TWI880627B - 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 are 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. (II)

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, its preparation method and use.

US Patent US17/289379 discloses a compound shown in Formula II (referred to as Compound II for short), 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.

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 states by carboxylesterase (carboxylesterase-1/carboxylesterase-2, CES-1/CES-2) in vivo. Compound V exerts anticoagulant effect by selectively inhibiting thrombin.

However, excessive inhibition of thrombin activity may cause bleeding risk, so 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,

In some embodiments of the present invention, 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 at 2θ=6.7±0.2°, 16.2±0.2°, 21.8±0.2°, and 22.9±0.2° in an X-ray diffraction pattern measured by Cu-Kα radiation; typically, it has diffraction peaks at 6.7±0.2°, 16.2±0.2°, 16.5±0.2°, 20.7±0.2°, and 21.8±0.2°. .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°, 27.5±0.2°.

In some embodiments of the present invention, the crystalline form A of the monohydrochloride salt of the compound of formula II has an X-ray diffraction pattern measured by Cu-Kα radiation substantially the same as that shown in FIG1.

In some embodiments of the present invention, the crystalline form A of the monohydrochloride salt of the compound of formula II has a DSC spectrum substantially the same as that shown in FIG2 or a TGA spectrum substantially the same as that shown in FIG3.

On the other hand, the present invention provides a crystalline form B of a monohydrochloride salt of a compound of formula II, which has diffraction peaks of 2θ=6.6±0.2°, 15.2±0.2°, 21.2±0.2°, and 21.6±0.2° in an 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°. 2±0.2°, 21.6±0.2°, 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°, 25.7±0.2°.

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

In some embodiments of the present invention, the crystalline form B of the monohydrochloride salt of the compound of formula II has a DSC spectrum substantially the same as that shown in FIG5 or a TGA spectrum substantially the same as that shown in FIG6.

The present invention also provides 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 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, it has 2θ=6.0±0.2°, 17.1±0.2°, 19.9±0.2°, 23.3±0.2°. 0.2°, 25.4±0.2°, 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°, 25.7±0.2°.

In some embodiments of the present invention, the dihydrochloride salt of the compound of formula II in Form I has an X-ray diffraction pattern measured by Cu-Kα radiation that is substantially the same as that of Form I in Figure 7.

In some embodiments of the present invention, 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.

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 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, it has 2θ=6.0±0.2°, 13.4±0.2°, 17.1±0.2°, 20.0±0.2°, and 22.8±0.2°. ±0.2°, 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°, 30.2±0.2°.

In some embodiments of the present invention, the dihydrochloride salt of the compound of formula II has an X-ray diffraction pattern measured by Cu-Kα radiation that is substantially the same as that of the dihydrochloride salt of the compound of formula II shown in Figure 7.

In some embodiments of the present invention, 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.

The present invention also provides a crystalline form III of the dihydrochloride salt of the compound of formula II, which has diffraction peaks of 2θ=6.0±0.2°, 17.2±0.2°, 20.1±0.2°, and 22.0±0.2° in an X-ray diffraction (XRD) pattern measured by Cu-Kα radiation; typically, it has diffraction peaks of 2θ=6.0±0.2°, 13.5±0.2°, 17.2±0.2°, 20.1±0.2°, and 22.0±0.2°. 2°, 21.2±0.2°, 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 of the present invention, the dihydrochloride salt of the compound of formula II, Form III, has an X-ray diffraction pattern measured by Cu-Kα radiation that is substantially the same as that of Form III shown in Figure 7.

In some embodiments of the present invention, the crystalline form III of the dihydrochloride salt of the compound of formula II has a DSC spectrum substantially the same as shown in Figure 12 or a TGA spectrum substantially the same as shown in Figure 13.

This article 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 this 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 1:1~2:1, 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 1.1:1~1.5:1.

In some embodiments of this invention, in the preparation method of Form A, 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 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 of this invention, in the preparation method of Form B, in step (b), the acid-base feed ratio of HCl and the free base of the compound of Formula II is 1:1~2:1, for example, 11:1, 1.3:1, 1.4:1, 1.5:1, 1.6, 1.7:1, 1.8:1, 1.9:1, 2:1, preferably 1.1:1~1.5:1.

In some embodiments of this invention, in the preparation method of Form B, 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 room temperature for 24 hours.

