WO2023040876A1 - Azaaromatic compound, polymorph of pharmaceutically acceptable salt thereof, pharmaceutical composition and application - Google Patents

Abstract

An azaaromatic compound, a polymorph of a pharmaceutically acceptable salt thereof, a pharmaceutical composition comprising same, a preparation method therefor, and an application of the polymorph or the pharmaceutical composition comprising same in the preparation of a medicine for preventing or treating cancer or immune-related diseases mediated by an adenosine A2A receptor and/or an adenosine A2B receptor.

Description

Polymorphs, pharmaceutical compositions and applications of azaaromatic compounds and pharmaceutically acceptable salts thereof

related application

This application requires an application filed on September 15, 2021. The application number is No. 202111080394.2, and the name is "Polymorphic Forms, Pharmaceutical Compositions and Applications of Azaaromatic Compounds and Their Pharmaceutically Acceptable Salts". Priority of Patent Application, which is hereby incorporated by reference in its entirety.

technical field

This application relates to the field of medical technology, in particular to an azaaromatic compound 3-(4-amino-6-(1-((1-ethyl-1H-pyrazol-3-yl)methyl)-1H -1,2,3-triazol-4-yl)-5-fluoropyrimidin-2-yl)-2-methylbenzonitrile polymorphs or pharmaceutically acceptable salts thereof, and polymorphs comprising the polymorph The pharmaceutical composition of the crystal form, the preparation method and application of the polymorph and the pharmaceutical composition.

Background technique

Adenosine (Adenosine) is an endogenous nucleoside throughout human cells, composed of adenine and ribose, which is widely distributed inside and outside cells. Adenosine participates in a variety of physiological and biochemical functions in the body. For example, adenosine can directly enter the myocardium and be phosphorylated to generate adenosine triphosphate (ATP), which participates in the energy metabolism of the myocardium. In the central nervous system (Central Nervous System, CNS), adenosine controls the release of neurotransmitters and the response of postsynaptic neurons, and plays an important role in regulating movement, protecting neurons, affecting sleep and wakefulness, etc. . In pathological conditions, the extracellular adenosine concentration increases significantly under tumor or hypoxic conditions. Adenosine can play an important role in tumor immunosuppression by promoting tumor angiogenesis, proliferation, development and tumor migration.

Adenosine receptor (Adenosine Receptor, AR) belongs to G protein-coupled receptor (Guanosine-binding Protein Coupled Receptor, GPCR) family, and its endogenous ligand is adenosine. The currently known adenosine receptors are composed of four subtype receptors, A ₁ , A _2A , A _2B and A ₃ . Among them, the combination of adenosine with _A1 or _A3 receptors can inhibit the production of cyclic adenosine monophosphate (cAMP); while the combination with _A2A or _A2B receptors can activate adenosine activating enzyme, and then up-regulate the level of cAMP, and play a further role. Physiological regulation.

A ₁ and A ₃ receptors are mainly expressed in the central nervous system, while A _2A and A _2B adenosine receptors are expressed in both the central nervous system and the peripheral system. In the tumor microenvironment, two adenosine receptors, _A2A and _A2B , are widely expressed in immune cells and have strong immunosuppressive functions. The increase of extracellular adenosine concentration is one of the important mechanisms of immune escape of tumor cells, and its concentration level is jointly determined by the level of ATP and the expression levels of CD39 and CD73. The increase in extracellular adenosine concentration is related to the release of large amounts of ATP by cell death or hypoxia in the tumor microenvironment, and its concentration can reach 10-20 times that of normal tissues. Adenosine binds to adenosine receptors in the tumor microenvironment, which can inhibit anti-tumor responses, such as inhibiting the function of CD8+ T cells, enhancing the function of immunosuppressive regulatory T cells, and inhibiting the function of antigen-presenting cells through dendritic cells wait. Recent studies have shown that binding to _A2A receptors can also inhibit the tumor-killing effect of natural killer cells. Further studies have shown that _A2A adenosine receptor antagonists can improve the activity and killing ability of dendritic antigen-presenting cells, T cells and natural killer cells, and inhibit regulatory T cells (T-regs), bone marrow-derived suppressive Cells (MDSCs) and tumor-associated macrophages (TAMs) eliminate tumor immune tolerance and promote the occurrence of tumor immune responses, which in turn lead to suppressed tumor growth and prolong the survival of mice. In addition, _A2B receptors have also been reported to promote tumor migration in mouse melanoma and triple-negative breast cancer models, so _A2B receptor antagonists are also effective targets for cancer therapy. Therefore, blocking the activation of adenosine signaling pathway to reduce or relieve immunosuppression and enhance the anti-tumor function of immune cells—especially T cells is considered to be one of the effective methods for cancer treatment. The _A2A / _A2B dual receptor antagonist is used to simultaneously block the activation of these two receptors. Mechanistically speaking, it is regulating different immune cell populations and comprehensively blocking the immune response brought about by adenosine in the microenvironment. Inhibitory effect has far-reaching clinical application value for tumor therapy.

In order to better meet market demand, further develop compounds with A _2A /A _2B receptor antagonistic activity, pharmaceutically acceptable salts and polymorphs thereof to facilitate further drug development.

Contents of the invention

The present application provides a polymorph of an azaaromatic compound represented by formula (X) or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing it, a preparation method or application thereof,

The pharmaceutically acceptable salt is selected from: hydrochloride, sulfate, citrate, fumarate or succinate.

In some embodiments, the polymorphs of the compound of Formula X or the polymorphs of the pharmaceutically acceptable salt of the compound of Formula X are each independently in an anhydrous form, a hydrate form, or a solvate form.

Each of the X-ray powder diffraction patterns involved in the present application can be obtained independently using Cu-Kα radiation.

In the first aspect of the present application, the type I crystal form of the compound represented by formula (X) is provided, that is, the free base crystal form I, and its X-ray powder diffraction pattern has a characteristic diffraction peak at the following 2θ (°) angle: 7.57 ± 0.2 , 13.41±0.2, 14.64±0.2, 19.81±0.2, and 24.43±0.2.

In some embodiments, the X-ray powder diffraction pattern of the free base crystal form I further includes one or two characteristic diffraction peaks selected from the following 2θ (°) angles: 24.87±0.2 and 27.46±0.2.

In some embodiments, the X-ray powder diffraction pattern of the free base crystal form I further includes 2 or more characteristic diffraction peaks selected from the following 2θ (°) angles: 8.56±0.2, 15.40±0.2, 18.55 ±0.2, 21.25±0.2, 23.05±0.2, 23.59±0.2, and 25.60±0.2.

In some embodiments, the X-ray powder diffraction pattern of the free base crystal form I has characteristic diffraction peaks at the following 2θ (°) angles: 7.57±0.2, 8.56±0.2, 10.60±0.2, 11.80±0.2, 13.41±0.2 0.2, 14.16±0.2, 14.64±0.2, 15.40±0.2, 16.63±0.2, 17.23±0.2, 18.55±0.2, 19.81±0.2, 21.25±0.2, 22.18±0.2, 22.46±0.2, 23.05±0.2, 23.59±0.2, 24.43±0.2, 24.87±0.2, 25.60±0.2, 26.03±0.2, and 27.46±0.2.

In some embodiments, the X-ray powder diffraction pattern of the free base crystal form I has diffraction peaks at the 2θ (°) values shown in Table 1, and the relative intensities of each diffraction peak are shown in Table 1. Among them, the definition of each relative intensity symbol is shown in Table 12.

Table 1

2θ(°)	I/I 0	2θ(°)	I/I 0	2θ(°)	I/I 0
7.57	vs.	16.63	W	23.59	m
8.56	m	17.23	W	24.43	S
10.60	W	18.55	m	24.87	S
11.80	W	19.81	S	25.60	m
13.41	S	21.25	m	26.03	W
14.16	W	22.18	W	27.46	S
14.64	S	22.46	W	-	-
15.40	m	23.05	m	-	-

In some embodiments, the X-ray powder diffraction pattern of the free base crystalline form I is substantially as shown in FIG. 1 .

In some embodiments, the X-ray powder diffraction pattern of the free base crystalline form I is shown in FIG. 1 .

In some embodiments, the differential scanning calorimetry curve of the free base crystalline form I has an endothermic peak at 179.10±3°C.

In some embodiments, the differential scanning calorimetry curve of the free base crystalline form I has an endothermic peak at 179.10±2°C.

In some embodiments, the differential scanning calorimetry curve of the free base crystalline form I has an endothermic peak at 179.10±0.5°C.

In some embodiments, the differential scanning calorimetry curve of the free base Form I is substantially as shown in FIG. 2 .

In some embodiments, the differential scanning calorimetry curve of the free base crystalline form I is shown in FIG. 2 .

In the second aspect of the present application, a method for preparing free base crystal form I is provided, comprising the following steps:

The compound of formula (X) is mixed with the I-3 solvent, heated to dissolve at a temperature of 45°C to 85°C, cooled to 0°C to room temperature, separated to obtain the I-3 solid, and

drying the I-3 solid to obtain the free base crystal form I;

Wherein said 1-3 solvent is methanol or ethanol.

In some specific embodiments, the preparation method of free base crystal form I comprises the following steps:

The compound of formula (X) is mixed with the I-3 solvent, heated to dissolve at a temperature of 45°C to 55°C, cooled to 0°C to 4°C, separated to obtain the I-3 solid, and

drying the I-3 solid to obtain the free base crystal form I;

Wherein said 1-3 solvent is methanol or ethanol.

In some specific embodiments, the preparation method of free base crystal form I comprises the following steps:

Mixing the compound of formula (X) with the solvent I-3A, heating to dissolve at a temperature of 70° C. to 80° C., cooling to room temperature, and separating to obtain the I-3 solid, and

drying the I-3 solid to obtain the free base crystal form I;

Wherein said I-3A solvent is ethanol.

In some specific embodiments, the preparation method of free base crystal form I comprises the following steps:

The compound of formula (X) is mixed with the I-3B solvent, heated to dissolve at a temperature of 60° C. to 70° C., cooled to room temperature, and separated to obtain the I-3 solid, and

drying the I-3 solid to obtain the free base crystal form I;

Wherein said I-3B solvent is methanol.

The third aspect of the present application provides the type IV crystal form of the compound of formula (X), that is, the free base crystal form IV, whose X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ (°) angles: 11.88±0.2, 14.17 ±0.2, 16.96±0.2, 22.63±0.2, 23.56±0.2, and 25.66±0.2.

In some embodiments, the X-ray powder diffraction pattern of the free base crystal form IV further includes at least one characteristic diffraction peak selected from the following 2θ (°) angles: 15.76±0.2 and 26.08±0.2.

In some embodiments, the X-ray powder diffraction pattern of the free base crystal form IV further includes at least one characteristic diffraction peak selected from the following 2θ (°) angles: 15.76±0.2, 26.08±0.2 and 37.48±0.2.

In some embodiments, the X-ray powder diffraction pattern of the free base crystal form IV further includes 2 or more characteristic diffraction peaks selected from the following 2θ (°) angles: 14.89±0.2, 21.73±0.2, 24.43 ±0.2, 26.65±0.2, 27.39±0.2, 28.42±0.2, and 30.24±0.2.

