Crystalline form of quinazoline compound and preparation method thereof

Abstract

The application discloses a crystal form of a quinazoline compound and a preparation method of the crystal form, and particularly discloses a crystal form of a compound of formula (I) and a preparation method and application of the crystal form.

Description

[0001] This application claims priority to:

[0002] CN202211169562.X, September 23, 2022. TECHNICAL FIELD

[0003] The application discloses a crystal form of a quinazoline compound and a preparation method thereof, and specifically discloses a crystal form of a compound of formula (I) and a preparation method and application thereof. BACKGROUND

[0004] RAS protein is a guanine nucleotide binding protein with guanosine triphosphatase (GTPase) activity, mainly including three subtypes, KRAS, NRAS and HRAS. As a binary molecular switch for GDP / GTP cycle control, RAS protein can cycle between active GTP binding state (GTP-RAS) and inactive GDP binding state (GDP-RAS). This cycle has important regulatory functions in cells, and is closely related to cell proliferation, survival, metabolism, migration, immunity and growth.

[0005] SOS1 (English full name Son of Sevenless 1) is a kind of GEF regulating the GDP / GTP cycle of RAS protein. After the cell surface receptor is activated and combined with intracellular Grb2, Grb2 recruits SOS1 to the cell membrane, and then SOS1 catalyzes the exchange of RAS-GDP / GTP, thereby activating the downstream signal pathway. Small molecule SOS1 inhibitors combined in the catalytic site can block the binding of SOS1 and RAS protein, thereby effectively reducing the abnormal activation of the RAS downstream signal pathway in cancer cells, and playing a role in treating cancer. At present, only the SOS1 small molecule inhibitor BI-1701963 (WO2018115380, WO2019122129) developed by Boehringer Ingelheim has entered the I phase clinical experiment. The SOS1 inhibitor developed by Bayer (WO2018172250, WO2019201848) is still in the preclinical research stage. In recent years, some studies have shown that RAS pathway drugs are prone to produce drug resistance in clinical application, and part of the drug resistance is caused by the negative feedback activation of the upstream RAS pathway after the inhibition of ERK phosphorylation. This negative feedback regulation mechanism is closely related to SOS1. Therefore, the development of SOS1 small molecule inhibitors has broad application prospects.

[0006] AMG-510 is an effective, orally bioavailable, selective KRAS G12C covalent inhibitor developed by Amgen, which is used for treating locally advanced or metastatic non-small cell lung cancer carrying KRAS G12C mutation. Its structure is as follows:

[0007] SUMMARY

[0008] The present application provides a crystalline form of the compound of formula (I), which has an X-ray powder diffraction (XRPD) pattern with characteristic diffraction peaks at the following 2Θ angles: 15.492±0.200°, 16.458±0.200°, 18.657±0.200° and 20.638±0.200°;

[0009]

[0010] In some embodiments of the present application, the X-ray powder diffraction pattern of the above-mentioned crystalline form A has characteristic diffraction peaks at the following 2Θ angles: 14.223±0.200°, 14.589±0.200°, 14.894±0.200°, 15.492±0.200°, 16.061±0.200°, 16.458±0.200°, 18.657±0.200° and 20.638±0.200°.

[0011] In some embodiments of the present application, the above-mentioned crystalline form A has an X-ray powder diffraction pattern, in terms of 2Θ angles, comprising at least 4, 5, 6, 7 or 8 characteristic diffraction peaks selected from the following: 14.223±0.200°, 14.589±0.200°, 14.894±0.200°, 15.492±0.200°, 16.061±0.200°, 16.458±0.200°, 18.657±0.200° and 20.638±0.200°.

[0012] In some embodiments of the present application, the X-ray powder diffraction pattern of the above-mentioned crystalline form A has characteristic diffraction peaks at the following 2Θ angles: 14.223±0.100°, 14.589±0.100°, 14.894±0.100°, 15.492±0.100°, 16.061±0.100°, 16.458±0.100°, 18.657±0.100° and 20.638±0.100°.

[0013] In some embodiments of the present application, the above-mentioned crystalline form A has an X-ray powder diffraction pattern, in terms of 2Θ angles, comprising at least 4, 5, 6, 7 or 8 characteristic diffraction peaks selected from the following: 14.223±0.100°, 14.589±0.100°, 14.894±0.100°, 15.492±0.100°, 16.061±0.100°, 16.458±0.100°, 18.657±0.100° and 20.638±0.100°.

[0014] In some embodiments of the application, the Form A has an X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2Θ angles: 7.736 ± 0.200°, 14.223 ± 0.200°, 14.589 ± 0.200°, 14.894 ± 0.200°, 15.492 ± 0.200°, 16.061 ± 0.200°, 16.458 ± 0.200°, 18.657 ± 0.200°, 19.407 ± 0.200°, 20.638 ± 0.200°, 21.810 ± 0.200°, and 22.836 ± 0.200°.

[0015] In some embodiments of the application, the Form A has an X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2Θ angles: 7.736 ± 0.100°, 14.223 ± 0.100°, 14.589 ± 0.100°, 14.894 ± 0.100°, 15.492 ± 0.100°, 16.061 ± 0.100°, 16.458 ± 0.100°, 18.657 ± 0.100°, 19.407 ± 0.100°, 20.638 ± 0.100°, 21.810 ± 0.100°, and 22.836 ± 0.100°.

[0016] In some embodiments of the application, the Form A has an X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2Θ angles: 7.736 ± 0.100°, 14.223 ± 0.100°, 14.589 ± 0.100°, 14.894 ± 0.100°, 15.492 ± 0.100°, 16.061 ± 0.100°, 16.458 ± 0.100°, 18.657 ± 0.100°, 19.407 ± 0.100°, 20.638 ± 0.100°, 21.810 ± 0.100°, and 22.836 ± 0.100°.

[0017] In some embodiments of the application, the Form A has an X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2Θ angles: 7.736 ± 0.100°, 14.223 ± 0.100°, 14.589 ± 0.100°, 14.894 ± 0.100°, 15.492 ± 0.100°, 16.061 ± 0.100°, 16.458 ± 0.100°, 18.657 ± 0.100°, 19.407 ± 0.100°, 20.638 ± 0.100°, 21.810 ± 0.100°, and 22.836 ± 0.100°.

[0018] In some embodiments of the application, the X-ray powder diffraction pattern of the Form A is characterized by peaks at 7.736 ± 0.200°, 9.070 ± 0.200°, 11.289 ± 0.200°, 11.678 ± 0.200°, 14.223 ± 0.200°, 14.589 ± 0.200°, 14.894 ± 0.200°, 15.492 ± 0.200°, 16.061 ± 0.200°, 16.458 ± 0.200°, 18.657 ± 0.200°, 19.407 ± 0.200°, 20.638 ± 0.200°, 21.085 ± 0.200°, 21.810 ± 0.200°, and 22.836 ± 0.200°.

[0019] In some embodiments of the application, the X-ray powder diffraction pattern of the Form A is characterized by peaks at 7.736 ± 0.100°, 9.070 ± 0.100°, 11.289 ± 0.100°, 11.678 ± 0.100°, 14.223 ± 0.100°, 14.589 ± 0.100°, 14.894 ± 0.100°, 15.492 ± 0.100°, 16.061 ± 0.100°, 16.458 ± 0.100°, 18.657 ± 0.100°, 19.407 ± 0.100°, 20.638 ± 0.100°, 21.085 ± 0.100°, 21.810 ± 0.100°, and 22.836 ± 0.100°.

[0020] In some embodiments of the application, the X-ray powder diffraction pattern of the Form A has characteristic diffraction peaks at the following 2Θ angles: 7.736 ± 0.200°, 8.475 ± 0.200°, 9.070 ± 0.200°, 11.289 ± 0.200°, 11.678 ± 0.200°, 12.363 ± 0.200°, 14.223 ± 0.200°, 14.589 ± 0.200°, 14.894 ± 0.200°, 15.492 ± 0.200°, 16.061 ± 0.200°, 16.458 ± 0.200°, 17.000 ± 0.200°, 18.657 ± 0.200°, 19.030 ± 0.200°, 19.407 ± 0.200°, 19.882 ± 0.200°, 20.638 ± 0.200°, 21.085 ± 0.200°, 21.810 ± 0.200°, 22.836 ± 0.200°, 23.717 ± 0.200°, 24.147 ± 0.200°, 24.693 ± 0.200°, 25.311 ± 0.200°, 26.802 ± 0.200°, 27.462 ± 0.200°, 28.537 ± 0.200°, and 31.264 ± 0.200°.

