Kα radiation.

[0057] In some embodiments of the present invention, crystalline Form A of the monohydrochloride salt of the compound of Formula II has substantially the same characterization data as set forth in Table 1.

[0058] [Table 1]

[0059] In some embodiments of the present invention, crystalline Form A of the monohydrochloride salt of the compound of Formula II has a DSC spectrum substantially the same as the DSC spectrum shown in FIG. 2 or a TGA spectrum substantially the same as the TGA spectrum shown in FIG. 3.

[0060] The present invention further provides crystalline Form B of the monohydrochloride salt of the compound of formula II.

[0061] In some embodiments of the present invention, crystalline Form B of the monohydrochloride salt of compound of Formula II has diffraction peaks at 6.6±0.2°, 15.2±0.2°, 21.2±0.2°, and 21.6±0.2° 2θ in an X-ray diffraction (XRD) spectrum measured using Cu-Kα radiation. Typically, it has diffraction peaks at 6.6±0.2°, 15.2±0.2°, 20.3±0.2°, 21.2±0.2°, 21.6±0.2°, and 23.7±0.2° 2θ. More typically, it has diffraction peaks at 2θ of 6.6±0.2°, 9.2±0.2°, 15.2±0.2°, 17.6±0.2°, 20.3±0.2°, 21.2±0.2°, 21.6±0.2°, 22.4±0.2°, 23.7±0.2°, and 25.7±0.2°.

[0062] In some embodiments of the present invention, crystalline form B of the monohydrochloride salt of compound of formula II has an X-ray diffraction spectrum substantially the same as the X-ray diffraction spectrum shown in FIG. 4, measured using Cu-Kα radiation.

[0063] In some embodiments of the present invention, crystalline form B of the monohydrochloride salt of the compound of formula II is has substantially the same characterization data as that shown in Table 2.

[0064] [Table 2]

[0065] In some embodiments of the present invention, crystalline Form B of the monohydrochloride salt of the compound of Formula II has a DSC spectrum substantially the same as the DSC spectrum displayed in FIG. 5 or a TGA spectrum substantially the same as the TGA spectrum displayed in FIG. 6.

[0066] The present invention further provides the dihydrochloride salt of the compound of formula II.

[0067] The present invention further provides crystalline Form I of the dihydrochloride salt of the compound of formula II.

[0068] In some embodiments of the present invention, crystalline Form I of the dihydrochloride salt of compound of Formula II has diffraction peaks at 6.0±0.2°, 17.1±0.2°, 19.9±0.2°, and 23.3±0.2° 2θ in an X-ray diffraction (XRD) spectrum measured using Cu-Kα radiation. Typically, it has diffraction peaks at 6.0±0.2°, 17.1±0.2°, 19.9±0.2°, 23.3±0.2°, 25.4±0.2°, and 25.7±0.2° 2θ. More typically, it has diffraction peaks at 2θ of 6.0±0.2°, 13.1±0.2°, 17.1±0.2°, 17.5±0.2°, 19.9±0.2°, 20.2±0.2°, 22.0±0.2°, 23.3±0.2°, 25.4±0.2°, and 25.7±0.2°.

[0069] In some embodiments of the present invention, crystalline Form I of the dihydrochloride salt of compound of Formula II has an X-ray diffraction spectrum substantially the same as the X-ray diffraction spectrum shown for Form I in FIG. 7, measured using Cu-Kα radiation.

[0070] In some embodiments of the present invention, crystalline Form I of the dihydrochloride salt of the compound of Formula II has characterization data substantially the same as the characterization data set forth in Table 3.

[0071] [Table 3]

[0072] In some embodiments of the present invention, crystalline Form I of the dihydrochloride salt of the compound of Formula II has a DSC spectrum substantially the same as the DSC spectrum shown in FIG. 8 or a TGA spectrum substantially the same as the TGA spectrum shown in FIG. 9.

[0073] In some embodiments of the present invention, the DSC spectrum of crystalline Form I of the dihydrochloride salt of compound of Formula II shows one endothermic peak at 139°C and one decomposition peak at 145°C, and the TGA spectrum shows a weight loss of about 0.6% by 80°C.

[0074] As can be seen from the DVS results of Form I, Form I exhibits water absorption of 28.8% / 49.9% at 80% RH / 90% humidity, respectively. The sample deliquesced under high humidity and transformed into amorphous form after the DVS test.

[0075] The physicochemical properties of crystalline form I are as follows:

[0076] [Table 4]

[0077] The solubility of crystalline Form I was determined as follows.

[0078] [Table 5]

[0079] The accelerated stability test data for Form I at T=25±2°C and humidity 60±5% are as follows:

[0080] [Table 6]

[0081] The present invention further provides crystalline Form II of the dihydrochloride salt of the compound of formula II.

[0082] In some embodiments of the present invention, crystalline Form II of the dihydrochloride salt of compound of Formula II has diffraction peaks at 6.0±0.2°, 17.1±0.2°, 20.0±0.2°, and 22.8±0.2° 2θ in an X-ray diffraction (XRD) spectrum measured using Cu-Kα radiation. Typically, it has diffraction peaks at 6.0±0.2°, 13.4±0.2°, 17.1±0.2°, 20.0±0.2°, 21.2±0.2°, and 22.8±0.2° 2θ. More typically, the values ​​were 6.0±0.2°, 9.9±0.2°, 13.4±0.2°, 17.1±0.2°, 20.0±0.2°, 20.5±0.2°, 21.2±0.2°, 22.8±0.2°, 26.9±0.2°, and 30.2±0. It has a diffraction peak at 2° 2θ.

