WO2018177276A1 - A crystal form of tubulin inhibitor - Google Patents

Abstract

The present invention provides an A crystal form of (3Z,6Z)-3-[((E)-3-(5-tert-butyl)-1H-imidazol-4-yl)methylene]-6-((E)-3-(3-fluorophenyl)-2-propenylidene)piperazine-2,5-dione (VDA-1). The A crystal form has a stable morphology, a definite melting point, and good chemical stability and is high-temperature resistant and suitable for pharmaceutical uses. The A crystal form can be used to treat a hyperproliferative disease.

Description

A crystal form of a tubulin inhibitor Technical field

The invention belongs to the technical field of medicine, in particular to a new crystal form of a tubulin inhibitor.

Background technique

As the main means of cancer treatment, anti-tumor drugs have made considerable contributions to prolonging the survival time of patients and improving their quality of life. Among them, drugs acting on microtubules (microtubule inhibitors) have a very important position in oncology drugs. However, the current clinical drugs are affected by the following unfavorable problems: poor water solubility, unfavorable drug administration, and easy to cause allergic reactions, serious toxic side effects, and acquired drug resistance, resulting in reduced efficacy, complicated chemical structure, and difficulty in synthesis. As a result, the sources are scarce, limiting their further use.

Chinese patent application CN106565686A discloses a novel small molecule tubulin inhibitor of simple structure. Since the drug molecules of different crystal forms have significant differences in solubility, melting point, dissolution rate, bioavailability, etc., preparation of a drug crystal form having a certain melting point, good chemical stability, high temperature resistance, etc. will be beneficial to the drug. Preparation and application of the formulation.

Summary of the invention

It is an object of the present invention to provide a novel crystalline form of a tubulin inhibitor, namely (3Z,6Z)-3-[((E)-3-(5-tert-butyl)-1H-imidazolyl-4 Form A of methylene]-6-((E)-3-(3-fluorophenyl)-2-propenylene)piperazine-2,5-dione (abbreviated as VDA-1) , VDA-1 has the structure shown in formula (I):

The characteristics of Form A of the VDA-1 are characterized by melting point, X-ray powder diffraction (XRD), differential scanning calorimetry (DSC), thermogravimetric analysis (TG), and infrared spectroscopy (IR). The crystalline form possesses the properties required to prepare a pharmaceutical formulation.

Specifically, when X-ray powder diffraction is performed with a Cu radiation source, the A crystal form includes characteristic diffraction peaks at 6.6 ± 0.2, 9.7 ± 0.2, and 16.0 ± 0.2 (°) at 2θ, and the relative of these peaks The intensity (I/I ₀ ) is greater than or equal to 30%. Further, the crystal may further comprise, in X-ray powder diffraction, at 13.4±0.2, 16.3±0.2, 16.6±0.2, 17.1±0.2, 18.1±0.2, 25.5±0.2, 25.9±0.2, 26.5 at 2θ. Characteristic diffraction peaks of ±0.2 (°), the relative intensities of these peaks are all greater than or equal to 16% (see Figure 1). Where "±0.2°" is the allowable measurement error range.

The Form A of the present invention can be characterized by an X-ray powder diffraction pattern. It is characterized in that its X-ray powder diffraction pattern has the characteristic peak represented by the above 2θ, as shown in Table 1, and its relative intensity is close to the following values.

Table 1

The term "proximity" as used herein refers to the uncertainty of the relative intensity measurement. Those skilled in the art understand that the uncertainty of relative intensity is highly dependent on the measurement conditions. The relative intensity value may be a value within a certain range, preferably a range selected from the range of ±25% of the value shown in Table 1, and more preferably a range of ±10% of the value shown in Table 1.

The above A crystal form has the X-ray powder diffraction pattern shown in Fig. 1.

