Crystal forms of HS378 and their manufacturing methods

Abstract

The present invention relates to crystalline form A and crystalline form B of compound of formula (I) (HS378) and their preparation methods. The crystalline form A and crystalline form B have good physical and chemical stability, regular crystal habit, good particle size uniformity, and good flowability. The preparation method of the crystalline form A and crystalline form B is simple, has a high impurity removal effect, is easy to filter, has a high yield, and can obtain samples with low impurities and high purity, so that the purification problem of difficult-to-remove impurities can be effectively solved. [Formula 1] JPEG2025516023000010.jpg48169

Description

Technical Field

[0001] The present invention relates to the field of chemical pharmaceuticals. More specifically, the present invention relates to crystalline form A and crystalline form B of (1R,2S,5S)-N-{(1S)-1-cyano-2-[(3S)-2-oxopyrrolidin-3-yl]-ethyl}-6,6-dimethyl-3-[3-methyl-N-(trifluoroacetyl)-L-valyl]-3-azabicyclo[3.1.0]hexane-2-carboxamide (HS378), and to methods for producing said crystalline forms and their pharmaceutical uses.

Background Art

[0002] In 2019, the novel coronavirus disease occurred globally and became a worldwide pandemic. The novel coronavirus is a highly infectious ribonucleic acid coronavirus that can cause life-threatening viral pneumonia in the worst cases. Similar to vaccines, antiviral therapy is an important part of medical countermeasures against the novel coronavirus disease. HS378 is an orally bioavailable main protease inhibitor of SARS-CoV-2 with in vitro antiviral activity against coronaviruses, excellent off-target selectivity, and in vivo safety. Oral activity was demonstrated in a mouse-adapted model of the novel coronavirus, and in a Phase 1 clinical trial with healthy human participants, oral plasma concentrations exceeding the in vitro antiviral cell effect were achieved.

[0003] The chemical name of the compound of formula (I) is (1R,2S,5S)-N-{(1S)-1-cyano-2-[(3S)-2-oxopyrrolidin-3-yl]-ethyl}-6,6-dimethyl-3-[3-methyl-N-(trifluoroacetyl)-L-valyl]-3-azabicyclo[3.1.0]hexane-2-carboxamide (named HS378 in the present application), and its structural formula is as follows.

Chem.

[0004] Currently, Pfizer's patent WO2021250648A discloses crystal forms 1, 2, 4, and 5 of HS378. Crystal form 2 is a methyl tert-butyl ether solvate, and crystal form 5 is amorphous. The production of crystal forms 1, 4, and 5 all use crystal form 2 as a substrate, and there are two impurities that are difficult to remove during the process. These two impurities have relative retention times of RRT = 0.96 and RRT = 1.04, and cannot be effectively removed by the crystallization processes of crystal forms 1, 2, 4, and 5. Therefore, it is necessary to develop new crystal forms and new processes. [Overview of the Initiative]

[0005] Therefore, in order to solve the problems that exist in the prior art described above, the present invention provides crystal forms A and B of HS378. The method for crystallizing crystal forms A and B can exhibit a superior impurity removal effect.

[0006] The powder X-ray diffraction pattern of crystal form A of HS378 according to the present invention (hereinafter referred to as crystal form A) has characteristic peaks at the following diffraction angles 2θ: 6.2±0.2°, 6.9±0.2°, 7.9±0.2°, 10.5±0.2°, 11.5±0.2°, 11.9±0.2°, 12.5±0.2°, 13.9±0.2°, 14.3±0.2°, 15.8±0.2°, 18.7±0.2°, 19.4±0.2°, 20.7±0.2°, and 23.3±0.2°.

[0007] Furthermore, the powder X-ray diffraction pattern of crystal form A of the present invention has the 2θ value, interplanar spacing d value, and relative intensity data shown in Table 1 below. [Table 1]

[0008] Without limitation, the powder X-ray diffraction pattern of crystal form A of the present invention is substantially as shown in Figure 1.

[0009] The differential scanning calorimetry (DSC) pattern of crystal form A according to the present invention has an endothermic peak with a peak value of 105.2°C.

[0010] While not limited to the present invention, crystal form A has a DSC pattern as shown in Figure 2.

[0011] The thermogravimetric analyzer (TGA) pattern of crystal form A according to the present invention exhibits weight loss.

[0012] While not limited to the present invention, crystal form A has a TGA pattern as shown in Figure 3.