In some embodiments, the X-ray powder diffraction pattern of the free base crystal form IV further includes at least one characteristic diffraction peak selected from the following 2θ (°) angles: 8.71±0.2, 19.93±0.2 and 31.78±0.2.

In some embodiments, the X-ray powder diffraction pattern of the free base crystal form IV has characteristic diffraction peaks at the following 2θ (°) angles: 8.71±0.2, 9.57±0.2, 11.88±0.2, 14.17±0.2, 14.89±0.2 0.2, 15.76±0.2, 16.96±0.2, 19.93±0.2, 21.73±0.2, 22.63±0.2, 23.56±0.2, 24.43±0.2, 25.66±0.2, 26.08±0.2, 26.65±0.2, 27.39±0.2, 28.42±0.2, 30.24±0.2, 31.78±0.2, and 33.15±0.2.

In some embodiments, the X-ray powder diffraction pattern of the free base crystal form IV has characteristic diffraction peaks at the following 2θ (°) angles: 8.71±0.2, 9.57±0.2, 11.88±0.2, 14.17±0.2, 14.89±0.2 0.2, 15.76±0.2, 16.96±0.2, 19.93±0.2, 21.73±0.2, 22.63±0.2, 23.56±0.2, 24.43±0.2, 25.66±0.2, 26.08±0.2, 26.65±0.2, 27.39±0.2, 28.42±0.2, 30.24±0.2, 31.78±0.2, 33.15±0.2, and 37.48±0.2.

In some embodiments, the X-ray powder diffraction pattern of the free base crystal form IV has diffraction peaks at the 2θ (°) values shown in Table 2, and the relative intensity of each diffraction peak is shown in Table 2. Among them, the definition of each relative intensity symbol is shown in Table 12.

Table 2

2θ(°)	I/I 0	2θ(°)	I/I 0	2θ(°)	I/I 0
8.71	m	19.93	m	26.65	m
9.57	W	21.73	m	27.39	m
11.88	vs.	22.63	S	28.42	m
14.17	vs.	23.56	vs.	30.24	m
14.89	m	24.43	m	31.78	m

15.76	S	25.66	vs.	33.15	W
16.96	vs.	26.08	S	37.48	S

In some embodiments, the X-ray powder diffraction pattern of the free base crystalline Form IV is substantially as shown in FIG. 4 .

In some embodiments, the X-ray powder diffraction pattern of the free base crystalline form IV is shown in FIG. 4 .

In some embodiments, the differential scanning calorimetry curve of the free base crystalline form IV has an endothermic peak at 170.43±3°C.

In some embodiments, the differential scanning calorimetry curve of the free base crystalline form IV has an endothermic peak at 170.43±2°C.

In some embodiments, the differential scanning calorimetry curve of the free base crystalline form IV has an endothermic peak at 170.43±0.5°C.

In some embodiments, the differential scanning calorimetry curve of the free base crystalline Form IV is substantially as shown in FIG. 5 .

In some embodiments, the differential scanning calorimetry curve of the free base crystalline Form IV is shown in FIG. 5 .

In the fourth aspect of the present application, a method for preparing free base crystal form IV is provided, comprising the following steps:

Mix the free base crystal form I described in the first aspect of the present application with the IV-1 solvent, suspend and shake at room temperature for 20 to 30 hours, and separate to obtain the IV-1 solid. The IV- 1. drying the solid to obtain the free base crystalline form IV; wherein the IV-1 solvent is isopropanol; or

Mix the free base crystal form I described in the first aspect of the present application with solvent IV-2, suspend and shake at room temperature for 160 to 170 hours, and separate to obtain solid IV-2. 2. Drying the solid to obtain the free base crystal form IV; wherein the IV-2 solvent is isopropanol or water.

In the fifth aspect of the present application, the V-type crystal form of the compound of formula (X) is provided, that is, the free base crystal form V, and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ (°) angles: 11.88±0.2, 16.36 ±0.2, 16.99±0.2, 22.99±0.2, 23.47±0.2, and 26.41±0.2.

In some embodiments, the X-ray powder diffraction pattern of the free base crystal form V has characteristic diffraction peaks at the following 2θ (°) angles: 11.88±0.2, 16.36±0.2, 16.99±0.2, 22.99±0.2, 23.47± 0.2, 26.41±0.2 and 37.48±0.2.

In some embodiments, the X-ray powder diffraction pattern of the free base crystal form V further includes at least one characteristic diffraction peak selected from the following 2θ (°) angles: 7.90±0.2, 13.71±0.2, 14.14±0.2, 15.31 ±0.2, 17.80±0.2, 18.76±0.2, 21.34±0.2, 24.82±0.2, 25.51±0.2, 27.88±0.2, and 30.46±0.2.

In some embodiments, the X-ray powder diffraction pattern of the free base crystal form V further includes at least one characteristic diffraction peak selected from the following 2θ (°) angles: 7.90±0.2, 13.71±0.2, 14.14±0.2, 15.31 ±0.2, 17.80±0.2, 18.76±0.2, 21.34±0.2, 24.82±0.2, 25.51±0.2, 27.88±0.2, 30.46±0.2, and 43.60±0.2.

In some embodiments, the X-ray powder diffraction pattern of the free base crystal form V further includes 2 or more characteristic diffraction peaks selected from the following 2θ (°) angles: 15.77±0.2, 20.03±0.2, 28.40 ±0.2, 30.85±0.2, 32.71±0.2, 33.15±0.2, and 34.62±0.2.

In some embodiments, the X-ray powder diffraction pattern of the free base crystal form V has characteristic diffraction peaks at the following 2θ (°) angles: 7.90±0.2, 8.62±0.2, 9.73±0.2, 11.88±0.2, 13.71±0.2 0.2, 14.14±0.2, 15.31±0.2, 15.77±0.2, 16.36±0.2, 16.99±0.2, 17.80±0.2, 18.76±0.2, 20.03±0.2, 21.34±0.2, 22.99±0.2, 23.47±0.2, 24.82±0.2, 25.51±0.2, 26.41±0.2, 27.88±0.2, 28.40±0.2, 30.46±0.2, 30.85±0.2, 32.71±0.2, 33.15±0.2, and 34.62±0.2.

In some embodiments, the X-ray powder diffraction pattern of the free base crystal form V has characteristic diffraction peaks at the following 2θ (°) angles: 7.90±0.2, 8.62±0.2, 9.73±0.2, 11.88±0.2, 13.71±0.2 0.2, 14.14±0.2, 15.31±0.2, 15.77±0.2, 16.36±0.2, 16.99±0.2, 17.80±0.2, 18.76±0.2, 20.03±0.2, 21.34±0.2, 22.99±0.2, 23.47±0.2, 24.82±0.2, 25.51±0.2, 26.41±0.2, 27.88±0.2, 28.40±0.2, 30.46±0.2, 30.85±0.2, 32.71±0.2, 33.15±0.2, 34.62±0.2, 37.48±0.2, and 43.60±0.2.

In some embodiments, the X-ray powder diffraction pattern of the free base crystal form V has diffraction peaks at the 2θ (°) values shown in Table 3, and the relative intensity of each diffraction peak is shown in Table 3. Among them, the definition of each relative intensity symbol is shown in Table 12.

table 3

2θ(°)	I/I 0	2θ(°)	I/I 0	2θ(°)	I/I 0
7.90	S	17.80	S	28.40	m
8.62	W	18.76	S	30.46	S
9.73	W	20.03	m	30.85	m

11.88	vs.	21.34	S	32.71	m
13.71	S	22.99	vs.	33.15	m
14.14	S	23.47	vs.	34.62	m
15.31	S	24.82	S	37.48	vs.
15.77	m	25.51	S	43.60	S
16.36	vs.	26.41	vs.	-	-
16.99	vs.	27.88	S	-	-

In some embodiments, the X-ray powder diffraction pattern of the free base Form V is substantially as shown in FIG. 6 .

In some embodiments, the X-ray powder diffraction pattern of the free base form V is shown in FIG. 6 .

In some embodiments, the differential scanning calorimetry curve of the free base crystal form V has an endothermic peak at 179.02±3°C.

In some embodiments, the differential scanning calorimetry curve of the free base crystalline form V has an endothermic peak at 179.02±2°C.

In some embodiments, the differential scanning calorimetry curve of the free base form V has an endothermic peak at 179.02±0.5°C.

In some embodiments, the differential scanning calorimetry curve of the free base Form V is substantially as shown in FIG. 7 .

In some embodiments, the differential scanning calorimetry curve of the free base Form V is shown in FIG. 7 .

In the sixth aspect of the present application, a method for preparing free base crystal form V is provided, comprising the following steps:

Mix the free base crystal form I described in the first aspect of the present application with the V-1 solvent, suspend and shake at room temperature for 160 to 170 hours, and separate to obtain the V-1 solid, and the V- 1. Dry the solid to obtain the free base crystal form V; wherein the V-1 solvent is methyl tert-butyl ether.

Mix and dissolve the compound of formula (X) described in the first aspect of the present application with the V-2 solvent, place it at room temperature, let the V-2 solvent evaporate to dryness, and obtain the free base crystal form V ; Wherein the V-2 solvent is acetone.

The seventh aspect of the present application provides the VIII crystal form of the compound of formula (X), that is, the free base crystal form VIII, whose X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ (°) angles: 6.40±0.2, 13.12 ±0.2 and 15.91±0.2.

In some embodiments, the X-ray powder diffraction pattern of the free base crystal form VIII further includes one or two characteristic diffraction peaks selected from the following 2θ (°) angles: 22.69±0.2 and 26.95±0.2.

In some embodiments, the X-ray powder diffraction pattern of the free base crystal form VIII further includes at least one characteristic diffraction peak selected from the following 2θ (°) angles: 11.62±0.2, 12.36±0.2, 16.62±0.2, 18.34±0.2, 18.88±0.2, 24.94±0.2, and 29.65±0.2.

In some embodiments, the X-ray powder diffraction pattern of the free base crystal form VIII further includes at least one characteristic diffraction peak selected from the following 2θ (°) angles: 11.62±0.2, 12.36±0.2, 16.62±0.2, 18.34±0.2, 18.88±0.2, 24.94±0.2, 29.65±0.2, and 37.48±0.2.

In some embodiments, the X-ray powder diffraction pattern of the free base crystal form VIII has characteristic diffraction peaks at the following 2θ (°) angles: 6.40±0.2, 11.62±0.2, 12.36±0.2, 13.12±0.2, 15.91±0.2 0.2, 16.62±0.2, 18.34±0.2, 18.88±0.2, 22.69±0.2, 24.94±0.2, 26.95±0.2, and 29.65±0.2.

In some embodiments, the X-ray powder diffraction pattern of the free base crystal form VIII has characteristic diffraction peaks at the following 2θ (°) angles: 6.40±0.2, 11.62±0.2, 12.36±0.2, 13.12±0.2, 15.91±0.2 0.2, 16.62±0.2, 18.34±0.2, 18.88±0.2, 22.69±0.2, 24.94±0.2, 26.95±0.2, 29.65±0.2, and 37.48±0.2.

In some embodiments, the X-ray powder diffraction pattern of the free base crystal form VIII has diffraction peaks at the 2θ (°) values shown in Table 4, and the relative intensities of each diffraction peak are shown in Table 4. Among them, the definition of each relative intensity symbol is shown in Table 12.