The present invention characterizes the Form A of VDA-1 using differential scanning calorimetry (DSC) techniques (see Figure 2), wherein the maximum endotherm with differential scanning calorimetry is 146.5 °C. The endothermic process appears as an endothermic peak on the DSC spectrum;

The invention uses thermogravimetric analysis technology to characterize the crystal form of VDA-1 (see Fig. 3), wherein the thermogravimetric diagram (TG) shows a weight loss of 1.0% at 127.1 °C, indicating that the adsorbed water is lost at this temperature. . The weight loss at 235.9 ° C exceeded 16%, indicating that the compound degraded as the temperature increased.

An infrared spectrum of Form A of the compound of the present invention is shown in Figure 4, wherein in 3228, 3064, 2966, 2911, 2732, 2589, 1707, 1686, 1664, 1639, 1597, 1499, 1411, 1375, 1350, 1272 1,149, 1049, 1022, 952, 937, 810, 773, 677, 443 (cm ^-1 ) have strong absorption peaks.

Another object of the present invention is to provide a process for the preparation of VDA-1 Form A.

It is still another object of the present invention to provide a pharmaceutical composition comprising the VDA-1 Form A.

A final object of the present invention is to provide the use of VDA-1 Form A for the preparation of a medicament for the treatment of hyperproliferative diseases.

According to one aspect of the present invention, the crude product of VDA-1 is first prepared, and then the crude material is crystallized by the recrystallization method of the present invention to obtain a new crystal form, and the melting point measurement, X-ray is performed on the crystal. The detection and analysis by powder diffraction, DSC, TG, IR, etc., confirmed that the obtained crystal is a novel crystal, which is referred to as a crystal form of VDA-1 in the present invention.

According to another aspect of the invention, a process for preparing the VDA-1A crystalline form comprises: adding a crude VDA-1 to ethyl acetate, dimethyl sulfoxide, N,N-dimethylformamide and tetrahydrofuran In a mixed solvent or added to a mixed solvent of C3-C4 alkyl ketone and tetrahydrofuran, heated to reflux until dissolved; after the solution is clarified, the temperature is lowered to precipitate a solid, and the solid is collected by filtration, and the collected solid is air-dried to obtain A crystal. type. The ketone is selected from the group consisting of acetone, methyl ethyl ketone and n-butyl ketone, etc., a preferred solvent mixture of ethyl acetate and tetrahydrofuran, a mixed solvent of acetone and tetrahydrofuran; ethyl acetate, dimethyl sulfoxide, N, N - the volume ratio (V / V) of dimethylformamide, ketone and tetrahydrofuran is 1:1 to 1:5; the ratio of the crude product to the solvent is 1 (g) by weight to volume ratio (W/V). : 5 to 30 (ml), preferably 1:10 (g/ml). The solution is preferably heated to 50 to 80 ° C, more preferably ethyl acetate, a mixed solvent of a C3-C4 alkyl ketone and a tetrahydrofuran, and heated to 60 ° C; according to this embodiment, the precipitation is carried out for 2 to 8 hours, more preferably 4 hours. The precipitation temperature is 0 to 40 ° C, preferably 5 to 15 ° C. After the solid solution is completely filtered, the drying temperature is 30 to 60 ° C, preferably 45 ° C.

According to still another aspect of the present invention, there is provided a pharmaceutical composition comprising Form A of Form V of the present invention, the pharmaceutical composition comprising the novel crystalline form compound and optionally a pharmaceutically acceptable carrier and/or form Agent.

The pharmaceutical composition can be further formulated into a form for administration according to a conventional formulation method, including an oral or parenteral administration form. In the form available for administration, a therapeutically effective amount of Form A of VDA-1 should be included. By "therapeutically effective amount" is meant that at this dose, the compounds of the invention are capable of ameliorating or alleviating the symptoms of the disease, or are capable of inhibiting or blocking the progression of the disease.

Those skilled in the art can readily select and determine the amount of various carriers and/or excipients based on experience and consideration of standard methods and references in the art.

According to still another aspect of the present invention, the A crystal form of VDA-1 can be used alone for the preparation of a medicament for treating a proliferative disease, or can be prepared in combination with other therapeutic agents to synergistically.

