Salt form and crystal form of pyridine amide compound, and preparation methods therefor

Abstract

Disclosed in the present invention are a salt form and a crystal form of a pyridine amide compound, and preparation methods therefor. Specifically, disclosed in the present invention is a salt form of a compound of formula (I), which is a compound of formula (II), a compound of formula (III) or a compound of formula (IV). The salt form and the crystal form provided in the present invention have excellent PARP1 inhibitory activity and selectivity, thereby avoiding anemia toxicity. Moreover, the compounds of the present invention have different distribution volumes (which can reduce the toxicity to neutrophils and platelets) and different trapping activities (which are helpful for adjusting the drug dosage). While maintaining the clinical efficacy of PARP1 / 2 inhibitors, the compounds reduce the hematotoxicity, making it possible to further combine highly selective PARP1 inhibitors with chemotherapeutic drugs.

Description

Salt form, crystal form of a class of pyridine amide compounds and preparation method thereof

[0001] This application claims priority to Chinese Patent Application No. 2024107943630, filed on June 19, 2024. This application incorporates the entirety of the aforementioned Chinese Patent Application. TECHNICAL FIELD

[0002] The present application relates to a salt form, crystal form of a class of pyridine amide compounds and a preparation method thereof. BACKGROUND

[0003] Poly ADP-ribose polymerase (PARP) is a family of proteins that can catalyze ADP-ribosylation. PARP1 is an important member of the family of proteins that has been most widely studied, is highly expressed in cells and responds quickly, can quickly catalyze and modify DNA repair factors, interacts with them to participate in various DNA repair processes. In normal cells, single-strand breaks in DNA can be repaired by base excision repair, and PARP1 uses NAD+ as a substrate, binds to the damage site through its zinc finger domain, changes its conformation, and catalyzes the transfer of ADP-ribosyl group, ultimately completing single-strand repair. In cancer cells with double-strand breaks (DSB), DNA breaks are mainly repaired by homologous recombination (HR), and BRCA1 and BRCA2 are key proteins that mediate HR. In cancer cells with BRCA1 / 2 mutations, inhibiting the function of PARP1 simultaneously causes obstacles in the main DNA repair pathway, ultimately leading to cell death, which is the currently fully verified synthetic lethal effect. Therefore, PARP1 has become a hot cancer treatment target.

[0004] Four small-molecule PARP inhibitors (PARPi) have been approved by the US FDA for the treatment of cancer, which are olaparib, niraparib, rucaparib and telazoparib. These drugs have shown good clinical treatment effects in the treatment of BRCA1 / 2 mutant ovarian cancer and / or breast cancer patients, and have shown very good prospects in other BRCA mutant cancer patients, including prostate cancer and pancreatic cancer.

[0005] On the other hand, there is a potential basis for the combination of chemotherapeutic drugs and PARP inhibitors. However, since chemotherapeutic drugs inhibit rapidly proliferating cells while also having strong hematotoxicity, the combination of the two can lead to the superposition of hematotoxicity, which greatly limits the combination of PARP inhibitors.

[0006] Studies have shown that anemia caused by PARP inhibitors can be mainly due to the inhibition of PARP2 (Farrés J, et al. Cell Death Differ. 2015 Jul;22(7):1144-57.). The currently marketed PARPi all have PARP1 / 2 inhibition activity, and if a PARP1 highly selective inhibitor can be developed to reduce the inhibition of PARP2, the anemia toxicity caused by the existing PARP1 / 2 inhibitor can be avoided. WO2021013735A1 discloses a series of PARP1 selective inhibitors, such as AZD5305.

[0007] Therefore, it is of great clinical application value to develop more highly selective PARP1 inhibitors, and there is a demand for such inhibitors in the art. SUMMARY

[0008] The technical problem to be solved by the present application is to overcome the defects of the prior art highly selective PARP1 inhibitors, and to provide a salt type and crystal form of a pyridine amide compound and a preparation method thereof.

[0009] The present application solves the above technical problems by the following technical solutions:

[0010] The present application provides a salt type of a compound of formula (I), which is a compound of formula (II), a compound of formula (III) or a compound of formula (IV);

[0011] The present application also provides a crystal form A of a compound of formula (I);

[0012] The crystal form A has diffraction peaks in the X-ray powder diffraction pattern expressed by 2θ angles using Cu-Kα radiation at the following positions: 9.738°±0.2°, 11.350°±0.2°, 12.504°±0.2°, 12.888°±0.2°, 14.438°±0.2°, 15.025°±0.2°, 15.369°±0.2°, 16.837°±0.2°.

[0013] In a certain aspect, the crystal form A has diffraction peaks in the X-ray powder diffraction pattern expressed by 2θ angles at the following positions: 9.738°±0.2°, 11.350°±0.2°, 12.504°±0.2°, 12.888°±0.2°, 14.438°±0.2°, 15.025°±0.2°, 15.369°±0.2°, 16.837°±0.2°, 17.813°±0.2°, 18.285°±0.2°.

[0014] In one aspect, the Form A has an X-ray powder diffraction pattern, expressed in terms of 2 theta, with diffraction peaks at the following positions: 9.738°±0.2°, 11.350°±0.2°, 12.504°±0.2°, 12.888°±0.2°, 14.438°±0.2°, 15.025°±0.2°, 15.369°±0.2°, 16.837°±0.2°, 17.813°±0.2°, 18.285°±0.2°, 18.512°±0.2°, 18.734°±0.2°, 19.591°±0.2°, 19.840°±0.2°, 20.162°±0.2°, 20.513°±0.2°, 21.197°±0.2°, 22.152°±0.2°, 22.398°±0.2°, 22.923°±0.2°, 25.274°±0.2°, 25.853°±0.2°, 25.980°±0.2°, 27.316°±0.2°.

[0015] In one aspect, the Form A has an X-ray powder diffraction pattern, expressed in terms of 2 theta, with diffraction peaks at the following positions: 9.738°±0.2°, 11.350°±0.2°, 12.504°±0.2°, 12.888°±0.2°, 14.438°±0.2°, 15.025°±0.2°, 15.369°±0.2°, 16.837°±0.2°, 17.813°±0.2°, 18.285°±0.2°, 18.512°±0.2°, 18.734°±0.2°, 19.591°±0.2°, 19.840°±0.2°, 20.162°±0.2°, 20.513°±0.2°, 21.197°±0.2°, 22.152°±0.2°, 22.398°±0.2°, 22.923°±0.2°, 25.274°±0.2°, 25.853°±0.2°, 25.980°±0.2°, 27.316°±0.2°.

[0016] In one aspect, the Form A has an X-ray powder diffraction pattern, expressed in terms of 2 theta, with diffraction peaks at the following positions: 9.738°±0.2°, 11.350°±0.2°, 12.504°±0.2°, 12.888°±0.2°, 14.438°±0.2°, 15.025°±0.2°, 15.369°±0.2°, 16.837°±0.2°, 17.813°±0.2°, 18.285°±0.2°, 18.512°±0.2°, 18.734°±0.2°, 19.591°±0.2°, 19.840°±0.2°, 20.162°±0.2°, 20.513°±0.2°, 21.197°±0.2°, 22.152°±0.2°, 22.398°±0.2°, 22.923°±0.2°, 25.274°±0.2°, 25.853°±0.2°, 25.980°±0.2°, 27.316°±0.2°.

[0017] In one aspect, the Form A has an X-ray powder diffraction pattern, expressed in terms of 2 theta, with diffraction peaks at the following positions: 9.738°±0.2°, 11.350°±0.2°, 12.504°±0.2°, 12.888°±0.2°, 14.438°±0.2°, 15.025°±0.2°, 15.369°±0.2°, 16.837°±0.2°, 17.813°±0.2°, 18.285°±0.2°, 18.512°±0.2°, 18.734°±0.2°, 19.591°±0.2°, 19.840°±0.2°, 20.162°±0.2°, 20.513°±0.2°, 21.197°±0.2°, 22.152°±0.2°, 22.398°±0.2°, 22.923°±0.2°, 25.274°±0.2°, 25.853°±0.2°, 25.980°±0.2°, 27.316°±0.2°.

[0018] The present application also provides a crystalline form of the compound of formula (II), which is Form I, Form II, Form III, Form IV, Form V, Form VI, Form VII, Form VIII, Form IX or Form X;

[0019] The Form I has an X-ray powder diffraction pattern, expressed in terms of 2 theta, with diffraction peaks at the following positions: 8.487°±0.2°, 9.663°±0.2°, 11.259°±0.2°, 12.684°±0.2°, 13.627°±0.2°, 13.994°±0.2°.

[0020] In one aspect, the Form I has an X-ray powder diffraction pattern, using Cu-Kalpharadiation, expressed in degrees 2-theta (2Q), with peaks at 8.487°±0.2°, 9.663°±0.2°, 11.259°±0.2°, 12.684°±0.2°, 13.627°±0.2°, 13.994°±0.2°, 16.128°±0.2°, 17.037°±0.2°, 17.989°±0.2°, 19.376°±0.2°, 20.642°±0.2°.

[0021] In one aspect, the Form I has an X-ray powder diffraction pattern, using Cu-Kalpharadiation, expressed in degrees 2-theta (2Q), with peaks at 8.487°±0.2°, 9.663°±0.2°, 11.259°±0.2°, 12.684°±0.2°, 13.627°±0.2°, 13.994°±0.2°, 16.128°±0.2°, 17.037°±0.2°, 17.989°±0.2°, 19.376°±0.2°, 20.642°±0.2°, 22.187°±0.2°, 22.749°±0.2°, 24.567°±0.2°, 26.171°±0.2°, 27.445°±0.2°.

[0022] In one aspect, the Form I has an X-ray powder diffraction pattern, using Cu-Kalpharadiation, expressed in degrees 2-theta (2Q), with peaks at 8.487°±0.2°, 9.663°±0.2°, 11.259°±0.2°, 12.684°±0.2°, 13.627°±0.2°, 13.994°±0.2°, 16.128°±0.2°, 17.037°±0.2°, 17.989°±0.2°, 19.376°±0.2°, 20.642°±0.2°, 22.187°±0.2°, 22.749°±0.2°, 24.567°±0.2°, 26.171°±0.2°, 27.445°±0.2°.

[0023] In one aspect, the Form I has an X-ray powder diffraction pattern, using Cu-Kalpharadiation, expressed in degrees 2-theta (2Q), with peaks at 8.487°±0.2°, 9.663°±0.2°, 11.259°±0.2°, 12.684°±0.2°, 13.627°±0.2°, 13.994°±0.2°, 16.128°±0.2°, 17.037°±0.2°, 17.989°±0.2°, 19.376°±0.2°, 20.642°±0.2°, 22.187°±0.2°, 22.749°±0.2°, 24.567°±0.2°, 26.171°±0.2°, 27.445°±0.2°.

[0024] In one aspect, the Form I has an X-ray powder diffraction pattern, using Cu-Kalpharadiation, expressed in degrees 2-theta (2Q), with peaks at 8.487°±0.2°, 9.663°±0.2°, 11.259°±0.2°, 12.684°±0.2°, 13.627°±0.2°, 13.994°±0.2°, 16.128°±0.2°, 17.037°±0.2°, 17.989°±0.2°, 19.376°±0.2°, 20.642°±0.2°, 22.187°±0.2°, 22.749°±0.2°, 24.567°±0.2°, 26.171°±0.2°, 27.445°±0.2°.

[0025] The Form II has an X-ray powder diffraction pattern, using Cu-Kalpharadiation, expressed in degrees 2-theta (2Q), with peaks at 8.262°±0.2°, 10.219°±0.2°, 11.445°±0.2°, 11.887°±0.2°, 12.352°±0.2°, 13.994°±0.2°, 14.536°±0.2°, 15.243°±0.2°, 15.457°±0.2°, 16.462°±0.2°, 17.115°±0.2°, 17.646°±0.2°, 17.831°±0.2°, 19.041°±0.2°.

