WO2022206800A1 - Tetrahydronaphthyridine compound crystal form and salt form, and preparation methods therefor - Google Patents

Abstract

A tetrahydronaphthyridine compound (I) crystal form and salt form and preparation methods therefor, and applications thereof in drugs for treating related diseases.

Description

Tetrahydronaphthyridine compound crystal form, salt form and preparation method thereof

This application claims the following priority:

CN202110343198.3, the application date is March 30, 2021;

CN202111015174.1, the application date is August 31, 2021;

CN2022102958433, the filing date is March 23, 2022.

technical field

The invention relates to a crystal form, a salt form of a tetrahydronaphthyridine compound and a preparation method thereof, and their application in medicines for treating related diseases.

Background technique

Programmed cell death 1 (PD-1), also known as CD279, is an important immunosuppressive molecule in the CD28/CTLA-4 receptor family. It is a membrane protein containing 268 amino acid residues and is widely expressed in T. On the surface of various immune cells such as cells, macrophages, and B cells, its ligands are PD-L1 and PD-L2. PD-L1 is a protein encoded by the CD274 gene and mainly expressed on the surface of tumor cells, dendritic cells and macrophages. After the combination of PD-1 and PD-L1, the PD-1/PD-L1 signaling pathway is activated, which in turn inhibits the activation of T cells, causing T cell inactivation and contributing to the immune escape of tumor cells. Another ligand of PD-1, PD-L2, is mainly expressed on the surface of dendritic cells, macrophages and B cells and is associated with inflammation and autoimmune diseases.

PD-1 negatively regulates immune responses by binding to its ligand PD-L1 and dephosphorylating multiple key molecules in the TCR signaling pathway. In healthy organisms, the activation of PD-1/PD-L1 signaling pathway can avoid the surrounding tissue damage caused by excessive immune response, thereby reducing the occurrence of autoimmune diseases. However, under the induction of the tumor microenvironment, the expressions of PD-1 and PD-L1 are abnormally increased, and tumor cells can successfully escape the recognition and detection of the immune system by the binding of these PD-L1 molecules to PD-1 on T cells. attack. PD-(L)1 mAb can block this "tumor immune escape mechanism" and restore the patient's own immune system's anti-cancer function.

Currently, the marketed drugs targeting the PD-1/PD-L1 pathway are monoclonal antibodies (mAbs). In 2014, the U.S. Food and Drug Administration (FDA) approved the first two monoclonal antibodies (Pembrolizumab and Nivolumab) to target the PD-1 molecule to the market. Over the next few years, three additional mAbs targeting the PD-L1 molecule (Atezolizumab, Durvalumab, and Avelumab) appeared on the market. All of the above are biological macromolecules, which have their own significant defects, such as being easily decomposed by proteases, poor in vivo stability, and need to be administered by injection; product quality is not easy to control, and production technology requirements are high; large-scale preparation and purification are difficult , high production cost and easy to produce immunogenicity. Small molecule PD-1/PD-L1 inhibitors have attracted more and more attention. Incyte's PD-L1 small molecule inhibitor INCB86550 (WO2018119263, WO2019191707) and Gilead's PD-L1 small molecule inhibitor GS-4224 (US20180305315, WO2019160882) ) has entered the clinical phase 2, and the small molecule PD-1/PD-L1 inhibitor of BMS benzyl phenyl ether (WO2015034820, WO2015160641) is in the preclinical research stage. Because small-molecule drugs can cross the cell membrane and act on intracellular targets, they have the advantages of convenient storage and transportation, low production cost, no immunogenicity, and usually oral administration. Therefore, research and development of small molecule blockers of PD-1/PD-L1 has broad application prospects.

SUMMARY OF THE INVENTION

The present invention also provides compounds of formula (II)

The present invention also provides Form A of the compound of formula (II), characterized in that its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 4.19±0.20°, 7.77±0.20° and 16.22±0.20°.

In some embodiments of the present invention, the above-mentioned A crystal form is characterized in that its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 4.19±0.20°, 7.77±0.20°, 8.71±0.20°, 9.89± 0.20° and 16.22±0.20°.

In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned A crystal form has characteristic diffraction peaks at the following 2θ angles: 4.19±0.20°, 7.77±0.20°, 8.71±0.20°, 9.89±0.20°, 16.22± 0.20°, 17.74±0.20°, 22.82±0.20° and 25.49±0.20°.

In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned Form A has characteristic diffraction peaks at the following 2θ angles: 4.19±0.20°, 7.77±0.20°, and/or 8.71±0.20°, and/or 9.89 ±0.20°, and/or 11.47°±0.20°, and/or 14.41°±0.20°, and/or 16.22±0.20°, and/or 17.74±0.20°, and/or 19.67°±0.20°, and/or 20.98°±0.20°, and/or 22.82±0.20°, and/or 25.49±0.20°, and/or 28.15°±0.20°.

In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned A crystal form has characteristic diffraction peaks at the following 2θ angles: 4.19±0.20°, 7.77±0.20°, 8.71±0.20°, 9.89±0.20°, 11.47° ±0.20°, 14.41°±0.20°, 16.22±0.20°, 17.74±0.20°, 19.67°±0.20°, 20.98°±0.20°, 22.82±0.20°, 25.49±0.20° and 28.15°±0.20°.

In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned A crystal form has characteristic diffraction peaks at the following 2θ angles: 4.19±0.10°, 7.77±0.10°, 8.71±0.10°, 9.89±0.10°, 11.47° ±0.10°, 14.41°±0.10°, 16.22±0.10°, 17.74±0.10°, 19.67°±0.10°, 20.98°±0.10°, 22.82±0.10°, 25.49±0.10° and 28.15°±0.10°.

In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned A crystal form has characteristic diffraction peaks at the following 2θ angles: 4.19°, 7.77°, 8.71°, 9.89°, 11.47°, 14.41°, 16.22°, 17.74° °, 19.67°, 20.98°, 22.82°, 25.49° and 28.15°.

In some embodiments of the present invention, the XRPD pattern of the above-mentioned crystal form A is shown in FIG. 1 .

In some embodiments of the present invention, the XRPD pattern of the above-mentioned crystal form A is substantially as shown in FIG. 1 .

In some schemes of the present invention, the XRPD spectrum analysis data of above-mentioned A crystal form is shown in Table 1:

Table 1 XRPD pattern analysis data of the crystal form of compound A of formula (II)

In some aspects of the present invention, the differential scanning calorimetry curve of the above-mentioned Form A has endothermic peaks onset at 31.2°C±5°C and 148.7°C±5°C.

In some embodiments of the present invention, the DSC spectrum of the above-mentioned A crystal form is shown in FIG. 2 .

In some embodiments of the present invention, the DSC spectrum of the above-mentioned Form A is substantially as shown in FIG. 2 .

In some embodiments of the present invention, the thermogravimetric analysis curve of the above-mentioned crystal form A has a weight loss of 1.77% at 130°C±3°C, and a weight loss of 3.32% at 160°C±3°C.

In some embodiments of the present invention, the TGA spectrum of the above-mentioned A crystal form is shown in FIG. 3 .

In some embodiments of the present invention, the TGA spectrum of the above-mentioned Form A is substantially as shown in FIG. 3 .

The present invention also provides the B crystal form of the compound of formula (I)

It is characterized in that its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 6.52±0.20°, 9.40±0.20° and 14.94±0.20°.

In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form B has characteristic diffraction peaks at the following 2θ angles: 6.52±0.20°, 9.40±0.20°, 14.94±0.20°, 16.10±0.20° and 22.86± 0.20°.

In some embodiments of the present invention, the X-ray powder diffraction pattern of the above crystal form B has characteristic diffraction peaks at the following 2θ angles: 6.52±0.20°, 9.40±0.20°, 12.93±0.20°, 14.94±0.20°, 16.10± 0.20°, 19.06±0.20°, 19.89±0.20° and 22.86±0.20°.

In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned Form B has characteristic diffraction peaks at the following 2θ angles: 6.52±0.20°, 9.40±0.20°, and/or 7.47±0.20°, and/or 11.29 ±0.20°, and/or 12.93±0.20°, and/or 13.90±0.20°, and/or 14.94±0.20°, and/or 16.10±0.20°, and/or 19.06±0.20°, and/or 19.89±0.20 °, and/or 21.13±0.20°, and/or 22.86±0.20°, and/or 24.55±0.20°, and/or 25.25±0.20°, and/or 25.83±0.20°, and/or 26.23±0.20°, and/or 27.45±0.20°, and/or 34.58±0.2°.

In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form B has characteristic diffraction peaks at the following 2θ angles: 6.52±0.20°, 7.47±0.20°, 9.40±0.20°, 11.29±0.20°, 12.93± 0.20°, 13.90±0.20°, 14.94±0.20°, 16.10±0.20°, 19.06±0.20°, 19.89±0.20°, 21.13±0.20°, 22.86±0.20°, 24.55±0.20°, 25.25±0.20°, 25.83± 0.20°, 26.23±0.20°, 27.45±0.20° and 34.58±0.20°.

In some embodiments of the present invention, the X-ray powder diffraction pattern of the above crystal form B has characteristic diffraction peaks at the following 2θ angles: 6.52±0.10°, 7.47±0.10°, 9.40±0.10°, 11.29±0.10°, 12.93± 0.10°, 13.90±0.10°, 14.94±0.10°, 16.10±0.10°, 19.06±0.10°, 19.89±0.10°, 21.13±0.10°, 22.86±0.10°, 24.55±0.10°, 25.25±0.10°, 25.83± 0.10°, 26.23±0.10°, 27.45±0.10° and 34.58±0.10°.