Advantageous Effects: The present invention produces (3Z,6Z)-3-[((E)-3-(5-tert-butyl)-1H-imidazolyl-4-yl)methylene]-6-(( E)-3-(3-fluorophenyl)-2-propenylene)piperazin-2,5-dione (VDA-1) Form A, which has a stable morphology and a defined melting point, chemically stable Good in properties and high in temperature resistance, the A crystal form of VDA-1 has the properties required for preparation of the preparation, and is convenient to store, simple in production operation, and easier to control in quality.

The crystalline form A of VDA-1 of the present invention is for treating a hyperproliferative disease. Preferably, the hyperproliferative disease is cancer, including but not limited to non-small cell lung cancer, colorectal cancer, refractory non-small cell lung cancer. , pancreatic cancer, ovarian cancer, breast cancer, glioma, brain cancer or neck cancer.

DRAWINGS

Figure 1 is an X-ray diffraction pattern of Form A of VDA-1;

Figure 2 is a DSC pattern of Form A of VDA-1;

Figure 3 is a TG map of Form A of VDA-1;

Figure 4 is an IR spectrum of Form A of VDA-1;

Figure 5 is an HPLC chromatogram of Form A of VDA-1.

detailed description

The invention is further illustrated by the following examples, without any limitation of the scope of the invention.

All materials and reagents are commercially available.

Preparation of crude VDA-1

Example 1

The preparation method of the crude VDA-1 includes the steps (1) to (3):

(1)

The synthesis of Compound A was prepared by referring to the method of Example 2 of Chinese Patent No. CN1684955.

(2)

To a 1 L dry single-mouth flask was placed 0.10 mol of 3-fluorobenzaldehyde, 0.11 mmol of formylmethylenetriphenylphosphorane and toluene (200 ml). The reaction was refluxed for 16 hours and then concentrated. The crude product was purified by EtOAc EtOAc EtOAc:EtOAc:

(3)

A 250 ml dry single-mouth flask was charged with DMF (100 ml), Intermediate A 5 mmol, 3-fluorobenzaldehyde 10 mmol, and dec. The reaction system was stirred at 25 to 30 ° C for 2 hours, cooled to room temperature, and then poured into ice water. The organic layer was washed three times with brine, dried over anhydrous sodium sulfate The crude product was washed with methylene chloride / methanol (10/1).

Preparation of VDA-1 crystal

Example 2.1

200 g of crude VDA-1 was added to the reaction flask, and 2400 ml of a mixed solvent of ethyl acetate and tetrahydrofuran (V/V = 1:5) was added, and the mixture was heated to reflux at 60 ° C under stirring. After dissolving, stirring for 10 min, and then cooling to 5-15 ° C, until the solid precipitated and stirred for 4 h, suction filtration, and the filter cake was rinsed with acetone. The filter cake was blast dried at 45 ° C and dried with phosphorus pentoxide. 166 g of a white solid were obtained, the yield was 83.0%, and the moisture was measured by Karl Fischer.

Example 2.2

200 g of crude VDA-1 was added to the reaction flask, and 1600 ml of a mixed solvent of acetone and water (V/V = 1:3) was added, and the mixture was heated to reflux at 60 ° C with stirring. After dissolving, stirring for 10 min, and then cooling to 5-15 ° C, until the solid precipitated and stirred for 4 h, suction filtration, and the filter cake was rinsed with acetone. The filter cake was blast dried at 45 ° C and dried with phosphorus pentoxide. The white solid was obtained in 148 g, yield 74.0%. The water was measured by Karl Fischer to be 0.3%.

Example 2.3 to 2.8

The method according to Example 2.1, wherein the solvent, the heating temperature, the precipitation temperature and time, and the drying temperature are as shown in Table 2, respectively.

Table 2

As a result, it was found that none of the examples 2.2 to 2.8 obtained a suitable crystal form.

Examples 2.9 to 2.11

In addition to the above processes, the effects of different solvents such as dimethyl sulfoxide, absolute ethanol, and methanol solvent on the crystal form of the product were also examined.