[0013] Combining DSC and TGA data reveals that crystal form A of the present invention is isobutyl acetate solvate.

[0014] While not limited to these, crystal form A of the present invention has a massive crystal habit, is nearly spherical, has low interparticle friction, and has good fluidity, as shown in the micrograph in Figure 7.

[0015] Another object of the present invention is to provide a method for producing the crystal form A, the method being (1) A step of mixing the crude product of the compound of formula (I) with a mixed solvent of alcohol and isobutyl acetate, (2) A step of stirring for 1 to 24 hours to crystallize, (3) The process includes the step of filtering to obtain crystalline form A.

[0016] Preferably, in step (1) above, the mass / volume ratio of the crude product of the compound of formula (I) to the mixed solvent is 1:5 to 1:40 in units of g / ml; the volume ratio (ml / ml) of the alcohol to isobutyl acetate is 1:7 to 1:100; the alcohol is a C1 to C2 alcohol; and the mixing temperature is 25 to 80°C.

[0017] Preferably, the temperature for stirring and crystallization in step (2) is 5 to 25°C.

[0018] The powder X-ray diffraction pattern of crystalline form B of the compound of formula (I) of the present invention (hereinafter referred to as crystalline form B) has peaks characteristic of the following diffraction angles 2θ: 6.7 ± 0.2°, 7.4 ± 0.2°, 9.3 ± 0.2°, 10.0 ± 0.2°, 10.9 ± 0.2°, 11.2 ± 0.2°, 15.8 ± 0.2°, 16.4 ± 0.2°, 17.4 ± 0.2°, 18.4 ± 0.2°, 19.4 ± 0.2°, 20.3 ± 0.2°, and 20.8 ± 0.2°.

[0019] Furthermore, the powder X-ray diffraction pattern of crystalline form B of the present invention has the 2θ values, interplanar spacing d values, and relative intensity data shown in Table 2 below.

Table 2

[0020] Without limitation, the powder X-ray diffraction pattern of crystalline form B of the present invention is substantially as shown in FIG. 4.

[0021] The differential scanning calorimetry (DSC) pattern of crystalline form B according to the present invention has an endothermic peak with a peak value of 94.4 °C.

[0022] Without limitation, crystalline form B of the present invention has a DSC pattern as shown in FIG. 5.

[0023] The thermogravimetric analyzer (TGA) pattern of crystalline form B according to the present invention shows a stepwise weight loss.

[0024] Without limitation, crystalline form B of the present invention has a TGA pattern as shown in FIG. 6.

[0025] Combining the DSC and TGA data indicates that crystalline form B of the present invention is an isopropyl alcohol solvate.

[0026] Without limitation, crystalline form B of the present invention is shown in the micrograph of FIG. 8. The crystal habit of crystalline form B is columnar, and its surface is smooth and has good fluidity.

[0027] Another object of the present invention is to provide a method for producing the crystal form B, the method being (1) A step of mixing the crude product of the compound of formula (I) with isopropyl alcohol, (2) A step of stirring for 1 to 24 hours to crystallize, (3) The process includes the step of filtering to obtain crystalline form B.

[0028] Preferably, in step (1) above, the mass / volume ratio of the crude product of the compound of formula (I) to isopropyl alcohol is 1:5 to 1:30 in units of g / ml; and the mixing temperature is 25 to 80°C.

[0029] Preferably, the temperature for stirring and crystallization in step (2) is 5 to 25°C.

[0030] The present invention also relates to a pharmaceutical composition comprising crystalline form A and / or crystalline form B of a compound of formula (I). The pharmaceutical composition comprises a therapeutically effective amount of crystalline form A and / or crystalline form B of a compound of formula (I) and one or more pharmaceutically acceptable carriers.

[0031] The present invention also relates to the use of crystalline form A, crystalline form B, or pharmaceutical compositions thereof of the compound of formula (I) in the manufacture of a drug for treating the novel coronavirus.