In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form B has characteristic diffraction peaks at the following 2θ angles: 6.52°, 7.47°, 9.40°, 11.29°, 12.93°, 13.90°, 14.94°, 16.10° °, 19.06°, 19.89°, 21.13°, 22.86°, 24.55°, 25.25°, 25.83°, 26.23°, 27.45° and 34.58°.

In some embodiments of the present invention, the XRPD pattern of the above-mentioned B crystal form is shown in FIG. 4 .

In some aspects of the present invention, the XRPD pattern of the above-mentioned Form B is substantially as shown in FIG. 4 .

In some schemes of the present invention, the XRPD spectrum analysis data of above-mentioned B crystal form is shown in Table 2:

Table 2 XRPD pattern analysis data of compound B crystal form of formula (I)

In some aspects of the present invention, the differential scanning calorimetry curve of the above-mentioned Form B has an onset of an endothermic peak at 145.2°C ± 5°C.

In some embodiments of the present invention, the DSC spectrum of the above-mentioned B crystal form is shown in FIG. 5 .

In some embodiments of the present invention, the DSC spectrum of the above-mentioned Form B is substantially as shown in FIG. 5 .

In some embodiments of the present invention, the thermogravimetric analysis curve of the above-mentioned crystal form B has a weight loss of 0.77% at 150°C±3°C.

In some embodiments of the present invention, the TGA spectrum of the above-mentioned B crystal form is shown in FIG. 6 .

In some embodiments of the present invention, the TGA pattern of the above-mentioned Form B is substantially as shown in FIG. 6 .

The present invention also provides a method for preparing the crystal form of compound B of formula (I), comprising:

(a) adding the compound of formula (I) to a solvent to form a suspension;

(b) The suspension is stirred at 35 to 45°C for 8 to 16 hours;

(c) drying after filtration for 8 to 16 hours;

Wherein, the solvent is tetrahydrofuran, ethanol, isopropanol, acetonitrile, methanol/water (1:1), ethanol/water (1:1), acetonitrile/water (1:1), isopropanol/water (1:1) :1) or water.

The present invention also provides the C crystal form of the compound of formula (I)

It is characterized in that its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 15.15±0.20°, 17.50±0.20° and 25.04±0.20°.

In some embodiments of the present invention, the above-mentioned crystal form C, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 8.75±0.20°, 15.15±0.20°, 17.50±0.20°, 22.94±0.20° and 25.04 ±0.20°.

In some embodiments of the present invention, the above crystal form C, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 8.75±0.20°, 15.15±0.20°, 17.50±0.20°, 18.77±0.20°, 22.94 ±0.20°, 25.04±0.20°, 26.20±0.20°, 37.65±0.20°.

In some embodiments of the present invention, the above-mentioned crystal form C, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 15.15±0.20°, 17.50±0.20°, and/or 8.39±0.20°, and/or 8.75±0.20°, and/or 11.32±0.20°, and/or 12.55±0.20°, and/or 13.74±0.20°, and/or 14.07±0.20°, and/or 17.03±0.20°, and/or 17.28± 0.20°, and/or 18.07±0.20°, and/or 18.77±0.20°, and/or 19.70±0.20°, and/or 20.35±0.20°, and/or 21.56±0.20°, and/or 21.83±0.20° , and/or 22.54±0.20°, and/or 22.94±0.20°, and/or 23.33±0.20°, and/or 23.74±0.20°, and/or 24.56±0.20°, and/or 25.04±0.20°, and /or 25.33±0.20°, and/or 26.20±0.20°, and/or 27.24±0.20°, and/or 27.93±0.20°, and/or 29.33±0.20°, and/or 30.75±0.20°, and/or 31.39±0.20°, and/or 32.44±0.20°, and/or 33.01±0.20°, and/or 34.07±0.20°, and/or 36.32±0.20°, and/or 37.65±0.20°, and/or 38.01± 0.2°.

In some embodiments of the present invention, the above crystal form C, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 8.39±0.20°, 11.32±0.20°, 12.55±0.20°, 13.74±0.20°, 14.07 ±0.20°, 15.15±0.20°, 17.03±0.20°, 17.50±0.20°, 18.07±0.20°, 18.77±0.20°, 19.70±0.20°, 20.35±0.20°, 21.56±0.20°, 22.54±0.20°, 23.33 ±0.20°, 23.74±0.20°, 24.56±0.20°, 25.04±0.20°, 26.20±0.20°, 27.24±0.20°, 27.93±0.20°, 29.33±0.20°, 30.75±0.20°, 31.39±0.20°, 33.01 ±0.20°, 34.07±0.20°, 36.32±0.20°, 37.65±0.20°, and 38.01±0.20°.

In some embodiments of the present invention, the above-mentioned crystal form C, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 8.39±0.10°, 8.75±0.10°, 11.32±0.10°, 12.55±0.10°, 13.74 ±0.10°, 14.07±0.10°, 15.15±0.10°, 17.03±0.10°, 17.28±0.10°, 17.50±0.10°, 18.07±0.10°, 18.77±0.10°, 19.70±0.10°, 20.35±0.10°, 21.56 ±0.10°, 21.83±0.10°, 22.54±0.10°, 22.94±0.10°, 23.33±0.10°, 23.74±0.10°, 24.56±0.10°, 25.04±0.10°, 25.33±0.10°, 26.20±0.10°, 27.24 ±0.10°, 27.93±0.10°, 29.33±0.10°, 30.75±0.10°, 31.39±0.10°, 33.01±0.10°, 34.07±0.10°, 36.32±0.10°, 37.65±0.10°, and 38.01±0.10°.

In some embodiments of the present invention, the above-mentioned crystal form C, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 8.39°, 8.75°, 11.32°, 12.55°, 13.74°, 14.07°, 15.15°, 17.03°, 17.28°, 17.50°, 18.07°, 18.77°, 19.70°, 20.35°, 21.56°, 21.83°, 22.54°, 22.94°, 23.33°, 23.74°, 24.56°, 25.04°, 25.33°, 26.20° , 27.24°, 27.93°, 29.33°, 30.75°, 31.39°, 32.44°, 33.01°, 34.07°, 36.32°, 37.65°, 38.01°.

In some embodiments of the present invention, the XRPD pattern of the above crystal form C is shown in FIG. 7 .

In some embodiments of the present invention, the XRPD pattern of the above-mentioned crystal form C is substantially as shown in FIG. 7 .

In some schemes of the present invention, the XRPD spectrum analysis data of above-mentioned C crystal form is shown in Table 3:

Table 3 XRPD pattern analysis data of compound C crystal form of formula (I)

In some aspects of the present invention, the differential scanning calorimetry curve of the above-mentioned crystal form C has an onset of an endothermic peak at 184.7°C ± 5°C.

In some embodiments of the present invention, the DSC spectrum of the above-mentioned C crystal form is shown in FIG. 8 .

In some embodiments of the present invention, the DSC spectrum of the above-mentioned crystal form C is substantially as shown in FIG. 8 .

In some embodiments of the present invention, the thermogravimetric analysis curve of the above-mentioned crystal form C has a weight loss of 0.69% at 150°C±3°C.

In some embodiments of the present invention, the TGA spectrum of the above crystal form C is shown in FIG. 9 .

In some embodiments of the present invention, the TGA pattern of the above-mentioned crystal form C is substantially as shown in FIG. 9 .

In some embodiments of the present invention, the DVS isotherm pattern of the above-mentioned crystal form C is substantially as shown in FIG. 10 .

The present invention also provides a method for preparing the crystal form of compound C of formula (I), comprising:

(a) adding the crystalline form of compound B of formula (I) to a solvent to form a suspension;

(b) The suspension is stirred at 25 to 45°C for 8 to 16 hours;

(c) drying after filtration for 8 to 16 hours;

Wherein, the solvent is methanol, acetone, isopropyl acetate, acetone:isopropyl acetate (2:1) and the like.

The present invention also provides a method for preparing the crystal form of compound C of formula (I), comprising:

(a) adding the B crystal form of the compound of formula (I) into a solvent to form a suspension;

(b) The suspension is stirred at 55 to 60° C. for 8 to 16 hours;

(c) drying after filtration for 8 to 16 hours;

Wherein, the solvent is acetone, isopropyl acetate and the like.

The present invention also provides the use of the above compound or crystal form A or crystal form B or crystal form C or the crystal form B or crystal form C prepared according to the above method in the preparation of antitumor drugs.

technical effect

The compound of the present invention has good PK property and oral absorption rate, stable crystal form and good druggability.

The compound of the present invention has a good inhibitory effect on the excessive activation of the PD-1/PD-L1 signaling pathway, thereby obtaining excellent tumor growth inhibitory activity.

Definition and Explanation

Unless otherwise specified, the following terms and phrases used herein are intended to have the following meanings. A particular phrase or term should not be considered indeterminate or unclear without a specific definition, but should be understood in its ordinary meaning. When a trade name appears herein, it is intended to refer to its corresponding trade or its active ingredient.

The intermediate compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by their combination with other chemical synthesis methods, and those skilled in the art. Well-known equivalents, preferred embodiments include, but are not limited to, the examples of the present invention.

