TW202434612A - A salt of a dipeptide compound, its preparation method and use - Google Patents

Abstract

The present invention discloses a salt of a dipeptide compound (II). More specifically, the present invention relates to a novel salt of the compound (II) and its crystal, and a method for preparing the salt and its crystal. Also provided is a pharmaceutical composition comprising a hydrochloride of the compound (II), and the use of the salt in preparing a drug for preventing and treating thrombin-mediated and/or thrombin-related diseases.

Description

A salt of a dipeptide compound, its preparation method and use

The present invention belongs to the field of pharmaceutical chemistry, and specifically relates to a salt of a dipeptide compound, a preparation method and use thereof.

U.S. Patent No. 17289379 discloses a compound of formula II (referred to as Compound II), which can be used to mediate the activity of trypsin-like serine proteases, and can be used as an anticoagulant and an agent for modulating or inhibiting the activity of trypsin-like serine proteases, thereby treating thromboembolic diseases and other related cardiovascular diseases. (II)

Blood has both a coagulation system and an anticoagulation system (fibrinolytic system). Under physiological conditions, the coagulation factors in the blood are continuously and limitedly activated, thereby generating thrombin and forming a trace amount of fibrin. After these fibrins are formed, they will be deposited in the inner membrane of the cardiovascular system, and the fibrinolytic system can dissolve a small amount of deposited fibrin in time after being activated. Therefore, the coagulation system and the anticoagulation system (fibrinolytic system) can ensure both the potential coagulation of blood and the fluidity of blood. When certain factors that induce thrombosis appear, the above dynamic balance is broken, thereby triggering a coagulation reaction. The coagulation reaction mainly includes the primary coagulation process and the secondary coagulation process. The secondary coagulation process involves a series of biochemical reactions (blood coagulation cascade), which includes the activation of inactive enzymes (or proenzymes) of blood coagulation factors to serine proteases (factor X to factor Xa), which can continue to activate subsequent coagulation factors (factor Xa activates factor II to form factor IIa), and finally convert soluble plasma protein fibrinogen into insoluble plasma protein. Fibrinogen thrombin is an extracellular insulin-like serine protease that plays a key role in the blood coagulation cascade and is the last enzyme in the process. If the activity of thrombin can be inhibited, the formation of thrombus can be blocked. After oral administration, compound II is degraded into active metabolite compound V via two intermediate forms by carboxylesterase (carboxylesterase-1/carboxylesterase-2, CES-1/CES-2) in vivo. Compound V exerts its anticoagulant effect by selectively inhibiting thrombin. (V)

However, excessive inhibition of thrombin activity may cause bleeding risk. Therefore, the dosage of anticoagulants needs to be precisely controlled clinically.

In one aspect, the present invention provides a crystalline form of a hydrochloride salt of a compound of formula II, (II)

In some embodiments herein, the stoichiometry of the compound of Formula II and HCl is 1:1 or 1:2.

On the other hand, the present invention provides a crystalline form A of a monohydrochloride salt of a compound of formula II, which has diffraction peaks of 2θ=6.7±0.2°, 16.2±0.2°, 21.8±0.2°, 22.9±0.2° in an X-ray diffraction pattern measured by Cu-Kα radiation; Typically, it has diffraction peaks of 6.7±0.2°, 16.2±0.2°, 16.5±0.2°, 20.7±0.2°, 21.8±0.2°, 22.9±0.2°; More typically, it has Diffraction peaks 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°.

In some embodiments herein, the crystalline form A of the monohydrochloride salt of the compound of formula II has an X-ray diffraction pattern measured using Cu-Kα radiation substantially the same as that shown in FIG. 1 .

In some embodiments herein, the crystalline form A of the monohydrochloride salt of the compound of formula II has a DSC spectrum substantially the same as shown in FIG. 2 or a TGA spectrum substantially the same as shown in FIG. 3 .

On the other hand, the present invention provides a crystalline form B of a monohydrochloride salt of a compound of formula II, wherein the X measured by Cu-Kα radiation is The X-ray diffraction (XRD) pattern has diffraction peaks of 2θ=6.6±0.2°, 15.2±0.2°, 21.2±0.2°, and 21.6±0.2°; typically, it has diffraction peaks of 2θ=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°; more typically, it has diffraction peaks of 2θ=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°.

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

In some embodiments herein, Form B of the monohydrochloride salt of the compound of Formula II has a DSC spectrum substantially the same as shown in FIG. 5 or a TGA spectrum substantially the same as shown in FIG. 6 .

Also provided herein is the dihydrochloride salt of the compound of Formula II.

On the other hand, the present invention provides a crystalline form I of the dihydrochloride salt of the compound of formula II, which has 2θ=6.0±0.2°, 17.1±0.2°, 19.9±0.2°, 23.3 in the X-ray diffraction (XRD) pattern measured by Cu-Kα radiation. ±0.2°; typically, it has a diffraction peak of 2θ＝6.0±0.2°, 17.1±0.2°, 19.9±0.2°, 23.3±0.2°, 25.4±0.2°, 25.7±0.2°; more typically, it has a diffraction peak of 2θ＝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°, 25.7±0.2°.

In some embodiments herein, the Form I of the dihydrochloride salt of the compound of Formula II has an X-ray diffraction pattern measured using Cu-Kα radiation that is substantially the same as that of Form I shown in FIG. 7 .

In some embodiments herein, the crystalline form I of the dihydrochloride salt of the compound of formula II has a DSC spectrum substantially the same as shown in FIG. 8 or a TGA spectrum substantially the same as shown in FIG. 9 .

On the other hand, the present invention also provides a crystalline form II of the dihydrochloride salt of the compound of formula II, which has an X value measured by Cu-Kα radiation of The X-ray diffraction (XRD) pattern has diffraction peaks of 2θ=6.0±0.2°, 17.1±0.2°, 20.0±0.2°, and 22.8±0.2°; typically, it has diffraction peaks of 2θ=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°; more typically, it has diffraction peaks of 2θ=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°.

In some embodiments herein, the dihydrochloride salt of the compound of formula II has a X-ray diffraction pattern measured using Cu-Kα radiation that is substantially the same as that of Form II shown in FIG. 7 .

In some embodiments herein, the crystalline form II of the dihydrochloride salt of the compound of formula II has a DSC spectrum substantially the same as shown in FIG. 10 or a TGA spectrum substantially the same as shown in FIG. 11 .

Also provided herein is a crystalline form III of the dihydrochloride salt of the compound of formula II, which has an X value measured by Cu-Kα radiation. The X-ray diffraction (XRD) pattern has diffraction peaks of 2θ=6.0±0.2°, 17.2±0.2°, 20.1±0.2°, and 22.0±0.2°; typically, it has diffraction peaks of 2θ=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°; more typically, it has diffraction peaks of 2θ=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°.