[0032] The two new crystal forms, crystal form A and crystal form B, produced by the manufacturing method of the present invention, possess good physical and chemical stability, regular crystal habit, good particle size uniformity, and good fluidity. The manufacturing method of the present invention successfully solves the shortcomings of existing patented technologies, such as the complexity of the crystallization process, particle refinement, and particle non-uniformity, and has excellent processability. Furthermore, this manufacturing method for new crystal forms can effectively remove impurities that are difficult to remove, yielding high-purity samples with fewer impurities, and can also be used as an intermediate to simplify the crystallization process of the final product. The crystallization process of the present invention has the advantages of being simple, easy to operate, low-pollution, and suitable for industrial production. [Brief explanation of the drawing]

[0033] [Figure 1] Figure 1 shows the powder X-ray diffraction pattern of crystal form A obtained in Example 1. [Figure 2] Figure 2 shows the DSC pattern of crystal form A obtained in Example 1. [Figure 3] Figure 3 shows the TGA pattern of crystal form A obtained in Example 1. [Figure 4] Figure 4 shows the powder X-ray diffraction pattern of crystal form B obtained in Example 5. [Figure 5] Figure 5 shows the DSC pattern of crystal form B obtained in Example 5. [Figure 6] Figure 6 shows the TGA pattern of crystal form B obtained in Example 5. [Figure 7] Figure 7 is a micrograph of crystal form A obtained in Example 1. [Figure 8] Figure 8 is a micrograph of crystal form B obtained in Example 5. [Figure 9] Figure 9 is a micrograph of crystal form 1 obtained in the manufacturing example. [Figure 10] Figure 10 is a micrograph of crystal form 2 obtained in the manufacturing example. [Figure 11] Figure 11 is a micrograph of crystal form 4 obtained in the manufacturing example. [Figure 12] Figure 12 is a micrograph of crystal form 5 obtained in the manufacturing example. [Modes for carrying out the invention]

[0034] The following examples are provided to further illustrate the present invention and are not intended to limit or restrict the scope of the invention.

[0035] The crude product of the compound of formula (I) can be used as a raw material for producing crystalline form A or crystalline form B in the method of the present invention. The crude product of the compound of formula (I) can be prepared by vacuum concentrating the eluent according to Example 13, page 129, step 8, line 27, disclosed in patent WO2021250648A1. The above sources include, but are not limited to, the above sources.

[0036] The present invention does not have any particular restrictions on the solvent used, and commercially available solvents can be used.

[0037] In the method of the present invention, unless otherwise specified, "stirring" can be any method commonly used in the art. For example, stirring methods include magnetic stirring and mechanical stirring, and the stirring speed is 150 to 300 rpm / min.

[0038] The powder X-ray diffraction apparatus and test conditions according to the present invention were as follows: X-ray diffraction apparatus model MiniFlex600Cu target, operating method: scanning speed 20° / min, scanning step width 0.02°.

[0039] The DSC test conditions according to the present invention were as follows: DSC detector model: NETZSCH DSC 214 Polyma, operation method: heating rate: 10°C / min, temperature range: 30~250°C.

[0040] The TGA test conditions according to the present invention were: TGA detector model: METTLER TOLEDO TGA2, operation method: heating rate: 10°C / min, temperature range: 35~300°C.

[0041] The OLYMPUS microscope according to the present invention was model CX31 (10 × 10).

[0042] The liquid-phase test conditions according to the present invention were as follows: Chromatography column Agilent Zorbax SB-18, 150 × 4.6 mm, 5 μm, mobile phase A: 0.1% phosphoric acid aqueous solution, mobile phase B: acetonitrile, detection wavelength: 205 nm, flow rate: 1.0 ml / min, injection volume: 5 μl, column temperature: 80 °C. Eluent gradient: [Table 3]

[0043] It should be emphasized that the meaning or intended scope of protection of numerical values ​​or numerical endpoints included in the proposed technical proposal of this invention is not limited to the numerical values ​​themselves. Those skilled in the art will understand that these numerical values ​​or numerical endpoints include acceptable error ranges widely accepted in the art, such as experimental errors, measurement errors, statistical errors, and random errors, and these error ranges are included within the scope of this invention.

[0044] [Example 1] 1 g of the crude product was added to 0.3 ml of methanol and 10 ml of isobutyl acetate, mixed well, heated to 40°C, slowly cooled to 25°C, stirred for 2 hours to crystallize, filtered, and 0.87 g of crystals with a purity of 99.86% was obtained. The obtained crystals were measured by powder X-ray diffraction (XRD) and confirmed to be crystal form A.

[0045] The powder X-ray diffraction pattern, DSC pattern, and TGA pattern of this crystal form are shown in Figures 1-3, and a micrograph is shown in Figure 7. In this invention, this is named crystal form A.

