Crystalline form of HS378 and its manufacturing method

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 the 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 has been demonstrated in a mouse-adapted model of the novel coronavirus, and in a Phase 1 clinical trial involving 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 HS378 crystalline form 1, crystalline form 2, crystalline form 4, and crystalline form 5. Crystal form 2 is a methyl tert-butyl ether solvate, and crystal form 5 is amorphous. The preparation of crystal form 1, crystal form 4, and crystal form 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 RRT=0.96 and RRT=1.04, and the crystallization process of crystal forms 1, 2, 4, and 5 cannot effectively remove them. Therefore, it is necessary to develop a new crystal form and a new process. Summary of the Invention

[0005] Therefore, in order to solve the above problems in the prior art, the present invention provides HS378 crystalline form A and crystalline form B. The crystallization method of crystalline form A and crystalline form B can exhibit a better impurity removal effect.

[0006] The powder X-ray diffraction pattern of crystalline form A of HS378 according to the present invention (hereinafter referred to as crystalline 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 crystalline form A of the present invention has 2θ values, d-spacing values, and relative intensity data as shown in Table 1 below. [Table 1]

[0008] Without being limited thereto, the powder X-ray diffraction pattern of the crystalline form A of the present invention is substantially as shown in FIG.

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

[0010] Without being limited thereto, crystalline form A of the present invention has a DSC pattern as shown in FIG.

[0011] The thermogravimetric analyzer (TGA) pattern of the crystalline form A of the present invention has a weight loss.

[0012] Without being limited thereto, the crystalline form A of the present invention has a TGA pattern as shown in FIG.

[0013] The combined DSC and TGA data indicate that crystalline form A of the present invention is an isobutyl acetate solvate.

[0014] Without being limited thereto, crystalline form A of the present invention has a blocky crystal habit, is nearly spherical, has small interparticle friction, and has good flowability, as shown in the micrograph of FIG.

[0015] Another object of the present invention is to provide a process for preparing said crystalline form A, comprising the steps of: (1) mixing the crude product of the compound of formula (I) with a mixed solvent of alcohol and isobutyl acetate; (2) Stirring for 1 to 24 hours to crystallize; (3) filtering to obtain crystalline form A.

[0016] Preferably, in the above step (1), 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 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 the stirring and crystallization in the step (2) is 5 to 25°C.

[0018] The powder X-ray diffraction pattern of crystalline form B of compound of formula (I) of the present invention (hereinafter referred to as crystalline form B) has 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°.

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

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

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

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

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

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

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

[0026] Without being limited thereto, crystalline form B of the present invention is shown in the micrograph of Figure 8. The crystal habit of crystalline form B is columnar, its surface is smooth, and its flowability is good.

[0027] Another object of the present invention is to provide a process for preparing said crystalline form B, comprising the steps of: (1) mixing a crude product of the compound of formula (I) with isopropyl alcohol; (2) Stirring for 1 to 24 hours to crystallize; (3) filtering to obtain crystalline form B.

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

[0029] Preferably, the temperature for the stirring and crystallization in the 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 the compound of formula (I), said pharmaceutical composition comprising a therapeutically effective amount of crystalline form A and / or crystalline form B of the compound of formula (I) and one or more pharma- ceutically acceptable carriers.

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

[0032] The two new crystal forms, crystal form A and crystal form B, produced by the production method of the present invention have good physical and chemical stability, regular crystal habit, good particle size uniformity, and good flowability. The production method of the present invention successfully solves the shortcomings of existing patented technologies such as the cumbersome crystallization process, fine particle size, and particle nonuniformity, and has excellent processability properties. In addition, the production method of this new crystal form can effectively remove impurities that are difficult to remove, and obtain samples with low impurities and high purity, which can also be used as intermediates 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, less polluting, and suitable for industrial production. [Brief description of the drawings]