[0026] In a certain embodiment, the Form II has an X-ray powder diffraction pattern, using Cu-Kalpharadiation, with diffraction peaks, in terms of 2 theta, at 8.262°±0.2°, 10.219°±0.2°, 11.445°±0.2°, 11.887°±0.2°, 12.352°±0.2°, 13.994°±0.2°, 14.536°±0.2°, 15.243°±0.2°, 15.457°±0.2°, 16.462°±0.2°, 17.115°±0.2°, 17.646°±0.2°, 17.831°±0.2°, 19.041°±0.2°, 19.522°±0.2°, 19.818°±0.2°, 20.295°±0.2°, 20.698°±0.2°, 21.389°±0.2°, 21.821°±0.2°, 22.423°±0.2°, 22.800°±0.2°, 23.485°±0.2°, 23.825°±0.2°, 24.822°±0.2°, 25.332°±0.2°, 25.739°±0.2°, 26.369°±0.2°, 26.936°±0.2°, 27.351°±0.2°, 28.038°±0.2°, 28.394°±0.2°, 29.003°±0.2°, 29.293°±0.2°.

[0027] In one embodiment, the Form II has an X-ray powder diffraction pattern, using Cu-Ka radiation, expressed in degrees 2-theta with reflection peaks at 8.262°±0.2°, 10.219°±0.2°, 11.445°±0.2°, 11.887°±0.2°, 12.352°±0.2°, 13.994°±0.2°, 14.536°±0.2°, 15.243°±0.2°, 15.457°±0.2°, 16.462°±0.2°, 17.115°±0.2°, 17.646°±0.2°, 17.831°±0.2°, 19.041°±0.2°, 19.522°±0.2°, 19.818°±0.2°, 20.295°±0.2°, 20.698°±0.2°, 21.389°±0.2°, 21.821°±0.2°, 22.423°±0.2°, 22.800°±0.2°, 23.485°±0.2°, 23.825°±0.2°, 24.822°±0.2°, 25.332°±0.2°, 25.739°±0.2°, 26.369°±0.2°, 26.936°±0.2°, 27.351°±0.2°, 28.038°±0.2°, 28.394°±0.2°, 29.003°±0.2°, 29.293°±0.2°, 29.865°±0.2°, 30.078°±0.2°, 30.612°±0.2°, 31.123°±0.2°, 31.440°±0.2°, 31.878°±0.2°, 32.066°±0.2°, 32.574°±0.2°, 33.187°±0.2°, 33.488°±0.2°, 33.841°±0.2°, 34.489°±0.2°, 35.106°±0.2°, 35.671°±0.2°, 36.108°±0.2°, 37.180°±0.2°, 37.660°±0.2°, 38.180°±0.2°, 38.573°±0.2°, 39.223°±0.2°.

[0028] In one embodiment, the Form II has an X-ray powder diffraction pattern, expressed in degrees 2-theta with reflection peaks as shown in the following table:

[0029] In one embodiment, the Form II has an X-ray powder diffraction pattern, expressed in degrees 2-theta with reflection peaks as shown in the following table:

[0030] In one aspect, the Form II has an X-ray powder diffraction pattern, expressed in terms of 2 theta (2Q) angles, substantially as shown in FIG. 5.

[0031] In one aspect, the Form II has a thermogravimetric analysis pattern with a weight loss of 9.7 ± 0.2% when heated to 114 °C ± 2 °C.

[0032] In one aspect, the Form II has a thermogravimetric analysis pattern substantially as shown in FIG. 6.

[0033] In one aspect, the Form II has a differential scanning calorimetry pattern with an endothermic peak at 108 °C ± 2 °C.

[0034] In one aspect, the Form II has a differential scanning calorimetry pattern with an endothermic peak at 108 °C ± 2 °C and a heat of fusion of 210 J / g.

[0035] In one aspect, the Form II has a differential scanning calorimetry pattern substantially as shown in FIG. 6.

[0036] In one aspect, the Form II is a hydrate of the compound of formula (II).

[0037] In one aspect, the Form II is a trihydrate of the compound of formula (II).

[0038] The Form III has an X-ray powder diffraction pattern, expressed in terms of 2 theta (2Q) angles, using Cu-Ka radiation, with peaks at 8.197° ± 0.2°, 10.619° ± 0.2°, 11.200° ± 0.2°, 12.114° ± 0.2°, 13.285° ± 0.2°, 13.581° ± 0.2°, 14.787° ± 0.2°.

[0039] In one aspect, the Form III has an X-ray powder diffraction pattern, expressed in terms of 2 theta (2Q) angles, using Cu-Ka radiation, with peaks at 8.197° ± 0.2°, 10.619° ± 0.2°, 11.200° ± 0.2°, 12.114° ± 0.2°, 13.285° ± 0.2°, 13.581° ± 0.2°, 14.787° ± 0.2°, 15.730° ± 0.2°, 16.943° ± 0.2°.

[0040] In one aspect, the Form III has an X-ray powder diffraction pattern, using Cu- K alpha radiation, expressed in degrees 2-theta with diffraction peaks at 8.197°±0.2°, 10.619°±0.2°, 11.200°±0.2°, 12.114°±0.2°, 13.285°±0.2°, 13.581°±0.2°, 14.787°±0.2°, 15.730°±0.2°, 16.943°±0.2°, 17.815°±0.2°, 20.459°±0.2°, 21.063°±0.2°, 21.371°±0.2°, 21.861°±0.2°, 22.567°±0.2°, 23.589°±0.2°, 24.865°±0.2°, 25.248°±0.2°, 26.057°±0.2°, 27.127°±0.2°, 29.226°±0.2°, 30.399°±0.2°, 31.800°±0.2°.

[0041] In one aspect, the Form III has an X-ray powder diffraction pattern, expressed in degrees 2-theta with diffraction peaks as shown in the following table:

[0042] In one aspect, the Form III has an X-ray powder diffraction pattern, expressed in degrees 2-theta with diffraction peaks as shown in the following table:

[0043] In one aspect, the Form III has an X-ray powder diffraction pattern substantially as shown in FIG. 7.

[0044] The Form IV has an X-ray powder diffraction pattern, using Cu-K alpha radiation, expressed in degrees 2-theta with diffraction peaks at 6.648°±0.2°, 7.725°±0.2°, 11.328°±0.2°, 12.101°±0.2°, 13.154°±0.2°, 13.647°±0.2°.

[0045] In one aspect, the Form IV has an X-ray powder diffraction pattern, using Cu-K alpha radiation, expressed in degrees 2-theta with diffraction peaks at 6.648°±0.2°, 7.725°±0.2°, 11.328°±0.2°, 12.101°±0.2°, 13.154°±0.2°, 13.647°±0.2°, 16.209°±0.2°, 19.662°±0.2°, 19.988°±0.2°.

[0046] In one aspect, the Form IV has an X-ray powder diffraction pattern, expressed in angles 2Θ, with diffraction peaks as set out in the following table:

[0047] In one aspect, the Form IV has an X-ray powder diffraction pattern, expressed in angles 2Θ, with diffraction peaks as set out in the following table:

[0048] In one aspect, the Form IV has an X-ray powder diffraction pattern, expressed in angles 2Θ, with diffraction peaks as set out in the following table:

[0049] In one aspect, the Form IV has an X-ray powder diffraction pattern, expressed in angles 2Θ, substantially as set out in Figure 8.

[0050] The Form V has an X-ray powder diffraction pattern, expressed in angles 2Θ, with diffraction peaks at 7.935°± 0.2°, 9.092°± 0.2°, 9.447°± 0.2°, 11.083°± 0.2°.

[0051] In one aspect, the Form V has an X-ray powder diffraction pattern, expressed in angles 2Θ, with diffraction peaks at 7.935°± 0.2°, 9.092°± 0.2°, 9.447°± 0.2°, 11.083°± 0.2°, 14.289°± 0.2°, 20.970°± 0.2°.

[0052] In one aspect, the Form V has an X-ray powder diffraction pattern, expressed in angles 2Θ, with diffraction peaks as set out in the following table:

[0053] In one aspect, the Form V has an X-ray powder diffraction pattern, expressed in angles 2Θ, with diffraction peaks as set out in the following table:

[0054] In one aspect, the Form V has an X-ray powder diffraction pattern, expressed in angles 2Θ, substantially as set out in Figure 9.

[0055] The X-ray powder diffraction pattern, in terms of 2 theta, of said crystalline Form VI has diffraction peaks at the following positions: 8.499°±0.2°, 9.674°±0.2°, 11.290°±0.2°, 13.982°±0.2°, 14.502°±0.2°, 14.910°±0.2°, 17.050°±0.2°, 17.995°±0.2°, using Cu-Ka radiation.

[0056] In a certain embodiment, the X-ray powder diffraction pattern, in terms of 2 theta, of said crystalline Form VI has diffraction peaks at the following positions: 8.499°±0.2°, 9.674°±0.2°, 11.290°±0.2°, 13.982°±0.2°, 14.502°±0.2°, 14.910°±0.2°, 17.050°±0.2°, 17.995°±0.2°, 19.102°±0.2°, 20.565°±0.2°, 20.733°±0.2°, using Cu-Ka radiation.

[0057] The X-ray powder diffraction pattern, in terms of 2 theta, of said crystalline Form VI has diffraction peaks at the following positions: 8.499°±0.2°, 9.674°±0.2°, 11.290°±0.2°, 13.982°±0.2°, 14.502°±0.2°, 14.910°±0.2°, 17.050°±0.2°, 17.995°±0.2°, 19.102°±0.2°, 20.565°±0.2°, 20.733°±0.2°, 21.158°±0.2°, 22.238°±0.2°, 22.729°±0.2°, 23.370°±0.2°, 24.585°±0.2°, 25.396°±0.2°, 28.831°±0.2°, using Cu-Ka radiation.

[0058] In a certain embodiment, the X-ray powder diffraction pattern, in terms of 2 theta, of said crystalline Form VI has diffraction peaks as shown in the following table:

[0059] In a certain embodiment, the X-ray powder diffraction pattern, in terms of 2 theta, of said crystalline Form VI has diffraction peaks as shown in the following table:

[0060] In a certain embodiment, the X-ray powder diffraction pattern, in terms of 2 theta, of said crystalline Form VI is substantially as shown in Figure 10.

[0061] The crystalline Form VII has an X-ray powder diffraction pattern, expressed in terms of 2-theta using Cu-Kalpharadiation, having diffraction peaks at the following positions: 7.720°±0.2°, 9.747°±0.2°, 11.885°±0.2°, 12.772°±0.2°, 13.392°±0.2°, 13.606°±0.2°.

[0062] In a certain embodiment, the crystalline Form VII has an X-ray powder diffraction pattern, expressed in terms of 2-theta using Cu-Kalpharadiation, having diffraction peaks at the following positions: 7.720°±0.2°, 9.747°±0.2°, 11.885°±0.2°, 12.772°±0.2°, 13.392°±0.2°, 13.606°±0.2°, 15.221°±0.2°, 15.496°±0.2°, 17.334°±0.2°, 18.018°±0.2°, 18.884°±0.2°.

[0063] In a certain embodiment, the crystalline Form VII has an X-ray powder diffraction pattern, expressed in terms of 2-theta using Cu-Kalpharadiation, having diffraction peaks at the following positions: 7.720°±0.2°, 9.747°±0.2°, 11.885°±0.2°, 12.772°±0.2°, 13.392°±0.2°, 13.606°±0.2°, 15.221°±0.2°, 15.496°±0.2°, 17.334°±0.2°, 18.018°±0.2°, 18.884°±0.2°, 19.866°±0.2°, 20.240°±0.2°, 20.825°±0.2°, 22.174°±0.2°, 22.672°±0.2°, 23.589°±0.2°, 24.748°±0.2°, 25.118°±0.2°, 26.232°±0.2°, 27.328°±0.2°, 28.580°±0.2°, 29.530°±0.2°, 30.857°±0.2°.