[0046] [Example 2] 1 g of the crude product was added to 0.1 ml of ethanol and 10 ml of isobutyl acetate, mixed well, heated to 80°C, slowly cooled to 15°C, stirred for 1 hour to crystallize, filtered, and 0.89 g of crystals with a purity of 99.78% was obtained. The crystals obtained were measured by powder X-ray diffraction (XRD) and confirmed to be crystal form A.

[0047] [Example 3] 1 g of the crude product was added to 0.2 ml of methanol and 4.8 ml of isobutyl acetate, mixed well, dissolved at 25°C, slowly cooled to 5°C, stirred for 24 hours to crystallize, filtered, and 0.85 g of crystals with a purity of 99.91% was obtained. The crystals obtained were measured by powder X-ray diffraction (XRD) and confirmed to be crystal form A.

[0048] [Example 4] 1 g of the crude product was added to 5 ml of methanol and 35 ml of isobutyl acetate, mixed well, dissolved at 25°C, slowly cooled to 5°C, stirred for 2 hours to crystallize, filtered, and 0.80 g of 100% pure crystals was obtained. The obtained crystals were measured by powder X-ray diffraction (XRD) and confirmed to be crystal form A.

[0049] [Example 5] 1 g of the crude product was added to 5 ml of isopropyl alcohol, mixed well, heated to 80°C, slowly cooled to 25°C, stirred for 3 hours to crystallize, filtered, and 0.87 g of crystals with a purity of 99.57% was obtained. The obtained crystals were measured by powder X-ray diffraction (XRD) and confirmed to be crystal form B.

[0050] The powder X-ray diffraction patterns, DSC patterns, and TGA patterns of this crystalline form are shown in Figures 4-6, and a micrograph is shown in Figure 8. In this invention, this is named crystalline form B.

[0051] [Example 6] 1 g of the crude product was added to 20 ml of isopropyl alcohol, mixed well, heated to 35°C, slowly cooled to 15°C, stirred for 1 hour to crystallize, filtered, and 0.84 g of crystals with a purity of 99.73% was obtained. The obtained crystals were measured by powder X-ray diffraction (XRD) and confirmed to be crystal form B.

[0052] [Example 7] 1 g of the crude product was added to 30 ml of isopropyl alcohol, mixed well, dissolved at 25°C, slowly cooled to 5°C, stirred for 24 hours to crystallize, filtered, and 0.82 g of crystals with a purity of 99.91% was obtained. The obtained crystals were measured by powder X-ray diffraction (XRD) and confirmed to be crystal form B.

[0053] [Manufacturing example] Preparation of crystal form 1: Crystal form 1 was prepared according to step 2 on page 135 of the patent disclosed in WO2021250648A.

[0054] Preparation of Crystal Form 2: Crystal Form 2 was prepared using ethyl acetate and methyl tert-butyl ether according to Example 13, step 8, line 28 on page 129, disclosed in patent WO2021250648A.

[0055] Preparation of crystal form 4: Crystal form 4 was prepared according to steps 1 and 2 on pages 151-152 disclosed in patent WO2021250648A.

[0056] Preparation of crystalline form 5: Crystalline form 5 was prepared by concentration using dichloromethane according to Example 96 on page 257 of the patent disclosed in WO2021250648A.

[0057] [Comparative Example 1] Table 1 shows the results of HPLC detection of the crystal form A obtained in Example 1 of the present invention, the crystal form B obtained in Example 5, and the samples and crude products of crystal forms 1, 2, 4, and 5 obtained in the production example. [Table 4]

[0058] Table 1 shows that the manufacturing methods for crystal forms A and B are superior to those for crystal forms 1, 2, 4, and 5 in terms of their effectiveness in removing impurities that are difficult to remove.

[0059] [Comparative Example 2] Table 2 shows the results of detecting samples of crystal form A obtained in Example 1, crystal form B obtained in Example 5, and crystal forms 1, 2, 4, and 5 obtained in the production example by HPLC and XRD, respectively, after being left at 60°C for 5 days and 10 days. [Table 5]

[0060] Table 2 shows that when left at 60°C for 10 days, crystal form A, crystal form B, and crystal form 1 This indicates that the crystalline form and liquid-phase HPLC data are more stable than crystalline form 2, crystalline form 4, and crystalline form 5.