In some embodiments herein, the dihydrochloride salt of the compound of Formula II in Form III has an X-ray diffraction pattern measured using Cu-Kα radiation that is substantially the same as that of Form III shown in FIG. 7 .

In some embodiments herein, Form III of the dihydrochloride salt of the compound of Formula II has a DSC spectrum substantially the same as shown in FIG. 12 or a TGA spectrum substantially the same as shown in FIG. 13 .

The present invention also provides a method for preparing a hydrochloride crystal form A, comprising the following steps: (c) dissolving the free base of the compound of formula II in a mixed solvent of methyl ethyl ketone/n-heptane, and stirring to obtain a clear solution; (d) slowly adding ethyl acetate solution of HCl to the solution, stirring, filtering and drying.

In some embodiments of the present invention, in the preparation method of Form A, in step (a), the volume ratio of methyl ethyl ketone/n-heptane can be 1:1; in step (b), the acid-base feed ratio of HCl and 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, 1.7:1, 1.8:1, 1.9:1, 2:1, preferably 1.1:1 to 1.5:1.

In some embodiments herein, in the method for preparing Form A, in step (a), the free base of the compound of Formula II is dissolved at room temperature, and in step (b), stirring is performed at room temperature for 24 hours.

On the other hand, the present invention provides a method for preparing a hydrochloride crystal form B, comprising the following steps: (a) dissolving the free base of the compound of formula II in ethyl acetate, and stirring to obtain a clear solution; (b) slowly adding ethyl acetate solution of HCl to the solution, stirring, filtering and drying.

In some embodiments herein, in the method for preparing Form B, in step (b), the acid-base feed ratio of HCl and the free base of the 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, 1.7:1, 1.8:1, 1.9:1, 2:1, preferably 1.1:1 to 1.5:1.

In some embodiments herein, in the method for preparing Form B, in step (a), the free base of the compound of Formula II is dissolved at room temperature, and in step (b), stirring is performed at room temperature for 24 hours.

On the other hand, the present invention provides a method for preparing dihydrochloride crystal form I, comprising the following steps: (a) dissolving the free base of the compound of formula II in ethyl acetate, and stirring to obtain a clear solution; (b) slowly adding ethyl acetate solution of HCl to the solution, stirring, filtering and drying.

In some embodiments herein, in the method for preparing Form I, in step (b), the acid-base feed ratio of HCl and 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.

In some embodiments herein, in the method for preparing Form I, in step (a), the free base of the compound of Formula II is dissolved at room temperature, and in step (b), the mixture is stirred at low temperature for 5 hours.

On the other hand, the present invention provides a method for preparing the dihydrochloride crystal form II, comprising the following steps: suspending the dihydrochloride crystal form I of the compound of formula II in a ketone solvent, slurrying and stirring, filtering and drying.

The ketone solvent of the crystal form II compound of formula II is one or more of acetone, butanone or methyl ethyl ketone.

In some embodiments of the present invention, in the preparation method of Form II, the slurrying and stirring is performed at room temperature to 50°C for 24 hours.

On the other hand, the present invention provides a method for preparing the dihydrochloride crystal form III, comprising the following steps: suspending the dihydrochloride crystal form I of the compound of formula II in methyl tert-butyl ether, slurrying and stirring, filtering and drying.

In some embodiments herein, in the method for preparing Form III, the slurrying and stirring is slurrying and stirring at room temperature to 50° C. for 24 hours.

Also provided herein is a pharmaceutical composition comprising a crystalline form of a hydrochloride of a compound of formula II or a monohydrochloride and/or dihydrochloride of a compound of formula II, and one or more pharmaceutically acceptable inert pharmaceutical excipients.

Also provided herein is a pharmaceutical composition comprising more than 95% (e.g., 95%, 96%, 97%, 98%, 99% or any value or range therebetween) of the dihydrochloride salt of the compound of formula II.

Also provided herein is a crystalline composition comprising more than 80% (e.g., 80%, 90%, 95% or any value or range therebetween) of the crystalline form I of the hydrochloride salt of the compound of formula II.

In some embodiments herein, the pharmaceutical compositions herein are formulated for oral administration.

In another aspect, the present invention provides the use of a crystalline form of the hydrochloride of the compound of formula II as defined above, a monohydrochloride, a dihydrochloride, a pharmaceutical composition, or a crystalline composition of the compound of formula II in the preparation of a medicament for the prevention and treatment of thrombin-mediated and thrombin-related diseases.

On the other hand, the present invention provides the use of a crystalline form of the hydrochloride of the compound of formula II as defined above, a monohydrochloride, a dihydrochloride, a pharmaceutical composition, or a crystalline composition of the compound of formula II in the preparation of a medicament for treating venous and/or arterial thrombotic diseases.

In the following description, certain specific details are included to provide a comprehensive understanding of each disclosed embodiment. However, a person skilled in the art will recognize that the embodiments can be implemented without using one or more of these specific details and using other methods, components, materials, etc.

Herein, the phrase "in some embodiments" means that at least one embodiment includes the specific reference elements, structures or features described in the embodiment. Therefore, the phrase "in some embodiments" appearing in different places throughout the specification does not necessarily refer to the same embodiment. In addition, the specific elements, structures or features can be combined in one or more embodiments in any appropriate manner.

In one aspect, the present invention provides hydrochlorides of the compound of formula II, including monohydrochlorides and dihydrochlorides. Specifically, the monohydrochloride of the compound of formula II has a chemical ratio of the compound of formula II: HCl of 1:1, and can exist in the form of anhydrous crystals, hydrated forms, and solvates. The dihydrochloride of the compound of formula II has a chemical ratio of the compound of formula II: HCl of 1:2, and can exist in the form of anhydrous crystals, hydrated forms, and solvates.

Herein, the hydrochloride of the compound of formula II is a crystalline salt, and compared with the free base, the hydrochloride of the compound of formula II has improved pharmaceutical properties, excellent storage properties (such as stability) and is easy to be formulated into a pharmaceutical composition such as a tablet or capsule, etc. For example, the free base of the compound of formula II is an oily substance, while the monohydrochloride or dihydrochloride of the compound of formula II is a crystalline compound; the free base of the compound of formula II is insoluble in water, while the monohydrochloride or dihydrochloride of the compound of formula II has excellent water solubility and inherent dissolution rate, and improves bioavailability, and these properties make it particularly suitable for use in medicine. Furthermore, certain process impurities that are difficult to remove in the free form are surprisingly present to a much smaller extent after conversion of the free form to the mono- or di-hydrochloride form, and the mono- or di-hydrochloride salt is easier to separate than the free base, so salt formation can provide a method for purification.

Any suitable method known in the art for separating the crystallized hydrochloride can be used. Suitably, the monohydrochloride or dihydrochloride of the compound of formula II obtained is collected by filtration separation. Preferably, the monohydrochloride or dihydrochloride of the compound of formula II obtained is collected by drying under vacuum.

Each crystalline form of the hydrochloride salt described herein is in substantially pure form.