[0083] In some embodiments of the present invention, crystalline Form II of the dihydrochloride salt of compound of Formula II has an X-ray diffraction spectrum substantially the same as the X-ray diffraction spectrum shown for Crystalline Form II in Figure 7, measured using Cu-Kα radiation.

[0084] In some embodiments of the present invention, crystalline Form II of the dihydrochloride salt of the compound of Formula II has substantially the same characterization data as set forth in Table 4.

[0085] [Table 7]

[0086] In some embodiments of the present invention, crystalline Form II of the dihydrochloride salt of the compound of Formula II has a DSC spectrum substantially the same as the DSC spectrum shown in FIG. 10 or a TGA spectrum substantially the same as the TGA spectrum shown in FIG. 11.

[0087] In some embodiments of the present invention, the DSC spectrum of crystalline Form II of the dihydrochloride salt of compound of Formula II shows one endothermic peak at 134°C and one decomposition peak at 142°C, and the TGA spectrum shows a weight loss of about 0.8% by 120°C.

[0088] The DSC plot of crystalline Form II has one broad endothermic peak before 100° C. Heating to 110° C. in the DSC did not change the crystalline form. In some embodiments of the present invention, when crystalline Form II is obtained by slurrying in acetone, about 0.2% acetone remains.

[0089] The present invention further provides crystalline Form III of the dihydrochloride salt of the compound of formula II.

[0090] In some embodiments of the present invention, crystalline Form III of the dihydrochloride salt of compound of Formula II has diffraction peaks at 6.0±0.2°, 17.2±0.2°, 20.1±0.2°, and 22.0±0.2° 2θ in an X-ray diffraction (XRD) spectrum measured using Cu-Kα radiation. Typically, it has diffraction peaks at 6.0±0.2°, 13.5±0.2°, 17.2±0.2°, 20.1±0.2°, 21.2±0.2°, and 22.0±0.2° 2θ. More typically, it has diffraction peaks at 2θ of 6.0±0.2°, 9.3±0.2°, 13.5±0.2°, 17.2±0.2°, 18.7±0.2°, 20.1±0.2°, 22.0±0.2°, 21.2±0.2°, 24.0±0.2°, and 26.4±0.2°.

[0091] In some embodiments of the present invention, crystalline Form III of the dihydrochloride salt of compound of Formula II has an X-ray diffraction spectrum substantially the same as the X-ray diffraction spectrum shown for Form III in Figure 7, measured using Cu-Kα radiation.

[0092] In some embodiments of the present invention, crystalline Form III of the dihydrochloride salt of the compound of Formula II has substantially the same characterization data as set forth in Table 5.

[0093] [Table 8]

[0094] In some embodiments of the present invention, crystalline Form III of the dihydrochloride salt of the compound of Formula II has a DSC spectrum substantially the same as the DSC spectrum displayed in FIG. 12 or a TGA spectrum substantially the same as the TGA spectrum displayed in FIG. 13.

[0095] In some embodiments of the present invention, the DSC spectrum of crystalline Form III of the dihydrochloride salt of compound of Formula II shows one endothermic peak at 138° C. and one decomposition peak at 145° C., and the TGA spectrum shows no significant weight loss up to 120° C. The DVS results show a water absorption of 28.8 / 50.0% at 80% RH / 90% humidity, respectively.

[0096] Crystalline Form III is subjected to mechanical milling for 2 minutes to convert it to Crystalline Form I. When Crystalline Form III is stored at 60°C under sealed conditions for 7 days, the purity decreases from 95.81% to 89.35% and it converts to Crystalline Form I.

[0097] In another aspect, the present invention provides a method for preparing each crystalline form of the hydrochloride salt of the compound of Formula II, comprising mixing the free form of the compound of Formula II with hydrogen chloride in a suitable organic solvent, and filtering and drying after the salt formation is complete. The method can also be performed by stirring a slurry of the hydrochloride salt of the particular compound of Formula II in a suitable organic solvent, and filtering and drying after the crystal transformation is complete. The solvent can be ethyl acetate, isopropyl acetate, methyl ethyl ketone, methyl t-butyl ether, toluene, cyclohexane, isopropanol, acetonitrile, n-heptane, or a mixture thereof.

[0098] In some embodiments of the present invention, the process for preparing crystalline form A of the monohydrochloride salt of compound of formula II comprises the steps of: Step (a) dissolving the free base of compound of formula II in a mixture of methyl ethyl ketone and n-heptane and stirring to obtain a clear solution; (b) slowly adding a solution of HCl in ethyl acetate to the solution, stirring, filtering and drying; Includes.