The chemical reactions of specific embodiments of the present invention are carried out in suitable solvents suitable for the chemical changes of the present invention and their required reagents and materials. In order to obtain the compounds of the present invention, it is sometimes necessary for those skilled in the art to modify or select the synthetic steps or reaction schemes on the basis of the existing embodiments.

The present invention will be specifically described below through examples, which do not imply any limitation to the present invention.

All solvents used in the present invention are commercially available and used without further purification.

The following abbreviations are used in the present invention: _CD3OD stands for deuterated methanol; _CDCl3 stands for deuterated chloroform.

软件命名，市售化合物采用供应商目录名称。 Compounds are manually or

Software naming, commercially available compounds use supplier catalog names.

The powder X-ray diffraction (X-ray powder diffractometer, XRPD) method of the present invention

Instrument model: PANalytical X'pert ³ X-ray diffractometer

Test method: About 10 mg of sample is evenly spread on a single crystal silicon sample pan for XRPD detection.

The detailed XRPD parameters are as follows:

K-Alphal

Ray source: K-Alphal

K-Alphal

Light tube voltage: 45kV, light tube current: 40mA

Measurement time: 46.665s

Scanning angle range: 3-40deg

Step width angle: 0.0263deg

Differential Thermal Analysis (Differential Scanning Calorimeter, DSC) method of the present invention

Instrument model: TA 2500 Differential Scanning Calorimeter

Test method: Take a sample (~1mg) and place it in a DSC aluminum pot for testing. Under the condition of 10ml/min _N2 , at a heating rate of 10°C/min, heat the sample from 25°C (room temperature) to 200°C (or 250°C).

Thermogravimetric Analysis (Thermal Gravimetric Analyzer, TGA) method of the present invention

Instrument Model: TA 5500 Thermogravimetric Analyzer

Test method: Take the sample (2~5mg) and put it in a TGA platinum pot for testing. Under the condition of 10ml/min _N2 , at a heating rate of 10℃/min, heat the sample from room temperature to 350℃ or lose 20% of the weight.

The dynamic vapor adsorption analysis (Dynamic Vapor Sorption, DVS) method of the present invention

Instrument model: SMS DVS Advantage dynamic vapor adsorption instrument

Test conditions: Take a sample (10-15 mg) and place it in the DVS sample tray for testing.

The detailed DVS parameters are as follows:

Temperature: 25℃

Balance: dm/dt=0.01%/min (minimum: 10min, longest: 180min)

Drying: 120min at 0%RH

RH (%) test rung: 10%

RH (%) test step range: 0%-90%-0%

The hygroscopicity evaluation is classified as follows:

Table 4

Hygroscopic classification ΔW% deliquescence Absorbs enough water to form a liquid Very hygroscopic ΔW%≥15% hygroscopic 15%>ΔW%≥2% slightly hygroscopic 2%>ΔW%≥0.2% No or almost no hygroscopicity ΔW%<0.2%

Note: ΔW% represents the hygroscopic weight gain of the test product at 25±1℃ and 80±2%RH.

Description of drawings

Fig. 1 is the XRPD spectrum of (II) compound A crystal form;

Fig. 2 is the DSC spectrogram of (II) compound A crystal form;

Fig. 3 is the TGA spectrum of (II) compound A crystal form;

Fig. 4 is the XRPD spectrum of (I) compound B crystal form;

Fig. 5 is the DSC spectrogram of (I) compound B crystal form;

Fig. 6 is the TGA spectrum of (I) compound B crystal form;

Fig. 7 is the XRPD spectrum of (I) compound C crystal form;

Fig. 8 is the DSC spectrogram of (I) compound C crystal form;

Fig. 9 is the TGA spectrogram of (I) compound C crystal form;

Figure 10 is the DVS isotherm spectrum of (I) Compound C crystal form.

Detailed ways

The present invention will be described in detail by the following examples, but it does not mean any unfavorable limitation of the present invention. The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments enumerated below, embodiments formed in combination with other chemical synthesis methods, and those well known to those skilled in the art Equivalent to alternatives, preferred embodiments include, but are not limited to, the embodiments of the present invention. It will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the present invention without departing from the spirit and scope of the invention.

Example 1: Preparation of compounds of formula (I) and their formates

Synthesis of compound 1:

first step

Compounds 1-1 (100 g, 484 mmol, 1 equiv), 1-2 (135.3 g, 533 mmol, 1.1 equiv) were dissolved in dioxane (1000 mL), and potassium acetate was added to the reaction solution (142.6 g, 1.45 mol, 3 equiv), dichlorobis(triphenylphosphine)palladium (10.2 g, 14.53 mmol, 0.03 equiv), stirred at 110°C for 16 hours under nitrogen protection. After the reaction was completed, filtered, the filtrate was added to water (800 mL), extracted with isopropyl acetate (500 mL×2), the organic phase was washed with saturated brine (800 mL×1), dried over anhydrous sodium sulfate, filtered, The residue concentrated under reduced pressure was diluted with isopropyl acetate (80 mL), followed by stirring at 20° C. for 30 minutes, the mixture was filtered, and the crude product of the filtrate concentrated under reduced pressure was prepared with (n-heptane:isopropyl acetate=30:1, 320 mL). ) at room temperature for 30 minutes, filtered, and the filter cake was dried to obtain compound 1-3.

MS-ESI calculated [M+H] ⁺ 254, found 254. ¹ H NMR (400 MHz, CD ₃ OD) δ = 7.07-7.01 (m, 1H), 7.00-6.76 (m, 2H), 1.37 (s, 12H).

second step

Compounds 1-3 (89.3 g, 352.2 mmol, 1 equiv), 1-4 (95.22 g, 352.2 mmol, 1 equiv) were dissolved in N,N-dimethylformamide (800 mL), water (100 mL), then potassium carbonate (146 g, 1.06 mol, 3 equiv.), (dichlorobis(triphenylphosphine)palladium (7.42 g, 10.57 mmol, 0.03 equiv.) were added to the reaction solution, under nitrogen protection for 85 Stir at ℃ for 4 hours. After the reaction is completed, filter, add the filtrate to aqueous hydrochloric acid (2 mol/L, 2400 mL), extract with methyl tert-butyl ether (1000 mL×2), and use saturated brine ( 1000 mL × 2) washed, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure and the residue was diluted with methyl tert-butyl ether (100 mL), and then 98% concentrated sulfuric acid was added dropwise to the stirring solution until The pH of the solution was adjusted to 1-2, then (methyl tert-butyl ether: isopropanol=10:1, 200 ml) was added at room temperature for 30 minutes and stirred at 20°C for 60 minutes, the mixture was filtered, and the filter cake was dried and added to the solution. Saturated aqueous sodium carbonate solution (1200 mL), extracted with dichloromethane (600 mL×2), the organic phase was washed with saturated brine (800 mL×2), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was used Isopropanol (70 mL) was stirred at room temperature for 60 minutes, and the filter cake was filtered and dried to obtain compound 1.

MS-ESI calculated [M+H] ⁺ 317, found 317. ¹ H NMR (400 MHz, CD ₃ OD) δ=7.76-7.68 (m, 1H), 7.30-7.22 (m, 2H), 7.12 (t, J=7.8 Hz, 1H), 6.90 (dd, J=1.3, 8.1Hz, 1H), 6.54 (dd, J=1.3, 7.5Hz, 1H).

Synthesis of compound 2:

first step

Compound 2-1 (100 g, 723.99 mmol, 1 equiv) was dissolved in toluene (1000 mL), then compound 2-2 (168.13 g, 1.45 mol, 160.13 mL, 2 equiv) was added, and stirred at 20°C for 15 minutes Then, phosphorus oxychloride (333.03 g, 2.17 mol, 201.84 mL, 3 equiv.) was added dropwise, and the mixture was heated to 100°C and stirred for 2 hours. After the reaction, phosphorus oxychloride and toluene were distilled under reduced pressure at 100 ° C until the reaction solution became very viscous, diluted with 1000 ml of ethyl acetate, slowly poured into 6 liters of water and stirred for 10 minutes, and sodium carbonate was added to adjust the pH. to 7, then extracted with ethyl acetate (1000 mL×3), the combined organic phases were washed with brine (1000 mL×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product by silica gel chromatography Chromatographic column (petroleum ether:ethyl acetate=20:1-2:1) to obtain compound 2-3.

MS-ESI calculated [M+H] ⁺ 237, found 237. ¹ H NMR (400 MHz, CDCl ₃ ) δ=9.76 (s, 1H), 8.84 (d, J=5.8 Hz, 1H), 8.48 (s, 1H), 8.09 (d, J=5.9 Hz, 1H), 4.61 (q, J=7.2 Hz, 2H), 1.53 (t, J=7.2 Hz, 3H).

second step

Compound 2-3 (40 g, 169.02 mmol, 1 equiv) was dissolved in methanol (400 mL), then methanesulfonic acid (97.47 g, 1.01 mol, 72.20 mL, 6 equiv) was slowly added dropwise. After heating to 85°C, the mixture was stirred for 3 hours. After the reaction was completed, it was cooled to room temperature, then solid sodium acetate was added at 0°C to adjust the pH to 7, diluted with 2000 ml of water, stirred at room temperature for 30 minutes, filtered, and the filter cake was dried to obtain compound 2-4.