As used herein, the term "substantially pure" means at least 95% pure, preferably 98% pure, wherein 95% pure means no more than 5%, and 98% pure means no more than 2% of any other form (other crystalline forms, amorphous forms, etc.) of the compound of Formula II is present.

In another aspect, provided herein is a crystalline form A of the monohydrochloride salt of the compound of formula II.

In some embodiments herein, the crystalline form A of the monohydrochloride of the compound of formula II has diffraction peaks of 2θ=6.7±0.2°, 16.2±0.2°, 21.8±0.2°, and 22.9±0.2° in the X-ray diffraction (XRD) pattern measured by Cu-Kα radiation; typically, it has diffraction peaks of 2θ=6.7±0.2°, 16.2±0.2°, 16.5±0.2°, 20. The diffraction peaks are 2θ＝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°, 27.5±0.2°.

In some embodiments herein, the crystalline form A of the monohydrochloride salt of the compound of formula II has an X-ray diffraction pattern measured using Cu-Kα radiation substantially the same as shown in FIG. 1 .

In some embodiments herein, the crystalline form A of the monohydrochloride salt of the compound of formula II has substantially the same characterization data as shown in Table 1.

Table 1 Characteristic data of hydrochloride form A Salt type TGA weight loss ( wt% ) DSC Onset/Peak ( ℃ ) Water adsorption ( wt% ) Room temperature stability ( 10 days) Hydrochloride Form A 0.5 52.9, 149.2 0.5 (slightly hygroscopic) White powder with unchanged crystal form (10 days)

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

Also provided herein is Form B of the monohydrochloride salt of the compound of Formula II.

In some embodiments herein, the crystalline form B of the monohydrochloride of the compound of formula II has diffraction peaks of 2θ=6.6±0.2°, 15.2±0.2°, 21.2±0.2°, and 21.6±0.2° in the X-ray diffraction (XRD) pattern measured by Cu-Kα radiation; typically, it has diffraction peaks of 2θ=6.6±0.2°, 15.2±0.2°, 20.3±0.2°, 21.6±0.2°. The diffraction peaks are 2θ＝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°.

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

In some embodiments herein, the crystalline form B of the monohydrochloride salt of the compound of formula II has substantially the same characterization data as shown in Table 2.

Table 2 Characteristic data of hydrochloride form B Salt type TGA weight loss ( wt% ) DSC Onset/Peak ( ℃ ) Water adsorption ( wt% ) Room temperature stability ( 10 days) Hydrochloride Form B 1.0 52.8, 137.2 3.0 (moisture absorption) White powder with unchanged crystal form (10 days)

In some embodiments herein, Form B of the monohydrochloride salt of the compound of Formula II has a DSC spectrum substantially the same as shown in FIG. 5 or a TGA spectrum substantially the same as shown in FIG. 6 .

Also provided herein is the dihydrochloride salt of the compound of Formula II.

Also provided herein is Form I of the dihydrochloride salt of the compound of Formula II.

In some embodiments herein, the crystalline form I of the dihydrochloride salt of the compound of formula II has diffraction peaks of 2θ=6.0±0.2°, 17.1±0.2°, 19.9±0.2°, and 23.3±0.2° in the X-ray diffraction (XRD) pattern measured by Cu-Kα radiation; typically, the crystalline form I has diffraction peaks of 2θ=6.0±0.2°, 17.1±0.2°, 19.9±0.2°, and 23.3±0.2°. ±0.2°, 25.4±0.2°, and 25.7±0.2°; more typically, it has diffraction peaks of 2θ＝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°.

In some embodiments herein, Form I of the dihydrochloride salt of the compound of Formula II has an X-ray diffraction pattern measured using Cu-Kα radiation that is substantially the same as that of Form I shown in FIG. 7 .

In some embodiments herein, the crystalline form I of the dihydrochloride salt of the compound of formula II has substantially the same characterization data as shown in Table 3.

Table 3 Summary of DSC and TGA characteristics of dihydrochloride form I Crystal form DSC Onset/Peak ( ° C ) , ΔH (J/g ) TGA weight loss ( wt% ) Dihydrochloride Form I Hydrate 139/145,33 ~ 0.6

In some embodiments herein, the crystalline form I of the dihydrochloride salt of the compound of formula II has a DSC spectrum substantially the same as shown in FIG8 or a TGA spectrum substantially the same as shown in FIG9.

In some embodiments herein, the DSC spectrum of the crystalline form I of the dihydrochloride salt of the compound of formula II shows an endothermic peak at 139°C and a decomposition peak at 145°C; the TGA spectrum shows a weight loss of about 0.6% before 80°C.

The DVS results of Form I show that Form I absorbs 28.8%/49.9% water at 80%RH/90% humidity. The sample deliquesces at high humidity and transforms into an amorphous form after DVS testing.

The physicochemical properties of Form I are as follows: Characteristics Off-white to white solid Hygroscopicity Hygroscopic pKa pKa1=4.94, pKa2 =5.54, pKa3=10.61 Distribution coefficient Log P＞3.38, Log D _7.4 = 2.06 Solubility It is very soluble in buffers of different pH values (1.2, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10.0, 11.0, 12.0 and 13.0), freely soluble in acetonitrile and dichloromethane, and almost insoluble or insoluble in n-heptane and N,N-diisopropylethylamine.

The solubility of Form I was determined as follows: No. Solvent Solubility (mg/mL) 1 EtOH >250 2 IPA ~167 3 NBA ~167 4 MEK ~1.6 5 EA <0.3 6 IPA ~0.5 7 Hept <0.3 8 Diox ~125 9 THF ~11.1 10 Tol <0.3 11 ACN >250 12 Water >250 13 MTBE <0.3 14 DMSO >250 15 AC >250

The accelerated stability test data of Form I at T = 25 ± 2°C and humidity 60 ± 5% are as follows: Testing cycle Moisture Hydrolyzed impurities Other single impurities Total impurities Assay (HPLC) 0 month 0.4% 0.19% ＜RL(0.05%) 0.42% 99.6% January 0.5% 0.27% ＜RL(0.05%) 0.54% 99.3% February 0.3% 0.28% ＜RL(0.05%) 0.56% 100.3% March 0.4% 0.45% ＜RL(0.05%) 0.72% 100.3% June 0.3% 0.51% ＜RL(0.05%) 1.02% 99.6%

Also provided herein is Form II of the dihydrochloride salt of the compound of Formula II.

In some embodiments herein, the crystalline form II of the dihydrochloride salt of the compound of formula II has diffraction peaks of 2θ=6.0±0.2°, 17.1±0.2°, 20.0±0.2°, and 22.8±0.2° in the X-ray diffraction (XRD) pattern measured by Cu-Kα radiation; typically, the crystalline form II has diffraction peaks of 2θ=6.0±0.2°, 13.4±0.2°, 17.1±0.2°, 20.0±0.2°, and 21.2±0 .2°, 22.8±0.2°; more typically, it has diffraction peaks of 2θ＝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°.