On the other hand, the present invention provides a method for preparing dihydrochloride salt form I, comprising the following steps: (a) dissolving the free base of the compound of formula II in ethyl acetate, 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 this 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 2:1~5:1, for example, 2.5:1, 3:1, 3.5:1, 4:1, 4.2:1, preferably 2.5:1~4:1.

In some embodiments of the present invention, in the preparation method of 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, this article provides a method for preparing 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 this 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, this article provides a method for preparing 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 of this invention, in the preparation method of Form III, the slurrying and stirring is performed at room temperature ~50°C for 24 hours.

The present invention also provides a pharmaceutical composition comprising a crystalline form of the hydrochloride of the compound of formula II or the monohydrochloride and/or dihydrochloride of the compound of formula II, and one or more pharmaceutically acceptable inert pharmaceutical excipients.

The present invention also provides 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.

The present invention also provides 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.

On the other hand, the present invention provides the use of the crystalline form of the hydrochloride of the compound of formula II defined above, the monohydrochloride, dihydrochloride, pharmaceutical composition, or crystalline composition of the compound of formula II in the preparation of drugs for the prevention and treatment of thrombin-mediated and thrombin-related diseases.

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

Figure 1 shows the XRD spectrum of the crystalline form A of the monohydrochloride salt of the compound of formula II.

Figure 2 shows the DSC spectrum of the crystalline form A of the monohydrochloride salt of the compound of formula II.

Figure 3 shows the TGA spectrum of the crystalline form A of the monohydrochloride salt of the compound of formula II.

Figure 4 shows the XRD spectrum of the crystalline form B of the monohydrochloride salt of the compound of formula II.

Figure 5 shows the DSC spectrum of the crystalline form B of the monohydrochloride salt of the compound of formula II.

Figure 6 shows the TGA spectrum of the crystalline form B of the monohydrochloride salt of the compound of formula II.

Figure 7 shows the XRD spectra of the dihydrochloride salt of the compound of formula II in form I, form II, and form III.

Figure 8 shows the DSC spectrum of Form I of the dihydrochloride salt of the compound of Formula II.

Figure 9 shows the TGA spectrum of the crystalline form I of the dihydrochloride salt of the compound of formula II.

Figure 10 shows the DSC spectrum of the dihydrochloride salt of the compound of formula II in form II.

Figure 11 shows the TGA spectrum of the dihydrochloride salt of the compound of formula II in form II.

Figure 12 shows the DSC spectrum of the dihydrochloride salt of the compound of formula II in form III.

Figure 13 shows the TGA spectrum of the dihydrochloride salt of the compound of formula II in form III.

Figure 14 shows the XRD spectrum of the crystalline form I of the dihydrochloride salt of the compound of formula II.

Figure 15 shows the XRD spectrum of the dihydrochloride salt of the compound of formula II in form II.

Figure 16 shows the XRD spectrum of the crystalline form III of the dihydrochloride salt of the compound of formula II.

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 embodiment 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, specific elements, structures or features can be combined in one or more embodiments in any appropriate manner.

On the one hand, 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 1:1 of the compound of formula II: HCl, and can exist in the form of anhydrous crystals, hydrated forms, and solvents. The dihydrochloride of the compound of formula II has a chemical ratio of 1:2 of the compound of formula II: HCl, and can exist in the form of anhydrous crystals, hydrated forms, and solvents.

In this context, 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 formulate into pharmaceutical compositions such as tablets or capsules. 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 intrinsic 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 is easier to separate than the free base, so salt formation can provide a purification method.

Any suitable method known in the art for isolating the crystallized hydrochloride may be used. Suitably, the monohydrochloride or dihydrochloride of the compound of formula II is collected by filtration separation. Preferably, the monohydrochloride or dihydrochloride of the compound of formula II is collected and dried under vacuum.

Each crystalline form of the hydrochloride 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.

On the other hand, the present invention provides a crystalline form A of the monohydrochloride salt of the compound of formula II.

In some embodiments of the present invention, the crystalline form A of the monohydrochloride salt of the compound of formula II has diffraction peaks at 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, the crystalline form A has diffraction peaks at 2θ=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 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 of the present invention, the crystalline form A of the monohydrochloride salt of the compound of formula II has an X-ray diffraction pattern measured by Cu-Kα radiation substantially the same as that shown in FIG1.

In some embodiments of the present invention, 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.

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

This article also provides the crystalline form B of the monohydrochloride salt of the compound of formula II.