[0064] In a certain embodiment, the crystalline Form VII has an X-ray powder diffraction pattern, expressed in terms of 2-theta having the following diffraction peaks:

[0065] In a certain embodiment, the crystalline Form VII has an X-ray powder diffraction pattern, expressed in terms of 2-theta having the following diffraction peaks:

[0066] In a certain embodiment, the crystalline Form VII has an X-ray powder diffraction pattern, expressed in terms of 2-theta having the following diffraction peaks:

[0067] The X-ray powder diffraction pattern, in terms of 2 theta, of said crystalline Form VIII has diffraction peaks at the following positions: 9.600° ± 0.2°, 10.615° ± 0.2°, 11.319° ± 0.2°, 12.717° ± 0.2°, 13.610° ± 0.2°, 15.399° ± 0.2°, 15.978° ± 0.2°, 18.161° ± 0.2°, 19.307° ± 0.2°, 20.431° ± 0.2°, 21.367° ± 0.2°, 22.253° ± 0.2°, 22.793° ± 0.2°, 23.452° ± 0.2°, 26.059° ± 0.2°, 26.801° ± 0.2°, 27.446° ± 0.2°, 28.235° ± 0.2°, 29.126° ± 0.2°, 31.110° ± 0.2°, 32.286° ± 0.2°, using Cu-Ka radiation.

[0068] In a certain embodiment, the X-ray powder diffraction pattern, in terms of 2 theta, of said crystalline Form VIII has diffraction peaks at the following positions: 9.600° ± 0.2°, 10.615° ± 0.2°, 11.319° ± 0.2°, 12.717° ± 0.2°, 13.610° ± 0.2°, 15.399° ± 0.2°, 15.978° ± 0.2°, 18.161° ± 0.2°, 19.307° ± 0.2°, 20.431° ± 0.2°, 21.367° ± 0.2°, 22.253° ± 0.2°, 22.793° ± 0.2°, 23.452° ± 0.2°, 26.059° ± 0.2°, 26.801° ± 0.2°, 27.446° ± 0.2°, 28.235° ± 0.2°, 29.126° ± 0.2°, 31.110° ± 0.2°, 32.286° ± 0.2°, using Cu-Ka radiation.

[0069] In a certain embodiment, the X-ray powder diffraction pattern, in terms of 2 theta, of said crystalline Form VIII has diffraction peaks at the following positions: 9.600° ± 0.2°, 10.615° ± 0.2°, 11.319° ± 0.2°, 12.717° ± 0.2°, 13.610° ± 0.2°, 15.399° ± 0.2°, 15.978° ± 0.2°, 18.161° ± 0.2°, 19.307° ± 0.2°, 20.431° ± 0.2°, 21.367° ± 0.2°, 22.253° ± 0.2°, 22.793° ± 0.2°, 23.452° ± 0.2°, 26.059° ± 0.2°, 26.801° ± 0.2°, 27.446° ± 0.2°, 28.235° ± 0.2°, 29.126° ± 0.2°, 31.110° ± 0.2°, 32.286° ± 0.2°, using Cu-Ka radiation.

[0070] In a certain embodiment, the X-ray powder diffraction pattern, in terms of 2 theta, of said crystalline Form VIII has diffraction peaks at the following positions:

[0071] In a certain embodiment, the X-ray powder diffraction pattern, in terms of 2 theta, of said crystalline Form VIII has diffraction peaks at the following positions:

[0072] In a certain embodiment, the X-ray powder diffraction pattern, in terms of 2 theta, of said crystalline Form VIII is substantially as shown in Figure 12.

[0073] The crystal form IX, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction patterns (denoted as 2θ) with diffraction peaks at the following positions: 10.509°±0.2°, 11.158°±0.2°, 12.607°±0.2°, 13.154°±0.2°, 13.795°±0.2°, and 15.847°±0.2°.

[0074] In one embodiment, the crystal form IX is subjected to Cu-Kα radiation, and the X-ray powder diffraction pattern, expressed in 2θ, has diffraction peaks at the following positions: 10.509°±0.2°, 11.158°±0.2°, 12.607°±0.2°, 13.154°±0.2°, 13.795°±0.2°, 15.847°±0.2°, 17.362°±0.2°, 19.819°±0.2°, and 21.062°±0.2°.

[0075] In one embodiment, the crystal form IX is subjected to Cu-Kα radiation, and the X-ray powder diffraction pattern, expressed in 2θ, has diffraction peaks at the following positions: 10.509°±0.2°, 11.158°±0.2°, 12.607°±0.2°, 13.154°±0.2°, 13.795°±0.2°, 15.847°±0.2°, 17.362°±0.2°, 19.819°±0.2°, 21.062°±0.2°, 22.516°±0.2°, 24.987°±0.2°, 25.957°±0.2°, and 26.509°±0.2°.

[0076] In one embodiment, the X-ray powder diffraction pattern of crystal form IX, expressed at a 2θ angle, has the diffraction peaks shown in the table below:

[0077] In one embodiment, the X-ray powder diffraction pattern of crystal form IX, expressed at a 2θ angle, has the diffraction peaks shown in the table below:

[0078] In one embodiment, the X-ray powder diffraction pattern of the crystal form IX, expressed in terms of a 2θ angle, is basically as shown in Figure 13.

[0079] The crystal form X, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction patterns (denoted as 2θ) with diffraction peaks at the following positions: 7.657°±0.2°, 9.825°±0.2°, 11.917°±0.2°, 13.536°±0.2°, 15.367°±0.2°, and 17.339°±0.2°.

[0080] In one aspect, the Form X has an X-ray powder diffraction pattern, as indicated by 2Q, with diffraction peaks at 7.657° ± 0.2°, 9.825° ± 0.2°, 11.917° ± 0.2°, 13.536° ± 0.2°, 15.367° ± 0.2°, 17.339° ± 0.2°, 17.958° ± 0.2°, 18.896° ± 0.2°, 19.938° ± 0.2°, 20.930° ± 0.2°, 22.257° ± 0.2°, using Cu-Ka radiation.

[0081] In one aspect, the Form X has an X-ray powder diffraction pattern, as indicated by 2Q, with diffraction peaks at 7.657° ± 0.2°, 9.825° ± 0.2°, 11.917° ± 0.2°, 13.536° ± 0.2°, 15.367° ± 0.2°, 17.339° ± 0.2°, 17.958° ± 0.2°, 18.896° ± 0.2°, 19.938° ± 0.2°, 20.930° ± 0.2°, 22.257° ± 0.2°, 22.813° ± 0.2°, 23.598° ± 0.2°, 24.087° ± 0.2°, 24.974° ± 0.2°, 27.090° ± 0.2°, using Cu-Ka radiation.

[0082] In one aspect, the Form X has an X-ray powder diffraction pattern, as indicated by 2Q, with diffraction peaks at 7.657° ± 0.2°, 9.825° ± 0.2°, 11.917° ± 0.2°, 13.536° ± 0.2°, 15.367° ± 0.2°, 17.339° ± 0.2°, 17.958° ± 0.2°, 18.896° ± 0.2°, 19.938° ± 0.2°, 20.930° ± 0.2°, 22.257° ± 0.2°, 22.813° ± 0.2°, 23.598° ± 0.2°, 24.087° ± 0.2°, 24.974° ± 0.2°, 27.090° ± 0.2°, using Cu-Ka radiation.

[0083] In one aspect, the Form X has an X-ray powder diffraction pattern, as indicated by 2Q, with diffraction peaks at 7.657° ± 0.2°, 9.825° ± 0.2°, 11.917° ± 0.2°, 13.536° ± 0.2°, 15.367° ± 0.2°, 17.339° ± 0.2°, 17.958° ± 0.2°, 18.896° ± 0.2°, 19.938° ± 0.2°, 20.930° ± 0.2°, 22.257° ± 0.2°, 22.813° ± 0.2°, 23.598° ± 0.2°, 24.087° ± 0.2°, 24.974° ± 0.2°, 27.090° ± 0.2°, using Cu-Ka radiation.

[0084] In one aspect, the Form X has an X-ray powder diffraction pattern, as indicated by 2Q, with diffraction peaks at 7.657° ± 0.2°, 9.825° ± 0.2°, 11.917° ± 0.2°, 13.536° ± 0.2°, 15.367° ± 0.2°, 17.339° ± 0.2°, 17.958° ± 0.2°, 18.896° ± 0.2°, 19.938° ± 0.2°, 20.930° ± 0.2°, 22.257° ± 0.2°, 22.813° ± 0.2°, 23.598° ± 0.2°, 24.087° ± 0.2°, 24.974° ± 0.2°, 27.090° ± 0.2°, using Cu-Ka radiation.

[0085] The present application also provides a crystalline form III-A of the compound of Formula (III);

[0086] The Form III-A has an X-ray powder diffraction pattern, as indicated by 2Q, with diffraction peaks at 5.356° ± 0.2°, 7.628° ± 0.2°, 10.731° ± 0.2°, 12.019° ± 0.2°, 12.591° ± 0.2°, 13.678° ± 0.2°, 13.912° ± 0.2°, 14.237° ± 0.2°, using Cu-Ka radiation.

[0087] In certain embodiments, the Form III-A has an X-ray powder diffraction pattern, using Cu-Ka radiation, with diffraction peaks, in terms of 2Q, at 5.356°±0.2°, 7.628°±0.2°, 10.731°±0.2°, 12.019°±0.2°, 12.591°±0.2°, 13.678°±0.2°, 13.912°±0.2°, 14.237°±0.2°, 15.229°±0.2°, 16.126°±0.2°, 16.938°±0.2°, 17.171°±0.2°, 17.531°±0.2°, 17.837°±0.2°, 18.223°±0.2°, 18.459°±0.2°, 19.965°±0.2°, 20.997°±0.2°, 21.497°±0.2°, 21.892°±0.2°, 22.237°±0.2°, 22.597°±0.2°, 22.847°±0.2°, 23.182°±0.2°, 23.488°±0.2°, 24.103°±0.2°, 24.883°±0.2°, 25.295°±0.2°, 25.585°±0.2°.

[0088] In one aspect, the Form III-A has an X-ray powder diffraction pattern, expressed in terms of 2-theta, having diffraction peaks at approximately 5.356°±0.2°, 7.628°±0.2°, 10.731°±0.2°, 12.019°±0.2°, 12.591°±0.2°, 13.678°±0.2°, 13.912°±0.2°, 14.237°±0.2°, 15.229°±0.2°, 16.126°±0.2°, 16.938°±0.2°, 17.171°±0.2°, 17.531°±0.2°, 17.837°±0.2°, 18.223°±0.2°, 18.459°±0.2°, 19.965°±0.2°, 20.997°±0.2°, 21.497°±0.2°, 21.892°±0.2°, 22.237°±0.2°, 22.597°±0.2°, 22.847°±0.2°, 23.182°±0.2°, 23.488°±0.2°, 24.103°±0.2°, 24.883°±0.2°, 25.295°±0.2°, 25.585°±0.2°, 26.091°±0.2°, 26.604°±0.2°, 26.959°±0.2°, 27.539°±0.2°, 27.989°±0.2°, 28.637°±0.2°, 29.137°±0.2°, 29.994°±0.2°, 30.348°±0.2°, 30.803°±0.2°, 31.256°±0.2°, 32.501°±0.2°, 33.073°±0.2°, 34.286°±0.2°, 35.467°±0.2°, using Cu-Kalpharadiation.

[0089] In one aspect, the Form III-A has an X-ray powder diffraction pattern, expressed in terms of 2-theta, having the following peaks:

[0090] In one aspect, the Form III-A has an X-ray powder diffraction pattern, expressed in terms of 2-theta, having the following peaks:

[0091] In one aspect, the Form III-A has an X-ray powder diffraction pattern substantially as shown in FIG. 16.