In some embodiments herein, the dihydrochloride salt of the compound of formula II has an X-ray diffraction pattern measured using Cu-Kα radiation that is substantially the same as that of Form II shown in FIG. 7 .

In some embodiments herein, the crystalline form II of the dihydrochloride salt of the compound of formula II has substantially the same characterization data as shown in Table 4.

Table 4 Summary of DSC and TGA characteristics of dihydrochloride form II Crystal form DSC Onset/Peak (°C ) , ΔH (J/g ) TGA weight loss (% ) Dihydrochloride crystal form II No crystal form 134/142,17 ~ 0.8

In some embodiments herein, the crystalline form II of the dihydrochloride salt of the compound of formula II has a DSC spectrum substantially the same as shown in FIG. 10 or a TGA spectrum substantially the same as shown in FIG. 11 .

In some embodiments herein, the DSC spectrum of the crystalline form II of the dihydrochloride salt of the compound of formula II shows an endothermic peak at 134°C and a decomposition peak at 142°C; the TGA spectrum shows a weight loss of about 0.8% before 120°C.

The DSC curve of Form II has a broad endothermic peak before 100°C. The DSC does not change the form when heated to 110°C. In some embodiments of the present invention, when Form II is slurried in acetone, about 0.2% acetone remains.

Also provided herein is Form III of the dihydrochloride salt of the compound of Formula II.

In some embodiments herein, the crystalline form III of the dihydrochloride salt of the compound of formula II has 2θ=6.0±0.2°, 17.2±0.2°, 20.1±0.2°, 22.0± Typically, it has diffraction peaks of 2θ=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°; more typically, it has diffraction peaks of 2θ=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°.

In some embodiments herein, the dihydrochloride salt of the compound of Formula II in Form III has an X-ray diffraction pattern measured using Cu-Kα radiation that is substantially the same as that of Form III shown in FIG. 7 .

In some embodiments herein, the crystalline form III of the dihydrochloride salt of the compound of formula II has substantially the same characterization data as shown in Table 5.

Table 5 Summary of DSC and TGA characteristics of dihydrochloride form III Crystal form DSC Onset/Peak ( °C ) , ΔH (J/g ) TGA weight loss (% ) Dihydrochloride crystal form II crystal form III anhydrous crystal form 138/145,26 ~ 0

In some embodiments herein, Form III of the dihydrochloride salt of the compound of Formula II has a DSC spectrum substantially the same as shown in FIG. 12 or a TGA spectrum substantially the same as shown in FIG. 13 .

In some embodiments of the present invention, the DSC spectrum of the crystalline form III of the dihydrochloride salt of the compound of formula II has an endothermic peak at 138°C and a decomposition peak at 145°C; the TGA spectrum has no obvious weight loss before 120°C. The DVS results show that the water absorption is 28.8/50.0% at 80%RH/90% humidity.

After mechanical grinding for 2 minutes, Form III was transformed into Form I. Form III was stored at 60°C/closed for 7 days, and its purity decreased from 95.81% to 89.35%, and it was transformed into Form I.

On the other hand, the present invention provides a method for preparing each crystal form of the hydrochloride of the compound of formula II, comprising mixing the free form of the compound of formula II with hydrogen chloride in an appropriate organic solvent, filtering and drying after salification. The method can also be stirring a slurry of the hydrochloride of the specific compound of formula II in an appropriate organic solvent, filtering and drying after crystallization. The solvent can be ethyl acetate, isopropyl acetate, methyl ethyl ketone, methyl tert-butyl ether, toluene, cyclohexane, isopropanol, acetonitrile, n-heptane or a mixed solvent of each solvent.

In some embodiments of the present invention, the preparation method of the hydrochloride crystal form A comprises the following steps: (a) dissolving the free base of the compound of formula II in a mixed solvent of methyl ethyl ketone/n-heptane, and stirring to obtain a clear solution; (b) slowly adding ethyl acetate solution of HCl to the solution, stirring, filtering and drying.

In some embodiments of the present invention, in the preparation method of Form A, in step (a), the volume ratio of methyl ethyl ketone/n-heptane can be 1:1; in step (b), the acid-base feed ratio of HCl and 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, 1.7:1, 1.8:1, 1.9:1, 2:1, preferably 1.1:1 to 1.5:1.

In some embodiments herein, in the method for preparing Form A, in step (a), the free base of the compound of Formula II is dissolved at room temperature, and in step (b), stirring is performed at room temperature for 24 hours.

In some embodiments of the present invention, a method for preparing a hydrochloride crystal form B comprises the following steps: (a) dissolving the free base of the compound of formula II in ethyl acetate, and stirring to obtain a clear solution; (b) slowly adding ethyl acetate solution of HCl to the solution, stirring, filtering and drying.

In some embodiments herein, in the method for preparing Form B, in step (b), the acid-base feed ratio of HCl and the free base of the 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, 1.7:1, 1.8:1, 1.9:1, 2:1, preferably 1.1:1 to 1.5:1.

In some embodiments herein, in the method for preparing Form B, in step (a), the free base of the compound of Formula II is dissolved at room temperature, and in step (b), stirring is performed at room temperature for 24 hours.

In the process of preparing monohydrochloride, if the acid-base feed ratio is too low, some free alkali cannot be completely converted into monohydrochloride. If the acid-base feed ratio is high, a mixture of monohydrochloride and dihydrochloride is produced. Therefore, the addition speed and amount of hydrochloric acid need to be strictly controlled. This brings difficulties to industrial production and product quality control.

In some embodiments of the present invention, the preparation method of dihydrochloride crystal form I comprises the following steps: (a) dissolving the free base of the compound of formula II in ethyl acetate, and stirring to obtain a clear solution; (b) slowly adding ethyl acetate solution of HCl to the solution, stirring, filtering and drying.

In some embodiments of the present invention, in the preparation method of Form I, in step (b), the acid-base feed ratio of HCl and the free base of the compound of Formula II is 2:1 to 5:1, such as 2.5:1, 3:1, 3.5:1, 4:1, 4.2:1, preferably 2.5:1 to 4:1. Excessive hydrochloric acid feed helps to completely convert the free base into dihydrochloride, which is simple to operate in industrial production, and the obtained Form I has less impurities and high purity.

In some embodiments herein, in the method for preparing Form I, in step (a), the free base of the compound of formula II is dissolved at room temperature, and in step (b), the mixture is stirred at low temperature for 5 hours.

In some embodiments of the present invention, the preparation method of the dihydrochloride crystal form II comprises the following steps: suspending the dihydrochloride crystal form I of the compound of formula II in a ketone solvent, slurrying and stirring, filtering and drying.

In some embodiments of the present invention, in the preparation method of Form II, the ketone solvent is one or more of acetone, butanone or methyl ethyl ketone, and the slurrying and stirring is slurrying and stirring at room temperature to 50°C for 24 hours.