Table 4

2θ(°)	I/I 0	2θ(°)	I/I 0	2θ(°)	I/I 0
6.40	vs.	16.62	m	26.95	m
11.62	m	18.34	m	29.65	m
12.36	m	18.88	m	37.48	m
13.12	vs.	22.69	m	-	-

15.91

S

24.94

m

-

-

In some embodiments, the X-ray powder diffraction pattern of the free base Form VIII is substantially as shown in FIG. 8 .

In some embodiments, the X-ray powder diffraction pattern of the free base form VIII is shown in FIG. 8 .

In some embodiments, the differential scanning calorimetry curve of the free base crystalline form VIII has an endothermic peak at 162.93±3°C.

In some embodiments, the differential scanning calorimetry curve of the free base crystalline form VIII has an endothermic peak at 162.93±2°C.

In some embodiments, the differential scanning calorimetry curve of the free base crystalline form VIII has an endothermic peak at 162.93±0.5°C.

In some embodiments, the differential scanning calorimetry curve of the free base Form VIII is substantially as shown in FIG. 9 .

In some embodiments, the differential scanning calorimetry curve of the free base crystalline Form VIII is shown in FIG. 9 .

The eighth aspect of the present application provides a method for preparing free base crystal form VIII, comprising the following steps:

Mix the free base crystal form I, free base crystal form IV or free base crystal form V described in the above aspects with the Z ₃ -1 solvent, suspend and shake at room temperature for 70 to 80 hours, and separate to obtain the second Z ₃ -1 solid, drying the Z ₃ -1 solid to obtain the free base crystal form VIII; wherein the Z ₃ -1 solvent is acetone or ethanol aqueous solution with a volume fraction of 50%.

In the ninth aspect of the present application, the hydrochloride salt crystal form I of the compound of formula (X) is provided, and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ (°) angles: 11.95±0.2, 24.37±0.2, 25.21± 0.2 and 26.35±0.2.

In some embodiments, the X-ray powder diffraction pattern of the hydrochloride salt form I also includes at least one characteristic diffraction peak selected from the following 2θ (°) angles: 8.83±0.2, 14.13±0.2, 15.33±0.2, 15.68±0.2, 18.86±0.2, 23.64±0.2, 27.67±0.2, and 28.57±0.2.

In some embodiments, the X-ray powder diffraction pattern of the hydrochloride salt form I has characteristic diffraction peaks at the following 2θ (°) angles: 8.83±0.2, 11.95±0.2, 14.13±0.2, 15.33±0.2, 15.68 ±0.2, 17.25±0.2, 18.86±0.2, 23.64±0.2, 24.37±0.2, 25.21±0.2, 26.35±0.2, 27.67±0.2, and 28.57±0.2.

In some embodiments, the X-ray powder diffraction pattern of the hydrochloride salt form I has diffraction peaks at the 2θ (°) values shown in Table 5, and the relative intensity of each diffraction peak is shown in Table 5. Among them, the definition of each relative intensity symbol is shown in Table 12.

table 5

2θ(°)	I/I 0	2θ(°)	I/I 0	2θ(°)	I/I 0
8.83	S	17.25	m	26.35	vs.
11.95	vs.	18.86	S	27.67	S
14.13	S	23.64	S	28.57	S
15.33	S	24.37	vs.	-	-
15.68	S	25.21	vs.	-	-

In some embodiments, the X-ray powder diffraction pattern of the hydrochloride salt form I is substantially as shown in FIG. 10 .

In some embodiments, the X-ray powder diffraction pattern of the hydrochloride salt form I is shown in FIG. 10 .

In the tenth aspect of the present application, there is provided sulfate crystal form I of the compound of formula (X),

Its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ (°) angles: 8.22±0.2, 11.29±0.2, 14.14±0.2, 16.96±0.2, 18.13±0.2, 22.27±0.2, 23.86±0.2 and 27.79±0.2 .

In some embodiments, the X-ray powder diffraction pattern of the sulfate crystal form I further includes 2 or more characteristic diffraction peaks selected from the following 2θ (°) angles: 6.54±0.2 and 11.64±0.2.

In some embodiments, the X-ray powder diffraction pattern of the sulfate crystal form I further includes 2 or more characteristic diffraction peaks selected from the following 2θ (°) angles: 6.54±0.2, 11.64±0.2 and 37.48 ±0.2.

In some embodiments, the X-ray powder diffraction pattern of the sulfate crystal form I has characteristic diffraction peaks at the following 2θ (°) angles: 6.54±0.2, 8.22±0.2, 11.29±0.2, 11.64±0.2, 14.14±0.2 0.2, 16.96±0.2, 18.13±0.2, 22.27±0.2, 23.86±0.2, and 27.79±0.2.

In some embodiments, the X-ray powder diffraction pattern of the sulfate crystal form I has characteristic diffraction peaks at the following 2θ (°) angles: 6.54±0.2, 8.22±0.2, 11.29±0.2, 11.64±0.2, 14.14±0.2 0.2, 16.96±0.2, 18.13±0.2, 22.27±0.2, 23.86±0.2, 27.79±0.2, and 37.48±0.2.

In some embodiments, the X-ray powder diffraction pattern of the sulfate salt form I has diffraction peaks at the 2θ (°) values shown in Table 6, and the relative intensity of each diffraction peak is shown in Table 6. Among them, the definition of each relative intensity symbol is shown in Table 12.

Table 6

2θ(°)	I/I 0	2θ(°)	I/I 0	2θ(°)	I/I 0
6.54	m	14.14	S	23.86	S
8.22	S	16.96	vs.	27.79	S
11.29	S	18.13	S	37.48	m
11.64	m	22.27	S	-	-

In some embodiments, the X-ray powder diffraction pattern of the sulfate salt form I is substantially as shown in FIG. 11 .

In some embodiments, the X-ray powder diffraction pattern of the sulfate crystalline form I is shown in FIG. 11 .

In the eleventh aspect of the present application, the citrate crystal form I of the compound of formula (X) is provided, and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ (°) angles: 12.55±0.2, 14.50±0.2, 21.55 ±0.2 and 22.75±0.2.

In some embodiments, the X-ray powder diffraction pattern of the citrate crystal form I has characteristic diffraction peaks at the following 2θ (°) angles: 12.55±0.2, 14.50±0.2, 21.55±0.2, 22.75±0.2, 37.51 ±0.2 and 43.63±0.2.

In some embodiments, the X-ray powder diffraction pattern of the citrate crystal form I further includes at least one characteristic diffraction peak selected from the following 2θ (°) angles: 5.38±0.2, 9.78±0.2 and 13.63±0.2.

In some embodiments, the X-ray powder diffraction pattern of the citrate crystal form I has characteristic diffraction peaks at the following 2θ (°) angles: 5.38±0.2, 9.78±0.2, 12.55±0.2, 13.63±0.2, 14.50 ±0.2, 21.55±0.2, and 22.75±0.2.

In some embodiments, the X-ray powder diffraction pattern of the citrate crystal form I has characteristic diffraction peaks at the following 2θ (°) angles: 5.38±0.2, 9.78±0.2, 12.55±0.2, 13.63±0.2, 14.50 ±0.2, 21.55±0.2, 22.75±0.2, 37.51±0.2, and 43.63±0.2.

In some embodiments, the X-ray powder diffraction pattern of the citrate crystal form I has diffraction peaks at the 2θ (°) values shown in Table 7, and the relative intensity of each diffraction peak is shown in Table 7. Among them, the definition of each relative intensity symbol is shown in Table 12.

Table 7

2θ(°)	I/I 0	2θ(°)	I/I 0	2θ(°)	I/I 0
5.38	S	13.63	S	22.75	vs.
9.78	S	14.50	vs.	37.51	vs.
12.55	vs.	21.55	vs.	43.63	vs.

In some embodiments, the X-ray powder diffraction pattern of the citrate salt form I is substantially as shown in FIG. 12 .

In some embodiments, the X-ray powder diffraction pattern of the citrate salt form I is shown in FIG. 12 .

In the twelfth aspect of the present application, the fumarate salt crystal form I of the compound of formula (X) is provided, and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ (°) angles: 7.12 ± 0.2, 11.71 ± 0.2 and 16.90±0.2.

In some embodiments, the X-ray powder diffraction pattern of the fumarate salt form I also includes at least one characteristic diffraction peak selected from the following 2θ (°) angles: 23.26±0.2, 23.65±0.2, 24.64±0.2 and 25.54±0.2.

In some embodiments, the X-ray powder diffraction pattern of the fumarate salt form I also includes at least one characteristic diffraction peak selected from the following 2θ (°) angles: 8.02±0.2, 15.69±0.2, 18.22±0.2 and 26.44±0.2.

In some embodiments, the X-ray powder diffraction pattern of the fumarate salt form I also includes at least one characteristic diffraction peak selected from the following 2θ (°) angles: 8.02±0.2, 15.69±0.2, 18.22±0.2 , 26.44±0.2 and 37.87±0.2.

In some embodiments, the X-ray powder diffraction pattern of the fumarate salt form I also includes 2 or more characteristic diffraction peaks selected from the following 2θ (°) angles: 10.48±0.2, 13.80±0.2 , 22.00±0.2 and 22.42±0.2.

In some embodiments, the X-ray powder diffraction pattern of the fumarate salt form I has characteristic diffraction peaks at the following 2θ (°) angles: 7.12±0.2, 8.02±0.2, 10.48±0.2, 11.71±0.2 .

In some embodiments, the X-ray powder diffraction pattern of the fumarate salt form I has characteristic diffraction peaks at the following 2θ (°) angles: 7.12±0.2, 8.02±0.2, 10.48±0.2, 11.71±0.2 , 13.80±0.2, 15.69±0.2, 16.90±0.2, 18.22±0.2, 22.00±0.2, 22.42±0.2, 23.26±0.2, 23.65±0.2, 24.64±0.2, 25.54±0.2, 26.44±0.2, and 37.87±0.2.

In some embodiments, the X-ray powder diffraction pattern of the fumarate salt form I has diffraction peaks at the 2θ (°) values shown in Table 8, and the relative intensity of each diffraction peak is shown in Table 8. Among them, the definition of each relative intensity symbol is shown in Table 12.

Table 8

2θ(°)	I/I 0	2θ(°)	I/I 0	2θ(°)	I/I 0
7.12	vs.	16.90	vs.	24.64	vs.
8.02	S	18.22	S	25.54	vs.
10.48	m	22.00	m	26.44	S
11.71	vs.	22.42	m	37.87	S
13.80	m	23.26	vs.	-	-
15.69	S	23.65	vs.	-	-

In some embodiments, the X-ray powder diffraction pattern of the fumarate salt form I is substantially as shown in FIG. 13 .

In some embodiments, the X-ray powder diffraction pattern of the fumarate salt form I is shown in FIG. 13 .

In the thirteenth aspect of the present application, the succinate salt crystal form I of the compound of formula (X) is provided, and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ (°) angles: 13.78±0.2, 14.14±0.2 and 22.12 ±0.2.

In some embodiments, the X-ray powder diffraction pattern of succinate salt form I has characteristic diffraction peaks at the following 2θ (°) angles: 13.78±0.2, 14.14±0.2, 22.12±0.2 and 37.84±0.2.