[0092] The present application also provides a crystalline Form IV-A of the compound of formula (IV);

[0093] The X-ray powder diffraction pattern of the crystalline Form IV-A, in terms of 2Q using Cu-Kalpharadiation, has diffraction peaks at the following positions: 5.937°±0.2°, 9.115°±0.2°, 10.105°±0.2°, 11.734°±0.2°, 11.855°±0.2°, 12.790°±0.2°, 13.841°±0.2°, 14.955°±0.2°.

[0094] In a certain embodiment, the crystalline Form IV-A, has an X-ray powder diffraction pattern in terms of 2Q using Cu-Kalpharadiation, with diffraction peaks at the following positions: 5.937°±0.2°, 9.115°±0.2°, 10.105°±0.2°, 11.734°±0.2°, 11.855°±0.2°, 12.790°±0.2°, 13.841°±0.2°, 14.955°±0.2°, 15.313°±0.2°, 15.989°±0.2°, 17.356°±0.2°, 17.796°±0.2°, 18.774°±0.2°, 19.307°±0.2°, 19.515°±0.2°, 20.015°±0.2°, 21.640°±0.2°, 21.942°±0.2°, 22.964°±0.2°.

[0095] In one aspect, the Form IV-A has an X-ray powder diffraction pattern, expressed in terms of 2-theta, having diffraction peaks at approximately 5.937°±0.2°, 9.115°±0.2°, 10.105°±0.2°, 11.734°±0.2°, 11.855°±0.2°, 12.790°±0.2°, 13.841°±0.2°, 14.955°±0.2°, 15.313°±0.2°, 15.989°±0.2°, 17.356°±0.2°, 17.796°±0.2°, 18.774°±0.2°, 19.307°±0.2°, 19.515°±0.2°, 20.015°±0.2°, 21.640°±0.2°, 21.942°±0.2°, 22.964°±0.2°, 23.228°±0.2°, 23.781°±0.2°, 24.302°±0.2°, 25.697°±0.2°, 26.191°±0.2°, 26.872°±0.2°, 27.979°±0.2°, 28.632°±0.2°, 28.960°±0.2°, 29.843°±0.2°, 30.845°±0.2°, 31.716°±0.2°, 32.707°±0.2°, 33.815°±0.2°, 35.594°±0.2°, 36.992°±0.2°, 39.202°±0.2°, using Cu-Ka radiation.

[0096] In one aspect, the Form IV-A has an X-ray powder diffraction pattern, expressed in terms of 2-theta, having the following diffraction peaks:

[0097] In one aspect, the Form IV-A has an X-ray powder diffraction pattern, expressed in terms of 2-theta, having the following diffraction peaks:

[0098] In one aspect, the Form IV-A has an X-ray powder diffraction pattern substantially as shown in Figure 18.

[0099] The present application also provides a preparation method of the Form A of the compound of formula (I), comprising the following steps:

[0100] The isopropyl acetate is added to the DMSO solution of the compound of formula (I) at 50-70°C (e.g. 60°C), and then the temperature is lowered to 15-35°C to crystallize.

[0101] In one aspect, the temperature is lowered to 20-30°C.

[0102] In a certain embodiment, the mass to volume ratio of the compound of formula (I) and DMSO in the DMSO solution of the compound of formula (I) is 0.8-1.2 g / 10 ml, for example 1 g / 10 ml.

[0103] In a certain embodiment, the mass to volume ratio of the compound of formula (I) and isopropyl acetate is 0.8-1.2 g / 15 ml, for example 1 g / 15 ml.

[0104] The present application also provides a preparation method of the crystal form of the compound of formula (II), which comprises the following steps: beating up the compound of formula (II) in a solvent, and crystallizing.

[0105] When the solvent is n-heptane, the crystal form of the compound of formula (II) is crystal form I.

[0106] When the solvent is acetone / water with a volume ratio of (8-10):1, the crystal form of the compound of formula (II) is crystal form II.

[0107] When the solvent is methanol, the crystal form of the compound of formula (II) is crystal form III.

[0108] When the solvent is 1,4-dioxane, the crystal form of the compound of formula (II) is crystal form V.

[0109] When the solvent is isopropyl ester, the crystal form of the compound of formula (II) is crystal form VI.

[0110] When the solvent is isopropyl acetate, the crystal form of the compound of formula (II) is crystal form VII.

[0111] When the solvent is toluene, the crystal form of the compound of formula (II) is crystal form VIII.

[0112] When the solvent is DMF / EA=1:1 (volume ratio), the crystal form of the compound of formula (II) is crystal form IX.

[0113] In a certain embodiment, the beating up temperature is 10-30°C, for example 20-30°C.

[0114] In a certain embodiment, the stirring speed during the beating up is 300-700 rpm, for example 500 rpm.

[0115] In a certain embodiment, the mass to volume ratio of the compound of formula (II) and the solvent is (25-35) g / L.

[0116] In a certain embodiment, the beating up time is 3-7 days, for example 5 days.

[0117] In a certain embodiment, the crystallization is centrifugation treatment.

[0118] In an embodiment, the preparation of the compound of formula (II) also comprises the following post-treatment step: after crystallization, the solid is collected and the solvent is evaporated at 30-50℃ under vacuum.

[0119] The present application also provides a method for preparing a compound of formula (II) in a crystal form, which comprises the following steps: slurrying an acetonitrile solution of the compound of formula (II) and crystallizing.

[0120] When the slurry temperature is 10-30℃, the crystal form of the compound of formula (II) is crystal form IV.

[0121] When the slurry temperature is 40-60℃, the crystal form of the compound of formula (II) is crystal form X.

[0122] In an embodiment, the stirring speed during the slurry is 300-700rpm, for example 500rpm.

[0123] In an embodiment, the mass / volume ratio of the compound of formula (II) and the acetonitrile is (25-35)g / L.

[0124] In an embodiment, the slurry time is 3-7 days, for example 5 days.

[0125] In an embodiment, the crystallization is centrifugation.

[0126] In an embodiment, the preparation also comprises the following post-treatment step: after crystallization, the solid is collected and the solvent is evaporated at 30-50℃ under vacuum.

[0127] The present application also provides a method for preparing a crystal form III-A of a compound of formula (III), which comprises the following steps:

[0128] After the compound of formula (I) is reacted with p-toluenesulfonic acid in DMSO, isopropyl acetate is added after the temperature is raised to 50-80℃, and the temperature is lowered to 15-30℃ to crystallize.

[0129] In an embodiment, the temperature is raised to 60-70℃.

[0130] In an embodiment, the reaction temperature of the compound of formula (I) and the p-toluenesulfonic acid is 15-40℃, for example 20-30℃.

[0131] In an embodiment, the reaction time of the compound of formula (I) and the p-toluenesulfonic acid is 0.2-1h, for example 0.5h.

[0132] In an embodiment, the molar ratio of the compound of formula (I) and the p-toluenesulfonic acid is 1:(1-1.5), for example 1:1.05.

[0133] In an embodiment, the mass to volume ratio of the compound of formula (I) and the DMSO is 0.8-1.2 g / 20 ml, for example 1 g / 20 ml.

[0134] In an embodiment, the mass to volume ratio of the compound of formula (I) and the isopropyl acetate is 0.8-1.2 g / 20 ml, for example 1 g / 20 ml.

[0135] The present application also provides a preparation method of the crystalline form IV-A of the compound of formula (IV), comprising the following steps:

[0136] After the compound of formula (I) is reacted with maleic acid in DMSO, isopropyl acetate is added after the temperature is raised to 50-80℃, and the temperature is lowered to 15-30℃ for crystallization.

[0137] In an embodiment, the temperature is raised to 60-70℃.

[0138] In an embodiment, the temperature for the reaction of the compound of formula (I) and the maleic acid is 15-40℃, for example 20-30℃.

[0139] In an embodiment, the reaction time of the compound of formula (I) and the maleic acid is 0.2-1 h, for example 0.5 h.

[0140] In an embodiment, the molar ratio of the compound of formula (I) and the maleic acid is 1:(1-1.5), for example 1:1.05.

[0141] In an embodiment, the mass to volume ratio of the compound of formula (I) and the DMSO in the DMSO solution of the compound of formula (I) is 0.8-1.2 g / 20 ml, for example 1 g / 20 ml.

[0142] In an embodiment, the mass to volume ratio of the compound of formula (I) and the isopropyl acetate is 0.8-1.2 g / 20 ml, for example 1 g / 20 ml.

[0143] The present application also provides a pharmaceutical composition comprising one or more of the aforementioned compound of formula (II), compound of formula (III), compound of formula (IV), crystalline form of any one of the aforementioned compound of formula (I), compound of formula (II), compound of formula (III) or compound of formula (IV), and a pharmaceutically acceptable excipient.

[0144] The present invention also provides the use of the aforementioned pharmaceutical compositions, compounds of formula (II), (III), and (IV), as well as the crystal form of any of the aforementioned compounds of formula (I), (II), (III), or (IV), in the preparation of medicaments for treating and / or preventing PARP-mediated diseases. The present invention also provides the use of the aforementioned pharmaceutical compositions, compounds of formula (II), (III), and (IV), as well as the crystal form of any of the aforementioned compounds of formula (I), (II), (III), or (IV), as PARP inhibitors.

[0145] Preferably, the PARP is PARP1.

[0146] In one embodiment, the present invention is used to treat and / or prevent cancer, ischemic diseases, and neurodegenerative diseases.

[0147] In one scenario, the cancer lacks the HR-dependent DNADSB repair pathway.

[0148] In one embodiment, the cancer has a BRCA1 or BRCA2 defective phenotype.

[0149] In one embodiment, the present invention is used to treat and / or prevent the following cancers: breast cancer, ovarian cancer, pancreatic cancer, prostate cancer, blood cancer, gastrointestinal cancer, and lung cancer.

[0150] The positive and progressive effects of this invention are as follows: the crystal forms of compounds of formula (I), (II), (III), and (IV) provided by this invention exhibit excellent PARP1 inhibitory activity and selectivity, thereby avoiding anemia toxicity. Simultaneously, the compounds of this invention possess different volumes of distribution (reducing neutrophil and platelet toxicity) and different trapping activities (facilitating dosage adjustment), maintaining the clinical efficacy of PARP1 / 2 inhibitors while reducing hematologic toxicity, thus enabling further combination therapy of highly selective PARP1 inhibitors with chemotherapeutic drugs. Attached Figure Description

[0151] Figure 1 shows the 1H NMR spectrum of compound (I).

[0152] Figure 2 shows the 1H NMR spectrum of compound (II).

[0153] Figure 3 shows the XRPD spectrum of crystal form A of compound (I).

[0154] Figure 4 shows the XRPD spectrum of crystal form I of compound (II).

[0155] Figure 5 shows the XRPD spectrum of crystal form II of compound (II).