MS-ESI calculated [M+H] ⁺ 219, found 219. ¹ H NMR (400 MHz, CDCl ₃ ) δ=9.65 (s, 1H), 8.70 (d, J=5.7 Hz, 1H), 8.04 (d, J=5.7 Hz, 1H), 7.76 (s, 1H), 4.19 (s, 3H), 4.13 (s, 3H).

third step

Compound 2-4 (25 g, 114.57 mmol, 1 equiv) was dissolved in methanol (250 mL), then sulfuric acid (34.40 g, 343.71 mmol, 18.70 mL, 98% purity, 3 equiv) was slowly added dropwise, followed by nitrogen Wet palladium carbon (2.5 g, 11.46 mmol, 10% purity, 0.1 equiv) was added under protection, and the reaction solution was replaced with hydrogen several times, and then reacted at 20° C. for 5 hours under a hydrogen (50 psi) environment. After completion of the reaction, filter and concentrate the filtrate under reduced pressure to obtain compound 2-5.

MS-ESI calculated [M+H] ⁺ 223, found 223.

the fourth step

Compound 2-5 (40 g, 62.44 mmol, 1 equiv, sulfate, crude) was dissolved in dichloromethane (300 mL), and 1.8-diazabicyclo[5.4.0]undecane was slowly added dropwise -7-ene (38.02 g, 249.75 mmol, 37.65 mL, 2 equiv), then compound 2-6 (23.94 g, 137.36 mmol, 26.17 mL, 1.1 equiv) was added, and after stirring at 20°C for 0.5 hours, added Sodium borohydride acetate (79.40 g, 374.63 mmol, 3 equiv) was stirred at 20°C for 1.5 hours. After the reaction was completed, the reaction solution was poured into 600 mL of water, and the pH was adjusted to 7 with sodium bicarbonate, extracted with dichloromethane (300 mL×3), and the combined organic phases were washed with saturated brine (300 mL×2). , and dried over anhydrous sodium sulfate. The crude product obtained by filtration and concentration under reduced pressure was subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 10:1-2:1) and preparative high performance liquid chromatography (chromatographic column: Kromasil Eternity XT 250 × 80 mm × 10 μm; flow Phase: mobile phase A: 10 mmol aqueous ammonium bicarbonate solution; mobile phase B: acetonitrile; B%: 50%-80%, 20 minutes) purification and isolation of compound 2.

MS-ESI calculated [M+H] ⁺ 381, found 381. ¹ H NMR (400MHz, CD ₃ OD) δ=7.48(s, 1H), 3.88(s, 3H), 3.86(s, 3H), 3.79(t, J=5.9Hz, 2H), 3.68(s, 2H) ), 2.84-2.76(m, 2H), 2.73-2.68(m, 2H), 2.66(t, J=5.9Hz, 2H), 0.82(s, 9H), 0.01(br s, 6H).

Synthesis of compound 5:

first step

Compound 5-1 (200 g, 980.27 mmol) was dissolved in ethylene glycol dimethyl ether (600 mL) and toluene (600 mL), and cuprous iodide (37.34 g, 196.05 mmol) was added with stirring, Then add potassium iodide (325.45 g, 1.96 mol), set up a tail gas absorption device after nitrogen replacement three times, heat the reaction solution to 60 ° C, after the internal temperature rises to 60 ° C, start to slowly drip isoamyl nitrite (660 ml, 4.90 mol), the reaction solution was reacted at room temperature for 2 hours. After the completion of the reaction, the reaction solution was filtered through celite, the filtrate was poured into ice (1000 mL), and after stirring for 20 minutes, saturated ammonium chloride solution (1000 mL) was added, and the mixture was stirred for 30 minutes. mL) extraction and separation, the organic phase was washed with saturated sodium sulfite (2000 mL), washed with saturated brine (2000 mL), dried over sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was stirred with 120 mL of methyl tert-butyl ether and 120 mL of n-heptane, filtered, and the filter cake was dried to obtain compound 5-2.

MS-ESI calculated [M+H] ⁺ 315, found 315.

second step

Compound 5-2 (60 g, 190.53 mmol) was dissolved in 2-methyltetrahydrofuran (800 mL), nitrogen was replaced three times, the temperature was lowered to -40°C, and isopropyl was slowly added dropwise after the internal temperature rose to -40°C magnesium chloride and lithium chloride (1.3 mol/L, 1 mL), stirred at -40 °C for 0.5 hours, and added dropwise N,N-dimethylamide (73.3 mL, 952.67 mmol), and the reaction solution was reacted at room temperature for 3 Hour. After the completion of the reaction, the reaction solution was poured into glacial hydrochloric acid (1 mol/L, 800 mL) to quench, extracted and separated, the aqueous phase was extracted with methyl tert-butyl ether (400 mL×2), the organic phases were combined and saturated brine was used. (400 mL×2) washed, dried over sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 5-3.

third step

Compound 5-4 (10.59 g, 85.71 mmol) was dissolved in 2-methyltetrahydrofuran (150 mL), potassium carbonate (11.85 g, 85.71 mmol) was added, followed by compound 5-3 (15.5 g, 71.42 mmol) ), stirred for 0.5 hours and then added sodium borohydride acetate (22.71 g, 107.13 mmol) in portions. The reaction solution was reacted at 20°C for 1 hour. The reaction solution was poured into glacial hydrochloric acid (1 mol/L, 200 mL) to quench, and the liquid was extracted and separated. The aqueous phase was adjusted to pH=8 with solid sodium carbonate, extracted with ethyl acetate (100 mL×2), and the organic phase was added with saturated common salt. Washed with water (100 mL×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was subjected to preparative high performance liquid chromatography (chromatographic column: Xtimate C18 250 × 50 mm × 10 microns; mobile phase: mobile phase A: 0.05% ammonia solution by volume; mobile phase B: acetonitrile; B%: 15%-45%, 18 minutes) to isolate compound 5.

MS-ESI calculated [M+H] ⁺ 288, found 288. ¹ H NMR (400 MHz, CD ₃ OD) δ=8.23-8.19 (m, 1H), 4.08-4.04 (m, 1H), 4.01-3.99 (m, 3H), 3.78 (s, 2H), 3.73-3.69 ( m, H), 3.26-3.24 (m, 3H), 3.19-3.15 (m, 2H).

Synthesis of compounds of formula (I):

first step

At 20-25°C, 14 liters of anhydrous 2-methyltetrahydrofuran was added to the reaction kettle, and then 974.32 g of compound 2 (1.05 equiv.) and 720.00 g of compound 1 (1.00 equiv.) were added. Nitrogen was replaced for 5 to 10 minutes, then the temperature of the reactor was adjusted to 0 to 5°C, and 6.67 liters of tetrahydrofuran solution of bis(trimethylsilyl) lithium amide (1 mol/liter, 3 equivalents) were slowly added dropwise to the reactor in batches After the dropwise addition, the mixture was stirred at 0 to 5°C for 1 hour. At 0 ~ 5 ℃, the reaction solution was slowly pumped into a reactor containing 14 liters of saturated ammonium chloride aqueous solution in batches, and the liquids were separated, and the aqueous phase and the organic phase were collected respectively, washed with 14 liters of saturated brine, and the water was collected separately. Phase and organic phase, the organic phase was dried, filtered and concentrated to give a solid crude product. At 20-25° C., 4.2 liters of methanol was added to the reaction kettle, and then the solid crude product was added to continue stirring for 30 minutes. After filtration, the filter cake was washed twice with methanol, and the filter cake was vacuum-dried to constant weight to obtain compound 3.

MS-ESI calculated [M+H] ⁺ 666, found 666. ¹ H NMR (400 MHz, CDCl ₃ ) δ=10.64-10.90 (m, 1H), 8.63 (m, 1H), 7.50-7.68 (m, 2H) 7.33 (t, J=7.95Hz, 1H) 7.12-7.20 ( m,2H)6.94(m,1H)3.90(s,3H)3.67-3.83(m,4H)2.61-2.85(m,6H)0.83(s,9H)0.00(s,6H).

second step

At 20 ~ 25 ℃, 13 liters of anhydrous dioxane was added to the reaction kettle, and then 1306.10 grams of compound 3 (1.00 equivalents), 598.03 grams of double pinacol boronate (1.24 equivalents), 577.81 grams were added successively Potassium acetate (3.10 equiv.) and 13.77 g of dichlorobis(triphenylphosphine) palladium (0.01 equiv.) were added to the reactor, replaced with nitrogen for 5 to 10 minutes, and then the temperature of the reactor was adjusted to 100 to 110° C. Stir at 110°C for 12 hours to obtain crude compound 4, which is directly used in the next step. The temperature of the reaction kettle was adjusted to 20-25°C, 3.25 liters of water was added to the reaction kettle, and then 577.81 g of compound 5 (1.01 equiv.) and 790.97 g of potassium carbonate (3.01 equiv.) were added to the reaction kettle. Nitrogen replacement was carried out for 5 to 10 minutes, 13.72 g of dichlorobis(triphenylphosphine) palladium (0.01 equivalent) was added to the reaction kettle, and then the temperature of the reaction kettle was adjusted to 70 to 75 ° C and stirred for 12 hours at 70 to 75 ° C. . The reaction solution was cooled to room temperature, filtered, the filter cake was washed with 13 liters of ethyl acetate, 9.75 liters of water was added to the combined filtrate, and the layers were separated. The organic phase. The combined organic phases were washed with 13 liters of 10% citric acid aqueous solution, and the layers were separated. The aqueous and organic phases were collected separately, and the aqueous phase was extracted twice with 13 liters of ethyl acetate. The organic phases were combined, washed with 6.5 liters of water, separated, and the aqueous and organic phases were collected separately. Combine the aqueous phases, adjust the pH to 9 with 4 mol/L aqueous sodium hydroxide solution, add 13 liters of ethyl acetate, and separate the layers. The aqueous phase and the organic phase were collected separately, the organic phases were combined, washed with 13 liters of saturated brine, washed twice, and the organic phase was collected. The organic phase was dried, filtered and concentrated to crude. At 20-25°C, the crude product was dissolved in 18 liters of ethyl acetate and poured into the reaction kettle, and the palladium-removed modified silica gel was added to the reaction kettle, and stirred at 65 to 70°C for 16 hours, and then the reaction solution was filtered, The filter cake was washed with 36 liters of ethyl acetate, the washing liquid was spin-dried and combined with the filtrate, and then the palladium removal operation was repeated once, and the filtrate was concentrated to obtain the crude product of compound 6.