In some embodiments, the X-ray powder diffraction pattern of the succinate salt form I further includes at least one characteristic diffraction peak selected from the following 2θ (°) angles: 4.87±0.2, 9.31±0.2, 18.16±0.2, 20.71±0.2 and 26.83±0.2.

In some embodiments, the X-ray powder diffraction pattern of the succinate salt form I has characteristic diffraction peaks at the following 2θ (°) angles: 4.87±0.2, 9.31±0.2, 13.78±0.2, 14.14±0.2, 15.69 ±0.2, 18.16±0.2, 20.71±0.2, 22.12±0.2, and 26.83±0.2.

In some embodiments, the X-ray powder diffraction pattern of the succinate salt form I has characteristic diffraction peaks at the following 2θ (°) angles:

4.87±0.2, 9.31±0.2, 13.78±0.2, 14.14±0.2, 15.69±0.2, 18.16±0.2, 20.71±0.2, 22.12±0.2, 26.83±0.2, 37.84±0.2, and 44.07±0.2.

In some embodiments, the X-ray powder diffraction pattern of the succinate salt form I has diffraction peaks at the 2θ (°) values shown in Table 9, and the relative intensity of each diffraction peak is shown in Table 9. Among them, the definition of each relative intensity symbol is shown in Table 12.

Table 9

2θ(°)	I/I 0	2θ(°)	I/I 0	2θ(°)	I/I 0
4.87	S	15.69	m	26.83	S
9.31	S	18.16	S	37.84	vs.
13.78	vs.	20.71	S	44.07	m
14.14	vs.	22.12	S	-	-

In some embodiments, the X-ray powder diffraction pattern of the succinic salt form I is substantially as shown in FIG. 14 .

In some embodiments, the X-ray powder diffraction pattern of the succinic salt crystal form I is shown in FIG. 14 .

The fourteenth aspect of the present application provides the succinate crystal form II of the compound of formula (X), whose X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ (°) angles: 12.49±0.2 and 14.50±0.2.

In some embodiments, the X-ray powder diffraction pattern of the succinate crystal form II also includes at least one characteristic diffraction peak selected from the following 2θ (°) angles: 11.86±0.2, 15.22±0.2, 16.90±0.2, 21.64±0.2, 22.87±0.2, and 25.75±0.2.

In some embodiments, the X-ray powder diffraction pattern of the succinate crystal form II also includes at least one characteristic diffraction peak selected from the following 2θ (°) angles: 11.86±0.2, 15.22±0.2, 16.90±0.2, 21.64±0.2, 22.87±0.2, 25.75±0.2, and 37.84±0.2.

In some embodiments, the X-ray powder diffraction pattern of the succinate crystal form II has characteristic diffraction peaks at the following 2θ (°) angles: 5.35±0.2, 11.86±0.2, 12.49±0.2, 14.50±0.2, 15.22 ±0.2, 16.90±0.2, 21.64±0.2, 22.87±0.2, and 25.75±0.2.

In some embodiments, the X-ray powder diffraction pattern of the succinate crystal form II has characteristic diffraction peaks at the following 2θ (°) angles: 5.35±0.2, 11.86±0.2, 12.49±0.2, 14.50±0.2, 15.22 ±0.2, 16.90±0.2, 21.64±0.2, 22.87±0.2, 25.75±0.2, and 37.84±0.2.

In some embodiments, the X-ray powder diffraction pattern of the succinate salt form II has diffraction peaks at the 2θ (°) values shown in Table 10, and the relative intensity of each diffraction peak is shown in Table 10. Among them, the definition of each relative intensity symbol is shown in Table 12.

Table 10

2θ(°)	I/I 0	2θ(°)	I/I 0	2θ(°)	I/I 0
5.35	m	15.22	S	25.75	S
11.86	S	16.90	S	37.84	S
12.49	vs.	21.64	S	-	-
14.50	vs.	22.87	S	-	-

In some embodiments, the X-ray powder diffraction pattern of the succinic salt form II is substantially as shown in FIG. 15 .

In some embodiments, the X-ray powder diffraction pattern of the succinate salt form II is shown in FIG. 15 .

In the fifteenth aspect of the present application, there is provided a method for preparing a polymorphic form of a pharmaceutically acceptable salt of a compound of formula (X), comprising the following steps:

Mix the above-mentioned compound of formula (X) with the acid according to the molar ratio of acid (hydrogen ion) and compound of formula (X) of 1.1 to 1.5:1, add 1 to 3 mL of solvent, and heat the solution with ultrasonic to make the solution clear. React at 50°C for 3 to 5 hours, lower the temperature or add anti-solvent to precipitate the polymorph in solid form.

In some embodiments, the acid is hydrochloric acid, sulfuric acid, citric acid, fumaric acid, or succinic acid.

In some embodiments, the solvent is ethanol, ethyl acetate, or acetone.

In some embodiments, the anti-solvent is n-heptane.

The sixteenth aspect of the present application provides a pharmaceutical composition, comprising:

(a) a polymorphic form of a compound of formula (X) as described above, or a pharmaceutically acceptable salt thereof; and (b) a pharmaceutically acceptable carrier.

The seventeenth aspect of the present application provides free base crystal form I, free base crystal form IV, free base crystal form V, free base crystal form VIII, or hydrochloride crystal form of the compound of formula (X) as described above I, sulfate salt crystal form I, citrate crystal form I, fumarate salt crystal form I, succinate salt crystal form I, succinate crystal form II in the preparation of _A2A receptor or _A2B receptor inhibitors Applications.

In the eighteenth aspect of the present application, polymorphs of the above-mentioned compound of formula (X) or a pharmaceutically acceptable salt thereof are provided, such as free base crystal form I, free base crystal form IV, free base crystal form V, Free base form VIII, or its hydrochloride form I, sulfate form I, citrate form I, fumarate form I, succinate form I, succinate form II, Or the use of the pharmaceutical composition described in the sixteenth aspect of the present application in the preparation of drugs for preventing or treating diseases mediated by adenosine A _2A receptors and/or adenosine A _2B receptors.

The present application provides a method for preventing or treating diseases mediated by adenosine A _2A receptors and/or adenosine A _2B receptors, comprising administering to patients in need thereof a therapeutically effective amount of A polymorphic form of the compound of formula (X) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described in the above embodiments.

The application provides the polymorphic form of the compound of formula (X) or its pharmaceutically acceptable salt as described in the above embodiment, or the pharmaceutical composition as described in the above embodiment for the prevention or treatment of adenosine A _2A receptor and/or adenosine A _2B receptor-mediated diseases.

In some embodiments, the disease is cancer or an immune-related disease.

In some embodiments, the cancer is selected from the group consisting of prostate cancer, colon cancer, rectal cancer, pancreatic cancer, cervical cancer, stomach cancer, endometrial cancer, brain cancer, liver cancer, bladder cancer, ovarian cancer, testicular cancer, head cancer , neck cancer, melanoma, basal cancer, mesothelial lining cancer, white blood cell cancer, esophageal cancer, breast cancer, muscle cancer, connective tissue cancer, small cell lung cancer, non-small cell lung cancer, adrenal gland cancer, thyroid cancer, kidney cancer cancer and bone cancer; or malignant glioma, mesothelioma, renal cell carcinoma, gastric cancer, sarcoma, choriocarcinoma, basal cell carcinoma of the skin, and testicular seminoma.

In some embodiments, the immune-related disease is selected from rheumatoid arthritis, renal failure, lupus, asthma, psoriasis, colitis, pancreatitis, allergies, fibrosis, anemic fibromyalgia, Alzheimer's Haimer's disease, congestive heart failure, stroke, aortic stenosis, arteriosclerosis, osteoporosis, Parkinson's disease, infection, Crohn's disease, ulcerative colitis, allergic contact dermatitis, and others Eczema, systemic sclerosis, and multiple sclerosis.

The compound of formula (X) of the present application, the polymorphic form of the compound of formula X, the polymorphic form of the pharmaceutically acceptable salt of the compound of formula X, or the polymorphic form comprising the compound of formula (X) and the compound of formula (X) Or the pharmaceutical composition of the polymorphic form of the pharmaceutically acceptable salt of the compound of formula X has good pharmacological activity of inhibiting _A2A receptor and/or _A2B receptor. In addition, the polymorphic form of the compound of formula (X) and the polymorphic form of the pharmaceutically acceptable salt of the compound of formula X have good stability, so they have the potential to be developed into medicines.

It should be understood that within the scope of the present application, the above-mentioned technical features of the present application and the technical features specifically described in the following (such as the embodiments) can be combined with each other to form a new technical solution. Due to space limitations, we will not repeat them here.

Description of drawings

In order to more clearly illustrate the technical solutions in the embodiments of the present application or the conventional technology, the following will briefly introduce the accompanying drawings that need to be used in the description of the embodiments or the traditional technology. Obviously, the accompanying drawings in the following description are only the present invention For the embodiments of the application, those skilled in the art can also obtain other drawings based on the disclosed drawings without creative effort.

Fig. 1 is the XPRD spectrum of the free base crystal form I of the compound of formula X prepared in Example 2 (using Cu-Kα radiation, the abscissa is angle 2θ (°), and the ordinate is SQR (counts)), wherein SQR represents intensity square root of

Fig. 2 is the DSC spectrum of the free base crystal form I of the compound of formula X prepared in Example 2 (abscissa is temperature (°C), ordinate is heat flow rate (W/g));

Fig. 3 is the TGA spectrum of the free base crystal form I of the compound of formula X prepared in Example 2 (the abscissa is temperature (°C), and the ordinate is weight change (%));

Figure 4 is the XPRD spectrum of the free base crystal form IV of the compound of formula X prepared in Example 3 (using Cu-Kα radiation, the abscissa is the angle 2θ (°), and the ordinate is SQR (counts)), wherein SQR represents the intensity square root of

Fig. 5 is the DSC spectrum of the free base crystal form IV of the compound of formula X prepared in Example 3 (the abscissa is temperature (°C), and the ordinate is heat flow rate (W/g));

Fig. 6 is the XPRD spectrum of the free base crystal form V of the compound of formula X prepared in Example 4 (using Cu-Kα radiation, the abscissa is the angle 2θ (°), and the ordinate is the intensity (counts));

Figure 7 is the DSC spectrum of the free base crystal form V of the compound of formula X prepared in Example 4 (the abscissa is temperature (°C), and the ordinate is heat flow rate (W/g));

Fig. 8 is the XPRD spectrum of the free base crystal form VIII of the compound of formula X prepared in Example 5 (using Cu-Kα radiation, the abscissa is the angle 2θ (°), and the ordinate is the intensity (counts));

Figure 9 is the DSC spectrum of the free base crystal form VIII of the compound of formula X prepared in Example 5 (the abscissa is temperature (°C), and the ordinate is heat flow rate (W/g));

Figure 10 is the XPRD spectrum of the hydrochloride salt form I of the compound of formula X prepared in Example 6 (using Cu-Kα radiation, the abscissa is the angle 2θ (°), and the ordinate is the intensity (counts));

Figure 11 is the XPRD spectrum of the sulfate crystal form I of the compound of formula X prepared in Example 7 (using Cu-Kα radiation, the abscissa is the angle 2θ (°), and the ordinate is the intensity (counts));

Figure 12 is the XPRD spectrum of the citrate crystal form I of the compound of formula X prepared in Example 8 (using Cu-Kα radiation, the abscissa is the angle 2θ (°), and the ordinate is the intensity (counts));

Figure 13 is the XPRD spectrum of the fumarate salt form I of the compound of formula X prepared in Example 9 (using Cu-Kα radiation, the abscissa is the angle 2θ (°), and the ordinate is the intensity (counts));

Figure 14 is the XPRD spectrum of the succinate salt form I of the compound of formula X prepared in Example 10 (using Cu-Kα radiation, the abscissa is the angle 2θ (°), and the ordinate is the intensity (counts));

Fig. 15 is the XPRD spectrum of the succinate crystal form II of the compound of formula X prepared in Example 11 (using Cu-Kα radiation, the abscissa is the angle 2θ (°), and the ordinate is the intensity (counts)).