[0033] [Figure 1] FIG. 1 is a powder X-ray diffraction pattern of crystalline form A obtained in Example 1. [Diagram 2] FIG. 2 is a DSC pattern of the crystalline form A obtained in Example 1. [Diagram 3] FIG. 3 is a TGA pattern of the crystalline form A obtained in Example 1. [Figure 4] FIG. 4 is a powder X-ray diffraction pattern of crystalline form B obtained in Example 5. [Diagram 5] FIG. 5 is a DSC pattern of crystalline form B obtained in Example 5. [Figure 6] FIG. 6 is a TGA pattern of crystalline form B obtained in Example 5. [Figure 7] FIG. 7 is a micrograph of crystalline form A obtained in Example 1. [Figure 8] FIG. 8 is a micrograph of crystalline form B obtained in Example 5. [Figure 9] FIG. 9 is a micrograph of crystalline form 1 obtained in Preparation Example. [Figure 10] FIG. 10 is a micrograph of crystalline form 2 obtained in Preparation Example. [Figure 11] FIG. 11 is a micrograph of crystalline form 4 obtained in Preparation Example. [Figure 12] FIG. 12 is a micrograph of crystalline form 5 obtained in Preparation Example. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0034] The following examples are provided to further illustrate the present invention and are not intended to restrict or limit 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 step 8, line 27, page 129, Example 13, disclosed in patent WO2021250648A1. The above sources include, but are not limited to:

[0036] The solvent used in the present invention is not particularly limited, and any commercially available solvent can be used.

[0037] Unless otherwise specified, the "stirring" in the method of the present invention can be a method commonly used in the technical field. For example, the stirring method can be magnetic stirring, mechanical stirring, etc., 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: X-ray diffraction apparatus model MiniFlex600Cu target, operation method: scanning speed 20° / min, scanning step width 0.02°.

[0039] The DSC test conditions according to the present invention were: DSC detector model: NETZSCH DSC 214 Polymer; 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 a model CX31 (10x10).

[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 in water, Mobile phase B: acetonitrile, Detection wavelength: 205 nm, Flow rate: 1.0 ml / min, Injection volume: 5 μl, Column temperature: 80° C. Elution gradient: [Table 3]

[0043] It should be emphasized that the meaning or intended protection range of the numerical values ​​or numerical end points included in the technical solution of the present invention is not limited to the numerical values ​​themselves. Those skilled in the art can understand that the numerical values ​​or numerical end points include acceptable error ranges widely accepted in the art, such as experimental error, measurement error, statistical error, random error, etc., and these error ranges are included in the scope of the present 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 thoroughly, heated to 40°C, slowly cooled to 25°C, stirred for 2 hours to crystallize, and filtered to obtain 0.87 g of crystals with a purity of 99.86%. The crystals obtained were analyzed by powder X-ray diffraction pattern (XRD), and were confirmed to be crystalline form A.

[0045] The powder X-ray diffraction pattern, DSC pattern and TGA pattern of this crystal form are shown in Figures 1 to 3, respectively, and a micrograph is shown in Figure 7. In the present invention, this is designated as 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 thoroughly, heated to 80°C, slowly cooled to 15°C, stirred for 1 hour to crystallize, and filtered to obtain 0.89 g of crystals with a purity of 99.78%. The crystals obtained were analyzed by powder X-ray diffraction pattern (XRD), and were confirmed to be crystalline 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 thoroughly, dissolved at 25°C, cooled slowly to 5°C, stirred for 24 hours to crystallize, and filtered to obtain 0.85 g of crystals with a purity of 99.91%. The crystals obtained were analyzed by powder X-ray diffraction pattern (XRD) and confirmed to be crystalline 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 thoroughly, dissolved at 25°C, cooled slowly to 5°C, stirred for 2 hours to crystallize, and filtered to obtain 0.80 g of crystals with 100% purity. The crystals obtained were analyzed by powder X-ray diffraction pattern (XRD) and confirmed to be crystalline form A.

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

[0050] The powder X-ray diffraction pattern, DSC pattern and TGA pattern of this crystal form are shown in Figures 4 to 6, respectively, and a micrograph is shown in Figure 8. In the present invention, this is designated as crystal form B.

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

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

[0053] [Production Example] Preparation of crystalline form 1: Crystalline form 1 was prepared according to step 2 on page 135 of patent WO2021250648A.

[0054] Preparation of crystalline form 2: According to Example 13, page 129, step 8, line 28 of Patent WO2021250648A, crystalline form 2 was prepared using ethyl acetate and methyl tert-butyl ether.

[0055] Preparation of crystalline form 4: Crystalline form 4 was prepared according to steps 1 to 2 on pages 151 to 152 of patent WO2021250648A.