Add 14.6 liters of acetone to the reaction kettle, pour the above crude product into complete dissolution, then dissolve 607.95 grams of anhydrous oxalic acid (1.70 equivalents) in 2.92 liters of acetone, slowly add it to the above solution in batches, and finally add 5.84 liters Acetonitrile, stirred at 20-25°C for 1 hour, filtered, the filter cake was washed with 14.6 liters of acetonitrile, and the filter cake was vacuum dried to constant weight to obtain the oxalate of compound 6.

MS-ESI calculated [M+H] ⁺ 793, found 793. ¹ H NMR (400MHz, CD ₃ OD) δ=8.57-8.67(m, 1H), 8.48(s, 1H), 7.82(s, 1H), 7.72(m, 1H), 7.58(s, 1H), 7.48 (br d, J＝13.08Hz, 2H), 7.18(s, 1H), 4.72(s, 2H), 4.56-4.66(m, 4H), 4.39(br t, J＝5.26Hz, 1H), 4.19- 4.28(m, 2H), 4.05-4.14(m, 8H), 3.67(br t, J=6.17Hz, 2H), 3.43-3.50(m, 2H), 3.36(s, 3H), 3.11(s, 2H ), 0.92(s, 8H) 0.13(s, 6H).

third step

At 20～25 ℃, 7.5 liters of anhydrous tetrahydrofuran and 22.5 liters of water were added to the reaction kettle, then 3000.00 g of compound 6 oxalate salt (1.00 equiv.) and 1349.50 g of anhydrous oxalic acid (5.00 equiv.) were added and heated at 20～25 ℃ under stirring for 12 hours. The reaction solution was extracted twice with 30 liters of methyl tert-butyl ether, and the aqueous phase and the organic phase were collected separately. The organic phases were combined, washed with 15 liters of water, and the layers were separated, and the aqueous and organic phases were collected separately. Combine the aqueous phases, adjust the pH to 9 with 4 mol/L aqueous sodium hydroxide solution, add 45 liters of 2-methyltetrahydrofuran, separate the layers, collect the aqueous phase and the organic phase, and add 15 liters of 2-methyltetrahydrofuran to the aqueous phase. , the liquids were separated, the aqueous phase and the organic phase were collected separately, the organic phases were combined, washed twice with 30 liters of saturated brine, and the organic phase was collected. The organic phase was dried, filtered and concentrated to crude. At 20-25 ℃, the above solid was dissolved in 14 liters of anhydrous tetrahydrofuran and 6 liters of ethyl acetate and added to the reaction kettle, then the modified silica gel except for palladium was added to the reaction kettle, and at 20 to 25 ℃, stirred for 12 After 1 hour, the reaction solution was filtered, spin-dried, and the filtrate was concentrated to obtain a crude product, which was vacuum-dried to constant weight to obtain the compound of formula (I).

MS-ESI calculated [M+H] ⁺ 679, found 679. ¹ H NMR (400 MHz, CD ₃ OD) δ=8.60 (m, 1H), 8.40 (s, 1H), 7.64-7.76 (m, 2H), 7.54 (t, J=7.64 Hz, 1H), 7.40-7.50 (m,2H),7.15(m,1H),4.07-4.13(m,1H),4.03(d,J=12.96Hz,6H),3.89(s,2H),3.74-3.83(m,6H), 3.26(s, 3H), 3.19-3.24(m, 2H), 2.86(m, 4H), 2.76(t, J=5.87Hz, 2H).

Synthesis of formate salts of compounds of formula (I):

The crude compound 6 (68 mg, 85.66 μmol) was dissolved in dichloromethane (3 mL), hydrogen chloride in ethyl acetate (4 mol/L, 1.5 mL) was added, and the mixture was stirred at room temperature at 25° C. for 0.5 hour. After the reaction was completed, it was concentrated. The crude product was subjected to preparative high performance liquid chromatography (chromatographic column: Phenomenex Gemini-NX C18 75 × 30 mm × 3 microns; mobile phase: mobile phase A: 0.225% formic acid aqueous solution by volume; mobile phase B: acetonitrile; B%: 10%- 40%, 7 minutes) to isolate the formate salt of the compound of formula (I). MS-ESI calculated [M+H] ⁺ 679, found 679. ¹ H NMR (400MHz, CD ₃ OD) δ=8.58-8.65(m, 1H), 8.49(s, 1H) 8.43(br s, 1H), 7.70-7.78(m, 2H), 7.59(t, J= 7.63Hz, 1H), 7.45-7.53(m, 2H), 7.18(dd, J=7.63, 1.38Hz, 1H), 4.52(s, 2H), 4.30-4.41(m, 3H), 4.12(s, 3H) ),4.06(s,3H),3.94-4.01(m,4H),3.81-3.87(m,2H),3.35-3.37(m,3H),3.02-3.07(m,2H),2.91(q,J =5.38Hz, 4H).

Example 2: Preparation of compound B crystal form of formula (I)

100 mg of the compound of formula (I) was weighed into a 4.0 ml glass vial, and an appropriate amount of ethanol was added to form a suspension. After adding the magneton, the above suspension sample was placed on a magnetic heating stirrer (40°C) for testing, stirred overnight at 40°C and centrifuged, and the residual solid sample was placed in a vacuum drying oven (30°C) to dry overnight to obtain Form B of the compound of formula (I).

100 mg of the compound of formula (I) was weighed into a 4.0 ml glass vial, and an appropriate amount of isopropanol was added to form a suspension. After adding the magneton, the above suspension sample was placed on a magnetic heating stirrer (40°C) for testing, stirred overnight at 40°C and centrifuged, and the residual solid sample was placed in a vacuum drying oven (30°C) to dry overnight to obtain Form B of the compound of formula (I).

100 mg of the compound of formula (I) was weighed into a 4.0 ml glass vial, and an appropriate amount of acetonitrile was added to form a suspension. After adding the magneton, the above suspension sample was placed on a magnetic heating stirrer (40°C) for testing, stirred overnight at 40°C and centrifuged, and the residual solid sample was placed in a vacuum drying oven (30°C) to dry overnight to obtain Form B of the compound of formula (I).

100 mg of the compound of formula (I) was weighed into a 4.0 ml glass vial, and an appropriate amount of methanol/water (1:1) was added to form a suspension. After adding the magneton, the above suspension sample was placed on a magnetic heating stirrer (40°C) for testing, stirred overnight at 40°C and centrifuged, and the residual solid sample was placed in a vacuum drying oven (30°C) to dry overnight to obtain Form B of the compound of formula (I).

100 mg of the compound of formula (I) was weighed into a 4.0 ml glass vial, and an appropriate amount of acetonitrile/water (1:1) was added to form a suspension. After adding the magneton, the above suspension sample was placed on a magnetic heating stirrer (40°C) for testing, stirred overnight at 40°C and centrifuged, and the residual solid sample was placed in a vacuum drying oven (30°C) to dry overnight to obtain Form B of the compound of formula (I).

100 mg of the compound of formula (I) was weighed into a 4.0 ml glass vial, and an appropriate amount of isopropanol/water (1:1) was added to form a suspension. After adding the magneton, the above suspension sample was placed on a magnetic heating stirrer (40°C) for testing, stirred overnight at 40°C and centrifuged, and the residual solid sample was placed in a vacuum drying oven (30°C) to dry overnight to obtain Form B of the compound of formula (I).

9 liters of acetone was added to the reaction kettle, and then the crude product of the compound of formula (I) was added, and the mixture was stirred at 20-25° C. for 24 hours. Filtration, the filter cake was washed twice with 1.8 liters of acetone, and the filter cake was vacuum-dried to constant weight to obtain the B crystal form of the compound of formula (I).

MS-ESI calculated [M+H] ⁺ 679, found 679. ¹ H NMR (400 MHz, CD ₃ OD) δ=8.60 (m, 1H), 8.40 (s, 1H), 7.64-7.76 (m, 2H), 7.54 (t, J=7.64 Hz, 1H), 7.40-7.50 (m,2H),7.15(m,1H),4.07-4.13(m,1H),4.03(d,J=12.96Hz,6H),3.89(s,2H),3.74-3.83(m,6H), 3.26(s, 3H), 3.19-3.24(m, 2H), 2.86(m, 4H), 2.76(t, J=5.87Hz, 2H).