[0021] In some embodiments of the application, the X-ray powder diffraction pattern of the Form A has characteristic diffraction peaks at the following 2Θ angles: 7.736 ± 0.200°, 8.475 ± 0.200°, 9.070 ± 0.200°, 11.289 ± 0.200°, 11.678 ± 0.200°, 12.363 ± 0.200°, 14.223 ± 0.200°, 14.589 ± 0.200°, 14.894 ± 0.200°, 15.492 ± 0.200°, 16.061 ± 0.200°, 16.458 ± 0.200°, 17.000 ± 0.200°, 18.657 ± 0.200°, 19.030 ± 0.200°, 19.407 ± 0.200°, 19.882 ± 0.200°, 20.638 ± 0.200°, 21.085 ± 0.200°, 21.810 ± 0.200°, 22.836 ± 0.200°, 23.717 ± 0.200°, 24.147 ± 0.200°, 24.693 ± 0.200°, 25.311 ± 0.200°, 26.802 ± 0.200°, 27.462 ± 0.200°, 28.537 ± 0.200°, 30.090 ± 0.200°, 31.264 ± 0.200°, 32.346 ± 0.200°, 33.429 ± 0.200°, 35.334 ± 0.200°, and 36.567 ± 0.200°.

[0022] In some embodiments of the application, the X-ray powder diffraction pattern of the Form A is characterized by characteristic diffraction peaks at the following 2Θ angles: 7.736 ± 0.100°, 8.475 ± 0.100°, 9.070 ± 0.100°, 11.289 ± 0.100°, 11.678 ± 0.100°, 12.363 ± 0.100°, 14.223 ± 0.100°, 14.589 ± 0.100°, 14.894 ± 0.100°, 15.492 ± 0.100°, 16.061 ± 0.100°, 16.458 ± 0.100°, 17.000 ± 0.100°, 18.657 ± 0.100°, 19.030 ± 0.100°, 19.407 ± 0.100°, 19.882 ± 0.100°, 20.638 ± 0.100°, 21.085 ± 0.100°, 21.810 ± 0.100°, 22.836 ± 0.100°, 23.717 ± 0.100°, 24.147 ± 0.100°, 24.693 ± 0.100°, 25.311 ± 0.100°, 26.802 ± 0.100°, 27.462 ± 0.100°, 28.537 ± 0.100°, and 31.264 ± 0.100°.

[0023] In some embodiments of the application, the X-ray powder diffraction pattern of the Form A is characterized by characteristic diffraction peaks at the following 2Θ angles: 7.736 ± 0.100°, 8.475 ± 0.100°, 9.070 ± 0.100°, 11.289 ± 0.100°, 11.678 ± 0.100°, 12.363 ± 0.100°, 14.223 ± 0.100°, 14.589 ± 0.100°, 14.894 ± 0.100°, 15.492 ± 0.100°, 16.061 ± 0.100°, 16.458 ± 0.100°, 17.000 ± 0.100°, 18.657 ± 0.100°, 19.030 ± 0.100°, 19.407 ± 0.100°, 19.882 ± 0.100°, 20.638 ± 0.100°, 21.085 ± 0.100°, 21.810 ± 0.100°, 22.836 ± 0.100°, 23.717 ± 0.100°, 24.147 ± 0.100°, 24.693 ± 0.100°, 25.311 ± 0.100°, 26.802 ± 0.100°, 27.462 ± 0.100°, 28.537 ± 0.100°, 30.090 ± 0.100°, 31.264 ± 0.100°, 32.346 ± 0.100°, 33.429 ± 0.100°, 35.334 ± 0.100°, and 36.567 ± 0.100°.

[0024] In some embodiments of the application, the X-ray powder diffraction pattern of the Form A is characterized by peaks at 2-theta values of 7.736°, 8.475°, 9.070°, 11.289°, 11.678°, 12.363°, 14.223°, 14.589°, 14.894°, 15.492°, 16.061°, 16.458°, 17.000°, 18.657°, 19.030°, 19.407°, 19.882°, 20.638°, 21.085°, 21.810°, 22.836°, 23.717°, 24.147°, 24.693°, 25.311°, 26.802°, 27.462°, 28.537°, 30.090°, 31.264°, 32.346°, 33.429°, 35.334°, and 36.567°.

[0025] In some embodiments of the application, the X-ray powder diffraction pattern of the Form A is characterized by peaks at 2-theta values of 7.736°, 8.475°, 9.070°, 11.289°, 11.678°, 12.363°, 14.223°, 14.589°, 14.894°, 15.492°, 16.061°, 16.458°, 17.000°, 18.657°, 19.030°, 19.407°, 19.882°, 20.638°, 21.085°, 21.810°, 22.836°, 23.717°, 24.147°, 24.693°, 25.311°, 26.802°, 27.462°, 28.537°, 30.090°, 31.264°, 32.346°, 33.429°, 35.334°, and 36.567°.

[0026] In some embodiments of the application, the X-ray powder diffraction pattern of the Form A has characteristic diffraction peaks at the following 2 theta angles: 15.492+0.200°, 16.458+0.200°, 18.657+0.200°, and / or 7.736+0.200°, and / or 8.475+0.200°, and / or 9.070+0.200°, and / or 11.289+0.200°, and / or 11.678+0.200°, and / or 12.363+0.200°, and / or 14.223+0.200°, and / or 14.589+0.200°, and / or 14.894+0.200°, and / or 16.061+0.200°, and / or 17.000+0.200°, and / or 19.030+0.200°, and / or 19.407+0.200°, and / or 19.882+0.200°, and / or 20.638+0.200°, and / or 21.085+0.200°, and / or 21.810+0.200°, and / or 22.836+0.200°, and / or 23.717+0.200°, and / or 24.147+0.200°, and / or 24.693+0.200°, and / or 25.311+0.200°, and / or 26.802+0.200°, and / or 27.462+0.200°, and / or 28.537+0.200°, and / or 30.090+0.200°, and / or 31.264+0.200°, and / or 32.346+0.200°, and / or 33.429+0.200°, and / or 35.334+0.200°, and / or 36.567+0.200°.

[0027] In some embodiments of the application, the X-ray powder diffraction pattern of the Form A crystalline form described above has characteristic diffraction peaks at the following 2-theta angles: 15.492 ± 0.100°, 16.458 ± 0.100°, 18.657 ± 0.100°, and / or 7.736 ± 0.100°, and / or 8.475 ± 0.100°, and / or 9.070 ± 0.100°, and / or 11.289 ± 0.100°, and / or 11.678 ± 0.100°, and / or 12.363 ± 0.100°, and / or 14.223 ± 0.100°, and / or 14.589 ± 0.100°, and / or 14.894 ± 0.100°, and / or 16.061 ± 0.100°, and / or 17.000 ± 0.100°, and / or 19.030 ± 0.100°, and / or 19.407 ± 0.100°, and / or 19.882 ± 0.100°, and / or 20.638 ± 0.100°, and / or 21.085 ± 0.100°, and / or 21.810 ± 0.100°, and / or 22.836 ± 0.100°, and / or 23.717 ± 0.100°, and / or 24.147 ± 0.100°, and / or 24.693 ± 0.100°, and / or 25.311 ± 0.100°, and / or 26.802 ± 0.100°, and / or 27.462 ± 0.100°, and / or 28.537 ± 0.100°, and / or 30.090 ± 0.100°, and / or 31.264 ± 0.100°, and / or 32.346 ± 0.100°, and / or 33.429 ± 0.100°, and / or 35.334 ± 0.100°, and / or 36.567 ± 0.100°.

[0028] In some embodiments of the application, the XRPD pattern of the Form A crystalline form described above is substantially as shown in Figure 1. Figure 1

[0029] In some embodiments of the application, the XRPD pattern of the Form A crystalline form described above is substantially as shown in Figure 1.

[0030] Table 1. XRPD pattern data for Form A of the compound of formula (I)

[0031]

[0032]

[0033] In some embodiments of the application, the Form A crystalline form described above has a differential scanning calorimetry curve with an endothermic peak having an onset at 169.0 ± 5 °C.

[0034] ​In some embodiments of the application, the Form A has a differential scanning calorimetry curve with endothermic peaks at 31.6±5°C and 169.0±5°C.

[0035] In some embodiments of the application, the Form A has a DSC pattern substantially as shown in FIG. 2. Figure 2

[0036] In some embodiments of the application, the Form A has a thermogravimetric analysis curve with 1.48% weight loss at 160.0±3°C.

[0037] In some embodiments of the application, the Form A has a TGA pattern substantially as shown in FIG. 4. Figure 3

[0038] The present application also provides a Form B of the compound of formula (I), which has an X-ray powder diffraction (XRPD) pattern with characteristic diffraction peaks at the following 2θ angles: 5.932±0.200°, 12.435±0.200°, 14.091±0.200° and 16.496±0.200°;

[0039]

[0040] In some embodiments of the application, the Form B has an X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2θ angles: 5.932±0.200°, 8.551±0.200°, 10.855±0.200°, 12.435±0.200°, 14.091±0.200°, 15.171±0.200°, 16.496±0.200° and 23.682±0.200°.