[0099] In some embodiments of the present invention, in the method for preparing crystalline form A, in step (a), the volume ratio of methyl ethyl ketone / n-heptane may be 1:1. In step (b), the acid-base ratio of HCl to the free base of the compound of Formula II may be 1:1 to 2:1, for example, 1.3:1, 1.4:1, 1.5:1, 1.6:1, 1.7:1, 1.8:1, 1.9:1, 1.10:1, 1.11:1, 1.12:1, 1.13:1, 1.14:1, 1.15:1, 1.16:1, 1.17:1, 1.18:1, 1.19:1, 1.20:1, 1.21:1, 1.22:1, 1.23:1, 1.24:1, :1, 1.8:1, 1.9:1, 2:1, preferably 1.1:1 to 1.5:1.

[0100] In some embodiments of the present invention, the method for preparing crystalline form A comprises dissolving the free base of the compound of formula II at room temperature in step (a), and stirring at room temperature for 24 hours in step (b).

[0101] In some embodiments of the present invention, the process for preparing crystalline form B of the monohydrochloride salt of compound of formula II comprises: (a) dissolving the free base of compound of formula II in ethyl acetate and stirring to obtain a clear solution; (b) slowly adding a solution of HCl in ethyl acetate to the solution, stirring, filtering and drying; Includes.

[0102] In some embodiments of the present invention, in the method for preparing crystalline form B, in step (b), the acid-base charge ratio of HCl to the free base of compound of Formula II is 1:1 to 2:1, for example, 1.1:1, 1.3:1, 1.4:1, 1.5:1, 1.6:1, 1.7:1, 1.8:1, 1.9:1, 2:1, preferably 1.1:1 to 1.5:1.

[0103] In some embodiments of the present invention, the method for preparing crystalline form B comprises dissolving the free base of the compound of formula II at room temperature in step (a), and stirring at room temperature for 24 hours in step (b).

[0104] In the preparation of the monohydrochloride, if the acid-base ratio is too low, some of the free base will not be completely converted to the monohydrochloride, while if the ratio is too high, a mixture of the monohydrochloride and the dihydrochloride will be produced, so the rate and amount of hydrochloric acid added must be strictly controlled, which causes difficulties in industrial production and product quality control.

[0105] In some embodiments of the present invention, the process for preparing crystalline Form I of the dihydrochloride salt of compound of Formula II comprises: (a) dissolving the free base of compound of formula II in ethyl acetate and stirring to obtain a clear solution; (b) slowly adding a solution of HCl in ethyl acetate to the solution, stirring, filtering and drying; Includes.

[0106] In some embodiments of the present invention, in step (b) of the method for preparing Crystalline Form I, the acid-base ratio of HCl to the free base of the compound of Formula II is 2:1 to 5:1, for example, 2.5:1, 3:1, 3.5:1, 4:1, or 4.2:1, preferably 2.5:1 to 4:1. The addition of excess hydrochloric acid helps to completely convert the free base to the dihydrochloride salt, which is convenient for industrial handling, and the resulting Crystalline Form I has few impurities and high purity.

[0107] In some embodiments of the present invention, the method for preparing crystalline Form I comprises dissolving the free base of the compound of Formula II at room temperature in step (a), and stirring at low temperature for 5 hours in step (b).

[0108] In some embodiments of the present invention, the method for preparing crystalline Form II of the dihydrochloride salt of compound of Formula II includes suspending crystalline Form I of the dihydrochloride salt of compound of Formula II in a ketone solvent. The method includes the steps of: slurried, filtered, and dried.

[0109] In some embodiments of the present invention, in the method for preparing crystalline Form II, the ketone solvent is one or more of acetone, butanone, and methyl ethyl ketone, and the slurrying is carried out at room temperature to 50°C for 24 hours.

[0110] In some embodiments of the present invention, the process for preparing crystalline Form III of the dihydrochloride salt of compound of Formula II comprises suspending and slurrying crystalline Form I of the dihydrochloride salt of compound of Formula II in methyl t-butyl ether, filtering, and drying.

[0111] In some embodiments of the present invention, in the process for preparing crystalline Form III, the slurrying is carried out at room temperature for 24 hours.

[0112] According to another aspect, the present invention provides a pharmaceutical composition comprising a hydrochloride salt of a compound of Formula II as defined herein (e.g., the monohydrochloride salt of a compound of Formula II or the dihydrochloride salt of a compound of Formula II), and a pharmaceutically acceptable inactive excipient.

[0113] In some embodiments of the present invention, the pharmaceutical compositions are formulated as dosage forms for oral administration such as, for example, tablets, capsules, powders, granules, liquids (e.g., oral liquids, suspensions, emulsions, etc.), or syrups, which may contain pharmaceutically acceptable excipients such as lubricants, binders, disintegrants, fillers, dispersants, emulsifiers, stabilizers, etc.

[0114] For example, for injections, the compound of the present invention can be mixed or dissolved in physiological saline, and the pH can be adjusted to the most stable state by adding an appropriate dilute acid, alkali, or buffer salt. Additionally, antioxidants or metal chelating agents may be added. The solution is sterilized by filtration and filled into sterile ampoules under aseptic conditions.

[0115] For example, for tablets, the compound of the present invention and additives (e.g., microcrystalline cellulose, sodium carboxymethyl starch, corn starch, magnesium stearate, talc, etc.) can be thoroughly mixed, passed through a sieve, and formed into tablets using a tablet press. For example, for hard capsules, the compound of the present invention can be passed through a sieve and mixed with additives / excipients (e.g., dry starch, magnesium stearate, etc.), and the mixture can be filled into hard gelatin capsules using an appropriate device.