In some embodiments of the present invention, the preparation method of the dihydrochloride crystal form III comprises the following steps: suspending the dihydrochloride crystal form I of the compound of formula II in methyl tert-butyl ether, slurrying and stirring, filtering and drying.

In some embodiments of the present invention, in the preparation method of Form III, slurrying and stirring are performed at room temperature for 24 hours.

In another aspect, provided herein is a pharmaceutical composition comprising a hydrochloride of a compound of formula II as defined herein (such as a monohydrochloride of a compound of formula II or a dihydrochloride of a compound of formula II), and a pharmaceutically acceptable inert pharmaceutical excipient.

In some embodiments herein, the pharmaceutical composition is formulated into a dosage form for oral administration such as tablets, capsules, powders, granules, liquid preparations (e.g., oral solutions, suspensions, emulsions, etc.) or syrups, etc., which may contain pharmaceutically acceptable excipients such as lubricants, binders, disintegrants, fillers, dispersants, emulsifiers, stabilizers, etc.

For example, the injection can be prepared by mixing or dissolving the compound of the present invention in physiological saline, adding appropriate dilute acid or alkali or buffer salt to adjust the pH to the most stable state, and can also include antioxidants or metal chelators. The solution is sterilized by filtration and filled into sterile ampoules under sterile conditions.

For example, tablets can be prepared by fully mixing the compound of the present invention with excipients (such as microcrystalline cellulose, sodium carboxymethyl starch, corn starch, magnesium stearate, talc, etc.), sieving, and pressing on a tablet press. For example, hard capsules can be prepared by sieving the compound of the present invention, mixing it with excipients/formulators (such as dry starch, magnesium stearate, etc.), and filling the mixture into hard gelatin capsules using appropriate equipment.

The suspension can be prepared by sieving the compound of the present invention and mixing it with excipients (such as sodium carboxymethylcellulose and syrup) to form a uniform paste. Sometimes, pigments, benzoic acid, etc. can be diluted with a portion of purified water and added to the paste under stirring, and then sufficient water is added to produce the required volume. Other methods for preparing pharmaceutical compositions, excipients, etc. can be referred to Remington's Pharmaceutical Sciences, 18th edition, Alfonso R. Gennaro (1990), publisher: Mack Publishing Company and its revised editions.

The pharmaceutical composition herein may be a solid, semisolid or liquid preparation prepared by conventional techniques known per se. Such methods include mixing, dissolving, granulating, grinding, emulsifying, embedding, spray drying or freeze drying. The active ingredient in such a composition may account for 0.1% to 99.9% of the weight of the formulation. The carrier, diluent or excipient used in the composition is compatible with the active ingredient and is pharmaceutically acceptable.

In another aspect, the present invention provides the use of the hydrochloride salt of the compound of formula II in inhibiting thrombin and in preventing and treating thrombin-mediated and/or thrombin-related diseases.

As described in the Examples section herein, the hydrochloride of a representative compound of formula II has a significant antithrombotic effect in an animal model. The present invention relates to the use of the compounds of the present invention, such as the monohydrochloride of the compound of formula II or the dihydrochloride of the compound of formula II, for the prevention and treatment of thrombotic diseases, especially venous or arterial thromboembolism, for example: deep venous thromboembolism of the lower limbs, reocclusion after bypass surgery or angioplasty, peripheral arterial disease obstruction, pulmonary embolism, disseminated intravascular coagulation, coronary thromboembolism, stroke and occlusion of shunts or stents, etc.

This article relates to the use of the compound of the present invention, such as the monohydrochloride of the compound of formula II or the dihydrochloride of the compound of formula II, for preventing and treating stroke, pulmonary embolism, myocardial or cerebral infarction, atrial fibrillation and arrhythmia caused by thrombosis.

The present invention relates to the use of the compound of the present invention, such as the monohydrochloride of the compound of formula II or the dihydrochloride of the compound of formula II, for preventing and treating atherosclerotic diseases such as coronary artery disease, cerebral arterial disease and peripheral arterial disease.

The compounds described herein can also be used as anticoagulants in extracorporeal blood lines. In certain embodiments, the present invention provides a method for inhibiting coagulation in an extracorporeal or ex vivo blood line, the method comprising adding an effective amount of a compound of the present invention to the extracorporeal or ex vivo blood line.

The compounds described herein (e.g., monohydrochloride of the compound of formula II or dihydrochloride of the compound of formula II) can also be used in combination therapy with thrombolytic agents, for example, to reduce reperfusion time and prolong reocclusion time. In addition, the compounds described in the present invention can be used to prevent re-thrombosis after microsurgery. The compounds described in the present invention can also be effective in anticoagulant therapy for hemodialysis and disseminated intravascular coagulation. The compounds described in the present invention can also be used for ex vivo storage of blood, plasma and other blood products.

The compounds herein can be administered orally. The specific dosage of administration depends on the specific circumstances of the case, including the form of administration, the rate of administration, and the condition to be treated. The typical oral daily dose to produce an effect can be between about 0.01 mg/kg and about 1000 mg/kg (calculated as the free base of the compound of formula II). It can be a single dose per day, or multiple doses 2 to 4 times per day. The dosage and mode of administration can be adjusted according to the age and weight of the patient and the severity of the disease to be treated.

Herein, the compound of formula II was synthesized using the method described in US62751984.

In this paper, X-ray diffraction (XRD) was measured using the following method:

(1) The X-ray powder diffraction data of the samples were collected under ambient conditions using a Brooke D2 X-ray powder diffractometer with an X-ray emitter power of 300 W. There was no background signal on the sample stage, the step rate was 0.15 s/step, the total number of steps was 1837, the step length was 2θ = 0.02°, the voltage was 30 kV, and the current was 10 mA. 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) The XRPD analysis was performed using a PANalytical Empyrean X-ray powder diffractometer equipped with a PIXcel1D detector. In the XRPD analysis, the 2θ scanning angle of the sample was from 3° to 40°, the scanning step was 0.013°, and the light tube voltage and light tube current were 45 KV and 40 mA, respectively.

In this paper, differential scanning calorimetry (DSC) was measured using the following method: A TA Discovery series differential scanning calorimeter (DSC) was used to collect thermal data of the sample. Several milligrams of sample were weighed in a Tzero aluminum pan and sealed with a Tzero sealing lid. The pan was heated under _N2 protection at a heating rate of 10°C/min.

In this paper, thermogravimetric analysis (TGA) was measured using the following method: Thermogravimetric data of the samples were collected using a TA Discovery series thermogravimeter (TGA). A few milligrams of sample were placed in a Tzero aluminum pan and heated from room temperature to the target temperature under N2 protection at a heating rate of 10°C/min.