[0041] In some embodiments of the application, the Form B has an X-ray powder diffraction pattern, in terms of 2θ angles, comprising at least 4, 5, 6, 7 or 8 characteristic diffraction peaks selected from 5.932±0.200°, 8.551±0.200°, 10.855±0.200°, 12.435±0.200°, 14.091±0.200°, 15.171±0.200°, 16.496±0.200° and 23.682±0.200°.

[0042] ​​In some embodiments of the application, the Form B has an X-ray powder diffraction pattern with characteristic peaks at 2-theta = 5.932 ± 0.200°, 6.751 ± 0.200°, 8.551 ± 0.200°, 9.098 ± 0.200°, 10.855 ± 0.200°, 12.435 ± 0.200°, 14.091 ± 0.200°, 15.171 ± 0.200°, 16.496 ± 0.200°, 19.193 ± 0.200°, 20.501 ± 0.200°, and 23.682 ± 0.200°.

[0043] In some embodiments of the application, the Form B has an X-ray powder diffraction pattern with characteristic peaks at 2-theta = 5.932 ± 0.200°, 6.751 ± 0.200°, 8.551 ± 0.200°, 9.098 ± 0.200°, 10.855 ± 0.200°, 12.435 ± 0.200°, 14.091 ± 0.200°, 15.171 ± 0.200°, 16.496 ± 0.200°, 19.193 ± 0.200°, 20.501 ± 0.200°, and 23.682 ± 0.200°.

[0044] In some embodiments of the application, the Form B has an X-ray powder diffraction pattern with characteristic peaks at 2-theta = 5.932 ± 0.200°, 6.751 ± 0.200°, 8.551 ± 0.200°, 9.098 ± 0.200°, 10.186 ± 0.200°, 10.855 ± 0.200°, 11.710 ± 0.200°, 12.435 ± 0.200°, 14.091 ± 0.200°, 15.171 ± 0.200°, 16.049 ± 0.200°, 16.496 ± 0.200°, 17.438 ± 0.200°, 18.658 ± 0.200°, 19.193 ± 0.200°, 20.501 ± 0.200°, 21.241 ± 0.200°, 21.977 ± 0.200°, 23.682 ± 0.200°, 26.127 ± 0.200°, 26.981 ± 0.200°, and 29.033 ± 0.200°.

[0045] In some embodiments of the application, the X-ray powder diffraction pattern of the B crystalline form described above has characteristic diffraction peaks at the following 2Θ angles: 5.932°, 6.751°, 8.551°, 9.098°, 10.186°, 10.855°, 11.710°, 12.435°, 14.091°, 15.171°, 16.049°, 16.496°, 17.438°, 18.658°, 19.193°, 20.501°, 21.241°, 21.977°, 23.682°, 26.127°, 26.981°, and 29.033°.

[0046] In some embodiments of the application, the XRPD pattern of the B crystalline form described above has characteristic diffraction peaks at the following 2Θ angles: 5.932±0.200°, 12.435±0.200°, 14.091±0.200°, and / or 6.751±0.200°, and / or 8.551±0.200°, and / or 9.098±0.200°, and / or 10.186±0.200°, and / or 10.855±0.200°, and / or 11.710±0.200°, and / or 12.435±0.200°, and / or 14.091±0.200°, and / or 15.171±0.200°, and / or 16.049±0.200°, and / or 16.496±0.200°, and / or 17.438±0.200°, and / or 18.658±0.200°, and / or 19.193±0.200°, and / or 20.501±0.200°, and / or 21.241±0.200°, and / or 21.977±0.200°, and / or 23.682±0.200°, and / or 26.127±0.200°, and / or 26.981±0.200°, and / or 29.033±0.200°.

[0047] In some embodiments of the application, the XRPD pattern of the B crystalline form described above is substantially as shown in Figure 2. Figure 4

[0048] In some embodiments of the application, the XRPD pattern of the B crystalline form described above is substantially as shown in Figure 2.

[0049] Table 2. XRPD pattern resolution data for the B crystalline form of the compound of formula (I)

[0050]

[0051] In some embodiments of the application, the B crystalline form described above has a differential scanning calorimetry curve with endothermic peaks at 64.5±5°C and 121.7±5°C.

[0052] ​In some embodiments of the application, the B crystal form has a DSC pattern substantially as shown in Figure 5

[0053] In some embodiments of the application, the B crystal form has a TGA pattern substantially as shown in

[0054] In some embodiments of the application, the B crystal form has a TGA pattern substantially as shown in Figure 6

[0055] The present application also provides a C crystal form of the compound of formula (I), which has an X-ray powder diffraction (XRPD) pattern with characteristic diffraction peaks at the following 2θ angles: 13.546±0.200°, 14.908±0.200°, 15.539±0.200°, 18.230±0.200° and 22.932±0.200°;

[0056]

[0057] In some embodiments of the application, the C crystal form has an X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2θ angles: 13.546±0.200°, 14.908±0.200°, 15.539±0.200°, 16.401±0.200°, 18.230±0.200°, 18.699±0.200°, 20.226±0.200° and 22.932±0.200°.

[0058] In some embodiments of the application, the C crystal form has an X-ray powder diffraction pattern with at least 4, 5, 6, 7 or 8 characteristic diffraction peaks selected from the group consisting of: 13.546±0.200°, 14.908±0.200°, 15.539±0.200°, 16.401±0.200°, 18.230±0.200°, 18.699±0.200°, 20.226±0.200° and 22.932±0.200°.

[0059] In some embodiments of the application, the C crystal form has an X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2θ angles: 7.410±0.200°, 8.474±0.200°, 13.546±0.200°, 14.122±0.200°, 14.908±0.200°, 15.539±0.200°, 16.401±0.200°, 18.230±0.200°, 18.699±0.200°, 19.665±0.200°, 20.226±0.200° and 22.932±0.200°.

[0060] ​​In some embodiments of the application, the Form C has an X-ray powder diffraction pattern comprising at least 8, 9, 10, 11, or 12 peaks selected from the group consisting of 7.410±0.200°, 8.474±0.200°, 13.546±0.200°, 14.122±0.200°, 14.908±0.200°, 15.539±0.200°, 16.401±0.200°, 18.230±0.200°, 18.699±0.200°, 19.665±0.200°, 20.226±0.200°, and 22.932±0.200°, when the pattern is measured using the Cu K alpha 1 bandpass.

[0061] In some embodiments of the application, the Form C has an X-ray powder diffraction pattern comprising peaks at 7.410±0.200°, 8.474±0.200°, 9.093±0.200°, 11.284±0.200°, 13.546±0.200°, 14.122±0.200°, 14.908±0.200°, 15.539±0.200°, 16.401±0.200°, 18.230±0.200°, 18.699±0.200°, 18.911±0.200°, 19.665±0.200°, 20.226±0.200°, 21.508±0.200°, and 22.932±0.200°.

[0062] In some embodiments of the application, the Form C has an X-ray powder diffraction pattern comprising peaks at 7.410±0.100°, 8.474±0.100°, 9.093±0.100°, 11.284±0.100°, 13.546±0.100°, 14.122±0.100°, 14.908±0.100°, 15.539±0.100°, 16.401±0.100°, 18.230±0.100°, 18.699±0.100°, 18.911±0.100°, 19.665±0.100°, 20.226±0.100°, 21.508±0.100°, and 22.932±0.100°.

[0063] In some embodiments of the application, the X-ray powder diffraction pattern of the crystalline form C has characteristic diffraction peaks at the following 2Θ angles: 7.410 ± 0.100°, 8.474 ± 0.100°, 9.093 ± 0.100°, 11.284 ± 0.100°, 11.439 ± 0.100°, 13.546 ± 0.100°, 14.122 ± 0.100°, 14.908 ± 0.100°, 15.539 ± 0.100°, 16.401 ± 0.100°, 16.743 ± 0.100°, 17.169 ± 0.100°, 18.230 ± 0.100°, 18.699 ± 0.100°, 18.911 ± 0.100°, 19.665 ± 0.100°, 20.226 ± 0.100°, 21.508 ± 0.100°, 22.932 ± 0.100°, 23.907 ± 0.100°, 25.111 ± 0.100°, 27.317 ± 0.100°, 29.113 ± 0.100°, and 31.325 ± 0.100°.

[0064] In some embodiments of the application, the X-ray powder diffraction pattern of the crystalline form C has characteristic diffraction peaks at the following 2Θ angles: 7.410 ± 0.100°, 8.474 ± 0.100°, 9.093 ± 0.100°, 11.284 ± 0.100°, 11.439 ± 0.100°, 13.546 ± 0.100°, 14.122 ± 0.100°, 14.908 ± 0.100°, 15.539 ± 0.100°, 16.401 ± 0.100°, 16.743 ± 0.100°, 17.169 ± 0.100°, 18.230 ± 0.100°, 18.699 ± 0.100°, 18.911 ± 0.100°, 19.665 ± 0.100°, 20.226 ± 0.100°, 21.508 ± 0.100°, 22.932 ± 0.100°, 23.907 ± 0.100°, 25.111 ± 0.100°, 27.317 ± 0.100°, 29.113 ± 0.100°, and 31.325 ± 0.100°.