The preparation process of Examples 2.9 to 2.11 is as follows:

table 3

The water and solvent contents of the crystal form of the compound VDA-1 obtained in Example 2.1 and the samples obtained in Examples 2.9 to 2.11 were measured by a Karl Fischer analyzer. The results are shown in Table 4:

Table 4

The samples of Examples 2.9 and 2.11 had high solvent residues, which affected the quality. The sample of Example 2.10 was hygroscopic and was also a challenge to the formulation process. Thus the crystalline form obtained in Example 2.1, the VDA-1A crystalline form, is the predominant crystalline form.

Example 3 Detection of VDA-1 Form A

Test conditions for the sample of the example:

3.1XRD:

Testing equipment: Empyean X-ray diffractometer

Detection conditions: Cu target Kα ray, voltage 40kV, current 40mA, divergence slit 1/32°, anti-scatter slit 1/16°, anti-scatter slit 7.5mm, 2θ range: 3° to 50°, step size 0.02 °, each step of stay time 40S.

Test basis: People's Republic of China (2015 edition four) 0451 X-ray powder diffraction method

Test results: as shown in Figure 1.

3.2DSC:

Testing equipment: Germany NETZSCH DSC 214 differential scanning calorimeter

Test conditions: atmosphere: N2, 40ml/min

Scanning procedure: The temperature was raised from room temperature to 10 ° C / min to 250 ° C, and the temperature rise curve was recorded.

Test sample quality: Sample 1: 2.48mg (using aluminum sample tray)

Test basis: General rules for thermal analysis methods of JY/T 014-1996

Test results: as shown in Figure 2.

3.3TG:

Testing equipment: Germany NETZSCH company TG209 thermogravimetric analyzer

Test conditions: atmosphere: air, 20ml / min;

Scanning procedure: room temperature to 800 ° C, heating rate: 10 ° C / min.

Test basis: General rules for thermal analysis JY/T 014-1996

Test results: as shown in Figure 3.

3.4 Infrared spectrum:

Testing equipment: FT-IR NICOLET6700 (USA)

Detection conditions: potassium bromide tableting method

Test basis: GB/T 6040-2002 General rules for infrared spectrum analysis

Test results: as shown in Figure 4.

3.5 HPLC

Testing equipment: Agilent 1260series (USA)

Detection conditions:

Column: Waters Sunfire C18

Mobile phase A: water-mobile phase acetonitrile B (80:20)

Column temperature: 40 ° C detection wavelength: 390 nm.

Test basis: "Chinese Pharmacopoeia" two appendix VD high performance liquid chromatography

Test results: as shown in Figure 5.

Investigation on the stability of VDA-1 crystal form A

Example 4

The stability of the crystal form A obtained in Example 2.1 was examined (10-day accelerated test), and the crystal form A was placed at 60 ° C, humidity of 92.5%, and light conditions, and the results are shown in the following table.

Table 5: Results of investigation of factors affecting crystal form A

The water, purity, maximum single and total impurities of Form A were compared with the data for 0 days. As a result, the purity of Form A was slightly lowered under light conditions, and Form A was stable under other conditions.

Preparation of injection of VDA-1 Form A

Example 5.1

Take polysorbate-80, add 0.5% dried needle with activated carbon and VDA-1 crystal form A obtained in Example 2.1, heat to 50 ° C, stir at constant temperature for 30 min, heat and filter, to pass the clarity and color inspection. .

Take the prescribed amount of polysorbate-80, while adding 13% hydrochloric acid solution, stirring, so that the measured pH value is in the range of 3.5-3.9, adding VDA-1 crystal form A, stirring to dissolve VDA-1, the solution is passed through The microporous membrane was filtered under positive pressure until the solution was clear. The content was determined to be 99.81% and the pH was 3.74. After passing the test, the solution was filled in an antibiotic-controlled bottle in a clean condition, 0.5 ml per bottle (including VDA-1). ), after passing nitrogen, press the rubber plug, roll the aluminum cover, that is.