It should be noted that in X-ray diffraction spectra, the diffraction spectrum obtained from a crystalline compound is often characteristic for a specific crystal form, and the relative intensity of the spectral band (especially at low angles) may change due to the preferred orientation effect caused by differences in crystallization conditions, particle size and other measurement conditions. Therefore, the relative intensity of the diffraction peak is not characteristic for the crystal form being targeted. When judging whether it is the same as a known crystal form, more attention should be paid to the relative position of the peak rather than their relative intensity. In addition, for any given crystal form, there may be slight errors in the position of the peak, which is also well known in the field of crystallography. For example, due to changes in temperature, sample movement, or instrument calibration when analyzing the sample, the position of the peak can move, and the measurement error of the 2θ value is sometimes about ± 0.2°. Therefore, this error should be taken into account when determining the structure of each crystal form. In XRD spectra, the peak position is usually represented by the 2θ angle or the interplanar distance d, and there is a simple conversion relationship between the two: d = λ/2sinθ, where d represents the interplanar distance, λ represents the wavelength of the incident X-ray, and θ is the diffraction angle. For the same crystal form of the same compound, the peak position of the XRD spectrum is similar overall, and the relative intensity error may be large. It should also be pointed out that in the identification of mixtures, some diffraction lines may be missing due to factors such as decreased content. At this time, there is no need to rely on all the bands observed in high-purity samples, and even one band may be characteristic of a given crystal.

It should be noted that DSC measures the transition temperature when a crystal absorbs or releases heat due to changes in its crystal structure or melting of the crystal. For the same type of crystal form of the same compound, in consecutive analyses, the thermal transition temperature and melting point errors are typically within about 3°C. When we say that a compound has a given DSC peak or melting point, this refers to the DSC peak or melting point ±3°C. DSC provides an auxiliary method for distinguishing different crystal forms. Different crystal forms can be identified based on their different transition temperature characteristics.

Herein, the term "pharmaceutical composition" refers to a preparation comprising the active compound of the present application and a carrier, excipient and/or medium generally accepted in the art for delivering the biologically active compound to an organism (e.g., a human). The purpose of the pharmaceutical composition is to facilitate administration of the compound of the present application to an organism.

Herein, the term "pharmaceutically acceptable inert pharmaceutical excipients" includes but is not limited to any carrier, excipient, medium, glidant, sweetener, diluent, preservative, dye/colorant, taste enhancer, surfactant, wetting agent, dispersant, disintegrant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier that can be used for humans or animals (such as livestock).

All solvents used in this article were commercially available and used without further purification.

Herein, room temperature refers to 25°C.

Herein, low temperature refers to -10 to 25°C.

In this article, the abbreviations have the following meanings: EtOH Ethanol IPA Isopropyl alcohol NBA Butanol MEK Butanone EA Ethyl acetate IPA Isopropyl acetate ACN Acetonitrile THF Tetrahydrofuran MTBE Methyl tert-butyl ether Diox 1,4-Dioxane Tol Toluene DMSO Dimethyl sulfoxide Hept n-Heptane _H2O water AC Acetic acid DVS Dynamic Vapor Sorption Analysis

In this article, the calculation method of the product content is as follows: content = (100% - weight loss - residue) X (100% - total impurities) * 100%.

The purity of the product was calculated by HPLC area normalization method.

The following is an illustrative example of the content of this article, but the specific examples do not limit the scope of this article in any way. Example 1: Preparation of Monohydrochloride Crystal Form A

Dissolve 300 mg of free base in 3 mL of methyl ethyl ketone/n-heptane (1:1, v/v), and stir at room temperature to obtain a clear solution. Slowly add HCl in ethyl acetate (acid-base ratio 2:1) to the solution, stir at room temperature for 24 hours, and generate a white suspension. Separate the solid by filtration, and vacuum dry at 35 °C to obtain 285 mg of monohydrochloride crystal form A sample, with a yield of about 88.6%. Example 2: Preparation of monohydrochloride crystal form A

Dissolve 300 mg of free base in 3 mL of methyl ethyl ketone/n-heptane (1:1, v/v), and stir at room temperature to obtain a clear solution. Slowly add HCl in ethyl acetate (acid-base feed ratio 1.3:1) to the solution, stir at room temperature for 24 hours to generate a white suspension, separate the solid by filtration, and vacuum dry at 35 °C to obtain 241 mg of monohydrochloride crystal form A sample, with a yield of about 75.4%. Example 3: Preparation of monohydrochloride crystal form B

Dissolve 300 mg of free base in 3 mL of ethyl acetate and stir at room temperature to obtain a clear solution. Slowly add HCl in ethyl acetate (acid-base feed ratio 2:1) to the solution and stir at room temperature for 24 hours to generate a white suspension. The solid is separated by filtration and vacuum dried at 35 °C to obtain 223 mg of monohydrochloride crystal form B sample, with a yield of about 68.4%, 0.6% water, 3.77% hydrolyzed impurities, 4.5% total impurities, 95.6% HPLC purity, and 95.2% content. Example 4: Preparation of dihydrochloride crystal form I

Dissolve 1.75 kg of free base in 7 L of ethyl acetate and stir at room temperature to obtain a clear solution. Slowly add HCl in ethyl acetate (acid-base ratio 3:1) to the solution at low temperature and stir at room temperature for 5 hours to generate a white suspension. The solid is separated by filtration and vacuum dried at 40 °C to obtain 1.76 kg of dihydrochloride crystal form I sample with a yield of about 89.4% and HPLC purity of 99.6%. Example 5: Preparation of dihydrochloride crystal form II

The crystal form I (180 mg) obtained in Example 1 was suspended in methyl ethyl ketone (3 mL), slurried and stirred at room temperature for 24 h, the solid was separated by filtration, and vacuum dried at 35 °C to obtain 136 mg of the dihydrochloride crystal form II sample, with a yield of about 75.6%. Example 6: Preparation of dihydrochloride crystal form III

The crystal form I (180 mg) obtained in Example 1 was suspended in methyl tert-butyl ether (3 mL), slurried and stirred at room temperature for 24 h, the solid was separated by filtration, and vacuum dried at 35 °C to obtain 166 mg of the dihydrochloride crystal form III sample, with a yield of about 92.2%. Example 7: Rat deep vein thrombosis model

Experimental animals: SD rats (weight 220 ~ 250 g, sex: male)

Solution preparation: Weigh an appropriate amount of Form I, dissolve it in a solvent (0.5% (W/V) Natrosol ^TM 250 HX solution containing 0.1% (W/V) L(+)-tartaric acid) to prepare the required concentration, stir evenly, and administer according to the predetermined dose.