[0116] To prepare a suspension, the compound of the present invention can be passed through a sieve and mixed with additives (e.g., sodium carboxymethylcellulose and syrup) to form a uniform paste. Optionally, dyes, benzoic acid, etc. can be diluted with a portion of purified water and added to the paste with stirring, followed by adding sufficient water to make the desired volume. For other pharmaceutical composition preparation methods and additives, see Remington's Pharmaceutical Sciences, 18th edition, Alfonso R. Gennaro (1990), Publisher: Mack Publishing Company, and its revised editions.

[0117] The pharmaceutical compositions of the present invention may be solid, semi-solid, or liquid preparations prepared by known conventional techniques. These methods include, for example, mixing, dissolving, granulating, pulverizing, emulsifying, embedding, spray-drying, or lyophilizing. In such compositions, the active ingredient may account for 0.1% to 99.9% by weight of the formulation. Furthermore, the carrier, diluent, or excipient used in the composition must be pharmaceutically compatible with the active ingredient and pharmaceutically acceptable.

[0118] According to another aspect, the present disclosure provides the use of the hydrochloride salt of the compound of formula II in the inhibition of thrombin and the prevention and treatment of thrombin-mediated and / or thrombin-related diseases.

[0119] As described in the Examples section of the present specification, the hydrochloride salt of a representative compound of formula II showed significant antithrombotic activity in animal models. The present invention relates to the use of compounds according to the present invention, such as the monohydrochloride salt of compound of formula II or the dihydrochloride salt of compound of formula II, for the prevention and treatment of thrombotic diseases, particularly venous or arterial thrombotic diseases such as deep vein thromboembolism of the lower extremities, reocclusion after bypass surgery or angioplasty, peripheral occlusive arterial disease, pulmonary embolism, disseminated intravascular coagulation, coronary thromboembolism, stroke, and shunt or stent occlusion.

[0120] The present invention relates to the use of compounds according to the present invention, such as the monohydrochloride salt of compound of formula II or the dihydrochloride salt of compound of formula II, for the prevention and treatment of stroke, pulmonary embolism, myocardial or cerebral infarction, atrial fibrillation, and arrhythmias resulting from the formation of thrombi.

[0121] The present invention relates to the use of compounds according to the present invention, such as the monohydrochloride salt of compound of formula II or the dihydrochloride salt of compound of formula II, in the prevention and treatment of atherosclerotic diseases, such as coronary artery disease, cerebral artery disease, and peripheral artery disease.

[0122] The compounds described herein can also be used as anticoagulants in in vitro blood circuits. In some embodiments, the present invention provides a method for inhibiting coagulation in an in vitro or ex vivo blood circuit, the method comprising adding an effective amount of a compound of the present invention to an in vitro or ex vivo blood circuit.

[0123] The compounds described herein (e.g., the monohydrochloride salt of the compound of Formula II or the dihydrochloride salt of the compound of Formula II) can also be used in combination therapy with thrombolytic agents, for example, to shorten reperfusion time and prolong reocclusion time. The compounds described herein are also useful for preventing thrombus reformation after microsurgery. The compounds described herein are also effective in anticoagulant therapy for hemodialysis and disseminated intravascular coagulation. The compounds described herein are also useful for ex vivo storage of blood, plasma, and other blood products.

[0124] The compounds of the present invention can be administered orally. The specific dosage will depend on the specific circumstances of the case, including the dosage form, the rate of administration, and the condition being treated. A typical daily oral dosage that produces therapeutic effects can range from about 0.01 mg / kg to about 1000 mg / kg (based on the free base of the compound of Formula II). Administration can be in a single dose once daily or in divided doses two to four times daily. The dosage and administration method can be adjusted depending on the age and weight of the patient and the severity of the disease being treated.

[0125] The compounds of formula II herein are synthesized according to the method described in US Pat. No. 6,275,1984.

[0126] In this specification, X-ray diffraction (XRD) is measured by the following method.

[0127] (1) A Bruker D2 X-ray powder diffractometer was used to collect X-ray powder diffraction data of the sample under ambient conditions. The X-ray generator output was 300 W. The sample stage had no background signal, the step speed was 0.15 s / step, and the total number of steps was 1. The X-ray tube used a Cu target (Kα) with a Kα2 / Kα1 intensity ratio of 0.50 (1.54439 Å / 1.5406 Å). (2) PANalytical Empyrean equipped with a PIXcel 1D detector The X-ray powder diffractometer was used for the analysis. The 2θ scanning angle of the sample was 3° to 40°, the scanning step width was 0.013°, and the tube voltage and current were 45 kV and 40 mA, respectively.

[0128] In this specification, differential scanning calorimetry (DSC) is measured as follows: A TA Discovery series differential scanning calorimeter (DSC) is used to collect the thermal data of the sample. A few mg of sample is weighed into a Tzero aluminum pan and sealed with a Tzero sealing cap. Under N2 protection, the sample is heated at a heating rate of 10°C / min.