[0065] In some embodiments of the application, the X-ray powder diffraction pattern of the Form C has characteristic diffraction peaks at the following 2Θ angles: 7.410°, 8.474°, 9.093°, 11.284°, 11.439°, 13.546°, 14.122°, 14.908°, 15.539°, 16.401°, 16.743°, 17.169°, 18.230°, 18.699°, 18.911°, 19.665°, 20.226°, 21.508°, 22.932°, 23.907°, 25.111°, 27.317°, 29.113°, and 31.325°.

[0066] In some embodiments of the application, the X-ray powder diffraction pattern of the Form C has characteristic diffraction peaks at the following 2Θ angles: 7.410±0.200°, 8.474±0.200°, 9.093±0.200°, 11.284±0.200°, 11.439±0.200°, 13.546±0.200°, 14.122±0.200°, 14.908±0.200°, 15.539±0.200°, 16.401±0.200°, 16.743±0.200°, 17.169±0.200°, 18.230±0.200°, 18.699±0.200°, 18.911±0.200°, 19.665±0.200°, 20.226±0.200°, 21.508±0.200°, 22.932±0.200°, 23.907±0.200°, 25.111±0.200°, 27.317±0.200°, 29.113±0.200°, and 31.325±0.200°.

[0067] In some embodiments of the application, the X-ray powder diffraction pattern of the crystalline Form C has characteristic diffraction peaks at the following 2Θ angles: 7.410 ± 0.100°, 8.474 ± 0.100°, 9.093 ± 0.100°, 11.284 ± 0.100°, 11.439 ± 0.100°, 13.546 ± 0.100°, 14.122 ± 0.100°, 14.908 ± 0.100°, 15.539 ± 0.100°, 16.401 ± 0.100°, 16.743 ± 0.100°, 17.169 ± 0.100°, 18.230 ± 0.100°, 18.699 ± 0.100°, 18.911 ± 0.100°, 19.665 ± 0.100°, 20.226 ± 0.100°, 21.508 ± 0.100°, 22.932 ± 0.100°, 23.907 ± 0.100°, 25.111 ± 0.100°, 27.317 ± 0.100°, 29.113 ± 0.100°, and 31.325 ± 0.100°.

[0068] In some embodiments of the application, the X-ray powder diffraction pattern of the crystalline Form C has characteristic diffraction peaks at the following 2Θ angles: 13.546 ± 0.200°, 14.908 ± 0.200°, 15.539 ± 0.200°, and / or 7.410 ± 0.200°, and / or 8.474 ± 0.200°, and / or 9.093 ± 0.200°, and / or 11.284 ± 0.200°, and / or 11.439 ± 0.200°, and / or 14.122 ± 0.200°, and / or 16.401 ± 0.200°, and / or 16.743 ± 0.200°, and / or 17.169 ± 0.200°, and / or 18.230 ± 0.200°, and / or 18.699 ± 0.200°, and / or 18.911 ± 0.200°, and / or 19.665 ± 0.200°, and / or 20.226 ± 0.200°, and / or 21.508 ± 0.200°, and / or 22.932 ± 0.200°, and / or 23.907 ± 0.200°, and / or 25.111 ± 0.200°, and / or 27.317 ± 0.200°, and / or 29.113 ± 0.200°, and / or 31.325 ± 0.200°.

[0069] In some embodiments of the application, the X-ray powder diffraction pattern of the crystalline form C has characteristic peaks at 2-theta values of 13.546±0.100°, 14.908±0.100°, 15.539±0.100°, and / or 7.410±0.100°, and / or 8.474±0.100°, and / or 9.093±0.100°, and / or 11.284±0.100°, and / or 11.439±0.100°, and / or 14.122±0.100°, and / or 16.401±0.100°, and / or 16.743±0.100°, and / or 17.169±0.100°, and / or 18.230±0.100°, and / or 18.699±0.100°, and / or 18.911±0.100°, and / or 19.665±0.100°, and / or 20.226±0.100°, and / or 21.508±0.100°, and / or 22.932±0.100°, and / or 23.907±0.100°, and / or 25.111±0.100°, and / or 27.317±0.100°, and / or 29.113±0.100°, and / or 31.325±0.100°.

[0070] In some embodiments of the application, the XRPD pattern of the crystalline form C is substantially as shown in Figure 2. Figure 7

[0071] In some embodiments of the application, the XRPD pattern of the crystalline form C has the analytical data as shown in Table 3.

[0072] Table 3. XRPD pattern analytical data of the crystalline form C of the compound of formula (I)

[0073]

[0074] In some embodiments of the application, the crystalline form C has a differential scanning calorimetry curve with an endothermic peak starting point at 169.2±5°C.

[0075] In some embodiments of the application, the DSC pattern of the crystalline form C is substantially as shown in Figure 4. Figure 8

[0076] In some embodiments of the application, the crystalline form C has a thermogravimetric analysis curve with a weight loss of 2.00% at 130.0±3°C.

[0077] In some embodiments of the application, the TGA pattern of the crystalline form C is substantially as shown in Figure 6. Figure 9

[0078] The present application also provides use of the crystalline form of the compound of formula (I) in the preparation of a medicament for treating KRAS mutant solid tumor diseases.​​​

[0079] The present application also provides the use of the A crystal form of the compound of formula (I) in the manufacture of a medicament for treating KRAS mutant solid tumor diseases.

[0080] The present application also provides the following biological test method for the crystal form of the compound of formula (I) above:

[0081] Test method 1: in vivo efficacy evaluation of the compound in a Miapaca2 nude mouse xenograft model

[0082] Cell culture:

[0083] Human pancreatic cancer cells (Miapaca2) were cultured in vitro in a monolayer adherent manner, and the culture conditions were as follows: DMEM medium containing 10% fetal bovine serum, 37°C, 5% CO2 incubator. The cells were routinely digested and passaged with trypsin-EDTA twice to three times a week. When the cell saturation degree was 80%-90% and the number reached the requirement, the cells were collected, counted, and inoculated.

[0084] Experimental animals:

[0085] Balb / c nude mice, female, 6-7 weeks old, purchased from Shanghai Xipu-Bike Experimental Animal Co., Ltd.

[0086] Model preparation:

[0087] 0.2 mL (5 x 10 6 Miapaca2 cells (mixed with Matrigel at a volume ratio of 1:1) were subcutaneously inoculated on the right back of each mouse, and when the average tumor volume reached 118 mm 3 , the mice were grouped and administered.

[0088] Tumor measurement and experimental indicators:

[0089] The tumor diameter was measured twice a week with a vernier caliper, and the tumor volume was measured in cubic millimeters, calculated by the following formula: V = 0.5a x b 2 , where a and b are the long diameter and short diameter of the tumor, respectively. The antitumor effect of the test compound was evaluated by using TGI (%). TGI (%) reflects the tumor growth inhibition rate. TGI (%) = [1 - (average tumor volume at the end of administration in a certain treatment group - average tumor volume at the start of administration in the treatment group) / (average tumor volume at the end of treatment in the solvent control group - average tumor volume at the start of treatment in the solvent control group)] x 100%.

[0090] Experimental conclusion: the compound of the present application combined with AMG-510 exhibits excellent antitumor effect in a Miapaca2 nude mouse xenograft model.

[0091] Technical effects

[0092] The compound of the present application can better inhibit the activity of SOS1; also has obvious p-ERK proliferation inhibition activity on DLD-1 cells, and has good pharmacokinetic properties (including good oral bioavailability, oral exposure, half-life and clearance rate, etc.); the compound has no significant inhibition effect on hERG potassium ion channel, and has high safety; the compound of the present application has excellent tumor inhibition effect on human pancreatic cancer Miapaca-2 xenograft tumor model. The crystal form of the compound of formula (I) is easy to obtain, and has good physical stability and chemical stability, and has high industrial application value and economic value.

[0093] Definitions and Descriptions

[0094] The following terms and phrases used herein are intended to have the following meanings unless otherwise indicated. A particular phrase or term should not be construed as indefinite or unclear unless specifically defined, but should be construed in accordance with the ordinary meaning. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredient.

[0095] It must be noted that, as used herein and in the appended claims, the singular form "a", "an", and "the" include plural references unless the context clearly dictates otherwise. Thus, for example, reference to "a compound" includes a single or a plurality of compounds; and so forth.