[0156] Figure 6 is a DSC and TGA pattern of the crystalline Form II of the compound of formula (II)

[0157] Figure 7 is an XRPD pattern of the crystalline Form III of the compound of formula (II)

[0158] Figure 8 is an XRPD pattern of the crystalline Form IV of the compound of formula (II)

[0159] Figure 9 is an XRPD pattern of the crystalline Form V of the compound of formula (II)

[0160] Figure 10 is an XRPD pattern of the crystalline Form VI of the compound of formula (II)

[0161] Figure 11 is an XRPD pattern of the crystalline Form VII of the compound of formula (II)

[0162] Figure 12 is an XRPD pattern of the crystalline Form VIII of the compound of formula (II)

[0163] Figure 13 is an XRPD pattern of the crystalline Form IX of the compound of formula (II)

[0164] Figure 14 is an XRPD pattern of the crystalline Form X of the compound of formula (II)

[0165] Figure 15 is a nuclear magnetic resonance hydrogen spectrum of the compound of formula (III)

[0166] Figure 16 is an XRPD pattern of the crystalline Form III-A of the compound of formula (III)

[0167] Figure 17 is a nuclear magnetic resonance hydrogen spectrum of the compound of formula (IV)

[0168] Figure 18 is an XRPD pattern of the crystalline Form IV-A of the compound of formula (IV)

[0169] Figure 19 is the effect of the compound on the change in tumor volume on MDA-MB-436 mouse subcutaneous transplanted tumors

[0170] Figure 20 is the induction of apoptosis of DLD-1 BRCA2- / - cells by the compound in the first experiment (ns, no statistically significant difference; *, p < 0.05, statistically significant difference; **, p < 0.01, statistically significant difference)

[0171] Figure 21 is the induction of apoptosis of DLD-1 BRCA2- / - cells by the compound in the second experiment (ns, no statistically significant difference; *, p < 0.05, statistically significant difference; **, p < 0.01, statistically significant difference)

[0172] Figure 22 shows the induction of apoptosis in DLD-1 BRCA2- / - cells by the compounds in the third experiment (ns, not statistically significant; *, p < 0.05, statistically significant; **, p < 0.01, statistically significant)

[0173] Figure 23 shows the expression of cleaved Caspase-3 and Caspase-3 proteins in tumour tissue

[0174] Figure 24 shows the effect of the compound of formula (I) and the reference AZD5305 on Cleaved Caspase-3 protein in tumour tissue at the same dose (*, p < 0.05 by one-way ANOVA, no statistically significant difference between the other groups)

[0175] Figure 25 shows the effect of the compound of formula (I) and the reference AZD5305 on Cleaved Caspase-3 protein in tumour tissue at the same dose (*, p < 0.05 by one-way ANOVA, no statistically significant difference between the other groups) DETAILED DESCRIPTION

[0176] The application is further illustrated by way of the following examples without, however, being limited thereto.

[0177] Instruments and analytical methods

[0178] The instruments used are listed below

[0179] 1.1 Detection methods

[0180] 1.1.1 X-ray powder diffraction analysis (XRPD)

[0181] The light source is Cu K at 40 KV / 40 mA, the scanning mode is theta-theta, the scanning angle range is 4° to 40°, the step is 0.01° and the scanning speed is 0.1 s / step.

[0182] 1.1.2 Differential scanning calorimetry analysis (DSC)

[0183] The sample is weighed in an appropriate amount and placed in an open aluminium pan in a nitrogen stream (50 mL / min) environment. The sample is equilibrated at 31 °C and then heated from 31 °C to 300 °C at a rate of 10 °C / min.

[0184] 1.1.3 Thermogravimetric analysis (TGA)

[0185] An appropriate amount of sample was weighed into a platinum sample pan and heated from 31 °C to 300 °C at a rate of 10 °C / min under a nitrogen stream (20 mL / min) and a balance nitrogen stream (40 mL / min).

[0186] 1.1.4 Purity HPLC analysis method

[0187] 1.1.5 Chloride content determination method

[0188] The following table shows the English / abbreviation of the solvents involved in the present application and their Chinese names:

[0189] Example 1 Synthesis of compound of formula (I)

[0190] Synthesis of compound 1

[0191] Into a 100 L glass reactor, 25.00 L of dioxane was added, and then 2.50 kg of 2-fluoro-3-bromopyridine, 3.97 kg of N-Boc piperazine (1.5 ep), 13.90 kg of cesium carbonate, 0.98 kg of Xant-Phos, and 0.78 kg of Pd2(dba)3 were added in sequence under stirring. After the addition was completed, the temperature was raised to 100-105 °C, and the reaction was carried out for 6 h. After sampling, TLC showed that the starting material was completely reacted, and the reaction was completed.

[0192] The reaction solution was filtered through diatomite, and the filter cake was washed with 5*5 L of ethyl acetate to obtain filtrate 1. The filter cake was slurried at 20-30 °C for 1 h with 18 L of ethyl acetate, and then filtered to obtain filtrate 2. Filtrate 1 and filtrate 2 were combined, and concentrated at 50 °C under reduced pressure to 18-20 kg. The concentrated solution was added to a 100 L reactor. The organic phase was washed twice with 2*14 L of saturated aqueous ammonium chloride solution, and concentrated to dryness at 50 °C under reduced pressure. Compound 1 4.75 kg was obtained, which was directly used in the next step.

[0193] 1 H NMR (400 MHz, CD3Cl) δ = 7.910-7.90 (m, 1H), δ = 7.50-7.47 (m, 2H), δ = 3.39-3.37 (m, 4H), δ = 3.30-3.27 (m, 4H), δ = 1.43 (s, 9H).

[0194] Synthesis of compound 2

[0195] Into a 100 L glass reactor, 17 L of DMF was added, and 4.00 kg of compound 1 was added under stirring, and the temperature was lowered to 10-15 °C. 4.56 kg of NBS (1.8 ep) was added in batches, and the temperature was controlled at 10-25 °C. After the addition was completed, it was stirred at 20-30 °C for 3 h. Sampling, TLC showed that compound 1 was completely reacted, and the reaction was completed.

[0196] Into the reaction solution, 25 L of ethyl acetate and 15 L of 10% sodium chloride aqueous solution were added, and stirred for 15 min. After standing and layering, the aqueous phase was extracted with 20 L of ethyl acetate. The combined organic phase was washed once with 15 L of 10% sodium chloride aqueous solution. The organic phase was separated, and washed once with 15 L of 10% sodium sulfite aqueous solution. The organic phase was separated, and washed twice with 2*15 L of 10% sodium chloride aqueous solution. The organic phase was separated, and concentrated to dryness at 50 °C under reduced pressure. Compound 2 5.69 kg was obtained, which was directly used in the next step.

[0197] 1H NMR (400 MHz, CD3Cl) δ = 7.25 (S, 1H), δ = 7.13-7.09 (m, 1H), δ = 3.58-3.55 (t, 4H), δ = 3.02-3.00 (t, 4H), δ = 1.46 (s, 9H).

[0198] Synthesis of compound 3

[0199] Into a 50 L high-pressure reactor, 25.60 L of dioxane and 10.20 L of methanol were added, and 5.12 kg of compound 2 was added under stirring, and 4.31 kg of triethylamine and 415 g of Pd(dppf)Cl2 were added in sequence. After the addition was completed, it was replaced with nitrogen for three times, and then replaced with carbon monoxide for two times. Carbon monoxide was filled to control the pressure at 1.2-1.3 MPa, and the temperature was raised to 100-105 °C for 8 h. Sampling, TLC showed that compound 2 was completely reacted, and the reaction was completed.

[0200] The reaction solution was removed, concentrated at 50°C under reduced pressure to 18-22 kg, and the concentrated solution was filtered through diatomite. The filter cake was washed with 10 L of ethyl acetate. The filtrate was removed and concentrated at 50°C under reduced pressure to dryness. The concentrate was dissolved in 2.5 L of dichloromethane and loaded into a glass column pre-loaded with 9 kg of 100-200 mesh silica gel. Eluent (PE:EA = 3:1) was added to start column chromatography. TLC was used to detect the end of column chromatography. The eluent was concentrated to dryness to obtain 3.07 kg of crude product. The crude product was dissolved in 9 L of methyl tert-butyl ether and added to a 50 L reaction kettle. 9 L of n-hexane was added, and stirring was performed at 20-30°C for 3 h. Filtration was performed, and the filter cake was dissolved in 4 L of methyl tert-butyl ether and added to a 20 L reaction kettle. 4 L of n-hexane was added, and stirring was performed at 20-30°C for 4 h. Filtration was performed, and the filter cake was rinsed with 3 L of n-hexane. The filter cake was placed in a forced air oven at 50°C to dry. A total of 1.63 kg of compound 3 was obtained, with a yield of 35%.

[0201] 1 H NMR (400 MHz, MeOD) δ = 7.9310-7.9075 (m, 1H), δ = 7.5026-7.4567 (m, 1H), δ = 3.8984 (s, 3H), δ = 3.5999-3.5769 (m, 4H), δ = 3.3138-3.2075 (m, 4H), δ = 1.4739 (s, 9H).

[0202] Synthesis of compound 4

[0203] A 20 L glass reaction kettle was charged with 2.20 L of tetrahydrofuran and 2.20 kg of compound 3, and stirring was performed while the temperature was lowered to 15-20°C. Lithium hydroxide solution (0.31 kg of lithium hydroxide dissolved in 4.40 L of purified water) was added dropwise while the temperature was controlled at 15-25°C. After the addition was completed, the reaction was allowed to proceed at 20-30°C for 0.5 h, and a sample was taken; HPLC showed that compound 3 was <1.00%, and the reaction was completed. Hydrochloric acid solution (1.71 kg of concentrated hydrochloric acid dissolved in 1.45 L of purified water) was added dropwise while the temperature was controlled at 20-30°C and stirring was performed; after the addition was completed, stirring was performed at 20-30°C for 1 h, and filtration was performed. The filter cake was washed with 4.40 L of purified water, and the filter cake was dried at 50°C under vacuum for 48 h. The crude compound 4 was weighed, and the yield was 2.17 kg, which was 102%.

[0204] A 100 L glass reaction kettle was charged with 63.00 L of dichloromethane and 2.10 kg of compound 4, and stirring was performed at 20-30°C until the solution was clear. The solution was washed with 20.00 L of 10% sodium chloride aqueous solution, and the organic layer was separated. Compound 4 was obtained by concentration under reduced pressure, and the yield was 2.02 kg, which was 96%.

[0205] 1H NMR (400 MHz, DMSO) δ = 13.0362 (s, 1H), δ = 7.9177-7.8980 (m, 1H), δ = 7.5676-7.5208 (m, 1H), δ = 3.4887-3.3646 (m, 4H), δ = 3.1777-3.1530 (m, 4H), δ = 1.4255 (s, 9H).

[0206] Synthesis of compound 5

[0207] Into a 100 L glass reactor, add 18.50 kg DMF, 1.95 kg compound 4, 2.34 kg DIEA, 507 g deuterated methylamine hydrochloride, respectively; stir, slowly add 2.73 kg HATU in batches, control the temperature at 20-30 °C. After addition, keep the temperature at 20-30 °C for 0.5 h, then take a sample; HPLC shows that compound 4 < 1.00%, the reaction is completed. Add 40.00 L purified water to the reaction solution, extract with 29.00 L ethyl acetate; separate the organic layer for use, add 20.00 L ethyl acetate to the water layer for extraction; separate the water layer and discard, combine the two times of organic layer extraction, wash twice with 2 x 20.00 L purified water; separate the organic layer, wash once with 20.00 L saturated sodium bicarbonate solution, and then wash once with 20.00 L saturated sodium chloride solution. Separate the organic layer, concentrate under reduced pressure at 45-50 °C until no distillate is obtained, to obtain 1.77 kg of compound 5, yield: 86%.

[0208] 1 H NMR (400 MHz, DMSO) δ = 8.4062 (s, 1H), δ = 7.8689-7.8496 (m, 1H), δ = 7.6118-7.5652 (m, 1H), δ = 3.4916-3.3477 (m, 4H), δ = 3.1286-3.1040 (m, 4H), δ = 1.4278 (s, 9H).

[0209] Synthesis of compound 6

[0210] Into a 20 L glass reactor, add 6.50 L dichloromethane, 1.60 kg compound 5, 160 g activated carbon, stir at 20-30 °C for 0.5-1 h, filter, and reserve the filtrate. Into a 50 L glass reactor, add 11.20 L hydrogen chloride ethyl acetate solution (4 mol / L), cool to 10-15 °C and stir; add the above filtrate dropwise, control the temperature at 10-20 °C, after addition, keep the temperature at 15-20 °C for 0.5 h, then take a sample; HPLC shows that compound 5 < 1.00%, the reaction is completed. Filter, wash the filter cake with 3.00 L dichloromethane, and dry the filter cake at 40 °C under vacuum for 16 h, to obtain 1.35 kg of compound 6, yield: 104%.

[0211] 1 H NMR (400 MHz, MeOD) δ = 7.9516-7.9287 (m, 1H), δ = 7.6741-7.6283 (m, 1H), δ = 3.5367-3.5112 (m, 4H), δ = 3.4694-3.4444 (m, 4H); 13 C NMR (400 MHz, MeOD) δ: 47.7607-47.7180, 44.6403, 26.0427, 25.8357, 25.6250, 25.4195.