[0061] [Comparative Example 3] Microscopic images of crystal form A obtained in Example 1 of the present invention, crystal form B obtained in Example 5, and crystal forms 1, 2, 4, and 5 obtained in the manufacturing example are shown in Figures 7 to 12, respectively.

[0062] Crystals of crystal form A have uniform, consistent particle size, are nearly circular, have a small specific surface area, low intercrystalline friction, and good particle fluidity. Crystal form B has uniform, consistent particle size, is massive, has a small aspect ratio, a smooth crystal surface, and good particle fluidity. Crystal form 1 is needle-shaped, has a large aspect ratio, and low fluidity. Crystal form 2 is conical, has a large aspect ratio, and low fluidity. Crystal form 4 has uniform particle size, but the particle diameter is small, it is prone to aggregation, difficult to filter, has a large amount of mother liquor residue, and is difficult to calcin. Crystal form 5 has non-uniform particles, poor fluidity, and is relatively brittle, making it unsuitable for subsequent processing. Therefore, crystal forms A and B have superior crystal habit, more uniform particles, and are easier to store, transport, and process.

Claims

1. A crystalline compound of formula (I) having crystal form A, 【Chemistry 1】 The powder X-ray diffraction pattern of the crystalline compound of formula (I) having crystal form A is characterized by having characteristic peaks at the following diffraction angles 2θ: 6.2±0.2°, 6.9±0.2°, 7.9±0.2°, 10.5±0.2°, 11.5±0.2°, 11.9±0.2°, 12.5±0.2°, 13.9±0.2°, 14.3±0.2°, 15.8±0.2°, 18.7±0.2°, 19.4±0.2°, 20.7±0.2°, and 23.3±0.2°.

2. A method for producing a crystalline compound of formula (I) having the crystalline form A described in claim 1, (1) A step of mixing the crude product of the compound of formula (I) with a mixed solvent of alcohol and isobutyl acetate, (2) A step of stirring for 1 to 24 hours to crystallize, (3) A method comprising the step of filtering to obtain a crystalline compound of formula (I) having crystalline form A.

3. The method according to claim 2, characterized in that, in step (1) above, the mass / volume ratio of the crude product of the compound of formula (I) to the mixed solvent is 1:5 to 1:40 in units of g / ml; the volume ratio (ml / ml) of the alcohol to isobutyl acetate is 1:7 to 1:100; the alcohol is a C1 to C2 alcohol; and the mixing temperature is 25 to 80°C.

4. The method according to claim 2, wherein the temperature of the stirring crystallization in step (2) is 5 to 25°C.

5. A crystalline compound of formula (I) having crystalline form B, 【Chemistry 2】 The powder X-ray diffraction pattern of the crystalline compound of formula (I) having crystal form B is characterized by having characteristic peaks at the following diffraction angles 2θ: 6.7±0.2°, 7.4±0.2°, 9.3±0.2°, 10.0±0.2°, 10.9±0.2°, 11.2±0.2°, 15.8±0.2°, 16.4±0.2°, 17.4±0.2°, 18.4±0.2°, 19.4±0.2°, 20.3±0.2°, and 20.8±0.2°.

6. A method for producing a crystalline compound of formula (I) having the crystalline form B described in claim 5, (1) A step of mixing the crude product of the compound of formula (I) with isopropyl alcohol, (2) A step of stirring for 1 to 24 hours to crystallize, (3) A method comprising the step of filtering to obtain a crystalline compound of formula (I) having crystalline form B.

7. The method according to claim 6, characterized in that, in step (1) above, the mass / volume ratio of the crude product of the compound of formula (I) to isopropyl alcohol is 1:5 to 1:30 in units of g / ml; and the temperature of the mixing is 25 to 80°C.

8. The method according to claim 6, characterized in that the temperature of the stirring crystallization in step (2) is 5 to 25°C.

9. A pharmaceutical composition comprising a crystalline compound of formula (I) having crystal form A as described in claim 1 and / or a crystalline compound of formula (I) having crystal form B as described in claim 5.

10. Use of a pharmaceutical composition comprising a crystalline compound of formula (I) having crystal form A as described in claim 1, a crystalline compound of formula (I) having crystal form B as described in claim 5, or a crystalline compound of formula (I) having crystal form A as described in claim 1 and / or a crystalline compound of formula (I) having crystal form B as described in claim 5, in the manufacture of a drug for treating the novel coronavirus.

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