[0096] The differential scanning calorimetry (DSC) of the crystal form of the present application has experimental errors and is slightly affected by the drying degree of the sample. The position and peak value of the endothermic peak may vary slightly between one machine and another machine and between one sample and another sample. The numerical value of the experimental error or difference may be less than or equal to 10°C, or less than or equal to 9°C, or less than or equal to 8°C, or less than or equal to 7°C, or less than or equal to 6°C, or 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. Therefore, the numerical value of the peak position or peak value of the DSC endothermic peak cannot be considered absolute.

[0097] The intermediate compounds of the present application can be prepared by various synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by the combination of the specific embodiments with other chemical synthesis methods, and the equivalent replacement methods well known to those skilled in the art, and the preferred embodiments include but are not limited to the examples of the present application.

[0098] The compounds of the present application can be confirmed by conventional methods well known to those skilled in the art. If the present application relates to the absolute configuration of a compound, the absolute configuration can be confirmed by conventional techniques in the art. For example, single crystal X-ray diffraction (SXRD), a single crystal is grown and diffracted intensity data is collected using a Bruker D8 venture diffractometer with Cu Kα radiation, scanning mode: After the relevant data is collected, the crystal structure is further resolved using the direct method (Shelxs97), and the absolute configuration can be confirmed.

[0099] The chemical reactions of the present application are performed in solvents appropriate to the reagents and materials employed and suitable for the chemical changes being effected. In the synthetic schemes and examples described below, all substituents unless otherwise indicated are as previously defined. The chemical reactions described in the schemes and examples are performed in any suitable order, unless otherwise indicated. Exemplary synthetic methods useful in preparing the compounds of the present application are shown in the following schemes and examples.

[0100] The present application will now be described in detail by way of Examples, which are not meant to limit the present application in any way.

[0101] All solvents used in the present application are commercially available and used without further purification.

[0102] The solvents used in the present application are commercially available.

[0103] The following abbreviations are used in the present application: Boc represents tert-butyloxycarbonyl; DCM represents dichloromethane; DMF represents N,N-dimethylformamide; THF represents tetrahydrofuran; DMSO represents dimethylsulfoxide; EtOH represents ethanol; MeOH represents methanol; ACN (MeCN) represents acetonitrile; EtOAc represents ethyl acetate; H2O represents water; Acetone represents acetone; IPAc represents isopropyl acetate; MTBE represents methyl tert-butyl ether; 1,4-Dioxane represents 1,4-dioxane; n-Heptane represents n-heptane; i-PrOAc represents isopropyl acetate; TEA represents triethylamine; DIPEA represents diisopropylethylamine; BID represents twice a day; QD represents once a day; p.o. represents per os.

[0104] The compounds are named by hand or The commercially available compounds are named by the supplier's catalog name.

[0105] Instruments and analytical methods of the present application

[0106] X-ray powder diffractometer (XRPD) method 1 of the present application, the test parameters are shown in Table 4.

[0107] Table 4 XRPD test parameters

[0108]

[0109]

[0110] The X-ray powder diffractometer (XRPD) method two of the present application, the test parameters are shown in Table 5.

[0111] Table 5 XRPD test parameters

[0112]

[0113] The Differential Scanning Calorimeter (DSC) method of the present application, the test parameters are shown in Table 6.

[0114] Table 6 DSC test parameters

[0115]

[0116] The Thermal Gravimetric Analyzer (TGA) method of the present application, the test parameters are shown in Table 7.

[0117] Table 7 TGA test parameters

[0118]

[0119]

[0120] The Dynamic Vapor Sorption (DVS) method of the present application, the test parameters are shown in Table 8.

[0121] Table 8 DVS test parameters

[0122] BRIEF DESCRIPTION OF DRAWINGS

[0123] Figure 1 : XRPD pattern of Form A of compound of formula (I) in Cu-Kα radiation.

[0124] Figure 2 : DSC pattern of Form A of compound of formula (I).

[0125] Figure 3 : TGA pattern of Form A of compound of formula (I).

[0126] Figure 4 : XRPD pattern of Form B of compound of formula (I) in Cu-Kα radiation.

[0127] Figure 5 DSC pattern of the Form B of the compound of formula (I).

[0128] Figure 6 TGA pattern of the Form B of the compound of formula (I).

[0129] Figure 7 XRPD pattern of the Form C of the compound of formula (I) using Cu-Ka radiation.

[0130] Figure 8 DSC pattern of the Form C of the compound of formula (I).

[0131] Figure 9 TGA pattern of the Form C of the compound of formula (I).

[0132] Figure 10 DVS pattern of the Form A of the compound of formula (I).

[0133] Figure 11 Single crystal X-ray diffraction (SXRD) stereographic ellipsoid plot of the compound of formula (I). DETAILED DESCRIPTION

[0134] In order to better understand the content of the present application, further description will be made in combination with specific examples below, but the specific examples are not intended to limit the content of the present application.

[0135] Example 1: Preparation of hydrochloride salt of the compound of formula (I)

[0136] Synthetic route:

[0137]

[0138] First step

[0139] Compound 1 (10.0 g, 49.9 mmol) was dissolved in methanol (100 mL), and formaldehyde aqueous solution (20.3 g, 250 mmol, purity: 37%), sodium cyanoborohydride (9.41 g, 150 mmol) and acetic acid (15.0 g, 250 mmol) were added, and the reaction was stirred at 25°C for 12 hours. Water (300 mL) was added to the reaction solution, and extraction was performed with ethyl acetate (200 mL x 3), and the organic phase was washed with saturated brine (100 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (dichloromethane / methanol, 1 / 0 to 10 / 1, V / V) to obtain compound 2. 1H NMR (400 MHz, CDC13) δ 4.21-4.18 (m, 1H), 3.82-3.79 (m, 1H), 3.13-3.06 (m, 1H), 2.74-2.70 (m, 1H), 2.60-2.52 (m, 1H), 2.24 (s, 3H), 2.11-2.07 (m, 1H), 1.93-1.86 (m, 1H), 1.45 (s, 9H), 1.29 (d, J = 7.2 Hz, 3H). MS-ESI calculated [M+H] + 215, found 215.

[0140] Second step

[0141] Compound 2 (7.25 g, 33.8 mmol) was dissolved in dioxane (50 mL), and hydrogen chloride / dioxane solution (4 M, 67.6 mL) was added dropwise. The reaction was stirred at 25 °C for 12 h. The reaction was concentrated under reduced pressure to obtain crude compound 3 hydrochloride salt. 1 H NMR (400 MHz, CDC13) δ 4.21-4.18 (m, 1H), 3.82-3.79 (m, 1H), 3.13-3.06 (m, 1H), 2.74-2.70 (m, 1H), 2.60-2.52 (m, 1H), 2.24 (s, 3H), 2.11-2.07 (m, 1H), 1.93-1.86 (m, 1H), 1.45 (s, 9H), 1.29 (d, J = 7.2 Hz, 3H). MS-ESI calculated [M+H]

[0142] Third step

[0143] Compound 3 hydrochloride salt (811 mg, 5.39 mmol) was dissolved in N,N- dimethylformamide (15 mL), and triethylamine (1.64 g, 16.2 mmol) and compound 4 (1.00 g, 2.69 mmol) were added dropwise. The reaction was stirred at 25 °C for 12 h. Filtration, concentration under reduced pressure, and purification by silica gel column chromatography (dichloromethane / methanol, 20 / 1 to 1 / 1, V / V) gave compound 5. MS-ESI calculated [M+H] + 347, found 347.

[0144] Fourth step

[0145] Compound 5 (254 mg, 733 μmol) was dissolved in dichloromethane (5 mL), compound 6 (333 mg, 1.10 mmol), 4-dimethylaminopyridine (8.96 mg, 73.3 μmol) and N, N-diisopropylethylamine (284 mg, 2.20 mmol) were added, and the reaction was stirred at 25 °C for 12 h. Water (50 mL) was added to the reaction, and the product was extracted with ethyl acetate (40 mL x 5), the organic phase was washed with saturated brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (dichloromethane / methanol, 1 / 0 ~ 10 / 1, V / V) to obtain compound 7. MS-ESI calculated value [M+H] + 613, found 613.