[0212] Synthesis of compound 7

[0213] To the reaction vessel was added acetic acid (30 kg) and stirred at 20-30 °C, then added (7.14 kg, 127.85 moles) of reduced iron powder, and then slowly added a solution of 5-bromo-2-cyano-3-nitropyridine (5.3 kg, 23.24 moles) in acetic acid (25 kg) dropwise (about 6 h). The reaction was stirred at 20-30 °C for 3 h, and then terminated. Centrifuged, the filter cake was stirred with THF (32 kg) for 3 h, then centrifuged, and the filter cake was stirred with THF (32 kg) again, centrifuged, and the filtrates were combined and concentrated under reduced pressure at 50 °C. The concentrate was slurried with purified water (40 kg) at 60-70 °C for 1 h, cooled to 20-30 °C, and centrifuged, and the filter cake was slurried with a mixture of dichloromethane (6.9 kg) and n-hexane (3.7 kg) at 20-30 °C for 1 h, centrifuged, and the filter cake was air-dried at 45-55 °C to give light yellow compound 7 (3.8 kg, 84% yield).

[0214] 1 H NMR (400 MHz, DMSO-d6): δ 7.93 (d, 1H), 7.45 (d, 1H), 6.54 (s, 2H).

[0215] Synthesis of compound 8

[0216] Into a reaction kettle, add tetrahydrofuran (33.5 kg), compound 7 (3.8 kg, 19.20 mol), DMAP (0.23 kg, 1.92 mol), pyridine (2.40 kg, 1.92 mol), and drop butyryl chloride (3.10 kg, 29.10 mol) under nitrogen protection, and then warm to 20-30 °C and stir for 2 h. Add potassium tert-butoxide (12.70 kg, 113.60 mol) in batches, warm to 55-65 °C, and stir for 3 h. Concentrate the reaction solution to remove the distillate under reduced pressure at 50-60 °C in the kettle. Add 20 kg of purified water and continue to concentrate to remove the distillate. Add 18 kg of purified water, and cool to 20-30 °C. Centrifuge and filter, slurry the filter cake with 19 kg of purified water at 10-20 °C for 1 h, centrifuge and filter, and dry the filter cake at 45-55 °C under air flow to obtain compound 8 (4.3 kg, yield 84%).

[0217] 1 H NMR (400 MHz, DMSO-d6) δ = 10.93 (s, 1H), δ = 8.54-8.33 (m, 1H), δ = 8.32-8.28 (m, 1H), δ = 4.31-4.28 (m, 2H), δ = 1.05-1.02 (m, 3H).

[0218] Synthesis of compound 9

[0219] Into a high-pressure reaction kettle, add 1,4-dioxane (21.7 kg) and methanol (5.5 kg), then add compound 8 (3.5 kg, 13.10 mol), triethylamine (3.9 kg, 39.20 mol), and [1,1-bis(diphenylphosphino)ferrocene]palladium dichloride dichloromethane (0.47 kg, 0.65 mol), and then stir the reaction solution at 80-90 °C under a carbon monoxide atmosphere at 1.0 MPa for 5-8 h. LCMS shows that the raw material is completely consumed. Cool the reaction solution, filter, slurry the filter cake with methanol (10.50 kg) at 10-20 °C for 1 h, filter, and dry the filter cake at 45-55 °C under air flow to obtain compound 9 (2.7 kg, yield 84%).

[0220] 1 H NMR (400 MHz, DMSO-d6) δ = 11.02 (H-15, s, 1H), δ = 8.35-8.34 (m, 1H), δ = 7.59-7.58 (m, 1H), δ = 4.67-4.66 (m, 2H), δ = 3.91-3.90 (m, 3H); δ = 1.06-1.02 (m, 3H).

[0221] Synthesis of compound 10

[0222] Into a reaction kettle, add tetrahydrofuran (52.5 kg), compound 9 (1.5 kg, 6.07 mol), and cool to -5-5 °C under nitrogen protection. Add lithium aluminum hydride solution (2.5 M / THF, 2.7 kg, 7.28 mol) dropwise, and stir the reaction mixture at 10-20 °C for 16 h after the temperature is raised. Cool the reaction mixture to 5-15 °C, and then add purified water (2.0 kg) dropwise to quench the reaction. Add 5% NaOH aqueous solution (15.0 kg), stir at 20-30 °C, and then separate the phases. Extract the aqueous phase with tetrahydrofuran (6 x 12.5 kg). Combine the organic phases, concentrate under reduced pressure at 50 °C, filter the concentrate, and then slurry the filter cake with purified water (3.0 kg) at 10-20 °C for 1 h. Filter, and then dry the filter cake at 45-55 °C under air flow to obtain compound 10 (0.87 kg, yield 65.41%).

[0223] 1 H NMR (400 MHz, DMSO) δ = 11.0182 (H-15, s, 1H), δ = 8.3393-8.3352 (m, 1H), δ = 7.5855-7.5838 (m, 1H), δ = 6.3741 (s, 2H), δ = 5.5126-5.4871 (m, 1H), δ = 4.6573-4.6466 (m, 2H), δ = 1.0580-1.0214 (m, 3H); 13 C NMR (400 MHz, DMSO) δ: 162.2565, 147.4702, 140.8701, 139.0095, 133.2015, 129.6908, 119.8844, 107.3262, 60.3807, 17.2246, 12.1835.

[0224] Synthesis of compound 11

[0225] Into a 100 L glass reaction kettle, add 15.00 L tetrahydrofuran, 1.00 kg compound 10, and 295 g N,N-diisopropylethylamine, and stir while cooling to 15-20 °C. Add 1.63 kg dichlorosulfoxide dropwise while controlling the temperature at 15-25 °C. After the addition is complete, stir the reaction mixture at 20-25 °C for 1 h, and then take a sample; HPLC shows that compound 10 < 1.00%, and the reaction is complete. Filter, wash the filter cake with 2.00 L tetrahydrofuran, and then dry the filter cake at 40 °C for 16 h. Weigh the dried filter cake to obtain 1.37 kg of crude compound 11.

[0226] Into a 20 L glass reaction kettle, add 13.00 L acetonitrile, and 1.37 kg compound 11, and stir while controlling the temperature at 20-30 °C for 3 h. Filter, dry the filter cake at 40 °C for 16 h, and then weigh the dried filter cake to obtain 1.23 kg of compound 11, with a yield of 98%.

[0227] 1 H NMR (400 MHz, DMSO) δ = 11.3723 (s, 1H), δ = 8.5037-8.4995 (m, 1H), δ = 7.7668-7.7628 (m, 1H), δ = 6.5683 (m, 3H), δ = 4.9629 (s, 2H), δ = 2.6225-2.5568 (m 2H), δ = 1.0733-1.0367 (m 3H).

[0228] Synthesis of compound of formula (I)

[0229] Into a 50L glass reactor, add 5.50L dimethyl sulfoxide, 1.29kg compound 6, 2.42kg N,N-diisopropyl ethylamine, 1.10kg compound 11, stir, heat to 65-75℃, sample after 2h, HPLC shows compound 11 <2.00%, reaction is completed. Into the reactor, add 8.80kg absolute ethanol, control temperature 60-75℃, after addition, cool to 20-30℃, stir for 16h, filter, filter cake is washed with 2.20L absolute ethanol. Filter dry, add filter cake into a 50L glass reactor, 8.80L dimethyl sulfoxide, heat to 90-100℃, stir for 1h, cool to 20-30℃, stir for 1h, filter, filter cake is washed with 4.40L isopropyl acetate, filter dry, filter cake is dried at 50-55℃ under air for 40h, stop drying, take out and weigh, to get 1.48kg compound of formula (I) crude product.

[0230] Add crude product into a 50L glass reactor, add 5.92L dimethyl sulfoxide, heat to 85-95℃, stir for 1.5h, cool to 20-30℃, stir for 1h, filter, filter cake is washed with 5.92L isopropyl acetate, filter dry, filter cake is dried at 50-55℃ under vacuum for 15h, take out and weigh, to get 1.29kg compound of formula (I), yield 73%.

[0231] 1 H NMR (400 MHz, DMSO) δ = 10.8994 (s, 1H), δ = 8.3778 (s, 1H), δ = 8.3219-8.3181 (d, 1H), δ = 7.8559-7.8332 (m, 1H), δ = 7.5916-7.5445 (m, 1H), δ = 7.5341-7.5308 (m, 1H), δ = 6.3355 (s, 1H), δ = 3.6465 (s, 2H), δ = 3.1766 (s, 4H), δ = 2.5739-2.5128 (m, 4H), δ = 1.0095-0.9730 (m, 3H).

[0232] The nuclear magnetic resonance spectrum of compound of formula (I) is shown in Figure 1.

[0233] Preparation and characterization of the compound of formula II

[0234] Into a reactor, 6.00 L of dimethyl sulfoxide, 1.20 kg of the compound of formula I were added, and the temperature was controlled at 20-30 °C and stirred. Dimethyl sulfoxide hydrochloride solution (267 g of concentrated hydrochloric acid dissolved in 2.40 L of dimethyl sulfoxide) was added dropwise. After the addition was completed, the temperature was controlled at 20-40 °C and stirred until the system was clear. The temperature was raised to 75-85 °C and stirred. 6.00 L of isopropyl acetate was added dropwise. After the addition was completed, the temperature was controlled at 75-85 °C. The cooling rate was controlled, and the temperature was reduced to 55-60 °C in 1.5-2.5 h. 9.60 L of isopropyl acetate was added dropwise, and the addition was completed in 20-40 min. After the addition was completed, the temperature was reduced to 20-30 °C and stirred for 12-15 h. Filtration was performed, and the filter cake was washed with 6.40 L of isopropyl acetate. The filter cake was dried, and the filter cake was vacuum dried at 50 °C for 24 h to obtain the compound of formula II.

[0235] 1H NMR (400 MHz, DMSO) δ = 11.87 (s, 1H), δ = 11.22 (s, 1H), δ = 8.71 (d, 1H), δ = 8.42 (s, 1H), δ = 7.87-7.89 (dd, 1H), δ = 7.74-7.76 (d, 1H), δ = 7.67-7.71 (dd, 1H), δ = 6.42-6.45 (s, 2H), δ = 4.54-4.58 (s, 2H), δ = 3.68-3.70 (s, 2H), δ = 3.34-3.44 (m, 6H), δ = 2.57-2.51 (q, 2H), δ = 1.03-0.97 (t, 3H);

[0236] The nuclear magnetic resonance spectrum is shown in Figure 2. The results show that the equivalent ratio of free base to hydrochloric acid in the compound of formula (II) is 1:1.

[0237] It was detected that the content of chloride ion in the compound of formula (II) was 7.3%, and the equivalent ratio of free base to hydrochloric acid was 1:1.

[0238] Example 3

[0239] Preparation and characterization of the compound of formula II

[0240] 1 g of the compound of formula (I) was weighed, 10 mL of DMSO was added, and the sample was dissolved at 60 °C under stirring. 15 mL of isopropyl acetate was added dropwise. After the addition was completed, the temperature was reduced to 20-30 °C and crystallization was performed for 1 h. Filtration was performed to obtain a solid, and the solvent was removed in a vacuum drying oven at 40 °C to obtain the crystal form A. The XRPD pattern of the crystal form A is shown in Figure 3, and the XRPD data are shown in the following table.