[0056] Preparation of crystalline form 5: According to Example 96 on page 257 of patent WO2021250648A, crystalline form 5 was prepared by concentrating with dichloromethane.

[0057] [Comparative Example 1] The results of HPLC detection of samples and crude products of crystalline form A obtained in Example 1, crystalline form B obtained in Example 5, and crystalline forms 1, 2, 4, and 5 obtained in Preparation Examples of the present invention are shown in Table 1. [Table 4]

[0058] From Table 1, it can be seen that the preparation methods of crystalline forms A and B are superior to crystalline forms 1, 2, 4, and 5 in terms of the effect of removing difficult-to-remove impurities.

[0059] [Comparative Example 2] Samples of crystalline form A obtained in Example 1, crystalline form B obtained in Example 5, and crystalline forms 1, 2, 4, and 5 obtained in Preparation Examples of the present invention were left at 60°C for 5 days and 10 days, respectively, and then detected by HPLC and XRD, respectively. The results are shown in Table 2. [Table 5]

[0060] From Table 2, when left at 60°C for 10 days, the crystal forms A, B and C were 1 The crystal form and liquid phase HPLC data show that the compound is more stable than the crystal form 2, the crystal form 4 and the crystal form 5.

[0061] [Comparative Example 3] Micrographs of samples of the crystal form A obtained in Example 1 of the present invention, the crystal form B obtained in Example 5, and the crystal forms 1, 2, 4, and 5 obtained in the Production Examples are shown in FIGS. 7 to 12, respectively.

[0062] The crystals of crystal form A are uniform in size, almost circular, have a small specific surface area, small intercrystalline friction, and good particle fluidity. The crystals of crystal form B are uniform in size, uniform in size, lumpy, have a small aspect ratio, smooth crystal surface, and good particle fluidity. The crystal form 1 is needle-shaped, has a large aspect ratio, and has low fluidity. The crystal form 2 is cone-shaped, has a large aspect ratio, and has low fluidity. The crystal form 4 has a uniform particle size, but is small in particle size, is prone to aggregation, is difficult to filter, has a large amount of mother liquor residue, and is difficult to calcinate. The crystal form 5 has non-uniform particles, poor fluidity, and is relatively brittle, making it unsuitable for subsequent processing. Therefore, the crystal forms A and B have better crystal habits, more uniform particles, and are easier to store, transport, and further process.

Claims

1. Crystalline form A of the compound of formula (I), 【Chemistry 1】 The powder X-ray diffraction pattern 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. The crystalline form A of claim 1, characterized in that the powder X-ray diffraction pattern is substantially as shown in Figure 1.

3. A process for preparing crystalline form A according to any one of claims 1 to 2, comprising the steps of: (1) mixing the crude product of the compound of formula (I) with a mixed solvent of alcohol and isobutyl acetate; (2) stirring for 1 to 24 hours to cause crystallization; (3) filtering to obtain crystalline form A.

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

5. The method according to claim 3, wherein the stirring crystallization temperature in the step (2) is 5 to 25°C.

6. Crystalline form B of the compound of formula (I), 【Chemistry 2】 The powder X-ray diffraction pattern 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°.

7. 7. The crystalline form B of claim 6, characterized in that the powder X-ray diffraction pattern is substantially as shown in FIG.

8. A process for preparing crystalline form B according to any one of claims 6 to 7, comprising the steps of: (1) mixing a crude product of the compound of formula (I) with isopropyl alcohol; (2) stirring for 1 to 24 hours to cause crystallization; (3) filtering to obtain crystalline form B.

9. The method according to claim 8, characterized in that in step (1), the mass / volume ratio of the crude product of the compound of formula (I) to isopropyl alcohol is 1:5 to 1:30 in g / ml; and the temperature of the mixing is 25 to 80° C.

10. The method according to claim 8, wherein the stirring crystallization temperature in the step (2) is 5 to 25°C.

11. A pharmaceutical composition comprising crystalline form A of the compound of formula (I) according to any one of claims 1-2 and / or crystalline form B of the compound of formula (I) according to any one of claims 6-7.

12. Use of crystalline form A of the compound of formula (I) as defined in any one of claims 1-2, crystalline form B of the compound of formula (I) as defined in any one of claims 6-7, or the pharmaceutical composition of claim 11 in the manufacture of a medicament for treating a new coronavirus.

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