[0129] In this specification, thermogravimetric analysis (TGA) is measured as follows: Thermogravimetric data of samples was collected using a TA Discovery series thermogravimetric analyzer (TGA). A few milligrams of sample was placed in a Tzero aluminum pan and heated from room temperature to the target temperature at a heating rate of 10°C / min under N2 protection.

[0130] While X-ray diffraction spectra obtained from crystalline compounds are characteristic of specific crystalline forms, the relative intensities of bands (especially those in the low-angle region) can vary due to preferred orientation effects resulting from differences in crystallization conditions, particle size, and other measurement conditions. Therefore, the relative intensities of diffraction peaks are not characteristic of the crystalline form of interest. When determining identity with known crystalline forms, the relative positions of peaks are more important than their relative intensities. Furthermore, it is well known in the field of crystallography that slight errors in peak positions may exist for any given crystalline form. For example, peak positions may shift due to temperature changes during sample analysis, sample movement, or instrument calibration, resulting in a measurement error of approximately ±0.2° in 2θ values. Therefore, this error must be taken into account when identifying the structure of various crystalline forms. In XRD spectra, peak positions are usually expressed in terms of 2θ angles or lattice spacing d, which can be converted to each other using the simple conversion formula d = λ / 2 sin θ, where d is the lattice spacing, λ is the wavelength of the incident X-rays, and θ is the diffraction angle. For the same crystalline form of the same compound, the peak positions in the XRD spectrum will be similar overall, but there may be large errors in the relative intensities. When identifying a mixture, some diffraction lines may not be detected due to factors such as a decrease in content. In this case, it is not necessary to rely on all bands observed in a high-purity sample; even a single band may be characteristic of a particular crystalline form.

[0131] DSC measures the transition temperatures at which a crystal absorbs or releases heat as its crystalline structure changes or melts. For consecutive analyses of the same crystalline form of the same compound, the error in thermal transition temperatures and melting points is typically within about 3°C. When a compound is described as exhibiting a specific DSC peak or melting point, this refers to ±3°C of that DSC peak or melting point. DSC provides an auxiliary means for distinguishing between different crystalline forms. Different crystalline forms can be distinguished based on their different transition temperatures.

[0132] As used herein, the term "pharmaceutical composition" refers to a formulation containing an active compound of the present invention and a carrier, excipient, and / or vehicle generally accepted in the art for delivering a biologically active compound into an organism (e.g., a human). The purpose of a pharmaceutical composition is to facilitate administration of a compound of the present invention to an organism.

[0133] As used herein, the term "pharmaceutically acceptable inactive excipient" refers to any carrier, excipient, vehicle, glidant, or formulation suitable for use in humans or animals (e.g., livestock). These include, but are not limited to, sweeteners, diluents, preservatives, dyes / colorants, flavor enhancers, surfactants, wetting agents, dispersing agents, disintegrating agents, suspending agents, stabilizers, isotonicity agents, solvents, or emulsifiers.

[0134] All solvents used herein are commercially available and are used without further purification.

[0135] In this specification, room temperature refers to 25°C.

[0136] In this specification, low temperature refers to a temperature between -10 and 25°C.

[0137] In this specification, the meanings of the abbreviations are as follows:

[0138] [Table 9]

[0139] In this specification, the calculation method for the content of the product is as follows. Content = (100% - weight loss - residue) x (100% - total impurities) x 100%

[0140] The purity of the product is calculated by HPLC relative area comparison method.

[0141] The present invention will be described below by way of example with reference to specific examples, but the scope of the present invention is not limited to the following specific examples.

[0142] Example 1: Preparation of Crystalline Form A of the Monohydrochloride Salt of Compound of Formula II 300 mg of the free base was dissolved in 3 mL of methyl ethyl ketone / n-heptane (1:1, v / v) and stirred at room temperature to obtain a clear solution. To this solution, HCl in ethyl acetate (acid-base ratio 2:1) was slowly added and stirred at room temperature for 24 hours to form a white suspension. The solid was separated by filtration and dried under vacuum at 35°C to obtain a 285 mg sample of crystalline Form A of the monohydrochloride salt of compound of Formula II (yield: approximately 88.6%).

[0143] Example 2: Preparation of Crystalline Form A of the Monohydrochloride Salt of Compound of Formula II 300 mg of the free base was dissolved in 3 mL of methyl ethyl ketone / n-heptane (1:1, v / v) and stirred at room temperature to obtain a clear solution. To this solution, a solution of HCl in ethyl acetate (acid-base ratio 1.3:1) was slowly added and stirred at room temperature for 24 hours to form a white suspension. The solid was separated by filtration and dried under vacuum at 35°C to obtain 241 mg of a sample of crystalline Form A of the monohydrochloride salt of compound of Formula II (yield: approximately 75.4%).

[0144] Example 3: Preparation of Crystalline Form B of the Monohydrochloride Salt of Compound of Formula II 300 mg of the free base was dissolved in 3 mL of ethyl acetate and stirred at room temperature to obtain a clear solution. To this solution, HCl in ethyl acetate (acid-base ratio 2:1) was slowly added and stirred at room temperature for 24 hours to form a white suspension. The solid was separated by filtration and dried under vacuum at 35°C to obtain a 223 mg sample of crystalline Form B of the monohydrochloride salt of compound of Formula II in approximately 68.4% yield, with a water content of 0.6%, a hydrolysis impurity content of 3.77%, and a total impurity content of 4.5%. HPLC purity was 95.6%, and the content was 95.2%.