Detailed ways

The polymorphic form of the compound of formula (X) and the pharmaceutically acceptable salt of the compound of formula (X), the pharmaceutical composition comprising the polymorphic form, the polymorphic form The preparation method of the compound and the pharmaceutical composition, and its application are further described in detail. This application can be embodied in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of this application more thorough and comprehensive.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the technical field to which this application belongs. The terms used herein in the specification of the application are only for the purpose of describing specific embodiments, and are not intended to limit the application.

In this application, "first aspect", "second aspect", "third aspect", "fourth aspect", "fifth aspect" and so on are used for descriptive purposes only, and cannot be understood as indicating or implying relative importance or Quantity, nor should it be understood as implying the importance or quantity of the indicated technical features. Moreover, "first", "second", "third", "fourth", "fifth" and so on are only for the purpose of non-exhaustive enumeration and description, and it should be understood that they do not constitute a closed limitation on the quantity.

In this application, the technical features described in open form include closed technical solutions consisting of the listed features, and open technical solutions including the listed features.

In this application, when it comes to a numerical interval, unless otherwise specified, the above numerical interval is considered continuous, and includes the minimum and maximum values of the range, and every value between such minimum and maximum values. Further, when a range refers to an integer, every integer between the minimum and maximum of the range is included. Furthermore, when multiple ranges are provided to describe a feature or characteristic, the ranges may be combined. In other words, unless otherwise indicated, all ranges disclosed herein are to be understood to encompass any and all subranges subsumed therein.

The percentage content involved in this application, unless otherwise specified, refers to mass percentage for solid-liquid mixing and solid-solid phase mixing, and refers to volume percentage for liquid-liquid phase mixing.

The percentage concentration involved in this application, unless otherwise specified, refers to the final concentration. The final concentration refers to the proportion of the added component in the system after the component is added.

The temperature parameters in this application, unless otherwise specified, are allowed to be treated at a constant temperature, and also allowed to be treated within a certain temperature range. The isothermal treatment allows the temperature to fluctuate within the precision of the instrument control.

Compound of the application

In the present application, the compound of formula (X) is 3-(4-amino-6-(1-((1-ethyl-1H-pyrazol-3-yl)methyl)-1H-1,2,3 -triazol-4-yl)-5-fluoropyrimidin-2-yl)-2-methylbenzonitrile, which has high inhibitory activity on adenosine A _2A receptors and/or adenosine A _2B receptors.

In the present application, the polymorphic form of the compound of formula (X) includes the polymorphic form of the free base of the compound of formula (X) and the polymorphic form of the pharmaceutically acceptable salt of the compound of formula (X). Wherein, the pharmaceutically acceptable salt is selected from hydrochloride, sulfate, citrate, fumarate or succinate. Polymorphs of the compound of formula (X) and its pharmaceutically acceptable salts include but are not limited to free base crystal form I, free base crystal form IV, free base crystal form V, free base crystal form of the compound of formula (X) VIII, Hydrochloride Form I, Sulfate Form I, Citrate Form I, Fumarate Form I, Succinate Form I, and Succinate Form II.

As used herein, "therapeutically effective amount" refers to the amount of the compound of the present application that will cause the individual's biological or medical response, such as reducing or inhibiting enzyme or protein activity or improving symptoms, relieving symptoms, slowing down or delaying disease progression or preventing diseases, etc. quantity.

As used herein, "pharmaceutically acceptable carrier" should be non-toxic and inert in principle. The form of the "pharmaceutically acceptable carrier" is not particularly limited, including but not limited to solid, semi-solid, liquid and the like. The pharmaceutically acceptable carrier should be compatible with the patient, preferably a mammal, more particularly a human. One of the roles of the pharmaceutically acceptable carrier is to be suitable for delivering the active agent to the target site of interest without terminating the activity of the agent. As used herein, the language "pharmaceutically acceptable carrier" includes buffers, sterile water for injection, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorbing agents compatible with pharmaceutical administration Retardants and the like. Each carrier is "pharmaceutically acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient.

As used herein, "patient" refers to an animal, preferably a mammal, more preferably a human. The term "mammal" refers to warm-blooded vertebrate mammals including, for example, cats, dogs, rabbits, bears, foxes, wolves, monkeys, deer, mice, pigs and humans.

As used herein, "treating" refers to alleviating, delaying progression, attenuating, preventing, or maintaining an existing disease or condition (eg, cancer). Treatment also includes curing, preventing its development, or alleviating to some extent one or more symptoms of a disease or disorder.

In this application, "using Cu-Kα radiation" means that the corresponding spectrum is obtained by using the Kα ray detection of the Cu target. When other methods are used for detection, each diffraction peak may have deviations within the acceptable range in the field, which should not be understood For the limitation of this application.

polymorph

Solids exist either in amorphous or crystalline form. In the case of crystalline forms, the molecules are positioned within three-dimensional lattice sites. When a compound crystallizes from a solution or slurry, it can crystallize in a different spatial lattice arrangement (a property known as "polymorphism"), forming crystals with different crystalline forms, which are referred to as Known as "polymorphs". Different polymorphs of a given substance may differ from each other in one or more physical properties such as solubility and dissolution rate, true specific gravity, crystalline form, packing mode, flowability and/or solid state stability.

crystallization

Production-scale crystallization can be accomplished by manipulating the solution such that the solubility limit of the compound is exceeded. This can be accomplished by various methods, for example, dissolving the compound at a relatively high temperature and then cooling the solution below the saturation limit; or reducing the volume of the liquid by boiling, evaporation at atmospheric pressure, vacuum drying, or by some other method; The solubility of a compound can be reduced by adding an anti-solvent or a solvent in which the compound has low solubility, or a mixture of such solvents. Another alternative is to adjust the pH to reduce solubility. For a detailed description of crystallization see Crystallization, Third Edition, J W Mullens, Butterworth-Heineman Ltd., 1993, ISBN 0750611294.

Identification and properties of crystal forms

After the preparation of the polymorphic form of the compound of formula (X), the applicant used the following various methods and instruments to study its properties.

X-ray powder diffraction (XRPD)

Methods of determining crystalline forms by X-ray powder diffraction are known in the art. XRPD can detect information such as crystal form changes, crystallinity, and crystal structure state, and is a common method for identifying crystal forms. The peak position of the XRPD pattern mainly depends on the structure of the crystal form, and the measurement of 2θ of the XRPD pattern may be slightly different between different instruments, so the value of 2θ cannot be regarded as absolute. According to the conditions of the instrument used in the test of this application, the diffraction peaks allow a certain error (such as ±0.2°). It can be understood that for different test instruments and test conditions, the error range is not absolute. The crystal form of the compound of formula X in the present application has a specific crystal form, and has specific characteristic peaks in the XRPD pattern.

In the present application, "the X-ray powder diffraction pattern has a characteristic diffraction peak at a specific 2θ (°) angle", which means that the peak value of the peak is within the indicated numerical range, the indicated numerical point, and the vicinity of the indicated numerical range or near the indicated numerical point. Due to different measurement factors such as measuring instruments and measurement conditions, the position of a certain peak or some peaks in the actually obtained X-ray powder diffraction diagram may be slightly shifted, that is, combined with the characteristic peaks indicated in this application or X There may be slight differences between X-ray powder diffraction patterns, but it is understood that for those skilled in the art, it is possible to identify slightly different characteristic peak combinations or whether X-ray powder diffraction patterns can substantially constitute The crystalline form described in this application. Therefore, those that are substantially identified as being consistent with the crystal forms of the invention should be considered within the scope of protection of the present application. For example, "the X-ray powder diffraction pattern has a peak at a diffraction angle of 7.57±0.2" means that the peak of the peak can be located within the range of 7.57±0.2 or near this range, as long as it does not affect the identification of the crystal form as a whole. Can. In addition, "±0.2" here only means the error of the peak in the diffraction angle position, and has nothing to do with the peak shape and peak width of the peak.

In this application, the "substantially" in "the X-ray powder diffraction pattern is basically characterized by a specific figure" should also be understood similarly, as long as it can be recognized as a whole that a certain X-ray powder diffraction pattern is consistent with the X-ray powder diffraction pattern described in this application. Figures constitute substantially the same, it should be considered to fall within the scope of protection of the present application.

It can be understood that the differential scanning calorimetry curve in the present application and the position of the endothermic peak shown therein should also be understood similarly, allowing slight discrepancies with the specific numerical values or specific numerical ranges or specific spectrograms disclosed in the present application. Inconsistent, but as long as part or all of the endothermic peak positions in the differential scanning calorimetry curve, or the entire curve is substantially consistent with the present application, it should be deemed to fall within the protection scope of the present application.

It can be understood that a similar understanding can also be made for other spectra characterizing crystal types.

Differential Scanning Calorimetry (DSC)

Also known as "differential calorimetry scanning analysis", it is a technique to measure the relationship between the energy difference and temperature between the measured substance and the reference substance during the heating process. The peak position, shape and number of peaks on the DSC spectrum are related to the properties of the substance, so it can be used to identify the substance qualitatively. This method is commonly used in the field to detect various parameters such as phase transition temperature, glass transition temperature, and heat of reaction of substances. The peak position of the DSC spectrum may vary slightly between different instruments, so the value of the peak position of the DSC endothermic peak cannot be regarded as absolute. According to the conditions of the equipment used in the test of this application, the value of the experimental error or difference may be less than or equal to 5°C, or less than or equal to 4°C, or less than or equal to 3°C, or less than or equal to 2°C, or less than or equal to 1°C.

Thermogravimetric Analysis (TGA)

TGA is a technique for measuring the quality of substances with temperature under program control. It is suitable for checking the loss of solvent in crystals or the process of sublimation and decomposition of samples. It can be speculated that crystals contain crystal water or crystallization solvents. The mass change shown by the TGA curve depends on many factors such as sample preparation and instrumentation; the mass change detected by TGA varies slightly between different instruments. According to the condition of the instrument used in the test of this application, the error of mass change is not absolute, and a certain error (such as ±0.1%) is allowed.

If simultaneous salt formation and crystallization are desired, addition of an appropriate acid or base may result in direct crystallization of the desired salt if the salt is less soluble in the reaction medium than the starting material. Likewise, completion of the synthesis reaction allows direct crystallization of the final product in a medium in which the final desired form is less soluble than the reactants.