Example 3: Study on the salt form of the compound of formula (I)

About 100 mg of the compound of formula (I) was taken, dissolved in 0.5 mL of solvent, and then acid (1.05 equiv.) was added slowly with stirring. After stirring at 25°C for 0.5 hours, a solid was precipitated and filtered. The residual solid sample was placed in a vacuum drying oven (30° C.) to dry overnight to obtain the salt of the compound of formula (I). The specific properties of the compounds are as follows:

table 5

Example 4: Preparation of compound of formula (II)

2.01 g of the compound of formula (I) was taken, dissolved in 20 mL of ethyl acetate, and then fumaric acid (675 mg, 2.01 equiv) was slowly added with stirring. The mixture was stirred at 25°C for 16 hours, the solid was precipitated, filtered, and the residual solid sample was placed in a vacuum drying oven (30°C) to dry overnight to obtain the compound of formula (II). ¹ H NMR (400 MHz, CD ₃ OD) δ=8.62 (dd, J=1.5, 8.2 Hz, 1H), 8.50 (s, 1H), 7.79 (s, 1H), 7.73 (dd, J=1.6, 7.7 Hz) ,1H),7.59(t,J＝7.7Hz,1H),7.54-7.43(m,2H),7.18(dd,J＝1.5,7.7Hz,1H),6.70(s,4H),4.67(s, 2H), 4.53(dd, J=6.4, 11.7Hz, 2H), 4.44-4.34(m, 1H), 4.23(s, 2H), 4.13(s, 4H), 4.07(s, 3H), 3.91(t , J=5.4Hz, 2H), 3.38 (s, 3H), 3.14 (t, J=5.5Hz, 2H), 3.01 (t, J=6.1Hz, 2H).

Example 5: Preparation of the crystal form of compound A of formula (II)

100 mg of the compound of formula (II) was weighed into a 4.0 ml glass vial, and an appropriate amount of acetone was added to form a suspension. After adding the magneton, the above suspension sample was placed on a magnetic heating stirrer (40°C) for testing, stirred overnight at 40°C and centrifuged, and the residual solid sample was placed in a vacuum drying oven (30°C) to dry overnight to obtain Form A of the compound of formula (II).

100 mg of the compound of formula (II) was weighed into a 4.0 ml glass vial, and an appropriate amount of ethanol was added to form a suspension. After adding the magneton, the above suspension sample was placed on a magnetic heating stirrer (40°C) for testing, stirred overnight at 40°C and centrifuged, and the residual solid sample was placed in a vacuum drying oven (30°C) to dry overnight to obtain Form A of the compound of formula (II).

100 mg of the compound of formula (II) fumarate was weighed into a 4.0 ml glass vial, and an appropriate amount of ethyl acetate was added to form a suspension. After adding the magneton, the above suspension sample was placed on a magnetic heating stirrer (40°C) for testing, stirred overnight at 40°C and centrifuged, and the residual solid sample was placed in a vacuum drying oven (30°C) to dry overnight to obtain Form A of the compound of formula (II).

100 mg of the compound of formula (II) was weighed into a 4.0 ml glass vial, and an appropriate amount of methyl tert-butyl ether was added to form a suspension. After adding the magneton, the above suspension sample was placed on a magnetic heating stirrer (40°C) for testing, stirred overnight at 40°C and centrifuged, and the residual solid sample was placed in a vacuum drying oven (30°C) to dry overnight to obtain Form A of the compound of formula (II).

100 mg of the compound of formula (II) was weighed into a 4.0 ml glass vial, and an appropriate amount of acetonitrile was added to form a suspension. After adding the magneton, the above suspension sample was placed on a magnetic heating stirrer (40°C) for testing, stirred overnight at 40°C and centrifuged, and the residual solid sample was placed in a vacuum drying oven (30°C) to dry overnight to obtain Form A of the compound of formula (II).

100 mg of the compound of formula (II) was weighed into a 4.0 ml glass vial, and an appropriate amount of n-heptane was added to form a suspension. After adding the magneton, the above suspension sample was placed on a magnetic heating stirrer (40°C) for testing, stirred overnight at 40°C and centrifuged, and the residual solid sample was placed in a vacuum drying oven (30°C) to dry overnight to obtain Form A of the compound of formula (II).

Example 6: Preparation of compound C crystal form of formula (I)

100 mg of compound B crystal form of formula (I) was weighed into a 4.0 ml glass vial, and an appropriate amount of methanol was added to form a suspension. After adding the magneton, the above suspension sample was placed on a magnetic heating stirrer (40°C) for testing, stirred overnight at 40°C and centrifuged, and the residual solid sample was placed in a vacuum drying oven (30°C) to dry overnight to obtain Form C of the compound of formula (I).

100 mg of compound B crystal form of formula (I) was weighed into a 4.0 ml glass vial, and an appropriate amount of acetone was added to form a suspension. After adding the magneton, the above suspension sample was placed on a magnetic heating stirrer (40°C) for testing, stirred overnight at 40°C and centrifuged, and the residual solid sample was placed in a vacuum drying oven (30°C) to dry overnight to obtain Form C of the compound of formula (I).

100 mg of compound B crystal form of formula (I) was weighed into a 4.0 ml glass vial, and an appropriate amount of isopropyl acetate was added to form a suspension. After adding the magneton, the above suspension sample was placed on a magnetic heating stirrer (40°C) for testing, stirred overnight at 40°C and centrifuged, and the residual solid sample was placed in a vacuum drying oven (30°C) to dry overnight to obtain Form C of the compound of formula (I).

10 liters of isopropyl acetate were added to the reaction kettle, and then 1.32 kg of the crystal form of compound B of formula (I) was added, and the mixture was stirred at 20 to 25° C. for 72 hours. Filtration, the filter cake was washed twice with 2 liters of acetone, and the filter cake was vacuum-dried to constant weight to obtain the C crystal form of the compound of formula (I).

MS-ESI calculated [M+H] ⁺ 679, found 679. ¹ H NMR (400MHz, CD ₃ OD) δ=8.60 (m, 1H), 8.40 (s, 1H), 7.65-7.77 (m, 2H), 7.54 (t, J=7.64Hz, 1H) 7.38-7.50 ( m, 2H), 7.15(m, 1H), 4.09(t, J=5.81Hz, 1H), 4.04(d, J=12.72Hz, 6H), 3.90(s, 2H), 3.74-3.83(m, 6H) ), 3.26(s, 3H), 3.19-3.25(m, 2H), 2.86(m, 4H), 2.76(t, J=5.87Hz, 2H).

Example 7: Solvent stability study of compound C crystal form of formula (I)

Weigh 100 mg of compound C of formula (I) crystal form into a 4.0 ml glass vial, add an appropriate amount of solvent to make it into a suspension (in order to ensure that the sample is suspended as much as possible, during the experiment, the compound amount and solvent will be adjusted according to the experimental phenomenon. amount or even change the container used in the experiment). After adding the magnetron, the above suspension sample was placed on a magnetic heating stirrer (40°C) for testing, stirred overnight at 40°C and centrifuged. (XRPD) to give a crystalline form of a solid. The test results are as follows:

Table 6

Conclusion: The crystal form of compound C of formula (I) has good stability in different solvents.

Example 8: Study on the hygroscopicity of the crystal form of compound C of formula (I)

Experimental Materials:

SMS DVS Advantage Dynamic Vapor Sorption Apparatus

experimental method:

10-30 mg of compound C crystal form of formula (I) was placed in a DVS sample tray for testing.

Experimental results:

The DVS spectrum of the crystal form of compound C of formula (I) is shown in Figure 10, ΔW=0.33%.

Experimental results:

The hygroscopic weight gain of the crystalline form of compound C of formula (I) at 25° C. and 80% RH is 0.33%, which is slightly hygroscopic.

Example 9: Solid stability test of compound B crystal form and C crystal form of formula (I)

According to "Guidelines for Stability Testing of Raw Materials and Preparations" (General Principles 9001 of Part Four of the Chinese Pharmacopoeia, 2015 edition), the crystal form A of Compound III was investigated at high temperature (60°C, open), high humidity (room temperature/relative humidity 92.5%, open). Stability under the conditions of illumination (total illuminance=1.2×10 ⁶ Lux·hr/near ultraviolet=200w·hr/m ² , open).

Weigh 10 mg of the compound B crystal form or C crystal form of the formula (I) respectively, place it at the bottom of the glass sample bottle, and spread it out into a thin layer. Samples placed under high temperature (60°C) and high humidity (relative humidity 92.5%RH) conditions are sealed with aluminum foil, and some small holes are made in the aluminum foil to ensure that the samples can be fully contacted with the ambient air, and placed in the corresponding In a constant temperature and humidity box; light samples (open, not covered with aluminum foil) and light reference (the whole sample bottle is covered with aluminum foil) are placed in the light box. Weigh 2 copies at each time point as the official test sample. In addition, about 50 mg of the B crystal form or C crystal form of the compound of formula (I) was weighed for XRPD test, and the sample bottle was wrapped with aluminum foil paper and punched with small holes, and also placed in the corresponding constant temperature and humidity box. Samples were taken on day 5 and day 10 for detection (XRPD), and the results were compared with the initial detection results on day 0. The solid stability results of the compound B crystal form or C crystal form test of the formula (I) are as follows:

Table 7

Conclusion: The crystal forms B and C of the compound of formula (I) have good stability under the conditions of high temperature, high humidity and strong light.