[0145] Example 4: Preparation of Crystalline Form I of the Dihydrochloride Salt of Compound of Formula II 1.75 kg of the free base was dissolved in 7 L of ethyl acetate and stirred at room temperature to obtain a clear solution. To this solution was slowly added a solution of HCl in ethyl acetate (acid-base ratio 3:1) at low temperature and stirred at room temperature for 5 hours to form a white suspension. The solid was separated by filtration and dried under vacuum at 40°C to obtain a 1.76 kg sample of crystalline Form I of the dihydrochloride salt of compound of Formula II in a yield of approximately 89.4% with an HPLC purity of 99.6%.

[0146] Example 5: Preparation of Crystalline Form II of the Dihydrochloride Salt of Compound of Formula II Crystalline Form I (180 mg) obtained in the example was suspended in methyl ethyl ketone (3 mL) and slurried at room temperature for 24 hours. The solid was separated by filtration and dried under vacuum at 35°C to obtain 136 mg of a sample of Crystalline Form II of the dihydrochloride salt of compound of Formula II, with a yield of about 75.6%.

[0147] Example 6: Preparation of Crystalline Form III of the Dihydrochloride Salt of Compound of Formula II Crystalline Form I (180 mg) obtained in the example was dissolved in methyl tert-butyl ether (3 mL). The solid was separated by filtration and dried under vacuum at 35° C. to give a 166 mg sample of crystalline Form III of the dihydrochloride salt of compound of Formula II, a yield of about 92.2%.

[0148] Example 7: Rat deep vein thrombosis model Test animals: SD rats (weight: 220-250 g, sex: male) Solution preparation: An appropriate amount of crystalline Form I was weighed and dissolved in a solvent (0.5% (W / V) Natrosol™ 250 HX solution containing 0.1% (W / V) L(+)-tartaric acid) to prepare a predetermined concentration, and the solution was stirred uniformly and administered at the planned dose.

[0149] Procedure: After three days of acclimation, rats were randomly divided into three groups (8-10 rats per group): model group, positive control group, test compound group, and bridging group. The model group was administered an equal volume of vehicle by oral gavage, the positive control group or test compound group by oral gavage with crystalline Form I, and the bridging group was administered crystalline Form I of active compound V intravenously. Approximately 45 minutes later, the rats were anesthetized and blood was collected, and model construction began. Each group of rats was anesthetized by intraperitoneal injection of urethane (20%, 5 mL / kg). The abdominal cavity was exposed, the vena cava was bluntly isolated, and two 8-10 cm long surgical sutures were placed under the vessel at 1 cm intervals. After the model preparation time for each group was completed, rabbit thromboplastin (0.02 mg / kg) was injected into the left femoral vein, and 10 seconds later, the two threads were tied. After 1 hour, the ligated area was cut open with ophthalmic scissors, and the thrombus was removed and weighed.

[0150] After oral administration of crystalline Form I to rats at 2.5, 5, 10, and 20 mg / kg, the blood concentrations of Compound V were 190.1, 181.3, 341.9, and 609.7 ng / mL, respectively, showing a nearly dose-dependent increase in blood concentration with increasing dose. At similar doses, the antithrombotic effect of Crystalline Form I was approximately equivalent to that of dabigatran etexilate. When Compound V was intravenously infused to rats at 0.1 mg / kg / h, the antithrombotic effect was approximately equivalent to that of oral administration of crystalline Form I at 5 mg / kg, and the blood concentrations of Compound V were also equivalent.

[0151] The results were as follows:

[0152] [Table 10]

[0153] Example 8: Rat Pharmacokinetic Studies Test animals: SD rats (weight: 200-300g, sex: half male and half female) Preparation of solution for oral gavage: An appropriate amount of crystalline Form I was weighed and dissolved in a solvent (0.5% (W / V) Natrosol™ 250 HX solution containing 0.1% (W / V) L(+)-tartaric acid) to prepare a predetermined concentration, and the solution was stirred uniformly and administered at the planned dose.

[0154] Procedure: After three days of acclimation, rats were randomly assigned to groups of eight rats, half male and half female. Oral gavage doses of 5, 10, and 20 mg / kg were administered. Blood samples were collected from the jugular vein at 5, 10, 20, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 12 hours, and 24 hours after administration. Blood concentrations were then measured by LC-MS / MS.

[0155] Data were processed using the Phoenix WinNonlin 6.4 non-compartmental model to calculate pharmacokinetic parameters after administration to rats. The pharmacokinetic parameters were calculated as t 1 / 2 , AUC 0-t , AUC 0-∞ , Cl / F or Cl ss / F or Cl , V z / F or V ss , MRT, C max , C 5min , T max , and C that reaches steady state when administered multiple times min , C avg , AUC 0-τ Includes:

[0156] The results were as follows:

[0157] [Table 11]

[0158] Various modifications and variations of the embodiments of the present invention can be made without departing from the scope of the present invention. Therefore, it should be understood that the embodiments of the present invention are not limited to the above exemplary embodiments, but should be controlled by the limitations set forth in the claims and their equivalents.