[0241] Preparation and characterization of the compound of formula II

[0242] A sample of 300 mg of the compound of formula (II) was slurried in 10 mL of n-heptane at room temperature. The sample was stirred at room temperature on a multi-point magnetic stirrer (500 rpm) and after 5 days the slurry was removed and centrifuged to collect the solid, which was dried in a vacuum oven at 40 °C to remove the solvent to give Form I. The XRPD pattern of Form I is shown in Figure 4 and the XRPD data are shown in the table below:

[0243] Example 5. Preparation and characterization of Form II of the compound of formula II

[0244] A sample of 300 mg of the compound of formula (II) was slurried in 10 mL of acetone / water (9:1 by volume) at room temperature. The sample was stirred at room temperature on a multi-point magnetic stirrer (500 rpm) and after 5 days the slurry was removed and centrifuged to collect the solid, which was dried in a vacuum oven at 40 °C to remove the solvent to give Form II. The XRPD pattern of Form II is shown in Figure 5 and the DSC and TGA patterns are shown in Figure 6. The XRPD data are shown in the table below:

[0245] Example 6. Preparation and characterization of Form III of the compound of formula II

[0246] A sample of 300 mg of the compound of formula (II) was slurried in 10 mL of methanol at room temperature. The sample was stirred at room temperature on a multi-point magnetic stirrer (500 rpm) and after 5 days the slurry was removed and centrifuged to collect the solid, which was dried in a vacuum oven at 40 °C to remove the solvent to give Form III. The XRPD pattern of Form III is shown in Figure 7. The XRPD data are shown in the table below:

[0247] Example 7. Preparation and characterization of Form IV of the compound of formula II

[0248] A sample of 300 mg of the compound of formula (II) was slurried in 10 mL of acetonitrile at room temperature. The sample was stirred at room temperature on a multi-point magnetic stirrer (500 rpm) and after 5 days the slurry was removed and centrifuged to collect the solid, which was dried in a vacuum oven at 40 °C to remove the solvent to give Form IV. The XRPD pattern of Form IV is shown in Figure 8. The XRPD data are shown in the table below:

[0249] Example 8. Preparation and characterization of Form V of the compound of formula II

[0250] A 300 mg sample of the compound of formula (II) was weighed into 10 mL of 1,4-dioxane and the sample was slurried at room temperature. The sample at room temperature was placed on a multi-point magnetic stirrer (500 rpm) and after 5 days of slurry, the sample was removed and centrifuged to collect the solid, which was dried in a vacuum oven at 40 °C to remove the solvent to obtain Form V. The XRPD pattern of Form V is shown in Figure 9. The XRPD data are shown in the table below:

[0251] Example 9. Preparation and characterization of Form VI of the compound of formula II

[0252] A 300 mg sample of the compound of formula (II) was weighed into 10 mL of 1,4-dioxane and the sample was slurried at room temperature. The sample at room temperature was placed on a multi-point magnetic stirrer (500 rpm) and after 5 days of slurry, the sample was removed and centrifuged to collect the solid, which was dried in a vacuum oven at 40 °C to remove the solvent to obtain Form V. The XRPD pattern of Form V is shown in Figure 9. The XRPD data are shown in the table below:

[0253] Example 10. Preparation and characterization of Form VII of the compound of formula II

[0254] A 300 mg sample of the compound of formula (II) was weighed into 10 mL of 1,4-dioxane and the sample was slurried at room temperature. The sample at room temperature was placed on a multi-point magnetic stirrer (500 rpm) and after 5 days of slurry, the sample was removed and centrifuged to collect the solid, which was dried in a vacuum oven at 40 °C to remove the solvent to obtain Form V. The XRPD pattern of Form V is shown in Figure 9. The XRPD data are shown in the table below:

[0255] Example 11. Preparation and characterization of Form VIII of the compound of formula II

[0256] A 300 mg sample of the compound of formula (II) was weighed into 10 mL of 1,4-dioxane and the sample was slurried at room temperature. The sample at room temperature was placed on a multi-point magnetic stirrer (500 rpm) and after 5 days of slurry, the sample was removed and centrifuged to collect the solid, which was dried in a vacuum oven at 40 °C to remove the solvent to obtain Form V. The XRPD pattern of Form V is shown in Figure 9. The XRPD data are shown in the table below:

[0257] Example 12. Preparation and characterization of Form IX of the compound of formula II

[0258] A sample of 300 mg of the compound of formula (II) was weighed into 10 mL of DMF / EA (1:1 by volume) and slurried at room temperature. The sample was stirred at room temperature on a multi-point magnetic stirrer (500 rpm) and after 5 days of slurry, the suspension sample was centrifuged, the solid was collected and the solvent was evaporated in a vacuum oven at 40 °C to obtain Form IX. The XRPD pattern of Form IX is shown in Figure 13. The XRPD data are shown in the following table:

[0259] Example 13. Preparation and characterization of Form X of the compound of formula II

[0260] A sample of 300 mg of the compound of formula (II) was weighed into 10 mL of DMF / EA (1:1 by volume) and slurried at room temperature. The sample was stirred at room temperature on a multi-point magnetic stirrer (500 rpm) and after 5 days of slurry, the suspension sample was centrifuged, the solid was collected and the solvent was evaporated in a vacuum oven at 40 °C to obtain Form IX. The XRPD pattern of Form IX is shown in Figure 13. The XRPD data are shown in the following table:

[0261] Example 22. Preparation and characterization of Form III-A of the compound of formula III

[0262] A sample of 1 g of the compound of formula (I) was weighed into 20 mL of DMSO and stirred to dissolve at room temperature. To the system, p-toluenesulfonic acid (1.05 eq) was added and a salt reaction was carried out at 20-30 °C for 0.5 h to obtain the compound of formula (III). Then, 20 mL of isopropyl acetate was slowly added dropwise at 60-70 °C, and after the addition was completed, the temperature was lowered to 20-30 °C to crystallize for 1 h. The solid was filtered and the solvent was evaporated in a vacuum oven at 40 °C to obtain Form A.

[0263] 1H NMR (400 MHz, DMSO) δ = 11.22-11.25 (s, 1H), δ = 9.98-10.02 (s, 1H), δ = 8.46-8.48 (s, 1H), δ = 8.41-8.43 (s, 1H), δ = 7.87-7.89 (dd, 1H), δ = 7.74-7.76 (d, 1H), δ = 7.67-7.71 (dd, 1H), δ = 7.48-7.50 (d, 2H), δ = 7.10-7.12 (d, 2H), δ = 6.42-6.45 (s, 2H), δ = 4.54-4.58 (s, 2H), δ = 3.40-3.49 (s, 2H), δ = 3.14-3.44 (m, 6H), δ = 2.57-2.51 (q, 2H), δ = 2.28 (s, 3H), δ = 1.03-0.97 (t, 3H);

[0264] The NMR chart of the compound of formula III is shown in Figure 15, and the results show that the equivalent ratio of free base to p-toluenesulfonic acid in the compound of formula (III) is 1:1.

[0265] The XRPD chart of crystal form III-A is shown in Figure 16, and the XRPD data is shown in the following table

[0266] Example 23. Preparation and characterization of crystal form IV-A of the compound of formula IV

[0267] A sample of 1 g of the compound of formula (I) was weighed, 20 ml of DMSO was added, and the sample was stirred to dissolve at room temperature. Maleic acid (1.05 eq) was added to the system, and a salt reaction was carried out at 20-30 °C for 0.5 h to obtain the compound of formula (IV). The temperature was raised to 60-70 °C, 20 ml of isopropyl acetate solvent was slowly added dropwise, and after the dropwise addition was completed, the temperature was lowered to 20-30 °C for crystallization for 1 h. The solid was filtered, and the solvent was dried in a vacuum drying oven at 40 °C to obtain crystal form A.

[0268] 1H NMR (400 MHz, DMSO) δ = 11.10-11.12 (s, 1H), δ = 8.41-8.43 (d, 2H), δ = 7.87-7.89 (dd, 1H), δ = 7.61-7.66 (dd, 1H), δ = 6.39-6.42 (s, 2H), δ = 6.15-6.18 (s, 2H), δ = 4.13-4.15 (s, 2H), δ = 3.32-3.34 (s, 2H), δ = 3.01-3.09 (m, 6H), δ = 2.57-2.51 (q, 2H), δ = 1.03-0.97 (t, 3H);

[0269] The NMR chart of the compound of formula IV is shown in Figure 17, and the results show that the equivalent ratio of free base to maleic acid in the compound of formula (IV) is 1:1.

[0270] The XRPD chart of crystal form IV-A is shown in Figure 18, and the XRPD data is shown in the following table

[0271] Example 24

[0272] Inhibition effect of the compound on PARP1 / PARP2 enzyme activity

[0273] PARP1 / PARP2 enzyme activity was detected by chemiluminescence method. First, histone (Active Motif, 81126) was incubated in a 384-well plate for 2 hours, and different dilutions of numbered compounds and PARP1 working solution (Abeam, ab279663) or PARP2 working solution (BPS, 80502) were added. Max control wells only added PARP1 working solution or PARP2 working solution, and Min control wells only added assay buffer. Incubate at room temperature for 15 minutes, add biotin-labeled substrate NAD + (BPS, 80610), incubate at room temperature for 2 hours. After the substrate is catalyzed by the enzyme, the ADP ribosylation group still has a biotin label on the histone. Streptavidin-HRP solution (Abeam, ab7403) is added to develop the biotin, and the value is read on the EnSight (PE) instrument. The fluorescence values of Max and Min wells are used to calculate the inhibition rate, and the analysis software GraphPad Prism 5 is used to fit the dose-effect curve to obtain the IC 50 value of each compound on enzyme activity.

[0274] The results are shown in Table 1 below. The compound of formula (I) has a dose-dependent inhibitory effect on PARP1 enzyme, and the inhibitory activity is significant, reaching the picomolar concentration level. Compared with the reference compounds AZD5305 and Olaparib, the activity is similar. In terms of PARP2 selectivity, the compound of formula (I) has similar selectivity to the reference AZD5305. In terms of PARP2 selectivity, the compound of formula (I) has similar selectivity to the positive control. Compared with the currently marketed PARPi Olaparib, the compound of formula (I) significantly improves the selectivity for PARP2 while maintaining the inhibition of PARP1 enzyme activity, with a 30-fold increase in selectivity.

[0275] Table 1 Inhibition of PARP1 / PARP2 enzyme activity by compounds

[0276] Example 25

[0277] Compound-induced DNA trapping ability of PARP1 / PARP2

[0278] DNA trapping ability of PARP1 / PARP2 was detected by HTRF (homogeneous time-resolved fluorescence). First, PARP1 (BPS, 80501) or PARP2 (BPS, 80502) was labeled by Mab anti GST-Tb crypate (cisbio, 61GSTTLA) and DNA damage labeling probe (Generay) was added with different concentrations of numbered compounds, 50 μM Olaparib was added to Max control wells and medium buffer was added to Min wells. Incubate at room temperature for 1 hour, add substrate NAD + (Sigma, 10127965001) for 10 minutes, PARP enzyme activity causes PARP to be released from the damaged DNA, at this time only the fluorescence signal with an emission wavelength of 615 nm can be detected, when PARP is inhibited, PARP is induced to bind to damaged DNA, causing energy transfer, two emission wavelengths can be detected, one is the fluorescence signal with an emission wavelength of 615 nm from the damaged DNA probe itself, and the other is the fluorescence signal with an emission wavelength of 665 nm from the energy transfer after PARP binds to damaged DNA, and the fluorescence ratio of 665 nm to 615 nm represents the amount of PARP DNA trapping complex. The fluorescence values of Max and Min wells were used to calculate the trapping ability induced by each compound, and the dose-effect curve was fitted by using analysis software GraphPad Prism 5, so as to obtain the EC 50 value (the required concentration to achieve 50% trapping) of each compound to induce PARP enzyme DNA trapping ability.

[0279] The results are shown in Table 2 below, the DNA trapping ability of the compound of formula (I) induced by PARP reaches the nanomolar concentration of the first digit, and the trapping ability is significant, which is similar to the activity of the reference compound AZD5305 and better than Olaparib. In terms of selectivity to PARP2, the selectivity of the compound of formula (I) reaches 100 times, which is better than the reference AZD5305. Compared with the currently marketed PARPi Olaparib, the compound of formula (I) significantly improves the activity on PARP1 and the selectivity to PARP2, the activity is improved by 3 times, and the selectivity is improved by 333 times.