In some embodiments herein, the crystalline form B of the monohydrochloride salt of the compound of formula II has diffraction peaks at 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, the crystalline form B has diffraction peaks at 2θ=6.6±0.2°, 15.2±0.2°, 21.2±0.2°, and 21.6±0.2°. ±0.2°, 21.6±0.2°, 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°, 25.7±0.2°.

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

In some embodiments of the present invention, 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.

In some embodiments of the present invention, the crystalline form B of the monohydrochloride salt of the compound of formula II has a DSC spectrum substantially the same as that shown in FIG5 or a TGA spectrum substantially the same as that shown in FIG6.

The present invention also provides the dihydrochloride salt of the compound of formula II.

This article also provides the crystalline 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 at 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 at 2θ=6.0±0.2°, 17.1±0.2°, 19.9±0.2°, and 23.3±0.2°. ±0.2°, 25.4±0.2°, 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°, 25.7±0.2°.

In some embodiments of the present invention, the dihydrochloride salt of the compound of formula II in form I has an X-ray diffraction pattern measured by Cu-Kα radiation that is substantially the same as that of form I in Figure 7.

In some embodiments of the present invention, 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.

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

In some embodiments of the present invention, 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 the sample transforms into an amorphous form after DVS testing.

The physicochemical properties of Form I are as follows:

The solubility of Form I was determined as follows:

The accelerated stability test data of Form I at T=25±2℃ and humidity 60±5% are as follows:

This article also provides the crystalline 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 at 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 at 2θ=6.0±0.2°, 13.4±0.2°, 17.1±0.2°, 20.0±0.2°, and 22.8±0.2°. ±0.2°, 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°, 30.2±0.2°.

In some embodiments of the present invention, the dihydrochloride salt of the compound of formula II has an X-ray diffraction pattern measured by Cu-Kα radiation that is substantially the same as that of the dihydrochloride salt of the compound of formula II shown in Figure 7.

In some embodiments of the present invention, 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.

In some embodiments of the present invention, 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 of the present invention, the DSC spectrum of the crystal 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.

There is a broad endothermic peak before 100°C in the DSC curve of Form II. The crystal form does not change when the DSC is heated to 110°C. In some embodiments of the present invention, when Form II is obtained by slurrying in acetone, there is about 0.2% acetone residue.

This article also provides the crystalline 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 diffraction peaks at 2θ=6.0±0.2°, 17.2±0.2°, 20.1±0.2°, and 22.0±0.2° in the X-ray diffraction (XRD) pattern measured by Cu-Kα radiation; typically, the crystalline form III has diffraction peaks at 2θ=6.0±0.2°, 13.5±0.2°, 17.2±0.2°, 20.1±0.2°, and 22.0±0.2°. ±0.2°, 21.2±0.2°, 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 of the present invention, the dihydrochloride salt of the compound of formula II in form III has an X-ray diffraction pattern measured by Cu-Kα radiation that is substantially the same as that of form III shown in FIG7 .

In some embodiments of the present invention, 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.

In some embodiments of the present invention, the crystalline form III of the dihydrochloride salt of the compound of formula II has a DSC spectrum substantially the same as that shown in FIG. 12 or a TGA spectrum substantially the same as that shown in FIG. 13.

In some embodiments of this invention, the DSC spectrum of the crystal 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 shows 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.

Crystal form III was transformed into crystal form I after mechanical grinding for 2 minutes. Crystal form III was stored at 60℃/closed mouth for 7 days, and its purity dropped from 95.81% to 89.35%, and it was transformed into crystal 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 a suitable organic solvent, filtering and drying after the salification is completed. The method may also be stirring a slurry of the hydrochloride of the specific compound of formula II in a suitable organic solvent, filtering and drying after the crystallization is completed. The solvent may be ethyl acetate, isopropyl acetate, methyl ethyl ketone, methyl tert-butyl ether, toluene, cyclohexane, isopropanol, acetonitrile, n-heptane or a mixed solvent of the solvents.

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 1:1~2:1, 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 1.1:1~1.5:1.

In some embodiments of this invention, in the preparation method of Form A, 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 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 of this invention, in the preparation method of Form B, in step (b), the acid-base feed ratio of HCl and the free base of the compound of Formula II is 1:1~2:1, for example, 11:1, 1.3:1, 1.4:1, 1.5:1, 1.6, 1.7:1, 1.8:1, 1.9:1, 2:1, preferably 1.1:1~1.5:1.

In some embodiments of this invention, in the preparation method of Form B, 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 room temperature for 24 hours.