Claims

【Request Item 1】 【Chemistry 1】 A crystalline form of the hydrochloride salt of the compound of formula II.

2. The stoichiometric ratio of the compound of formula II to HCl is 1:1 or 1:

2.

2. A crystalline form of the hydrochloride salt of the compound of formula II according to claim 1.

3. Crystalline Form A of the monohydrochloride salt of the compound of formula II, In the X-ray diffraction spectrum measured using Cu-Kα radiation, having diffraction peaks at 2θ of 6.7±0.2°, 16.2±0.2°, 21.8±0.2°, and 22.9±0.2°; or having diffraction peaks at 2θ of 6.7±0.2°, 16.2±0.2°, 16.5±0.2°, 20.7±0.2°, 21.8±0.2°, and 22.9±0.2°; or having diffraction peaks at 2θ of 6.7±0.2°, 7.8±0.2°, 16.2±0.2°, 16.5±0.2°, 19.4±0.2°, 20.7±0.2°, 21.8±0.2°, 22.9±0.2°, 23.8±0.2°, and 27.5±0.2°; Crystalline form A.

4. has an X-ray diffraction spectrum substantially the same as the X-ray diffraction spectrum shown in Figure 1, measured using Cu-Kα radiation; or has a DSC spectrum substantially the same as the DSC spectrum shown in Figure 2; or having a TGA spectrum substantially the same as the TGA spectrum shown in FIG. The crystalline form A according to claim 3.

5. Crystalline form B of the monohydrochloride salt of the compound of formula II, In the X-ray diffraction spectrum measured using Cu-Kα radiation, having diffraction peaks at 2θ of 6.6±0.2°, 15.2±0.2°, 21.2±0.2°, and 21.6±0.2°; or having diffraction peaks at 2θ of 6.6±0.2°, 15.2±0.2°, 20.3±0.2°, 21.2±0.2°, 21.6±0.2°, and 23.7±0.2°; or having diffraction peaks at 2θ of 6.6±0.2°, 9.2±0.2°, 15.2±0.2°, 17.6±0.2°, 20.3±0.2°, 21.2±0.2°, 21.6±0.2°, 22.4±0.2°, 23.7±0.2°, and 25.7±0.2°; Crystal form B.

6. has an X-ray diffraction spectrum substantially the same as the X-ray diffraction spectrum shown in Figure 4, measured using Cu-Kα radiation; or has a DSC spectrum substantially the same as the DSC spectrum shown in Figure 5; or having a TGA spectrum substantially the same as the TGA spectrum shown in FIG. Crystalline form B according to claim 5. 【Request Item 7】 【Chemistry 2】 The dihydrochloride salt of the compound of formula II.

8. Crystalline Form I of the dihydrochloride salt of the compound of formula II, In the X-ray diffraction spectrum measured using Cu-Kα radiation, having diffraction peaks at 2θ of 6.0±0.2°, 17.1±0.2°, 19.9±0.2°, and 23.3±0.2°; or having diffraction peaks at 2θ of 6.0±0.2°, 17.1±0.2°, 19.9±0.2°, 23.3±0.2°, 25.4±0.2°, and 25.7±0.2°; or having diffraction peaks at 2θ of 6.0±0.2°, 13.1±0.2°, 17.1±0.2°, 17.5±0.2°, 19.9±0.2°, 20.2±0.2°, 22.0±0.2°, 23.3±0.2°, 25.4±0.2°, and 25.7±0.2°; Crystalline form I.

9. has an X-ray diffraction spectrum measured using Cu-Kα radiation that is substantially the same as the X-ray diffraction spectrum shown for crystalline Form I in Figure 7; or has a DSC spectrum substantially the same as the DSC spectrum shown in Figure 8; or having a TGA spectrum substantially the same as the TGA spectrum shown in FIG. Crystalline form I according to claim 8.

10. Crystalline Form II of the dihydrochloride salt of the compound of formula II, having diffraction peaks at 2θ of 6.0±0.2°, 17.1±0.2°, 20.0±0.2°, and 22.8±0.2° in an X-ray diffraction spectrum measured using Cu—Kα radiation; or having diffraction peaks at 2θ of 6.0±0.2°, 13.4±0.2°, 17.1±0.2°, 20.0±0.2°, 21.2±0.2°, and 22.8±0.2°; or having diffraction peaks at 2θ of 6.0±0.2°, 9.9±0.2°, 13.4±0.2°, 17.1±0.2°, 20.0±0.2°, 20.5±0.2°, 21.2±0.2°, 22.8±0.2°, 26.9±0.2°, and 30.2±0.2°; Crystalline Form II.

11. has an X-ray diffraction spectrum measured using Cu-Kα radiation substantially the same as the X-ray diffraction spectrum shown for crystalline Form II in Figure 7; or has a DSC spectrum substantially the same as the DSC spectrum shown in Figure 10; or having a TGA spectrum substantially the same as the TGA spectrum shown in FIG.