[0146] Fifth step

[0147] Compound 7 (154 mg, 251 μmol) was dissolved in dimethyl sulfoxide (5 mL), compound 8 (93.2 mg, 377 μmol) and triethylamine (76.3 mg, 754 μmol) were added, and the reaction was stirred at 90 °C for 12 h. Water (50 mL) was added to the reaction, and the product was extracted with ethyl acetate (50 mL x 3), the organic phase was washed with saturated brine (50 mL x 5), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was separated and purified by preparative high performance liquid chromatography (column: Xtimate C18 150 x 40 mm x 5 μm; mobile phase: 0.05% hydrochloric acid in water-acetonitrile; gradient: acetonitrile 5% ~ 30%, 10 min) to obtain a hydrochloride salt of the compound of formula (I). 1 HNMR (400 MHz, CD3OD) δ 8.40 (s, 1H), 7.61 (t, J = 7.2 Hz, 1H), 7.42 (t, J = 6.8 Hz, 1H), 7.23 (t, J = 7.2 Hz, 1H), 7.16 (s, 1H), 6.01-5.99 (m, 1H), 4.93-4.86 (m, 1H) 4.68-4.22 (m, 1H), 4.04 (s, 3H), 3.62-3.55 (m, 4H), 3.23-3.19 (m, 1H), 2.99 (s, 3H), 2.62 (s, 3H), 1.74 (d, J = 6.8 Hz, 3H), 1.57-1.48 (m, 3H), 1.29 (s, 6H). MS-ESI calculated value [M+H] + 576, found 576.

[0148] Example 2: Preparation of a crystalline form of the compound of formula (I) A

[0149] Synthetic route:

[0150]

[0151] First step

[0152] The hydrochloride salt of the compound of formula (I) (2.50 g, 4.34 mmol) was dissolved in water (100 mL) and ethyl acetate (100 mL), and a saturated sodium carbonate solution was added to the solution until the pH was about 10. The organic phase was extracted with ethyl acetate (100 mL x 3), washed with saturated brine (100 mL x 1), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and freeze-dried to obtain the compound of formula (I). MS-ESI calculated value [M+H] + 576, found 576.

[0153] Second step

[0154] The compound of formula (I) (150 mg, 261 μmol) was added to n-heptane (1.5 mL), and the reaction solution was stirred at 50°C for 24 hours, then stirred at 25°C for 2 hours, filtered, and the filter cake was washed with n-heptane (3 mL) and dried at 50°C under vacuum to obtain the A crystal form of the compound of formula (I). 1 H NMR (400 MHz, CD3OD) δ 8.05 (s, 1H), 7.51 (t, J = 6.8 Hz, 1H), 7.35 (t, J = 6.8 Hz, 1H), 7.13 (t, J = 7.6 Hz, 1H), 7.09 (s, 1H), 5.88-5.78 (m, 1H), 4.79-4.00 (m, 2H), 3.94 (s, 3H), 3.48-3.32 (m, 1H), 2.95-2.76 (m, 2H), 2.41 (s, 3H), 2.32 (s, 3H), 2.29-2.21 (m, 1H), 2.14-2.00 (m, 1H), 1.63 (d, J = 6.8 Hz, 3H), 1.42 (s, 3H), 1.33-1.22 (m, 6H). MS-ESI calculated value [M+H] + 576, found 576. The XRPD of the A crystal form of the compound of formula (I) was tested using the corresponding method, and the XRPD, DSC, TGA, and DVS test results thereof are shown in Figure 1 、 Figure 2 、 Figure 3 and Figure 10 , respectively.

[0155] The n-heptane solvent in the second step described above was replaced with the solvents in Table 9, and the A crystal form of the compound of formula (I) was obtained. The experimental results are shown in Table 9.

[0156] Table 9 50°C suspension stirring test

[0157]

[0158] Example 3: Preparation of the B crystal form of the compound of formula (I)

[0159] The compound of formula (I) (150 mg, 261 μmol) was added to water (1.5 mL), the reaction solution was stirred at 50 °C for 24 hours, then stirred at 25 °C for 2 hours, solid precipitated in the reaction solution, filtered, the filter cake was washed with water (3 mL), and dried at 50 °C under vacuum to obtain the B crystal form of the compound of formula (I). 1 H NMR (400 MHz, CD3OD) δ 8.05 (s, 1H), 7.50 (t, J = 6.8 Hz, 1H), 7.35 (t, J = 6.8 Hz, 1H), 7.13 (t, J = 7.8 Hz, 1H), 7.09 (s, 1H), 5.87-5.79 (m, 1H), 4.72-4.03 (m, 2H), 3.94 (s, 3H), 3.50-3.32 (m, 1H), 2.92-2.76 (m, 2H), 2.41 (s, 3H), 2.32 (s, 3H), 2.29-2.23 (m, 1H), 2.13-2.01 (m, 1H), 1.62 (d, J = 7.2 Hz, 3H), 1.43 (s, 3H), 1.35-1.24 (m, 6H). MS-ESI calculated [M+H] + 576, found 576. The XRPD of the B crystal form of the compound of formula (I) was tested using the corresponding method two, and the results of its XRPD, DSC, and TGA detection were shown in Figure 4 、 Figure 5 and Figure 6 respectively.

[0160] Example 4: Preparation of the C crystal form of the compound of formula (I)

[0161] The compound of formula (I) (300 mg, 521 μmol) was dissolved in isopropyl acetate (1.5 mL), the reaction solution was stirred at 25 °C for 12 hours, solid precipitated in the reaction solution, filtered, the filter cake was dried at 150 °C in a blast drier to obtain the C crystal form of the compound of formula (I). 1H NMR (400 MHz, CD3OD) δ 8.04 (s, 1H), 7.49 (t, J = 6.8 Hz, 1H), 7.38-7.31 (m, 1H), 7.12 (t, J = 7.8 Hz, 1H), 7.09 (s, 1H), 5.86-5.78 (m, 1H), 4.61-3.98 (m, 2H), 3.94 (s, 3H), 3.48-3.32 (m, 1H), 2.92-2.85 (m, 1H), 2.83-2.75 (m, 1H), 2.41 (s, 3H), 2.31 (s, 3H), ·2.29-2.23 (m, 1H), 2.12-2.01 (m, 1H), 1.62 (d, J = 7.2 Hz, 3H), 1.43 (s, 3H), 1.33-1.27 (m, 6H). MS-ESI calculated [M+H] + 576, found 576. The XRPD of the crystal form of the compound of formula (I) C was tested according to the corresponding method two, and the XRPD, DSC, and TGA detection results thereof are shown in the following table 5, table 6, and table 7, respectively. Figure 7 、 Figure 8 and Figure 9

[0162] Example 5: Solid pre-stability test of the crystal form of the compound of formula (I) A

[0163] According to the Guiding Principles for Stability Test of Raw Materials and Preparations (Chinese Pharmacopoeia 2020 Edition General Rules 9001), the pre-stability of the crystal form of the compound of formula (I) A under high temperature (60°C, open), high humidity (25°C / relative humidity 92.5%, open), strong light (5000 lx and ultraviolet intensity 90 μw / cm 2 , open), and high temperature and high humidity conditions (40°C / relative humidity 75% and 60°C / relative humidity 75% or 30°C / relative humidity 65%, open) was investigated.

[0164] About 30 mg of the crystal form of the compound of formula (I) A was weighed into a dry and clean glass bottle, spread into a thin layer, and placed under the influence factor conditions and accelerated conditions as the test sample, and the sample was completely exposed. The samples under high temperature and high humidity were taken for analysis at 5 days and 10 days, and the samples under accelerated conditions were taken for analysis at 1 month, 2 months, and 3 months. The samples placed under light conditions were completely exposed at room temperature. The samples placed under different conditions were detected by XRPD, and the detection results were compared with the initial detection results at 0 days. The results are shown in the following table 10, and the results show that the crystal form of the compound of formula (I) A is a stable crystal form.

[0165] Table 10: Pre-stability crystal form results of the crystal form of the compound of formula (I) A

[0166] ​

[0167] Example 6: Hygroscopicity study of the compound of formula (I) A crystal form

[0168] Experimental materials:

[0169] Dynamic vapor sorption instrument

[0170] Experimental method:

[0171] About 10-20 mg of the compound of formula (I) A crystal form was weighed into a DVS sample pan for testing.

[0172] The hygroscopicity evaluation classification is shown in Table 11:

[0173] Table 11 Hygroscopicity evaluation classification table

[0174]

[0175]

[0176] Note: ΔW% represents the hygroscopic weight gain of the test product at 25±1°C and 80±2% RH.

[0177] Experimental results:

[0178] The DVS spectrum of the compound of formula (I) A crystal form is shown in Figure 10 The DVS results show that the sample has a hygroscopic weight gain of 0.3815% at 25°C / 80% RH, and the sample has slight hygroscopicity. After completing the DVS test (0-95-0% RH), the sample was taken out and exposed to air for XRPD testing, and the results showed that the crystal form did not change before and after the DVS test.

[0179] Experimental conclusion:

[0180] The compound of formula (I) A crystal form has slight hygroscopicity at 25±1°C and 80±2% RH, and the crystal form does not change before and after the crystal form.