Take the medicinal ethanol, distill it on a water bath, discard the initial fraction, and collect the condensate. Weigh 195g of condensate, add injection water to 1500g, add 0.3% of the needle with activated carbon and VDA-1 crystal form A, heat and reflux for 30min, then let cool, decarbonize, filter through 0.22um microporous membrane to clear solution Filled in antibiotic-controlled bottles at a level of 100 clean, 1.5 ml per bottle. Press the rubber plug, roll the aluminum cover, that is.

Example 5.2

Take polysorbate-80, add 0.5% dried needle with activated carbon and VDA-1 crystal form A obtained in Example 2.1, heat to 50 ° C, stir at constant temperature for 60 min, heat and filter, to pass the clarity and color inspection. .

Take the prescribed amount of polysorbate-80, while adding 13% hydrochloric acid solution, stirring, so that the measured pH value is in the range of 3.5-3.9, adding VDA-1 crystal form A, stirring to dissolve VDA-1, the solution is passed through The microporous membrane was filtered under positive pressure until the solution was clear. The content was determined to be 99.81% and the pH was 3.74. After passing the test, the solution was filled in an antibiotic-controlled bottle in a clean condition, 0.5 ml per bottle (including VDA-1). , pressure rubber plug, rolled aluminum cover, that is.

Take the medicinal ethanol, distill it on a water bath, discard the initial fraction, and collect the condensate. Weigh 195g of condensate, add injection water to 1500g, add 0.3% of the needle with activated carbon and VDA-1 crystal form A, heat and reflux for 30min, then let cool, decarbonize, filter through 0.22um microporous membrane to clear solution Filled in antibiotic-controlled bottles at a level of 100 clean, 1.5 ml per bottle. Press the rubber plug, roll the aluminum cover, that is.

Example 5.3

Take polysorbate-80, add 0.5% dried needle with activated carbon and VDA-1 crystal form A obtained in Example 2.1, heat to 40 ° C, stir at constant temperature for 30 min, heat and filter until the clarity and color are qualified. .

Take the prescribed amount of polysorbate-80, while stirring, add VDA-1 crystal form A, stir to dissolve VDA-1, and pass the solution through a microfiltration membrane for positive pressure filtration until the solution is clear, the determined content is 98.36. % and pH are 6.14. After passing the test, the solution is filled in an antibiotic-controlled bottle in a clean condition, 0.5 ml (including VDA-1) per bottle, and the rubber stopper and aluminum cover are obtained.

Take the medicinal ethanol, distill it on a water bath, discard the initial fraction, and collect the condensate. Weigh 195g of condensate, add injection water to 1500g, add 0.3% of the needle with activated carbon and VDA-1 crystal form A, heat and reflux for 30min, then let cool, decarbonize, filter through 0.22um microporous membrane to clear solution Filled in antibiotic-controlled bottles at a level of 100 clean, 1.5 ml per bottle. Press the rubber plug, roll the aluminum cover, that is.

Example 5.4

Take polysorbate-80, add 0.5% dried needle with activated carbon and VDA-1 crystal form A obtained in Example 2.1, heat to 60 ° C, stir at constant temperature for 60 min, heat and filter, to pass the clarity and color inspection. .

Take a prescription amount of polysorbate-80, add 50% citric acid solution while stirring, and make the measured pH value in the range of 4.0-5.0, add VDA-1 crystal form A, stir to dissolve VDA-1, The solution was subjected to positive pressure filtration through a microporous membrane until the solution was clear, and the content was determined to be 99.16% and the pH was 4.50. After passing the test, the solution was filled in an antibiotic-regulated bottle in a clean condition, 0.5 ml per bottle (including VDA- 1), press the rubber plug, roll the aluminum cover, that is.

Take the medicinal ethanol, distill it on a water bath, discard the initial fraction, and collect the condensate. Weigh 195g of condensate, add injection water to 1500g, add 0.3% of the needle with activated carbon and VDA-1 crystal form A, heat and reflux for 30min, then let cool, decarbonize, filter through 0.22um microporous membrane to clear solution Filled in antibiotic-controlled bottles at a level of 100 clean, 1.5 ml per bottle. Press the rubber plug, roll the aluminum cover, that is.