Optimization of crystallization may include seeding the crystallization medium with crystals of the desired form. Additionally, many crystallization methods use combinations of the above strategies. One example is to dissolve the compound of interest in a solvent at elevated temperature, followed by the addition of an appropriate volume of antisolvent in a controlled manner to bring the system just below saturation levels. At this point, seeds of the desired form can be added (and the integrity of the seeds maintained) and the system cooled to complete crystallization.

As used herein, the term "room temperature" generally refers to 4-30°C, preferably 20±5°C.

Identification and properties of polymorphs

X-ray powder diffraction

The polymorphic form of the compound of formula (X) in the present application has a specific crystal form, and has specific characteristic peaks in the X-ray powder diffraction pattern (XPRD). The XPRD spectrum was collected on an Equinox3000S/N X-ray powder diffraction analyzer, and the XPRD parameters are shown in Table 11 below.

Table 11

parameter	XPRD
X-ray source	Cu K(λ＝1.54056 Angstrom)
light pipe setting	40 kV, 30 mA
Detector	psd
Scanning range (2θ°)	0°～120°
Scan step size (2θ°)	0.03
scan rate	1 second/step

In the X-ray powder diffraction pattern, the position of each peak is determined by 2θ (°). It is understood that different instruments and/or conditions may result in slightly different data, with variations in the position and relative intensity of peaks. Those skilled in the art should understand that when determining the crystal form based on the XRPD pattern, the peaks at low diffraction angles and their intensity, peak shape integrity and other factors are relatively more meaningful. In this application, after further research, the applicant found that when using the above-mentioned instrument for testing, strong background peaks of a blank metal disk appeared near the two 2θ (°) values of 37 and 44 at high diffraction angles. For example, the peak near 37° in Figure 4 is presumed to be the peak produced by a blank metal disk (such as the peak at 37.48° in Table 2), and the peak near 37° and 43° in Figure 6 is speculated to be a blank metal disk The peaks produced (as the peaks at 37.48° and 43.60° in Table 3), the peaks near 37° in Figure 8 are speculated to be the peaks produced by blank metal disks (as the peaks at 37.48° in Table 4), Figure 11 The peak near 37 ° in the middle is presumed to be the peak produced by the blank metal disc (as the peak at 37.48 ° in Table 6), and the peak near 37 ° and 43 ° in Figure 12 is speculated to be the peak produced by the blank metal disc ( As the peak at 37.51° and 43.63° in Table 7), the peak near 37° in Figure 13 is speculated to be the peak produced by the blank metal disk (as the peak at 37.87° in Table 8), near 37° in Figure 14 The peak and the peak around 43 ° are speculated to be the peaks produced by the blank metal disc (such as the peaks at 37.84 ° and 44.07 ° in Table 9), and the peaks near 37 ° in Figure 15 are speculated to be the peaks produced by the blank metal disc (such as Peak at 37.84° in Table 10). The intensity division of the peaks only reflects the approximate size of the peaks at each position. In this application, each crystal form takes the diffraction peak with the highest peak height as the base peak, defines its relative intensity as 100%, and regards it as I ₀ , and uses the ratio of the peak height of the other peaks to the base peak peak height as its relative intensity. Intensity I/I ₀ , the division definition of the relative intensity of each peak is shown in Table 12 below.

Table 12

Relative Intensity I/I 0 (%)	definition
50～100	VS (very strong)
25～50	S (strong)
10～25	M (medium)
1～10	W (weak)

Differential Scanning Calorimetry (DSC)

The DSC spectrum was collected on a DISCOVERY DSC25 differential scanning calorimeter, and the test parameters are shown in Table 13 below.

Table 13

parameter	DSC
method	linear heating
sample tray	Aluminum pan, cover
temperature range	25°C–300°C
Scan rate (℃/min)	10
Protective gas	Nitrogen

It is understood that other values may be obtained when using other types of instruments having the same function as the above-mentioned instruments or using test conditions different from those used in this application, therefore, the quoted values should not be regarded as absolute values.

Due to the error of the instrument or the difference of the operator, those skilled in the art can understand that the above parameters used to characterize the physical properties of the crystal may have slight differences, so the above parameters are only used to assist in characterizing the polymorphs provided by the application , and should not be considered as a limitation on the polymorphic forms of the present application.

Reagents and Instruments

In this application, the structure and purity of the compounds are determined by nuclear magnetic resonance ( ¹ HNMR) and/or liquid chromatography-mass spectrometry (LC-MS). ¹ HNMR: Bruker AVANCE-400 nuclear magnetic analyzer, the internal standard is tetramethylsilane (TMS). LC-MS: Agilent 1200 HPLC System/6140 MS liquid mass spectrometry (manufacturer: Agilent), column WatersX-Bridge, 150×4.6 mm, 3.5 μm.

The starting materials known in the application can be adopted or synthesized according to methods known in the art, or can be obtained from ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Shaoyuan Chemical Technology (Accela ChemBio Inc) and Darui Chemicals Wait for the company to buy.

As used herein, DCM: dichloromethane, DMF: dimethylformamide, DMSO: dimethylsulfoxide, THF: tetrahydrofuran, EA: ethyl acetate, PE: petroleum ether, Pd(dppf)Cl2: [ ₁ ,1'-bis(diphenylphosphine)ferrocene]palladium dichloride, Pd(PPh ₃ ) ₂ Cl ₂ : bis(triphenylphosphine)palladium(II) chloride, DPPA: diphenylphosphoryl azide Esters, DBU: 1,8-diazabicycloundec-7-ene.

Preparation of Intermediate V1

2-Methyl-3-bromoxynil (15g, 76.51mmol), bis-pinacol borate (23.32g, 91.82mmol), potassium acetate (15.02g, 153.03mmol) and Pd(dppf)Cl ₂ (2.80g) was dissolved in a mixed solution of DMSO (20mL) and dioxane (100mL), and stirred at 100°C for 3.5 hours under nitrogen protection. After the reaction was complete, the solid product obtained by evaporating the solvent under reduced pressure was separated by column chromatography (EA: PE15%-40%) to obtain compound V1 (21.05 g, purity: 78.0%, yield: 100%). MS (ESI) 244.1 [M+H] ⁺ .

Preparation of Intermediates 1-5

step one:

Compound V2-1 (40g, 198.60mmol) and triisopropylsilylacetylene (38.03g 208.53mmol) were dissolved in THF (400.00mL), under the protection of argon, stirred and cooled to 0°C, CuI (3.78g , 19.86mmol), Pd(PPh ₃ ) ₂ Cl ₂ (6.97g, 9.93mmol), then Et ₃ N (60.17g, 595.79mmol) was slowly added dropwise. After completion of the dropwise addition, stir at 0°C for 2 hours, raise the temperature to room temperature and continue stirring for 6 hours, then cool down to 0°C, add NH ₃ /H ₂ O (120.00g, 2.61mol, 120.00mL, 37% purity) dropwise, and then stir at room temperature overnight. LC-MS detected that the reaction was complete, and compound V2-2 was obtained. MS (ESI) 347 [M+H].

Step two:

Add a large amount of water to the reaction solution in step 1, extract with DCM (200mL×3), dry over anhydrous sodium sulfate, concentrate under reduced pressure to leave about 40mL of DCM, a large amount of solid precipitates, stir and add 60mL of petroleum ether, and filter to obtain compound V2-3( 31 g), and the remaining mother liquor was separated and purified by silica gel column (DCM:PE=30%-80%) to obtain compound V2-3 (43 g). MS (ESI) 328 [M+H] ⁺ .

Step three:

Compound V2-3 (41g, 125.04mmol) and compound V1 (42.56g, 175.05mmol) were dissolved in a mixed solvent of dioxane (400mL) and water (60mL), stirred at room temperature under argon protection, and Na ₂ was added successively CO ₃ (26.51g, 250.08mmol) and Pd(dppf) ₂ Cl (4.57g, 6.25mmol), then warmed up to 100°C and stirred for 24 hours. The reaction solution was poured into ice water, extracted with DCM (300mL×2), dried over anhydrous sodium sulfate, concentrated to dryness under reduced pressure, separated and purified by silica gel column to obtain yellow solid V2-4 (44g, 107.69mmol, 86.12% yield). MS (ESI) 409.2 [M+H] ⁺ .

Step four:

Add compound V2-4 (44g, 107.69mmol) into 400mL of methanol, stir at room temperature and add NH ₄ F (47.60g, 1.40mol), then raise the temperature to 70°C and stir for 8 hours under the protection of argon, and add the reaction solution while stirring In a large amount of water, a large amount of solids were washed out, and the brown product V2-5 (27g, 107.04mmol, 99.40% yield) was obtained by filtration. MS (ESI) 253.1 [M+H] ⁺ .

Example 1

Preparation of Formula X Compounds

step one:

Compound 1H-pyrazole-3-carboxylic acid ethyl ester (25g, 178.39mmol) and cesium carbonate (58.12g, 178.39mmol) were dissolved in DMF (150mL), stirred at room temperature and added iodoethane (30.61g, 196.23mmol), Then the temperature was raised to 80°C and stirred overnight. The reaction solution was poured into water, extracted with EA (150mL×2), dried over anhydrous sodium sulfate, concentrated to dryness under reduced pressure, separated and purified by silica gel column (EA:PE=0～70%) to obtain white solid 1-ethylpyrazole-3 - Ethyl carboxylate (11.5 g, 68.37 mmol, 38.33% yield). MS (ESI) 169 [M+H] ⁺ .

Step two:

The compound 1-ethylpyrazole-3-carboxylic acid ethyl ester (11.5g, 68.37mmol) was dissolved in THF (150mL), the temperature was lowered to -10°C under the protection of argon, and LiAlH ₄ (3.89g, 102.56mmol) was added in batches ), and then stirred at about 0°C for 1 hour. 5.0 g of sodium sulfate decahydrate was added in batches to the reaction solution at 0° C., stirred at 0° C. for 1 hour, then slowly raised to room temperature and stirred for 1 hour. After filtration, the filtrate was concentrated to dryness under reduced pressure to obtain (1-ethylpyrazol-3-yl)methanol (7.8 g, 61.83 mmol, 90.43% yield) as a colorless oil. MS (ESI) 127 [M+H] ⁺ .

Step three:

The compound (1-ethylpyrazol-3-yl)methanol (7.8g, 61.83mmol) was dissolved in THF (60mL), and DPPA (18.04g, 74.19mmol) was added with stirring under argon-protected ice-bath conditions, and then slowly DBU (14.12g, 92.74mmol, 13.84mL) was added dropwise, and the temperature was controlled below 15°C. After the addition, the temperature was raised to 45°C and stirred overnight. Stir the reaction solution and add 200ml of water, extract with methyl tert-butyl ether (100mL×2), combine the organic phases, wash the organic phase with water (200ml), and directly use in the next reaction.

Step four:

Add compound V2 (15.52g, 61.52mmol) in THF (100mL) and methyl tert-butyl ether (200mL) solution to the organic phase of step 3, stir at room temperature under argon protection, add copper sulfate pentahydrate (153.60mg , 615.21μmol) was dissolved in water (20mL), sodium ascorbate (487.51mg, 2.46mmol) was dissolved in water (20mL), then heated to 60°C and stirred for 16 hours. The reaction solution was poured into water, extracted with dichloromethane (1000ml×2), dried over anhydrous sodium sulfate, concentrated to dryness under reduced pressure, separated and purified through silica gel column (PE:DCM (containing 2% triethylamine)=30%- 100%) to obtain compound X (19.00 g).