[0181] Example 7: Single crystal X-ray diffraction detection analysis of the compound of formula (I)

[0182] 1. Instrument parameters and data collection

[0183] Manufacturer: Bruker Company;

[0184] Instrument model: Bruker D8 VENTURE

[0185] X-ray source: high-intensity microfocus rotating anode light source, Cu target;

[0186] Power: 2.5 kW;

[0187] Tube voltage: 50 kV;

[0188] Tube current: 45mA;

[0189] Angle measuring instrument: Four-axis (Kappa, ω, 2θ, Angle measuring instrument;

[0190] Detector: Large-area photon type II detector, with an effective detector area of ​​14cm × 10cm, and the distance between the detector and the sample is automatically adjustable by the motor.

[0191] 2. Crystal culture

[0192] Take 10 mg of the A crystal form of compound (I) and add it to 2 mL of acetonitrile. Stir until the sample is completely dissolved. Place the supernatant in a 4 mL semi-sealed sample bottle and allow it to evaporate slowly at room temperature. After two weeks, colorless flaky crystals are obtained.

[0193] 3. Test Results

[0194] The basic structural information of this compound is: molecular formula 2(C 29 H 36 F3N5O4)·CH3CN, crystal system: orthorhombic, space group: P212121, wavelength: Cell parameters are α=β=γ=90°, unit cell volume

[0195] 4. Conclusion

[0196] Single-crystal data show that the single crystal is an acetonitrile compound of formula (I). Its single-crystal SXRD stereostructure ellipsoid diagram is attached. Figure 11 The results show that C12 and C18 in the figure are R, R configuration.

[0197] Biological activity:

[0198] Experiment Example 1: Experiment combining KRAS (G12C) and SOS1

[0199] Experimental principle:

[0200] The small molecule compound binds to the catalytic site of SOS1, inhibiting the binding of SOS1 to KRAS(G12C). When the binding of fluorescently labeled SOS1 protein to fluorescently labeled KRAS(G12C) protein is inhibited, the emitted fluorescence changes. By detecting the fluorescence change, the ability of the small molecule to prevent the binding of SOS1 to KRAS(G12C) can be tested. A homogeneous time-resolved fluorescence (HTRF) binding assay was used to detect the ability of the compound of this invention to inhibit the interaction between SOS1 and KRAS(G12C).

[0201] Experimental materials:

[0202] KRAS(G12C) protein was expressed and purified by Wuhan Pu Jian Biotechnology Co., Ltd., SOS1 exchange domin (564-1049) protein (Human recombinant) was purchased from Cytoskeleton, Mab Anti 6HIS-XL665 and Mab Anti GST-Eu cryptate were purchased from Cisbio. Multifunctional enzyme marker Nivo5 was purchased from PerkinElmer.

[0203] Experimental method:

[0204] 1X buffer preparation (freshly prepared): Hepes: 5mM; NaCl: 150mM; EDTA: 10mM; Igepal: 0.0025%; KF: 100mM; DTT: 1mM; BSA: 005%;

[0205] The test compound was diluted 5 times by gun to the 8th concentration, i.e. from 1mM to 0.064μM, with DMSO.

[0206] Each gradient of the test compound was diluted with 1X buffer to a working solution of 2% DMSO, 5μL / well was added to the corresponding well, and the corresponding concentration gradient was 20μM to 0.00128nM, and the double-hole experiment was placed. 1000 revolutions, centrifugation for 1 minute.

[0207] KRAS(G12C) (200nM) and Mab Anti GST-Eu cryptate (1ng / μL) mixed working solution was prepared with 1X buffer, and the mixed working solution was incubated at 25℃ for 5 minutes, 2.5μL / well was added to the corresponding well.

[0208] SOS1 (80nM) and Mab Anti 6HIS-XL665 (8g / μL) mixed working solution was prepared with 1X buffer, 2.5μL / well was added to the corresponding well, 2.5μL Mab Anti 6HIS-XL665 (8g / μL) diluent was added to the blank well, and the final concentration of the compound was 10μM to 0.64nM, KRAS(G12C)(500nM), MAb Anti GST-Eu cryptate(0.25ng / μL), SOS1(20nM), Mab Anti 6HIS-XL665(2g / μL), the reaction system was placed at 25℃ for 60 minutes. After the reaction was completed, HTRF was read by using multi-label analyzer.

[0209] Data analysis:

[0210] The raw data was converted into inhibition rate using the equation (Sample-Min) / (Max-Min) x 100%, and the IC 50 value was obtained by four-parameter curve fitting (obtained in GraphPad Prism, log(inhibitor) vs. response - Variable slope mode). The results of the test of the inhibitory activity of the compounds of the present application on the binding of KRAS(G12C) and SOS1 are shown in Table 12.

[0211] Table 12 Test results of IC 50 values of the compounds of the present application on the binding of KRAS(G12C) and SOS1

[0212]

[0213] Experimental conclusion: The compounds of the present application have significant inhibitory effect on the binding of KRAS(G12C) and SOS1.

[0214] Experimental Example 2: Test of 3D proliferation inhibitory activity of H358 cells

[0215] Experimental principle:

[0216] The KRAS signaling pathway is abnormally activated in H358 cells with KRAS(G12C) mutation. Small molecule SOS1 inhibitors reduce the GEF activity of SOS1 by inhibiting the binding of SOS1 and RAS protein, and reduce the proportion of activated RAS-GTP. Further down-regulate the phosphorylation level of the MEK / ERK pathway downstream of RAS, to achieve the effect of inhibiting cell proliferation. The small molecules are co-cultured with H358 cells in 3D space, and then the cell reading is used to indirectly reflect the proliferation inhibitory activity of SOS1 inhibitors on H358 cells.

[0217] Experimental materials:

[0218] RPMI1640 medium, fetal bovine serum, penicillin / streptomycin antibiotics were purchased from Vincents, low-melting agarose was purchased from Sigma. Almar blue reagent was purchased from Invitrogen. NCI-H358 cell line was purchased from Nanjing Kebai Biotechnology Co., Ltd. Nivo multi-label analyzer (PerkinElmer).

[0219] Experimental method:

[0220] H358 cells were seeded in 96-well U-bottom plates. First, low-melting agarose was prepared into a 2% stock solution. When used, the agarose stock solution was heated in a microwave oven until completely melted, and then placed in a 42°C water bath to keep the agarose liquid. The gel was added to the serum-containing medium to prepare a 0.6% gel concentration as the bottom gel, and 50 μL per well was added to the 96-well U-bottom plates. After the bottom gel solidified, 2% gel was added to the cell-containing medium to prepare a 0.4% cell-containing top gel, and 75 μL per well was added to the 96-well U-bottom plates containing the bottom gel, with a cell density of 3000 cells per well. After the top gel solidified, the cell plates were placed in a carbon dioxide incubator overnight for culture. 4 H358 cells were seeded in 96-well U-bottom plates. First, low-melting agarose was prepared into a 2% stock solution. When used, the agarose stock solution was heated in a microwave oven until completely melted, and then placed in a 42°C water bath to keep the agarose liquid. The gel was added to the serum-containing medium to prepare a 0.6% gel concentration as the bottom gel, and 50 μL per well was added to the 96-well U-bottom plates. After the bottom gel solidified, 2% gel was added to the cell-containing medium to prepare a 0.4% cell-containing top gel, and 75 μL per well was added to the 96-well U-bottom plates containing the bottom gel, with a cell density of 3000 cells per well. After the top gel solidified, the cell plates were placed in a carbon dioxide incubator overnight for culture.

[0221] On the day of compound addition, 85 μL of liquid medium was added to the cell-seeded 96-well U-bottom plates. The test compound was diluted 3-fold to the 9th concentration, i.e., from 6 mM to 0.9 μM, with double-replicate wells. 97 μL of medium was added to the intermediate plate, and then 2.5 μL of the gradient-diluted compound per well was transferred to the intermediate plate according to the corresponding positions, and 40 μL per well was transferred to the cell plate after mixing. The compound concentration range transferred to the cell plate was 30 μM to 4.5 nM. The cell plate was placed in a carbon dioxide incubator for 7 days of culture. On the 8th day, the test compound was diluted 3-fold to the 9th concentration, i.e., from 6 mM to 0.9 μM, with double-replicate wells. 198 μL of medium was added to the intermediate plate, and then 2 μL of the gradient-diluted compound per well was transferred to the first intermediate plate according to the corresponding positions, and 100 μL of medium was added to the second intermediate plate, and 100 μL of the mixed compound from the first intermediate plate was added, and 40 μL per well was transferred to the cell plate after mixing. The compound concentration range transferred to the cell plate was 30 μM to 4.5 nM. The cell plate was placed in a carbon dioxide incubator for another 7 days of culture. The compound was co-incubated with the cells for 14 days, and 20 μL of Almar blue detection reagent per well was added to the cell plate. The plate with the added dye was placed on a horizontal shaker for 15 minutes, and then the plate was incubated at room temperature for 5 hours to stabilize the luminescence signal. A multi-label analyzer was used for reading.