Operation procedure: Rats were adaptively raised for 3 days and randomly divided into three groups: model group, positive control group, test group and bridge group, with 8 to 10 rats in each group. The model group was given an equal amount of solvent by oral gavage, the positive control group or the test group was given by oral gavage, and the bridge group was given active compound V in crystal form I by intravenous administration. After about 45 minutes, anesthesia and blood sampling were performed to start modeling. Rats in each group were anesthetized with uratan (20%, 5 mL/kg) by intraperitoneal injection. The abdominal cavity was exposed, the caesarean vein was bluntly separated, and two surgical sutures with a length of 8 to 10 cm were placed under the blood vessels, with a distance of 1 cm between them. After the modeling time of each group was reached, rabbit thromboplastin (0.02 mg/kg) was injected into the left femoral vein. After 10 seconds, the two sutures were tied. After 1 hour, the knotted part was cut open with ophthalmic scissors, and the thrombus was removed and weighed.

After rats were orally administered 2.5, 5, 10 and 20 mg/kg of Form I, the blood concentrations of Compound V were 190.1, 181.3, 341.9 and 609.7 ng/mL, respectively, which was basically dose-dependent, and the blood concentration increased with the increase of the dose. At the same dose, the antithrombotic effects of Form I and dabigatran are basically equivalent. After intravenous infusion of Compound V to rats at 0.1 mg/kg/h, the antithrombotic effect was equivalent to that of oral administration of 5 mg/kg Form I, and the blood concentration of Compound V was equivalent.

result Group No. Group Dosage (mg/kg) Thrombus weight (mg) Thrombosis inhibition rate (%) Blood drug concentration (ng/mL) 1 Solvent group N/A 71.0±8.8 - 2 Dabigatran 5 21.4±8.2** 69.9 126.8±28.4 3 Crystalline Form I Group 1 2.5 51.9±8.3** 27.0 190.1±62.9 4 Crystalline Form I Group 2 5 32.1±5.7** 54.7 181.3±69.5 5 Form I Group 3 10 21.3±5.1** 70.0 341.9±46.4 6 Crystalline Form I Group 4 20 14.4±3.6** 79.7 609.7±174.0 7 Compound V 0.1 mg/kg/h 36.8±6.6** 48.2 174.2±51.8 **P＜0.01, compared with the vehicle group Example 8: Pharmacokinetic study in rats

Experimental animals: SD rats (weight 200-300 g, sex: half male and half female)

Preparation of solution for oral administration: Weigh an appropriate amount of Form I, dissolve it in a solvent (0.5% (W/V) Natrosol™ 250 HX solution containing 0.1% (W/V) L(+)-tartaric acid) to prepare the required concentration, stir evenly, and administer the drug according to the predetermined dose.

Operation procedure: Rats were adaptively raised for 3 days and randomly divided into groups of 8 rats each, half male and half female. The drug dosage was 5, 10, and 20 mg/kg by gavage. Blood was collected from the cervical vein at 5 min, 10 min, 20 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, 12 h, and 24 h after drug administration, and the blood drug concentration was tested by LC-MS/MS after processing.

Data processing Phoenix WinNonlin 6.4 non-compartmental model was used to calculate the pharmacokinetic parameters of rats after drug administration, including 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 , C _min , C _avg , AUC _0-τ , etc. after multiple drug administration to achieve steady state.

result: Dosage ( mg/kg ) gender _Tmax (h) t _1/2 (h) C _max (ng/mL) AUC _0-t (h ng/mL) AUC _0-∞ (h ·ng/mL) V _z /F (L/kg) Cl/F (L/h/kg) MRT _0-t (h) MRT _0-∞ (h) 5 female 0.59±0.29 1.5±0.26 522±170 850±226 854±224 13.9±6.75 6.24±1.98 1.6±0.34 1.7±0.38 male 0.66±0.23 1.4±0.32 344±163 604±182 606±181 18.1±6.08 8.81±2.50 1.6±0.20 1.7±0.21 Overall 0.62±0.24 1.5±0.27 433±181 727±231 730±231 16.0±6.37 7.53±2.50 1.6±0.26 1.7±0.28 10 female 0.75±0.29 2.6 1120±385 2060±783 1830 20.1 5.54 1.6±0.27 1.6 male 0.63±0.25 4.5±1.8 934±299 1460±324 1460±324 46.9±24.6 7.1±1.59 1.6±0.24 1.7±0.20 Overall 0.69±0.26 3.9±1.8 1030±335 1760±642 1590±346 38.0±23.6 6.58±1.53 1.6±0.24 1.7±0.26 20 female 0.50±0.0 4.0±0.35 2170±380 3840±446 3850±446 30.8±5.67 5.25±0.610 1.7±0.19 1.8±0.18 male 0.88±0.25 3.7±1.7 2230±531 4650±2440 4660±2440 26.6±17.0 4.98±1.79 1.8±0.33 1.8±0.32 Overall 0.69±0.26 3.9±1.2 2200±429 4250±1680 4250±1680 28.7±11.9 5.12±1.25 1.8±0.25 1.8±0.25

The embodiments of this invention can be modified and altered in many ways without departing from the scope of this invention. Therefore, it should be understood that the embodiments of this invention should not be limited to the exemplary embodiments described above, but should be controlled by the limitations set forth in the claims and any equivalent forms thereof.

without

Figure 1 shows the XRD spectrum of the crystalline form A of the monohydrochloride of the compound of formula II. Figure 2 shows the DSC spectrum of the crystalline form A of the monohydrochloride of the compound of formula II. Figure 3 shows the TGA spectrum of the crystalline form A of the monohydrochloride of the compound of formula II. Figure 4 shows the XRD spectrum of the crystalline form B of the monohydrochloride of the compound of formula II. Figure 5 shows the DSC spectrum of the crystalline form B of the monohydrochloride of the compound of formula II. Figure 6 shows the TGA spectrum of the crystalline form B of the monohydrochloride of the compound of formula II. Figure 7 shows the XRD spectrum of the crystalline form I, crystalline form II, and crystalline form III of the dihydrochloride of the compound of formula II. Figure 8 shows the DSC spectrum of the crystalline form I of the dihydrochloride of the compound of formula II. Figure 9 shows the TGA spectrum of the crystalline form I of the dihydrochloride of the compound of formula II. Figure 10 shows the DSC spectrum of the crystalline form II of the dihydrochloride of the compound of formula II. Figure 11 shows the TGA spectrum of the crystalline form II of the dihydrochloride of the compound of formula II. Figure 12 shows the DSC spectrum of the crystalline form III of the dihydrochloride of the compound of formula II. Figure 13 shows the TGA spectrum of the crystalline form III of the dihydrochloride of the compound of formula II. Figure 14 shows the XRD spectrum of the crystalline form I of the dihydrochloride of the compound of formula II. Figure 15 shows the XRD spectrum of the crystalline form II of the dihydrochloride of the compound of formula II. Figure 16 shows the XRD spectrum of the crystalline form III of the dihydrochloride salt of the compound of formula II.