The prepared injection was allowed to stand at 40 ° C for 7 days and 0 days, and the effect of the addition of acid and the addition of different acids, and the different processes of the same formulation, such as nitrogen and no nitrogen, on the stability of the injection of Form A were compared.

Examples 5.1 to 5.4 are shown in Table 7:

Table 7

Note: *pH value determination method: Take 0.5g of this product, add 4.5ml of water, stir for 15 minutes, measure according to law (Chinese Pharmacopoeia 2015 edition), stir the electrode for 15 minutes, and measure the pH value.

Y2: indicates the standard colorimetric liquid yellow No. 2 (in accordance with the Chinese Pharmacopoeia 2015 edition, supervised by Shanghai Pharmaceutical Inspection Institute)

It can be seen from Table 7 that no acid is added in Example 5.3. After 7 days at 40 °C, the related substances are significantly increased, the VDA-1 content is decreased, and other indexes are not significantly changed. 13% hydrochloric acid (Example 5.2) or 50% citric acid is added (implementation) The prescription of Example 5.4) is significantly better than the prescription without acid (Example 5.3), while the prescription for adjusting the pH with 13% hydrochloric acid is superior to the prescription for adjusting the pH with 50% citric acid. The same prescription process is different, such as implementation. In Examples 5.1 and 5.2, the content of nitrogen-passing sample and the related substances did not change much after being placed at 40 °C for 7 days. The content of nitrogen-free samples decreased and the related substances increased significantly. The stability of nitrogen-passing samples was better than that of nitrogen-free samples. 5.1 is the formulation and preparation process of VDA-1 injection (through nitrogen).

The above description of the preferred embodiments of the present invention is not intended to limit the invention, and various modifications and changes can be made by those skilled in the art without departing from the spirit of the invention. .

Claims (10)

1. Compound (3Z,6Z)-3-[((E)-3-(5-tert-butyl)-1H-imidazolyl-4-yl)methylene]-6-((E)- Form A of 3-(3-fluorophenyl)-2-propenylene)piperazin-2,5-dione, the X-ray powder diffraction pattern of Form A has a diffraction angle 2θ of 6.6 ° ± 0.2°, 9.7°±0.2°, 13.4°±0.2°, 16.0°±0.2°, 16.3°±0.2°, 16.6°±0.2°, 17.1°±0.2°, 18.1°±0.2°, 25.5°±0.2° Characteristic diffraction peaks at 25.9°±0.2° and 26.5°±0.2°,

   
2. The compound (3Z,6Z)-3-[((E)-3-(5-tert-butyl)-1H-imidazolyl-4-yl)methylene]-6-(() according to claim 1 Form A of E)-3-(3-fluorophenyl)-2-propenylene)piperazine-2,5-dione, characterized in that the crystal form A has the diffraction shown in the following table Angle 2θ, interplanar spacing d, and relative intensity I/I ₀ , and the relative intensity I/I _{0 has} an error of ±25%.