11. Crystalline Form II according to claim 10.

12. Crystalline Form III of the dihydrochloride salt of the compound of formula II, In the X-ray diffraction spectrum measured using Cu-Kα radiation, 6.0±0.2°, 17.2±0.2°, 20.1±0.2°, and 22.0±0. has a diffraction peak at 2° 2θ, or having diffraction peaks at 2θ of 6.0±0.2°, 13.5±0.2°, 17.2±0.2°, 20.1±0.2°, 21.2±0.2°, and 22.0±0.2°; or having diffraction peaks at 2θ of 6.0±0.2°, 9.3±0.2°, 13.5±0.2°, 17.2±0.2°, 18.7±0.2°, 20.1±0.2°, 22.0±0.2°, 21.2±0.2°, 24.0±0.2°, and 26.4±0.2°; Crystalline Form III.

13. has an X-ray diffraction spectrum measured using Cu-Kα radiation substantially the same as the X-ray diffraction spectrum shown for crystalline Form III in Figure 7; or has a DSC spectrum substantially the same as the DSC spectrum shown in Figure 12; or having a TGA spectrum substantially the same as the TGA spectrum shown in FIG.

13. Crystalline Form III according to claim 12.

14. 5. A process for preparing crystalline form A according to claim 3 or 4, comprising the steps of: Step (a) dissolving the free base of compound of formula II in a mixture of methyl ethyl ketone and n-heptane and stirring to obtain a clear solution; (b) slowly adding a solution of HCl in ethyl acetate to the solution, stirring, filtering and drying; the acid-base charge ratio of the HCl to the free base of the compound of Formula II is 1:1 to 2:1, preferably 1.1:1 to 1.5:1; Preferably, in step (a), the free base of the compound of formula II is dissolved at room temperature, and in step (b), stirring is carried out at room temperature for 24 hours. Method for preparing crystalline form A.

15. 7. A process for preparing crystalline form B according to claim 5 or 6, comprising the steps of: Step (a) dissolving the free base of compound of formula II in ethyl acetate and stirring to obtain a clear solution; (b) slowly adding a solution of HCl in ethyl acetate to the solution, stirring, filtering and drying; the acid-base charge ratio of the HCl to the free base of the compound of Formula II is 1:1 to 2:1, preferably 1.1:1 to 1.5:1; Preferably, in step (a), the free base of the compound of formula II is dissolved at room temperature, and in step (b), stirring is carried out at room temperature for 24 hours. Method for preparing crystalline form B.

16. 10. A process for preparing crystalline form I according to claim 8 or 9, comprising the steps of: Step (a) dissolving the free base of compound of formula II in ethyl acetate and stirring to obtain a clear solution; (b) slowly adding a solution of HCl in ethyl acetate to the solution, stirring, filtering and drying; the acid-base charge ratio of the HCl to the free base of the compound of Formula II is 2:1 to 5:1, preferably 2.5:1 to 4:1; Preferably, in step (a), the free base of the compound of formula II is dissolved at room temperature, and in step (b), stirring is carried out at low temperature for 5 hours. Method for preparing crystalline form I.

17. 12. A process for preparing crystalline form II according to claim 10 or 11, comprising the steps of: suspending and slurrying crystalline Form I of the dihydrochloride salt of compound of Formula II in a ketone solvent, filtering and drying; Preferably, the ketone solvent is one or more of acetone, butanone, and methyl ethyl ketone; Preferably, the slurrying is carried out at room temperature to 50°C for 24 hours. Method for preparing crystalline form II.

18. 14. A process for preparing crystalline form III according to claim 12 or 13, comprising: Crystalline Form I of the dihydrochloride salt of the compound of formula II was suspended in methyl tert-butyl ether. slurried, filtered and dried; Preferably, the slurrying is carried out at room temperature to 50°C for 24 hours. Method for preparing crystalline form III.

19. 1. A pharmaceutical composition comprising: a crystalline form of the hydrochloride salt of the compound of formula II according to claim 1 or 2, or a monohydrochloride and / or dihydrochloride salt of the compound of formula II according to any one of claims 3 to 13; one or more pharmaceutically acceptable inactive excipients; Preferably, the pharmaceutical composition is prepared for oral administration. Pharmaceutical compositions.

20. Contains 95% or more of the dihydrochloride salt of the compound of formula II; Pharmaceutical compositions.

21. containing 80% or more of crystalline form I of the hydrochloride salt of the compound of formula II; Crystalline composition.

22. Use of a crystalline form of the hydrochloride salt of the compound of formula II according to claim 1 or 2, a monohydrochloride salt, a dihydrochloride salt of the compound of formula II according to any one of claims 3 to 13, a pharmaceutical composition according to claim 19 or 20, or a crystalline composition according to claim 21 in the preparation of a medicament for the prevention and treatment of a disease, The disease is a thrombin-mediated disease and / or a thrombin-related disease, use.

23. Use of a crystalline form of the hydrochloride salt of compound of formula II according to claim 1 or 2, a monohydrochloride salt, a dihydrochloride salt of compound of formula II according to any one of claims 3 to 13, a pharmaceutical composition according to claim 19 or 20, or a crystalline composition according to claim 21 in the preparation of a medicament for treating a disease, the disease is a venous thrombotic disease and / or an arterial thrombotic disease; use.

Images