¹ H NMR (400MHz, DMSO-d ₆ )δ8.64(s, 1H), 7.91(d, J=7.0Hz, 1H), 7.82(d, J=7.8Hz, 1H), 7.67(d, J= 2.2Hz, 1H), 7.51(s, 2H), 7.44(t, J=7.8Hz, 1H), 6.23(d, J=2.2Hz, 1H), 5.60(s, 2H), 4.06(q, J= 7.3Hz, 2H), 2.61(s, 3H), 1.30(t, J=7.4Hz, 3H).

The obtained solid compound of formula X was sent to XRPD for detection, and its powder X-ray diffraction pattern had no characteristic peaks, and it was in an amorphous form.

Test Example 1: Inhibitory Activity of Formula X Compounds on _A2A Receptors and _A2B Receptors

CHO-K1/ADORA _2A /Gα15 (GenScript, M00246) and CHO-K1/ADORA _2B /Gα15 (GenScript, M00329) cells were cultured in Ham's F-12 (Gibco, 31765092) medium. The culture medium conditions are 10% FBS, 200 μg/mL Zeocin and 100 μg/mL Hygromycin B or 10% FBS, 400 μg/mL G418 and 100 μg/mL Hygromycin B. For specific culture conditions, please refer to the corresponding instructions. The screening steps are as follows:

(1) Adjust the cell density to 6×10 ⁵ cells/ml with serum-free medium.

(2) Add 5 μL of cell solution, 2.5 μL of NECA (Sigma, 119140-10MG) and 2.5 μL of compound solution to each well of a 384-well plate (Greiner Bio-One, 784075) in sequence, and the final concentration of NECA is 50 nM (CHO- K1/ADORA2A) or 10 nM (CHO-K1/ADORA2B), the final concentration of the compound is 3 μM and the initial dilution is 3 times downward.

(3) Place in a 37°C incubator and incubate for 30mins.

(4) Add 5 μL of cAMP-d ₂ and 5 μL of cAMP-ab (Cisbio, 62AM4PEB) in sequence.

(5) Place the 384-well plate at room temperature for 1 hour in the dark.

(6) Read the plate (Victor X5, PerkinElmer), XLfit nonlinear regression analysis data, and calculate the IC ₅₀ of the compound. The results are shown in Table 14 below.

Table 14. Inhibitory activity of compounds of formula X on _A2A receptors and _A2B receptors

Compound number	A 2A receptor (IC 50 /μM)	A 2B receptor (IC 50 /μM)
x	<0.001	0.001

It can be seen from Table 14 that the compound of the formula X in the embodiment of the present application has relatively high inhibitory activity on both the _A2A receptor and the _A2B receptor.

Test Example 2: In Vivo Pharmacokinetic Test

Applying LC/MS/MS method to measure the drug concentration of the compound of formula X of the application at different time points after intravenous administration and intragastric administration of mice, to study the pharmacokinetic behavior of the compound of formula X of the application in mice, and to evaluate its Pharmacokinetic profile.

Experimental program:

Experimental animals: healthy adult male ICR mice (weight 25g-40g, 12 mice, the mice in the intravenous injection group were free to drink water and food, and the intragastric administration group was fasted overnight, and free to drink water and food after 4 hours of administration), provided by Beijing Weiwei Provided by Tonglihua Laboratory Animal Technology Co., Ltd.;

Administration method and dose: select animals that meet the experimental requirements before administration, and weigh the marks. ICR mice were administered via tail vein (2mg/kg, 5% DMSO, pH4.5 20% Captisol) and intragastrically (10mg/kg, 5% DMSO, pH4.5 20% Captisol).

Blood sample collection: about 100 μL of blood was collected through the jugular vein at 0.083 hours, 0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 6 hours, 7.5 hours and 24 hours after administration for both intravenous and intragastric administration. The blood was transferred to a 1.5mLEP tube pre-added with EDTA2K, centrifuged for 4min (8000rpm, 4°C), and the plasma was separated. The whole process was completed within 15min after blood collection. All plasma samples need to be stored in a -20°C freezer until sample analysis.

Table 15 shows the pharmacokinetic properties parameters of the compound of formula X in the present application in mice in the form of intravenous administration.

Table 15. Pharmacokinetic parameters under intravenous administration

Compound number	Clearance rate CLz(mL/min/kg)	Area under the curve AUC 0-t (hr·ng/mL)
Compound of formula X	17.2	1918

Table 16 shows the parameters of pharmacokinetic properties of compound X of the present application in mice in the form of intragastric administration.

Table 16. Pharmacokinetic parameters under gastric administration

Compound number	C max (ng/mL)	Area under the curve AUClast (hr·ng/mL)
Compound of formula X	7060	8569

Test Example 3: In vivo drug efficacy experiment of the compound of formula X of the present application

Experimental scheme: This test example investigates the mouse melanoma cell line B16F10-OVA-hPD-L1 subcutaneously transplanted tumor mice, after administering the compound of formula X of this application through the oral administration route, test its effect on melanoma B16F10-OVA-hPD-L1 In vivo drug efficacy in L1 tumor-bearing mice.

Experimental materials: C57BL/6 mice (female); mouse melanoma B16F10-OVA-hPD-L1 cells (Shanghai Jiaotong University cell bank), cultured in vitro as a monolayer, and the culture conditions are DMEM medium containing 10% fetal bovine serum , cultured at 37 °C in a 5% CO ₂ incubator. Routine digestion with trypsin-EDTA was performed for passage. When the cells were in the exponential growth phase and the saturation was 80%-90%, the cells were harvested and counted.

Compound preparation: Measure the compound of formula X and add it into the solvent (40% sulfobutyl-β-cyclodextrin (captisol) in acetic acid buffer, pH 4.0) to prepare a sample with the target concentration. Experimental operation: resuspend the cells in phosphate buffered saline at a density of 5×10 ⁶ cells/mL. Subcutaneously inoculate 0.1 mL of PBS solution containing 5× ¹⁰⁵ B16F10-OVA-hPD-L1 cells on the right back of each mouse. On the day of inoculation, the mice were randomly divided into groups according to their body weight, with 10 mice in each group. The drug was administered twice a day for 19 days. Animals were weighed and their health monitored daily throughout the experimental period. Tumor diameters were measured twice a week with vernier calipers.

The formula for calculating the tumor volume V is: V=0.5×a×b ² , where a and b represent the long diameter and short diameter of the tumor, respectively.

The antitumor efficacy of compounds was evaluated by relative tumor proliferation rate T/C (%). Relative tumor proliferation rate T/C (%)=Vt/Vc×100% (Vt: average tumor volume of treatment group; Vc: average tumor volume of negative control group). Vt and Vc take the data of the same day. The results at the end of the administration are shown in Table 17.

Table 17. The relative tumor proliferation rate obtained in Test Example 3

The preparation of embodiment 2 free base crystal form I

method one

Weigh about 20mg of the compound of formula X (amorphous) prepared according to Example 1 into a glass vial, add 1mL of ethanol and heat to dissolve at about 50°C, take out the glass vial and let it cool down at room temperature. Then place it in the refrigerator to cool down to 0-4°C, wait for the solid to precipitate, discard the supernatant after centrifugation, put the solid in an oven to dry, and perform XPRD detection. The XPRD diagram of the obtained solid product is shown in Figure 1, which is defined as free base crystal form I in this application. The DSC spectrum of the free base crystal form I is shown in Figure 2, in which the endothermic peak is 179.10°C, and there is an endothermic peak at 104.32°C, and the TGA spectrum is shown in Figure 3, and the weight loss before 150°C is about 8.8%. Form I contains the solvent ethanol.

Method Two

Weigh about 20 mg of the compound of formula X (amorphous) prepared according to Example 1 into a glass vial, add 1 mL of methanol and heat to dissolve at about 50°C, take out the glass vial and let it cool slowly at room temperature until it reaches room temperature Then place it in the refrigerator to cool down to 0-4°C, wait for the solid to precipitate, discard the supernatant after centrifugation, dry the solid in an oven, and perform XPRD detection to confirm that the free base crystal form I is obtained.

method three

Weigh about 19 g of the compound of formula X (amorphous) prepared according to Example 1 into a glass vial, add 100 mL of ethanol, and heat to reflux (about 75° C.). After cooling down to room temperature, a solid precipitated out and was filtered to obtain a solid product. The obtained solid was detected by XPRD, and it was confirmed that the free base crystal form I was obtained.

Example 3 Preparation of free base crystal form IV

method one

Weigh about 20 mg of the compound of formula X (free base crystal form I) into a glass vial, add 1 mL of isopropanol, suspend and shake the glass vial at room temperature for 24 hours, centrifuge, discard the supernatant, and place the solid in Dry in an oven for XPRD detection. The XPRD diagram of the obtained solid product is shown in Figure 4, which is defined as free base crystal form IV in this application. The DSC spectrum of the free base crystal form IV is shown in Figure 5, which has an endothermic peak at 170.43°C.

Method Two

Weigh about 20 mg of the compound of formula X (free base crystal form I) into a glass vial, add 1 mL of isopropanol or water, suspend and shake the glass vial at room temperature for 7 days, centrifuge, discard the supernatant, and dissolve the solid Place it in an oven to dry, and perform XPRD detection to confirm that the free base crystal form IV is obtained.

The preparation of embodiment 4 free base crystal form V

method one

Weigh about 20 mg of the compound of formula X (amorphous) into a glass vial, add 1 mL of acetone to dissolve the compound, place the solution at room temperature for slow volatilization, take the volatilized solid powder for XPRD detection, and the XPRD diagram of the obtained solid product As shown in Figure 6, it is defined as free base crystal form V in this application. The DSC spectrum of the free base crystal form V is shown in Figure 7, and the free base crystal form V has an endothermic peak at 179.02°C.

Method Two

Weigh about 20 mg of the compound of formula X (free base crystal form I) into a glass vial, add 1 mL of methyl tert-butyl ether, suspend and shake the glass vial at room temperature for 7 days, centrifuge, discard the supernatant, and The solid was dried in an oven and subjected to XPRD detection, which confirmed that the free base crystal form V was obtained.

The preparation of embodiment 5 free base crystal form VIII

Weigh 10 mg of free base crystal form I, free base crystal form IV, and free base crystal form V, mix thoroughly, take 20 mg of them in a glass vial, add 1 mL of acetone or 50% ethanol, suspend and shake at room temperature for 3 days, The precipitate was collected by centrifugation and measured for XPRD after drying. The XPRD diagram of the obtained solid product is shown in Figure 8, which is defined as the free base crystal form VIII in this application. The DSC spectrum of the free base crystal form VIII is shown in Figure 9, and the free base crystal form VIII has an endothermic peak at 162.93°C.

The preparation of embodiment 6 hydrochloride crystal form I

Weigh 20mg of the compound of formula X (amorphous), add 1M hydrochloric acid solution according to the molar ratio of acid (hydrogen ion) and free base of 1.2:1, then add 2mL ethyl acetate or acetone, heat and sonicate to clarify, and keep warm at 50°C for reaction After 4 hours, the temperature was slowly lowered to precipitate solids, and the solids were collected by centrifugation. The obtained solid was used for XRPD test after the solvent was evaporated, and its X-ray powder diffraction pattern is shown in Figure 10, which is defined as hydrochloride crystal form I in this application.