During the preparation of monohydrochloride, if the acid-base feed ratio is too low, some free bases 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 obtaining a clear solution after stirring; (b) slowly adding ethyl acetate solution of HCl to the solution, stirring, filtering and drying.

In some embodiments of this 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 2:1~5:1, for example, 2.5:1, 3:1, 3.5:1, 4:1, 4.2:1, preferably 2.5:1~4:1. Excessive hydrochloric acid feed helps the free base to be completely converted into dihydrochloride, which is simple to operate in industrial production, and the obtained Form I has less impurities and high purity.

In some embodiments of this invention, in the preparation method of 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 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 this 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 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 this invention, in the preparation method of Form III, slurry is stirred at room temperature for 24 hours.

On the other hand, the present invention provides 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 of the present invention, the drug 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 formulations 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, for tablets, the compound of the present invention and excipients (such as microcrystalline cellulose, sodium carboxymethyl starch, corn starch, magnesium stearate, talc, etc.) can be fully mixed, sieved, and compressed on a tablet press. For example, for hard capsules, the compound of the present invention can be sieved, mixed with excipients/formulators (such as dry starch, magnesium stearate, etc.), and the mixture can be filled into hard gelatin capsules using appropriate equipment.

The suspension can be prepared by screening 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 refer 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 made 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 formula weight. The carrier, diluent or excipient used in the composition is compatible with the active ingredient and is pharmaceutically acceptable.

On the other hand, the present invention provides the use of the hydrochloride 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 of this article, 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 described in 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, such as deep vein thromboembolism of the lower limbs, re-occlusion 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 compounds described in 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.

This article relates to the use of the compounds described in 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 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 the compound of the present invention to the extracorporeal or ex vivo blood line.

The compounds described herein (such as the monohydrochloride of the compound of formula II or the dihydrochloride of the compound of formula II) can also be used in combination with thrombolytic agents, for example: to reduce reperfusion time and prolong reclosure 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 treatment of 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 the administration depends on the specific circumstances of the case, including the form of administration, the administration rate, 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 (based on the free base of the compound of formula II). It can be a single dose per day, or multiple doses 2 to 4 times a day. The dosage and administration method can be adjusted according to the patient's age and weight and the severity of the disease to be treated.

In this article, the compound of formula II was synthesized using the method described in US62/751984.

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

(1) A Brooke D2 X-ray powder diffractometer was used to collect X-ray powder diffraction data of the sample under ambient conditions. The X-ray emitter power was 300W. There was no background signal on the sample stage, the step rate was 0.15s/step, the total number of steps was 1837 steps, the step length was 2θ=0.02°, the voltage was 30kV, and the current was 10mA. The X-ray tube used a Cu target (Kα), and the Kα2/Kα1 intensity ratio was 0.50 (1.54439Å/1.5406Å).

(2) The analysis was performed using a PANalytical Empyrean X-ray powder diffractometer equipped with a PIXcellD 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 45KV and 40mA, 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. Heating was performed 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 the 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 bands (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 to which it is targeted. When judging whether it is the same as a known crystal form, more attention should be paid to the relative position of the peaks rather than their relative intensity. In addition, for any given crystal form, there may be slight errors in the position of the peaks, which is also well known in the field of crystallography. For example, due to changes in temperature during sample analysis, sample movement, or instrument calibration, 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 its 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 for 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 crystal form of the same compound, the thermal transition temperature and melting point errors are typically within about 3°C in consecutive analyses. 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 containing 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., 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 are commercially available and used without further purification.

In this article, room temperature refers to 25℃.

In this article, low temperature refers to -10~25℃.

In this article, the abbreviations have the following meanings:

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 description of the content of this article in conjunction with specific embodiments, but the specific embodiments do not limit the scope of this article in any way.

Example 1: Preparation of hydrochloride 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 and 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 285 mg of monohydrochloride crystal form A sample with a yield of about 88.6%.

Example 2: Preparation of hydrochloride 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 1.3:1) to the solution and 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 hydrochloride 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 ratio 2:1) to the solution and 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 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 to the solution at low temperature (acid-base ratio 3:1), stir at room temperature for 5 hours, and generate a white suspension. Separate the solid by filtration and vacuum dry 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 the example 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 the example 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 220~250g, 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 dosage.