[0222] Data analysis:

[0223] The raw data was converted into inhibition rate using the equation (Sample-Min) / (Max-Min) x 100%, and the IC 50 value was obtained by four-parameter curve fitting (obtained in the "log(inhibitor) vs. response - Variable slope" mode in GraphPad Prism).

[0224] Experimental conclusion: The compound can inhibit the proliferation of H358 cells under 3D conditions.

[0225] Experimental example 3: DLD-1 cell p-ERK proliferation inhibition activity test

[0226] Experimental materials:

[0227] DLD-1 cells were purchased from Nanjing Kebai; 1640 culture medium was purchased from Biological Industries; fetal bovine serum was purchased from Biosera; Advanced Phospho-ERK1 / 2 (THR202 / TYR204) KIT was purchased from Cisbio, and the ingredient table is shown in Table 13.

[0228] Table 13 Ingredient table of Advanced Phospho-ERK1 / 2 (THR202 / TYR204) KIT

[0229]

[0230] Experimental method:

[0231] DLD-1 cells were seeded in a transparent 96-well cell culture plate, 80 μL of cell suspension per well, containing 8000 DLD-1 cells per well, and the cell plate was placed in a carbon dioxide incubator and incubated at 37°C overnight;

[0232] The test compound was diluted to 2mM with 100% DMSO as the first concentration, and then 5-fold dilution was performed with a pipette to the eighth concentration, i.e. from 2mM to 0.026μM. 2μL of compound was taken and added to 78μL of cell starvation medium, mixed well, and then 20μL of compound solution was added to the corresponding cell plate well, and the cell plate was placed back into the carbon dioxide incubator for further incubation for 1 hour, at this time the compound concentration was 10μM to 0.128nM, and the DMSO concentration was 0.5%;

[0233] After the incubation was completed, the cell supernatant was discarded and 50μL of cell lysis solution was added to each well, and the incubation was carried out at room temperature for 30 minutes;

[0234] Phospho-ERK1 / 2 Eu Cryptate antibody and Phospho-ERK1 / 2 d2 antibody were diluted 20 times with Detection buffer;

[0235] Take 16 μL cell lysate supernatant per well into a new 384 white microplate, then add 2 μL Phospho-ERK1 / 2 Eu Cryptate antibody dilution and 2 μL Phospho-ERK1 / 2 d2 antibody dilution, and incubate at room temperature for 4 hours;

[0236] After incubation, use multi-label analyzer to read HTRF excitation: 320 nm, emission: 615 nm, 665 nm.

[0237] Data analysis:

[0238] Convert the raw data into inhibition rate by equation (Sample-Min) / (Max-Min)*100%, and the IC 50 value can be obtained by four-parameter curve fitting (obtained in GraphPad Prism, log(inhibitor) vs. response-Variable slope mode).

[0239] Max well: the positive control well is 1X lysate

[0240] Min well: the negative control well is 0.5% DMSO cell lysate

[0241] The test results of the inhibitory activity of the compound of the present application on DLD-1 cell p-ERK are shown in Table 14.

[0242] Table 14 IC 50 value test results of the compound of the present application on DLD-1 cell p-ERK proliferation

[0243]

[0244]

[0245] Experimental conclusion: the compound of the present application has a significant inhibitory effect on DLD-1 cell p-ERK proliferation.

[0246] Experimental example 4: compound pharmacokinetic evaluation

[0247] Experimental materials:

[0248] Balb / c mice (male, Beijing Vital River Laboratory Animal Technology Co., Ltd.)

[0249] Experimental operation:

[0250] The pharmacokinetic characteristics of the compounds in rodents after intravenous injection and oral administration are tested according to a standard protocol. In the experiment, the candidate compounds are prepared into clear solutions, and single intravenous injection and oral administration are given to mice. The injection and oral solvents are a mixed solvent prepared from 5% dimethyl sulfoxide, 5% solutol and 90% water. Four male Balb / c mice are used in this project, two mice are given intravenous injection at a dose of 10 mg / kg, and plasma samples are collected at 0.083, 0.25, 0.5, 1, 2, 4, 8 and 24 hours after administration. The other two mice are given oral gavage at a dose of 50 mg / kg, and plasma samples are collected at 0.25, 0.5, 1, 2, 4, 6, 8, 12 and 24 hours after administration. After blood sampling, the blood is placed on ice, and the plasma is centrifuged (centrifugation conditions: 6000g, 3 minutes, 2-8℃) within 1 hour. The plasma samples are stored in a -80℃ refrigerator before analysis. The blood drug concentration is quantitatively analyzed by LC-MS / MS analysis method, and the pharmacokinetic parameters such as peak concentration (C max ), clearance (CL), half-life (T 1 / 2 ), tissue distribution (Vdss), area under the curve (AUC 0-last ), bioavailability (F) and the like are calculated.

[0251] The experimental results are shown in Table 15:

[0252] Table 15 Pharmacokinetic test results of the compounds of the present application

[0253]

[0254] Experimental conclusion: The compounds of the present application have good pharmacokinetic properties, including good oral bioavailability, oral exposure, half-life and clearance, etc.

Claims

1. A crystalline form of the compound of formula (I) having an X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2Θ angles: 15.492 ± 0.200°, 16.458 ± 0.200°, 18.657 ± 0.200° and 20.638 ± 0.200°; 。 2. The crystalline form of claim 1 having an X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2Θ angles: 14.223 ± 0.200°, 14.589 ± 0.200°, 14.894 ± 0.200°, 15.492 ± 0.200°, 16.061 ± 0.200°, 16.458 ± 0.200°, 18.657 ± 0.200° and 20.638 ± 0.200°.

3. The crystalline form of claim 2 having an X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2Θ angles: 7.736 ± 0.200°, 14.223 ± 0.200°, 14.589 ± 0.200°, 14.894 ± 0.200°, 15.492 ± 0.200°, 16.061 ± 0.200°, 16.458 ± 0.200°, 18.657 ± 0.200°, 19.407 ± 0.200°, 20.638 ± 0.200°, 21.810 ± 0.200° and 22.836 ± 0.200°.

4. The crystalline form of claim 3 having an X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2Θ angles: 7.736 ± 0.100°, 9.070 ± 0.100°, 11.289 ± 0.100°, 11.678 ± 0.100°, 14.223 ± 0.100°, 14.589 ± 0.100°, 14.894 ± 0.100°, 15.492 ± 0.100°, 16.061 ± 0.100°, 16.458 ± 0.100°, 18.657 ± 0.100°, 19.407 ± 0.100°, 20.638 ± 0.100°, 21.085 ± 0.100°, 21.810 ± 0.100° and 22.836 ± 0.100°.

5. The A crystalline form of claim 4, having an X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2Θ angles: 7.736 ± 0.200°, 8.475 ± 0.200°, 9.070 ± 0.200°, 11.289 ± 0.200°, 11.678 ± 0.200°, 12.363 ± 0.200°, 14.223 ± 0.200°, 14.589 ± 0.200°, 14.894 ± 0.200°, 15.492 ± 0.200°, 16.061 ± 0.200°, 16.458 ± 0.200°, 17.000 ± 0.200°, 18.657 ± 0.200°, 19.030 ± 0.200°, 19.407 ± 0.200°, 19.882 ± 0.200°, 20.638 ± 0.200°, 21.085 ± 0.200°, 21.810 ± 0.200°, 22.836 ± 0.200°, 23.717 ± 0.200°, 24.147 ± 0.200°, 24.693 ± 0.200°, 25.311 ± 0.200°, 26.802 ± 0.200°, 27.462 ± 0.200°, 28.537 ± 0.200°, and 31.264 ± 0.200°.

6. The A crystalline form of claim 5, having an X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2Θ angles: 7.736°, 8.475°, 9.070°, 11.289°, 11.678°, 12.363°, 14.223°, 14.589°, 14.894°, 15.492°, 16.061°, 16.458°, 17.000°, 18.657°, 19.030°, 19.407°, 19.882°, 20.638°, 21.085°, 21.810°, 22.836°, 23.717°, 24.147°, 24.693°, 25.311°, 26.802°, 27.462°, 28.537°, and 31.264°.

7. The A crystalline form of the compound of formula (I) according to claim 1, having an XRPD pattern substantially as shown in Figure 1.

8. The A crystalline form according to any one of claims 1 to 7, having a differential scanning calorimetry curve with an endothermic peak starting at 169.0 ± 5 °C.

9. The A crystalline form according to claim 8, having a DSC pattern substantially as shown in Figure 2.

10. The A crystalline form according to any one of claims 1 to 7, having a thermogravimetric analysis curve with a weight loss of 1.48% at 160.0 ± 3 °C.

11. The A crystalline form according to claim 10, having a TGA pattern substantially as shown in Figure 3.

12. Use of the A crystalline form of the compound of formula (I) according to any one of claims 1 to 11 for the manufacture of a medicament for the treatment of KRAS mutant solid tumor disease.

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