The preparation of embodiment 7 sulfate crystal form I

Weigh 20mg of the compound of formula X (amorphous), add 0.5M sulfuric acid solution according to the molar ratio of acid (hydrogen ion) and free base of 1.2:1, then add 2mL ethyl acetate or acetone, heat and sonicate to clarify, and keep warm at 50°C After reacting for 4 hours, the temperature was lowered slowly to precipitate a solid, which was collected by centrifugation. The obtained solid was used for XRPD test after the solvent was evaporated, and its X-ray powder diffraction pattern is shown in Figure 11, which is defined as sulfate crystal form I in this application.

The preparation of embodiment 8 citrate crystal form I

Weigh 20 mg of the compound of formula X (amorphous), add 0.5 mol/L citric acid solution according to the molar ratio of acid (hydrogen ion) and free base of 1.2:1, then add 2 mL of acetone, heat and sonicate for clarification, and keep warm at 50 ° C for reaction After 4 hours, it was slowly lowered to room temperature, and the anti-solvent n-heptane was added to precipitate a solid, which was collected by centrifugation. The obtained solid was used for XRPD test after the solvent was evaporated, and its X-ray powder diffraction pattern is shown in Figure 12, which is defined as citrate crystal form I in this application.

The preparation of embodiment 9 fumarate crystal form I

Weigh 20mg of formula X compound (amorphous), add 0.25mol/L fumaric acid solution according to the ratio of acid (hydrogen ion) and free base molar ratio 1.2:1, then add 2mL ethyl acetate, heat and ultrasonically make clarification, 50 The temperature was kept at ℃ for 4 hours, and then slowly lowered to room temperature, the anti-solvent n-heptane was added to precipitate a solid, and the solid was collected by centrifugation. The obtained solid was used for XRPD test after the solvent was evaporated, and its X-ray powder diffraction pattern is shown in Figure 13, which is defined as fumarate salt crystal form I in this application.

Preparation of Example 10 Succinate Form I

Weigh 20 mg of the compound of formula X (amorphous), add 0.5 mol/L succinic acid solution according to the molar ratio of acid (hydrogen ion) and free base of 1.2:1, then add 2 mL of ethyl acetate, heat and sonicate to clarify, 50 °C The reaction was incubated for 4 hours, then slowly lowered to room temperature, and the anti-solvent n-heptane was added to precipitate a solid, which was collected by centrifugation. The obtained solid was used for XRPD test after the solvent was evaporated, and its X-ray powder diffraction pattern is shown in Figure 14, which is defined as succinate salt crystal form I in this application.

Example 11 Preparation of succinate crystal form II

Weigh 20 mg of the compound of formula X (amorphous), add 0.5 mol/L succinic acid solution according to the molar ratio of acid (hydrogen ion) and free base of 1.2:1, then add 2 mL of acetone, heat and sonicate to clarify, and keep warm at 50 ° C for reaction After 4 hours, it was slowly lowered to room temperature, and the anti-solvent n-heptane was added to precipitate a solid, which was collected by centrifugation. The obtained solid was evaporated to dryness and used for XRPD test. Its X-ray powder diffraction pattern is shown in FIG. 15 , which is defined as succinate crystal form II in this application.

The technical features of the above-mentioned embodiments can be combined arbitrarily. To make the description concise, all possible combinations of the technical features in the above-mentioned embodiments are not described. However, as long as there is no contradiction in the combination of these technical features, should be considered as within the scope of this specification.

The above-mentioned embodiments only express several implementation modes of the present application, which is convenient for a specific and detailed understanding of the technical solution of the present application, but should not be construed as limiting the protection scope of the invention patent. It should be noted that those skilled in the art can make several modifications and improvements without departing from the concept of the present application, and these all belong to the protection scope of the present application. It should be understood that technical solutions obtained by those skilled in the art through logical analysis, reasoning or limited experiments on the basis of the technical solutions provided in this application are within the protection scope of the appended claims of this application. Therefore, the scope of protection of the patent application should be based on the content of the appended claims, and the description and drawings can be used to interpret the content of the claims.

Claims (11)

1. A polymorphic form of a compound of formula X or a pharmaceutically acceptable salt thereof,

   

   The pharmaceutically acceptable salt is selected from: hydrochloride, sulfate, citrate, fumarate or succinate.
2. The polymorph according to claim 1, wherein the polymorph of the compound of formula X is selected from any group of:

   The free base crystal form I of the compound of formula X has a characteristic diffraction peak at the following 2θ (°) angles in its X-ray powder diffraction pattern:

   7.57±0.2, 13.41±0.2, 14.64±0.2, 19.81±0.2 and 24.43±0.2;

   The free base crystal form IV of the compound of formula X has a characteristic diffraction peak at the following 2θ (°) angles in its X-ray powder diffraction pattern:

   11.88±0.2, 14.17±0.2, 16.96±0.2, 22.63±0.2, 23.56±0.2 and 25.66±0.2;

   The free base crystal form V of the compound of formula X has a characteristic diffraction peak at the following 2θ (°) angles in its X-ray powder diffraction pattern:

   11.88±0.2, 16.36±0.2, 16.99±0.2, 22.99±0.2, 23.47±0.2, and 26.41±0.2; and

   The free base crystal form VIII of the compound of formula X has a characteristic diffraction peak at the following 2θ (°) angles in its X-ray powder diffraction pattern:

   6.40±0.2, 13.12±0.2, 15.91±0.2, 22.69±0.2, and 26.95±0.2.
3. The polymorph according to claim 2, wherein the polymorph of the compound of formula X is selected from any group of:

   The free base crystal form I of the compound of formula X, its X-ray powder diffraction pattern also includes 1 or 2 characteristic diffraction peaks selected from the following 2θ (°) angles: 24.87±0.2 and 27.46±0.2;

   The free base crystal form IV of the compound of formula X, its X-ray powder diffraction pattern also includes at least one characteristic diffraction peak selected from the following 2θ (°) angles: 15.76±0.2 and 26.08±0.2;

   The free base crystal form V of the compound of formula X, its X-ray powder diffraction pattern also includes at least one characteristic diffraction peak selected from the following 2θ (°) angles: 7.90±0.2, 13.71±0.2, 14.14±0.2, 15.31±0.2, 17.80±0.2, 18.76±0.2, 21.34±0.2, 24.82±0.2, 25.51±0.2, 27.88±0.2, and 30.46±0.2; and

   The free base crystal form VIII of the compound of formula X, its X-ray powder diffraction pattern also includes at least one characteristic diffraction peak selected from the following 2θ (°) angles: 11.62±0.2, 12.36±0.2, 16.62±0.2, 18.34±0.2, 18.88±0.2, 24.94±0.2, and 29.65±0.2.
4. The polymorph according to claim 2, wherein

   The X-ray powder diffraction spectrum of the free base crystal form I is basically as shown in Figure 1;

   The X-ray powder diffraction pattern of the free base crystal form IV is basically as shown in Figure 4;

   The X-ray powder diffraction pattern of the free base crystal form V is basically shown in Figure 6; or

   The X-ray powder diffraction pattern of the free base crystal form VIII is basically shown in FIG. 8 .
5. The polymorph according to claim 2, wherein

   The differential scanning calorimetry curve of the free base crystal form I has an endothermic peak at 179.10±3°C;

   The differential scanning calorimetry curve of the free base crystal form IV has an endothermic peak at 170.43±3°C;

   The differential scanning calorimetry curve of the free base crystal form V has an endothermic peak at 179.02±3°C; or

   The differential scanning calorimetry curve of the free base crystal form VIII has an endothermic peak at 162.93±3°C.
6. The polymorphic form according to claim 1, wherein the polymorphic form of the pharmaceutically acceptable salt of the compound of formula X is selected from any group of:

   The hydrochloride crystal form I of the compound of formula X has a characteristic diffraction peak at the following 2θ (°) angles in its X-ray powder diffraction pattern:

   11.95±0.2, 24.37±0.2, 25.21±0.2 and 26.35±0.2;

   The sulfate crystal form I of the compound of formula X has a characteristic diffraction peak at the following 2θ (°) angles in its X-ray powder diffraction pattern:

   8.22±0.2, 11.29±0.2, 14.14±0.2, 16.96±0.2, 18.13±0.2, 22.27±0.2, 23.86±0.2 and 27.79±0.2;

   The citrate crystal form I of the compound of formula X has a characteristic diffraction peak at the following 2θ (°) angles in its X-ray powder diffraction pattern:

   12.55±0.2, 14.50±0.2, 21.55±0.2 and 22.75±0.2;

   The fumarate crystal form I of the compound of formula X has a characteristic diffraction peak at the following 2θ (°) angles in its X-ray powder diffraction pattern:

   7.12±0.2, 11.71±0.2, 16.90±0.2, 23.26±0.2, 23.65±0.2, 24.64±0.2 and 25.54±0.2;

   The succinate crystal form I of the compound of formula X has a characteristic diffraction peak at the following 2θ (°) angles in its X-ray powder diffraction pattern:

   4.87±0.2, 9.31±0.2, 13.78±0.2, 14.14±0.2, 18.16±0.2, 20.71±0.2, 22.12±0.2, and 26.83±0.2; and

   The succinate crystal form II of the compound of formula X has characteristic diffraction peaks at the following 2θ (°) angles in its X-ray powder diffraction pattern:

   11.86±0.2, 12.49±0.2, 14.50±0.2, 15.22±0.2, 16.90±0.2, 21.64±0.2, 22.87±0.2, and 25.75±0.2.
7. The polymorph according to claim 6, wherein

   The ray powder diffraction pattern of the hydrochloride crystal form I is basically as shown in Figure 10:

   The X-ray powder diffraction spectrum of the sulfate crystal form I is basically as shown in Figure 11;

   The X-ray powder diffraction pattern of the citrate crystal form I is basically as shown in Figure 12;

   The X-ray powder diffraction pattern of the fumarate salt form I is basically as shown in Figure 13;

   The X-ray powder diffraction pattern of the succinate salt form I is basically shown in Figure 14; or

   The X-ray powder diffraction pattern of the succinate crystal form II is basically shown in FIG. 15 .
8. A pharmaceutical composition comprising:

   (a) the polymorphic form of any one of claims 1 to 7; and

   (b) A pharmaceutically acceptable carrier.
9. Use of the polymorphic form of any one of claims 1-7, or the pharmaceutical composition of claim 8 in the preparation of an _A2A receptor inhibitor or an _A2B receptor inhibitor.
10. The polymorphic form described in any one of claims 1-7, or the pharmaceutical composition described in claim 8 is used in the preparation of preventing or treating cancer mediated by adenosine A _2A receptors or adenosine A _2B receptors or Use in medicine for immune-related diseases.
11. A method for preventing or treating cancer or immune-related diseases mediated by adenosine _A2A receptors or adenosine _A2B receptors, comprising administering to a patient in need thereof a therapeutically effective amount of the The polymorphic form described in any one, or the pharmaceutical composition according to claim 8.

Images