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 8-10 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%, 5mL/kg) by intraperitoneal injection. The abdominal cavity was exposed, the caesarean vein was bluntly separated, and two surgical sutures of 8-10 cm in length 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 (002 mg/kg) was injected into the left femoral vein. After 10 seconds, the two lines 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. The concentrations were dose-dependent, and the blood concentrations 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 concentrations of Compound V were equivalent.

result

Example 8: Pharmacokinetic study in rats

Experimental animals: SD rats (weight 200~300g, 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 ^TM 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 dosage of drug was 5, 10, and 20 mg/kg by gavage. Blood was collected from the cervical vein at 5min, 10min, 20min, 30min, 1h, 2h, 4h, 6h, 8h, 12h, and 24h 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:

The embodiments of this document may be modified and altered in many ways without departing from the scope of this document. It should therefore be understood that the embodiments of this document should not be limited to the exemplary embodiments described above, but should be subject to the limitations set forth in the claims and any equivalent forms thereof.

Claims (29)

A crystalline form A of a hydrochloride salt of a compound of formula II, (II) having diffraction peaks at 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° 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°, 27.5±0.2° in an X-ray diffraction pattern measured using Cu—Kα radiation. The crystalline form A as described in claim 1 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 hydrochloride salt of a compound of formula II, (II) having diffraction peaks at 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°, 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°, 25.7±0.2° in an X-ray diffraction pattern measured using Cu—Kα radiation. The crystal form B as described in claim 3 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 crystalline form I of the dihydrochloride salt of a compound of formula II, (II) It has diffraction peaks of 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. ±0.2°, 23.3±0.2°, 25.4±0.2°, 25.7±0.2° in its X-ray diffraction pattern measured by Cu-Kα radiation. The crystal form I as described in claim 5 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, (II) It has diffraction peaks at 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° 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°, 30.2±0.2° in its X-ray diffraction pattern measured by Cu-Kα radiation. The crystal form II as described in claim 7 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, (II) It has diffraction peaks at 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° in its X-ray diffraction pattern measured by Cu-Kα radiation. The crystal form III as described in claim 9 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 the crystal form A as described in claim 1 or 2, 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, 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. The method for preparing Form A as described in claim 11 is characterized in that the acid-base feed ratio of HCl and the free base of the compound of formula II is 1.1:1 to 1.5:1. The method for preparing Form A as described in claim 11 is characterized in that 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 the crystal form B 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 ethyl acetate, stirring to obtain a clear solution; (b) slowly adding ethyl acetate solution of HCl 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. The method for preparing Form B as described in claim 14 is characterized in that the acid-base feed ratio of HCl and the free base of the compound of formula II is 1.1:1 to 1.5:1. The method for preparing Form B as described in claim 14 is characterized in that in step (a), the free base of the compound of formula II is dissolved at room temperature, and in step (b), it is stirred at room temperature for 24 hours. A method for preparing the crystalline form I 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, stirring to obtain a clear solution; (b) slowly adding ethyl acetate solution of HCl 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. The method for preparing Form I as described in claim 17 is characterized in that the acid-base feed ratio of HCl and the free base of the compound of formula II is 2.5:1 to 4:1. The method for preparing Form I as described in claim 17 is characterized in that 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 7 or 8, 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. The method for preparing Form II as described in claim 20 is characterized in that the ketone solvent is one or more of acetone, butanone or methyl ethyl ketone. The method for preparing Form II as described in claim 20 is characterized in that the slurrying and stirring is performed at room temperature to 50°C for 24 hours. A method for preparing the crystal form III as described in claim 9 or 10, 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. The method for preparing Form III as described in claim 23 is characterized in that the slurrying and stirring is performed 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 any one of claims 1 to 10, and one or more pharmaceutically acceptable inert pharmaceutical excipients. The pharmaceutical composition as described in claim 25 is characterized in that the pharmaceutical composition is formulated for oral administration. A crystalline composition comprising more than 80% of the crystalline form I of the hydrochloride salt of the compound of formula II as described in claim 5 or 6. Use of a crystalline form of the hydrochloride of a compound of formula II as described in any one of claims 1 to 10, a pharmaceutical composition as described in claim 25 or 26, or a crystalline composition as described in claim 27 in the preparation of a medicament for preventing and treating thrombin-mediated and/or thrombin-related diseases. Use of a crystalline form of the hydrochloride of a compound of formula II as described in any one of claims 1 to 10, a pharmaceutical composition as described in claim 25 or 26, or a crystalline composition as described in claim 27 in the preparation of a medicament for treating venous and/or arterial thrombotic diseases.