Experimental Example 1: PD-1/PD-L1 Homogeneous Time-Resolved Fluorescence Binding Experiment

Experimental principle:

Small molecule compounds can competitively inhibit the binding of PD-1 and PD-L1 by binding to PD-L1; when the PD-1 molecule as the donor is very close to the PD-L1 molecule as the acceptor, the donor molecule will The energy is transferred to the receptor molecule, which in turn causes the receptor molecule to emit fluorescence; by detecting the intensity of fluorescence, the ability of small molecules to prevent the binding of PD-L1 to PD-1 can be tested. Homogenouse Time-Resolved Fluorescence (HTRF) binding assay was used to detect the ability of the compounds of the present invention to inhibit the mutual binding of PD-1/PD-L1.

Experimental Materials:

PD-1/PD-L1TR-FRET detection kit was purchased from BPS Biosciences. Nivo Multilabel Analyzer (PerkinElmer).

experimental method:

Dilute PD1-Eu, Dye-labeled acceptor, PD-L1-biotin and test compound with the buffer in the kit. The compound to be tested was diluted 5 times to the 8th concentration with a row gun, that is, from 40 micromolar to 0.5 nanomolar, and the DMSO concentration was 4%, and a double-well experiment was set up. Add 5 micromolar inhibitor concentration gradients to the microplate, wherein the maximum (Max) signal well and the minimum (Min) signal well add 5 microliters of buffer containing 4% DMSO, 5 microliters of PD-L1-biotin (PD-L1-biotin) (60 nanomolar), add only 5 microliters of buffer to the smallest (Min) signal well, and incubate at 25 degrees for 20 minutes. After the incubation, 5 microliters of diluted PD1-Eu (10 nM) and 5 microliters of diluted dye-labeled acceptor (Dye-labeled acceptor) were added to each well. The reaction system was placed at 25 degrees for 90 minutes. After the reaction, the TR-FRET signal was read by a multi-label analyzer.

data analysis:

Using the equation (sample-Min)/(Max-Min)×100% to convert the raw data into inhibition rate, the IC ₅₀ value can be obtained by curve fitting with four parameters (log(inhibitor) vs.response in GraphPad Prism --Variable slope mode). Table 8 provides the inhibitory activity of compounds of the present invention on PD-1/PD-L1 binding.

Table 8 Test results of IC ₅₀ values of the compounds of the present invention for binding to PD-1/PD-L1

test compound _IC50 (nM) Formate salts of compounds of formula (I) 4.61

Experimental conclusion: The compound of formula (I) has a significant inhibitory effect on the binding of PD-1/PD-L1.

Experimental Example 2: Using MDA-MR-231 cells to detect the effect of compounds on the expression level of PD-L1

Experimental principle:

The use of a triple-negative breast cancer cell line (MDA-MB-231) is an indirect method to assess PD-L1 endocytosis. PD-L1 molecules on the cell surface can be degraded by lysosomal and proteasome pathways, and small molecule inhibitors are added to induce PD-L1 endocytosis. After co-incubating small molecules with MDA-MB-231 cells for 24 hours, flow cytometry (Fluorescence-activated Cell Sorting, FACS) was used to detect the content of PD-L1 on the cell surface, which can indirectly reflect the small molecule-induced PD-L1 endocytosis. Effect. Flow cytometry (FACS) was used to detect the effect of the compounds of the present invention on the expression level of PD-L1 in MDA-MR-231 cells.

Experimental Materials:

Phosphate buffered saline (DPBS), 1640 medium, penicillin-streptomycin, fetal bovine serum, non-essential amino acids, β-mercaptoethanol (2-ME), human interferon γ, LIVE/DEAD staining solution, staining solution (staining buffer), fixation buffer, 0.25% trypsin, EDTA, anti-human PD-L1 (Anti-human PD-L1), isotype control anti-human PD-L1 (Anti-human PD-L1Isotype) .

1640 complete medium configuration: 439.5 ml of 1640 medium was added with 50 ml of fetal bovine serum, 5 ml of non-essential amino acids, 5 ml of penicillin-streptomycin and 0.5 ml of β liter mercaptoethanol, and mixed.

10mM EDTA configuration: add 1ml of 0.5M EDTA to 49ml of DPBS and mix.

Experimental steps:

1) MDA-MB-231 cell counting and plating: remove the culture flask, remove the medium and rinse once with phosphate buffered saline (DPBS). After washing, add 3 ml of 0.25% trypsin to the culture flask and place it in a 37°C incubator for 1.5 minutes. Take out the culture flask and add 9 ml of 1640 complete medium to stop the reaction, transfer the cells to a 50 ml centrifuge tube, and centrifuge at 1000 rpm at 37°C for 5 min. Add an appropriate volume of culture medium to resuspend the cells according to the number of cells, and count with a cell counter. The cell concentration was adjusted to 5 x 10 ⁵ cells/ml with the medium. Plating: A volume of 200 microliters of cell suspension was added to each well of a 96-well plate, so that the number of cells in each well was 1×10 ⁵ . Incubate overnight in an incubator.

2) Drug incubation: prepare 100X compound diluent, and dilute the drug in a 5-fold gradient. Two microliters of each 100X compound solution was added to each well of cells. Incubate in a 37°C incubator for 24 hours. 3) PD-L1 cell staining and FACS detection: take out the culture plate and discard the supernatant medium. Wash once with 200 μl 1XPBS. Add 100 microliters of EDTA (final concentration of 10 mM) to 37°C for 10 min. After centrifugation at 1500 rpm for 5 min, wash once with 200 microliters of staining solution. Staining: Dilute anti-human PD-L1 (2 microliters per well) and LIVE/DEAD staining solution (1:1000) in staining solution, add 50 microliters to each well, and stain at 4°C for 30 min. Wash twice with 200 μl staining solution. Fixation: Add 100 microliters of fixative to each well, and fix at 4°C for 15 min. Wash twice with 200 μl staining solution. Resuspend cells in 150 µl. FACS detection. Table 9 shows the test results of the effect of the compounds of the present invention on the expression level of PD-L1 in MDA-MR-231 cells.

Table 9 Test results of the effect of the compounds of the present invention on the expression level of PD-L1 in MDA-MR-231 cells

test compound _IC50 (nM) Formate salts of compounds of formula (I) 5.85

Experimental conclusion: The compound of formula (I) has a significant inhibitory effect on the expression level of PD-L1 in MDA-MR-231 cells.

Experimental example 3: NFAT activity test

Experimental principle:

The engineered T cells express PD-1 molecule and T cell receptor (TCR) on the surface, and after co-culture with engineered antigen presenting cells (APC), the NFAT signaling pathway of T cells can be activated. Expression of PD-L1 molecules on APCs can effectively attenuate the NFAT signaling pathway in T cells; PD-L1 inhibitors can effectively block the PD-1/PD-L1 regulatory mechanism, thereby reversing the weakened NFAT signaling pathway. After pretreatment with APC, the small molecule was co-cultured with T cells, and then the expression of luciferase was detected to indirectly reflect the activation of the NFAT pathway in T cells.

Experimental Materials:

The PD-1/PD-L1NFAT detection kit was purchased from BPS Biosciences. Birght-Glo reagent was purchased from Promega. Nivo Multilabel Analyzer (PerkinElmer).

experimental method:

The TCR Activitor/PD-L1CHO cells with a growth confluence of 80% were spread into the plate at 35,000 cells per well and then placed in a 37°C cell incubator overnight; the compounds to be tested were diluted 5-fold to the 8th cell Concentrations, i.e. dilution from 20 micromolar to 0.25 nanomolar, DMSO concentration of 2%, set up double well experiments. Discard the supernatant of TCR Activitor/PD-L1CHO cells, add 50 microliters of compound working solution to each well, and incubate at 37°C for 30 minutes; after the incubation, add 50 microliters of PD-1/NFAT to each well at a density of 4×10 ⁵ /ml Reporter-Jurkat cell suspension was incubated at 37°C for 5 hours. After the incubation, 100 microliters of Bright-Glo was added to each well, and after mixing, the chemiluminescence signal was read using a Nivo multi-label analyzer.

data analysis:

Using the equation (sample-Min)/(Max-Min)×100% to convert the raw data into inhibition rate, the value of EC ₅₀ can be obtained by curve fitting with four parameters (log(inhibitor) vs.response in GraphPad Prism --Variable slope mode). Table 10 shows the test results of the degree of activation of the NFAT pathway in T cells by the compounds of the present invention.

Table 10 Test results of the compounds of the present invention on the degree of activation of NFAT pathway in T cells

test compound _EC50 (nM) Maximum effect (induction fold) Formate salts of compounds of formula (I) 41.45 5.05

Experimental conclusion: The compound of formula (I) can inhibit the interaction of PD-1/PD-L1 at the cellular level, thereby significantly activating the NFAT signaling pathway of T cells.

Experimental Example 4: Pharmacokinetic Testing

Objective: To study the pharmacokinetics of compounds in C57BL/6 mice

Experimental materials: C57BL/6 mice (male, 8 weeks old, body weight 25g-30g)

Experimental operation: The pharmacokinetic characteristics of rodents after intravenous (IV) and oral (PO) administration of the compounds were tested according to standard protocols. In the experiment, the candidate compounds were formulated into a 1 mg/ml clear solution and administered to mice after a single intravenous injection and Oral administration. Intravenous and oral vehicles are 5% DMSO/5% polyethylene glycol 15-hydroxystearate (Solutol)/90% aqueous solution. This project uses four male C57BL/6 mice, and two mice are administered intravenously at a dose of 1 mg/kg. 24h plasma samples, the other two mice were given oral gavage at a dose of 10 mg/kg, and the plasma samples were collected at 0.25, 0.5, 1, 2, 4, 6, 8, and 24 h after administration. Collect whole blood samples within 24 hours, centrifuge at 3000g for 15 minutes, separate the supernatant to obtain plasma samples, add acetonitrile solution containing internal standard to precipitate proteins, thoroughly mix and centrifuge to take the supernatant for injection, and quantify by LC-MS/MS analysis method Analyze blood drug concentrations and calculate pharmacokinetic parameters, such as peak concentration (C _max ), clearance (CL), half-life (T _1/2 ), tissue distribution (V _dss ), area under the drug-time curve (AUC _{0- last} ), bioavailability (F), etc.