Claims (23)

A crystalline form of a hydrochloride salt of a compound of formula II, (II). A crystalline form of the hydrochloride of the compound of formula II as described in claim 1, wherein the stoichiometric ratio of the compound of formula II and HCl is 1:1 or 1:2. A crystalline form A of a monohydrochloride salt of a compound of formula II, which has 2θ=6.7±0.2°, 16.2±0.2°, 21.8±0.2°, 22.9±0.2° or 6.7±0.2°, 16.2±0.2°, 16.5±0.2°, 20.7±0.2°, 21.8±0.2°, 22.9±0.2° in an X-ray diffraction pattern measured by Cu-Kα radiation. Diffraction peaks are 0.2° or 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°. The crystal form A as described in claim 3 has an X-ray diffraction spectrum measured by Cu-Kα radiation substantially the same as that shown in FIG1; or it has a DSC spectrum substantially the same as that shown in FIG2; or it has a TGA spectrum substantially the same as that shown in FIG3. A crystalline form B of a monohydrochloride salt of a compound of formula II, which has 2θ=6.6±0.2°, 15.2±0.2°, 21.2±0.2°, 21.6±0.2° or 6.6±0.2°, 15.2±0.2°, 20.3±0.2°, 21.2±0.2°, 21.6±0.2° in an X-ray diffraction pattern measured by Cu-Kα radiation. Diffraction peaks at 21.6±0.2°, 23.7±0.2° or 6.6±0.2°, 9.2±0.20°, 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°. The crystal form B as described in claim 5 has an X-ray diffraction spectrum measured by Cu-Kα radiation substantially the same as that shown in FIG4; or it has a DSC spectrum substantially the same as that shown in FIG5; or it has a TGA spectrum substantially the same as that shown in FIG6. A dihydrochloride of a compound of formula II; (II). A crystalline form I of the dihydrochloride salt of a compound of formula II, which has 2θ=6.0±0.2°, 17.1±0.2°, 19.9±0.2°, 23.3°± 0.2° or 6.0±0.2°, 17.1±0.2°, 19.9±0.2°, 23.3±0.2°, 25.4±0.2°, 25.7± 0.2° or 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. Diffraction peaks are 23.3±0.2°, 25.4±0.2°, and 25.7±0.2°. The crystal form I as described in claim 8 has an X-ray diffraction spectrum measured by Cu-Kα radiation that is substantially the same as that of the crystal form I shown in FIG7; or It has a DSC spectrum that is substantially the same as that shown in FIG8; or It has a TGA spectrum that is substantially the same as that shown in FIG9. A crystalline form II of the dihydrochloride salt of a compound of formula II, which has 2θ=6.0±0.2°, 17.1±0.2°, 20.0±0.2°, 22.8°±0.2° or 6.0±0.2°, 13.4±0.2°, 17.1±0.2°, 20.0±0.2°, 21.2±0.2°, 22.8±0.2° in an X-ray diffraction pattern measured by Cu-Kα radiation. Diffraction peaks of 0.2° or 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°. The crystal form II as described in claim 10 has an X-ray diffraction spectrum measured by Cu-Kα radiation that is substantially the same as that of the crystal form II shown in FIG7; or It has a DSC spectrum that is substantially the same as that shown in FIG10; or It has a TGA spectrum that is substantially the same as that shown in FIG11. A crystalline form III of the dihydrochloride salt of a compound of formula II, wherein the X measured by Cu-Kα radiation is The X-ray diffraction pattern has diffraction peaks of 2θ=6.0±0.2°, 17.2±0.2°, 20.1±0.2°, 22.0±0.2° or 6.0±0.2°, 13.5±0.2°, 17.2±0.2°, 20.1±0.2°, 21.2±0.2°, 22.0±0.2° or 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°, 26.4±0.2°. The crystal form III as described in claim 12 has an X-ray diffraction spectrum measured by Cu-Kα radiation that is substantially the same as that of the crystal form III shown in FIG7; or It has a DSC spectrum that is substantially the same as that shown in FIG12; or It has a TGA spectrum that is substantially the same as that shown in FIG13. A method for preparing form A as described in claim 3 or 4, characterized in that it comprises the following steps: (a) dissolving the free base of the compound of formula II in a mixed solvent of methyl ethyl ketone/n-heptane, and stirring to obtain a clear solution; (b) slowly adding ethyl acetate solution of HCl to the solution, stirring, and filtering to dryness; The acid-base feed ratio of the HCl and 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 step (b) is stirred at room temperature for 24 hours. A method for preparing form B as described in claim 5 or 6, characterized in that it comprises the following steps: (a) dissolving the free base of the compound of formula II in ethyl acetate and stirring to obtain a clear solution; (b) slowly adding HCl in ethyl acetate to the solution, stirring, filtering and drying; The acid-base feed ratio of the HCl and 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 performed at room temperature for 24 hours. A method for preparing form I as described in claim 8 or 9, characterized in that it comprises the following steps: (a) dissolving the free base of the compound of formula II in ethyl acetate and stirring to obtain a clear solution; (b) slowly adding HCl in ethyl acetate to the solution, stirring, filtering and drying; The acid-base feed ratio of the HCl and 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 performed at low temperature for 5 hours. A method for preparing the crystal form II as described in claim 10 or 11, characterized in that it comprises the following steps: suspending the crystal form I of the dihydrochloride salt of the compound of formula II in a ketone solvent, slurrying and stirring, filtering and drying; Preferably, the ketone solvent is one or more of acetone, butanone or methyl ethyl ketone; Preferably, the slurrying and stirring is slurrying and stirring at room temperature to 50°C for 24 hours. A method for preparing the crystal form III as described in claim 12 or 13, characterized in that it comprises the following steps: suspending the crystal form I of the dihydrochloride salt of the compound of formula II in methyl tert-butyl ether, slurrying and stirring, filtering and drying; Preferably, the slurrying and stirring is slurrying and stirring at room temperature to 50°C for 24 hours. A pharmaceutical composition comprising a crystalline form of the hydrochloride of the compound of formula II as described in claim 1 or 2 or the monohydrochloride and/or dihydrochloride of the compound of formula II as described in any one of claims 3 to 13, and one or more pharmaceutically acceptable inert pharmaceutical excipients; Preferably, the pharmaceutical composition is formulated for oral administration. A pharmaceutical composition comprising more than 95% of the dihydrochloride salt of the compound of formula II. A crystalline composition comprising more than 80% of the crystalline form I of the hydrochloride salt of the compound of formula II. Use of a crystalline form of the hydrochloride of the compound of formula II as described in claim 1 or 2, the monohydrochloride or dihydrochloride of the compound of formula II as described in any one of claims 3 to 13, the pharmaceutical composition as described in claim 19 or 20, or the crystalline composition as described in claim 21 in the preparation of a drug for preventing and treating thrombin-mediated and/or thrombin-related diseases. Use of a crystalline form of the hydrochloride of the compound of formula II as described in claim 1 or 2, the monohydrochloride or dihydrochloride of the compound of formula II as described in any one of claims 3 to 13, the pharmaceutical composition as described in claim 19 or 20, or the crystalline composition as described in claim 21 in the preparation of a drug for treating venous and/or arterial thrombotic diseases.