   
3. The compound (3Z,6Z)-3-[((E)-3-(5-tert-butyl)-1H-imidazolyl-4-yl)methylene]-6-(() according to claim 2 Form A of E)-3-(3-fluorophenyl)-2-propenylene)piperazine-2,5-dione, characterized in that the relative intensity I/I _{0 has} an error of ± 10%.
4. (3Z,6Z)-3-[((E)-3-(5-tert-butyl)-1H-imidazolyl-4-yl)methylene]-) according to any one of claims 1-3 Form A of 6-((E)-3-(3-fluorophenyl)-2-propenylene)piperazine-2,5-dione, characterized by a temperature-increasing DSC of the A crystal form The maximum endothermic transition of the curve is at 146.5 °C.
5. (3Z,6Z)-3-[((E)-3-(5-tert-butyl)-1H-imidazolyl-4-yl)methylene]-) according to any one of claims 1-3 Form A of 6-((E)-3-(3-fluorophenyl)-2-propenylene)piperazine-2,5-dione, characterized by thermogravimetric analysis of the crystal form The weight loss exceeded 6% at 236 °C.
6. (3Z,6Z)-3-[((E)-3-(5-tert-butyl)-1H-imidazolyl-4-yl)methylene]-) according to any one of claims 1-3 Form A of 6-((E)-3-(3-fluorophenyl)-2-propenylene)piperazine-2,5-dione, characterized by an infrared spectrum measured by KBr tableting , which ^{3228cm -1, 3064cm -1, 2966cm -1} , 2911cm -1, 2732cm -1, 2589cm -1, 1707cm -1, 1686cm -1, 1664cm -1, 1639cm -1, 1597cm -1, 1499cm -1 , 1411cm ^-1 , 1375cm ^-1 , 1350cm ^-1 , 1272cm ^-1 , 1149cm ^-1 , 1049cm ^-1 , 1022cm ^-1 , 952cm ^-1 , 937cm ^-1 , 810cm ^-1 , 773cm ^-1 , 677cm ^-1 , 443cm ^-1 has a strong absorption peak.
7. Preparation of (3Z,6Z)-3-[((E)-3-(5-tert-butyl)-1H-imidazolyl-4-yl)methylene]-6-(E) according to claim A method of crystal form A of 3-(3-fluorophenyl)-2-propenylene)piperazine-2,5-dione, comprising the steps of:

   1) Preparation of (3Z,6Z)-3-[((E)-3-(5-tert-butyl)-1H-imidazolyl-4-yl)methylene]-6-((E)-3- a crude product of (3-fluorophenyl)-2-propenylene)piperazine-2,5-dione (VDA-1);

   2) (3Z,6Z)-3-[((E)-3-(5-tert-butyl)-1H-imidazolyl-4-yl)methylene]-6-((E)-3- a crude product of (3-fluorophenyl)-2-propenylene)piperazine-2,5-dione is added to a solvent and heated to reflux until dissolved;

   Wherein the solvent is ethyl acetate, dimethyl sulfoxide or a mixed solvent of N,N-dimethylformamide and tetrahydrofuran; or a mixed solvent of a C ₃ -C ₄ alkyl ketone and tetrahydrofuran, The C ₃ -C ₄ alkyl ketone is selected from the group consisting of acetone, methyl ethyl ketone and n-butyl ketone;

   The volume ratio (V/V) of ethyl acetate, dimethyl sulfoxide, N,N-dimethylformamide, C ₃ -C ₄ alkyl ketone to tetrahydrofuran is 1:1 to 1:5;

   The ratio of the crude product to the solvent is 1 (g): 5 to 30 (ml), and the heating temperature is 50 to 80 ° C;

   3) After the solution is clarified, the temperature is lowered to precipitate a solid, and the solid is collected by filtration, and the collected solid is air-dried to obtain an A crystal form;

   The precipitation is carried out for 2 to 8 hours, the precipitation temperature is 0 to 40 ° C, and the filtration is completed and filtered, and the drying temperature is 30 to 60 ° C.
8. The preparation method according to claim 7, wherein the solvent in the step 2) is a mixed solvent of ethyl acetate and tetrahydrofuran, or a mixed solvent of acetone and tetrahydrofuran, and the ratio of the crude product to the solvent is a weight-to-volume ratio. 1 (g): 10 (ml), the heating temperature is 60 ° C;

   In step 3), the precipitation time is 4 hours, the precipitation temperature is 5 to 15 ° C, and the drying temperature is 45 ° C.
9. A pharmaceutical composition comprising (3Z,6Z)-3-[((E)-3-(5-tert-butyl)-1H-imidazolyl-4-yl)methylene]- Form A of 6-((E)-3-(3-fluorophenyl)-2-propenylene)piperazin-2,5-dione or a salt thereof, and a pharmaceutically acceptable carrier.
10. (3Z,6Z)-3-[((E)-3-(5-tert-butyl)-1H-imidazolyl-4-yl)methylene]-6-((E)- Use of Form A of 3-(3-fluorophenyl)-2-propenylene)piperazin-2,5-dione for the preparation of a medicament for the treatment of hyperproliferative diseases.

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