The pharmacokinetic-related parameters of the compounds of the present invention in mice are shown in Table 11 below.

Table 11 Pharmacokinetic test results

compound Formate salts of compounds of formula (I) Peak Concentration C _max (nM) 370 Clearance CL (mL/min/kg) 18.5 Tissue distribution V _dss (L/kg) 36.8 Half-life T _1/2 (IV,h) 26.3 Area under the drug-time curve AUC _0-last PO (nM.hr) 5753 Bioavailability F(%) 80.8

Experimental conclusion: The compound of formula (I) has good pharmacokinetic properties, including good oral bioavailability, oral exposure, half-life and clearance rate.

Experimental Example 5: Pharmacodynamic evaluation of compounds in C57BL/6-hPDL1 mouse colorectal cancer MC38-hPDL1 subcutaneous transplantation model

OBJECTIVE: To evaluate the anti-tumor effect of the compounds on mouse colorectal cancer cells MC38-hPDL1 transplanted into humanized mice C57BL/6-hPDL1.

experimental design:

Table 12

Remarks: G: Group; N: Number of animals; p.o.: oral administration; i.p.: intraperitoneal injection; BID: twice a day; Q3D: once every three days; QD: once a day.

Experimental animals:

Table 13

species mouse strain C57BL/6-hPDL1 level SPF grade Zhou age 4.29～5.71 gender female

experimental method:

1. Tumor Cell Inoculation

Experimental cells: mouse colon cancer cells MC38-hPDL1 were recovered, and the recovery time was Pn+6. On the day of inoculation, MC38-hPDL1 cells in logarithmic growth phase were collected, the culture medium was removed, and the cells were washed twice with PBS before inoculation (the survival rates of MC38-hPDL1 cells before and after tumor bearing were: 98.6% and 98.0%, respectively). Amount: 1×106/100μL/mouse, inoculation location: right forelimb of mice.

2. Group administration

On the 7th day after inoculation, when the average tumor volume reached 85.81 mm ³ , 48 mice were randomly divided into 6 groups of 8 mice according to the tumor volume. The day of grouping was defined as D0 day, and the administration started on D0 day.

3. Drug preparation

Dosing volume: adjusted according to mouse body weight (mice dosing volume = 10 μL/g × mouse body weight (g))

4. Experimental observation and data collection

After the start of administration, the tumor size was observed and the mice were weighed on days D0, D3, D5, D7, D10, D12, D14, D17, D19, D21, and D24. The tumor volume was calculated as follows: tumor volume (mm3)=0.5×(longer diameter of tumor×shorter diameter of tumor2).

5. Experiment end point

At the end of the experiment, the following indicators were analyzed: 1) tumor volume change (TGItv); 2) average body weight change; TGITV (relative tumor inhibition rate) calculation formula:

TGItv(%)=[1-(meanTVtn-meanTVt0)/(meanTVvn-mean TVv0)]×100%

meanTVtn: the mean tumor volume of a given group when measured on day n

meanTVt0: the mean tumor volume of a given group when measured on day 0

meanTVvn: mean tumor volume of the solvent control group when measured on day n

mean TVv0: mean tumor volume of the solvent control group measured on day 0

Experimental results:

The effects of the compounds of the present invention on the tumor-inhibitory efficacy evaluation (calculated based on the tumor volume on the 24th day after administration) and the average body weight of mice in the colorectal cancer MC38-hPDL1 subcutaneous transplantation model of C57BL/6-hPDL1 mice are respectively as follows: Tables 14 and 15 show:

Table 14

Remarks: Data are expressed as mean±SEM.

Table 15

G Average body weight (g) before administration (day 0) Average body weight on the 24th day of administration (g) Vehicle (blank group) 18.29±0.27 20.01±0.35 Formate salts of compounds of formula (I) 18.08±0.32 18.91±0.38 Formate salts of compounds of formula (I) 18.44±0.33 18.94±0.50 Formate salts of compounds of formula (I) 18.64±0.28 19.41±0.43 Formate salts of compounds of formula (I) 18.89±0.33 20.48±0.38

Remarks: Data are expressed as mean±SEM.

Experimental conclusion: The compound of the present invention has excellent tumor-inhibiting effect on the colorectal cancer MC38-hPDL1 subcutaneous transplantation model of C57BL/6-hPDL1 mice, and the animal body weight does not decrease significantly during the administration process, and the tolerance is good.

Claims (28)

Form A of the compound of formula (II)

Its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 4.19±0.20°, 7.77±0.20° and 16.22±0.20°. The crystal form A according to claim 1, wherein the X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 4.19±0.20°, 7.77±0.20°, 8.71±0.20°, 9.89±0.20° and 16.22±0.20 °. The crystal form A according to claim 2, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 4.19±0.20°, 7.77±0.20°, 8.71±0.20°, 9.89±0.20°, 16.22±0.20 °, 17.74±0.20°, 22.82±0.20° and 25.49±0.20°. The crystal form A according to claim 3, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 4.19°, 7.77°, 8.71°, 9.89°, 11.47°, 14.41°, 16.22°, 17.74° , 19.67°, 20.98°, 22.82°, 25.49° and 28.15°. Form A according to claim 4, its XRPD pattern is shown in Figure 1. The crystal form A according to any one of claims 1 to 5, wherein the differential scanning calorimetry curve has the onset points of endothermic peaks at 31.2°C±5°C and 148.7°C±5°C. Form A according to claim 6, its DSC spectrum is shown in Figure 2. The crystal form A according to any one of claims 1 to 5 has a thermogravimetric analysis curve of 1.77% weight loss at 130°C±3°C, and a weight loss of 3.32% at 160°C±3°C. Form A according to claim 8, its TGA spectrum is shown in Figure 3. Form B of the compound of formula (I)

Its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 6.52±0.20°, 9.40±0.20° and 14.94±0.20°. The crystal form B according to claim 10, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 6.52±0.20°, 9.40±0.20°, 14.94±0.20°, 16.10±0.20° and 22.86±0.20 °. The crystal form B according to claim 11, wherein the X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 6.52±0.20°, 9.40±0.20°, 12.93±0.20°, 14.94±0.20°, 16.10±0.20 °, 19.06±0.20°, 19.89±0.20° and 22.86±0.20°. The crystal form B according to claim 12, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 6.52°, 7.47°, 9.40°, 11.29°, 12.93°, 13.90°, 14.94°, 16.10° , 19.06°, 19.89°, 21.13°, 22.86°, 24.55°, 25.25°, 25.83°, 26.23°, 27.45° and 34.58°. The crystal form B according to claim 13, its XRPD pattern is shown in Figure 4. The crystal form B according to any one of claims 10 to 14, wherein the differential scanning calorimetry curve has an onset of an endothermic peak at 145.2°C±5°C. The crystal form B according to claim 15, its DSC spectrum is shown in Figure 5. According to the crystal form B according to any one of claims 10 to 14, the thermogravimetric analysis curve of the thermogravimetric analysis curve has a weight loss of 0.77% at 150°C±3°C. The crystal form B according to claim 17, its TGA spectrum is shown in Figure 6. Form C of the compound of formula (I)

Its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 15.15±0.20°, 17.50±0.20° and 25.04±0.20°. The crystal form C according to claim 19, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 8.75±0.20°, 15.15±0.20°, 17.50±0.20°, 22.94±0.20° and 25.04±0.20 °. The crystal form C according to claim 20, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 8.75±0.20°, 15.15±0.20°, 17.50±0.20°, 18.77±0.20°, 22.94±0.20 °, 25.04±0.20°, 26.20±0.20°, 37.65±0.20°. The crystal form C according to claim 21, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 8.39°, 8.75°, 11.32°, 12.55°, 13.74°, 14.07°, 15.15°, 17.03° , 17.28°, 17.50°, 18.07°, 18.77°, 19.70°, 20.35°, 21.56°, 21.83°, 22.54°, 22.94°, 23.33°, 23.74°, 24.56°, 25.04°, 25.33°, 26.20°, 27.24 °, 27.93°, 29.33°, 30.75°, 31.39°, 32.44°, 33.01°, 34.07°, 36.32°, 37.65°, 38.01°. The crystal form C according to claim 22, its XRPD pattern is shown in Figure 7. The crystal form C according to any one of claims 19 to 23, wherein the differential scanning calorimetry curve has an onset of an endothermic peak at 184.5°C±5°C. Form C according to claim 24, its DSC spectrum is shown in Figure 8. According to the crystal form C according to any one of claims 19 to 23, the thermogravimetric analysis curve of the thermogravimetric analysis curve has a weight loss of 0.69% at 150°C±3°C. The crystal form C according to claim 26, its TGA spectrum is shown in Figure 9. The crystal form A according to any one of claims 1 to 9 or the crystal form B according to any one of claims 10 to 18 or the crystal form C according to any one of claims 19 to 27 is used in the preparation of application in oncology drugs.

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