[0280] Table 2 DNA trapping ability of compounds induced by PARP

[0281] Example 26

[0282] Inhibition effect of compounds on PARP1 / PARP2 enzyme activity at the cellular level

[0283] The high content imaging method was used to detect the inhibitory effect of the compound on PARP1 / PARP2 enzyme activity at the cell level. A549 WT, PARP1-KO and PARP2-KO cell lines were constructed, and the cells were recovered. After the cell state was stable, a sufficient number of cells were collected, and 100 μL of cell suspension was seeded in a 96-well plate. The next day, an equal ratio dilution series of the numbered compound was added. The Max control well only added buffer, and the Min control well added 500 nM AZD5305 (A549 WT and PARP2-KO cell lines) or 1 μM Olaparib (PARP1-KO cell line). Incubation was performed for 1.5 hours (A549 WT and PARP2-KO cell lines) or 2 hours (PARP1-KO cell line). The supernatant was removed, and 0.4 mM H2O2 was added for incubation for 10 minutes (A549 WT and PARP2-KO cell lines) or 1.5 mM H2O2 for incubation for 15 minutes (PARP1-KO cell line) to cause massive damage to the cell DNA. The supernatant was removed, and 4% paraformaldehyde was used for fixation for 20 minutes. Then, 0.5% Triton X-100 was used for treatment for 20 minutes to increase the permeability of the cell membrane. After 3% BSA was incubated for 1 hour to avoid non-specific binding, 1:500 diluted primary antibody Poly(ADP-ribose) monoclonal antibody (CST, 83732S) was added, and after overnight incubation, 1:500 diluted secondary antibody Goat anti-Rabbit IgG, Alexa Fluor 488 (invitrogen, A-11034) was incubated for 1 hour. 50 μL of DAPI (invitrogen, R37606) was added to stain the cell nucleus for 30 minutes. After 2 times of PBS washing, the photograph was taken on the OPERETTA CLS TM (PE) in non-confocal mode under a 20x water objective. In the analysis of data, the DAPI-stained nucleus group was selected to distinguish the nucleus and cytoplasm, and the average intensity value of Alexa 488 in the nucleus of each well was calculated. The fluorescence values of the Max and Min wells were used to calculate the inhibition rate, and the analysis software GraphPad Prism 5 was used to fit the dose-effect curve to obtain the IC TM value of each compound on enzyme activity. 50 value of each compound on enzyme activity.

[0284] The results are shown in Table 3. Compound (I) showed dose-dependent inhibition of enzyme activity in both A549 WT and PARP2-KO cell lines, with significant inhibitory activity reaching single-digit nanomolar concentrations, indicating that the cellular enzyme activity mainly originates from PARP1. Compound (I) showed similar inhibitory activity against PARP1 compared to the reference AZD5305. Compound (I) exhibited relatively weak inhibitory activity in the A549 PARP1-KO cell line; no inhibition was detected at the highest concentration of 40 μM, indicating that after PARP1 knockout, compound (I) did not significantly inhibit PARylation at the cellular level, and its inhibitory effect on enzymes other than PARP1 was not significant. Compared to the reference AZD5305, compound (I) showed a selectivity fold increase of more than 6-fold, demonstrating better selectivity.

[0285] Table 3 shows the inhibitory effects of compounds on PARP1 / PARP2 enzyme-mediated parylation activity at the cellular level.

[0286] Example 27

[0287] Test of the antiproliferative activity of compounds against tumor cells

[0288] The anti-proliferative assay of the compounds against tumor cells was performed using the most widely used ATP concentration detection method. Both MDA-MB-231 and MDA-MB-436 cell lines were derived from ATCC, DLD-1 human colorectal adenocarcinoma epithelial cells, and DLD-1 BRCA2- / - Cells were obtained from Horizon. Cells were resuscitated and, once stabilized, collected with a viable cell count greater than 90%. 450-500 cells were seeded into 384-well plates. The next day, serially diluted, numbered compounds were added. Max wells received only buffer, and Min wells received 50 μM Laparib. Incubation was performed for 7 days. On day 8, [the following was added]... Reagent (Promega, G7573) was incubated at room temperature for 30 minutes, and values ​​were read on Envision (PE). Inhibition rates were calculated using the fluorescence values ​​of the Max and Min wells. Dose-response curves were fitted using GraphPad Prism 5 software to determine the IC50 of each compound on enzyme activity. 50 value.

[0289] The results are shown in Table 4 below. In the BRCA1 mutant MDA-MB-436 human triple negative breast cancer cell line and the BRCA1 normal MDA-MB-231 human triple negative breast cancer cell line, the compound of formula (I) showed dose-dependent anti-proliferative activity on the BRCA1 mutant MDA-MB-436, and the inhibitory activity on the proliferation of the cancer cells reached the single-digit nanomolar concentration, similar to the reference AZD5305. For the BRCA1 normal MDA-MB-231, the compound of formula (I) did not detect any activity of inhibiting cell proliferation at the highest concentration of 10 μM dose, consistent with the reference AZD5305, indicating that the compound of formula (I) is obviously selective on the BRCA1 mutant cell line and the BRCA1 normal cell, and the selectivity factor reaches 5,000 times. Compared with the marketed PARPi Olaparib, the anti-proliferative activity of the compound of formula (I) on the BRCA1 mutant MDA-MB-436 is increased by 154 times, and the selectivity factor of PARP2 is increased by 470 times, significantly improving the inhibitory activity of PARP1 and the selectivity of PARP2.

[0290] As shown in Table 5, in the BRCA2 mutant DLD-1 BRCA2 - / - and the BRCA2 normal DLD-1 human colorectal adenocarcinoma epithelial cell line, the compound of formula (I) showed dose-dependent anti-proliferative activity on the BRCA2 mutant cell line, and the inhibitory activity of the compound of formula (I) on the proliferation of the cancer cells reached the single-digit nanomolar concentration, similar to the reference AZD5305. For the BRCA2 normal DLD-1 cell line, the compound of formula (I) did not detect any activity of inhibiting cell proliferation at the highest concentration of 10 μM dose, consistent with the reference AZD5305, indicating that the compound of formula (I) is obviously selective on the BRCA2 mutant cell line and the BRCA2 normal cell, and the selectivity factor reaches several thousand times.

[0291] According to the anti-proliferative activity of the compound of formula (I) on the BRCA1 mutant and normal, BRCA2 mutant and normal cancer cell lines, it is shown that the compound of formula (I) has strong target cell anti-proliferative activity, and is used for clinical treatment with targeting and safety.

[0292] Table 4 Anti-proliferative activity of the compound on BRCA1 mutant and BRCA1 normal cancer cells

[0293] Table 5 Anti-proliferative activity of the compound on BRCA2 mutant and BRCA2 normal cancer cells

[0294] Comprehensive molecular level and cellular level enzymatic activity, induction of PARP capture ability of damaged DNA and anti-proliferative activity of cancer cells, the compound of formula (I) has excellent PARP1 selective inhibition activity and anti-proliferative ability of target cells, which is significantly better than the marketed PARPi Olaparib in terms of PARP1 activity and selectivity to PARP2, and compared with the reference compound AZD5305, the selectivity to PARP1 at the cellular level is significantly improved.

[0295] Selectivity of the compound of example 28 to PARP family PARP3, 5a, 6, 7, 11

[0296] The enzymatic activity of the PARP family was detected by chemiluminescence method. First, the histone (Active Motif, 81126) was incubated in a 384-well plate for 2 hours,

[0297] Different dilutions of the numbered compound and PARP3 working solution (BPS, 80503) were added, the Max control wells only added each PARP working solution, the Min control wells only added assay buffer, incubated at room temperature for 15 minutes, and then the biotin-labeled substrate NAD + (BPS, 80610) was added, and the PARP3 activating DNA (Generay) was incubated at room temperature for 2 hours. After the substrate was catalyzed by the enzyme, the ADP ribosylation group was combined to the histone which still had a biotin label. Streptavidin-HRP solution (Abeam, ab7403) was added to develop the biotin, and the value was read on the EnSight (PE) instrument.

[0298] Different dilutions of the numbered compound and PARP5a working solution (BPS, 80504) were added, the Max control wells only added each PARP working solution, the Min control wells only added assay buffer, incubated at room temperature for 15 minutes, and then the biotin-labeled substrate NAD + (BPS, 80610) was added, and the PARP3 activating DNA (Generay) was incubated at room temperature for 2 hours. After the substrate was catalyzed by the enzyme, the ADP ribosylation group was combined to the histone which still had a biotin label. Streptavidin-HRP solution (Abeam, ab7403) was added to develop the biotin, and the value was read on the EnSight (PE) instrument.

[0299] Different dilutions of the numbered compound and PARP6 working solution (BPS, 80506) were added, the Max control wells only added each PARP working solution, the Min control wells only added assay buffer, incubated at room temperature for 15 minutes, and then the biotin-labeled substrate NAD +(BPS, 80610), incubate at room temperature for 2 hours, the ADP-ribosylation group binds to the histone after the substrate is catalyzed by the enzyme, and the biotin label is still present, add Streptavidin-HRP solution (Abeam, ab7403) to develop the biotin, and read the value on the EnSight (PE) instrument.

[0300] Use PARP7 Chemiluminescent assay kit (BPS, 79729), add different dilutions of the numbered compound and PARP7 working solution (BPS, 80527), add only the respective PARP working solution to the Max control well, and add only the assay buffer to the Min control well, incubate at room temperature for 15 minutes, add the substrate mixture provided by the assay kit (BPS, 78371), incubate at room temperature for 1 hour, the ADP-ribosylation group binds to the histone after the substrate is catalyzed by the enzyme, and the biotin label is still present, add treptavidin-HRP solution (BPS, 80611) to develop the biotin, and read the value on the EnSight (PE) instrument.

[0301] Use PARP11 Chemiluminescent assay kit (BPS, 80561), add different dilutions of the numbered compound and PARP11 working solution (BPS, 80511), add only the respective PARP working solution to the Max control well, and add only the assay buffer to the Min control well, incubate at room temperature for 15 minutes, add the substrate mixture provided by the assay kit (BPS, 78371), incubate at room temperature for 1 hour, the ADP-ribosylation group binds to the histone after the substrate is catalyzed by the enzyme, and the biotin label is still present, add treptavidin-HRP solution (BPS, 80611) to develop the biotin, and read the value on the EnSight (PE) instrument.

[0302] The inhibition rate is calculated using the fluorescence values of the Max and Min wells, and the analysis software GraphPad Prism 5 is used to fit the dose-effect curve, so as to obtain the IC 50 value of each compound on enzyme activity.

[0303] The results are shown in Table 7 below, and the inhibitory effect of the compound of formula (I) on each PARP enzyme is weaker than the activity on PARP1 enzyme; the values in the brackets in Table 7 represent the ratio relative to the activity of PARP1 enzyme, and the selectivity of the compound of formula (I) on other enzymes is significantly improved compared with the marketed PARPi Olaparib, except that the selectivity on PARP11 is slightly weaker; the selectivity of the compound of formula (I) on the tested PARP enzymes is significantly improved compared with the reference AZD5305.

[0304] It is demonstrated that the compound of formula (I) is a PARP1 specific and selective inhibitor, with a selectivity over PARP3, PARP5a, PARP6, PARP7 and PARP11 superior to AZD5305, especially with a selectivity advantage over PARP11 particularly remarkable; with a selectivity over PARP3, PARP5a, PARP6, PARP7 superior to Olaparib, especially with a selectivity advantage over PARP3 particularly remarkable.

[0305] Table 7 Compound inhibition of PARP3 / PARP5a / PARP6 / PARP7 / PARP11 enzyme activity (nM)

[0306] Example 29 Compound of formula (I) induces PARP1 / 2 trapping of DNA in FP assay

[0307] To further explore the effect of the compound of formula (I) on PARP1-DNA and PARP2-DNA trapping, and the difference between the two, the method of fluorescence polarization (FP) was further evaluated. PARP1 / 2 enzymes bind to fluorescently labeled DNA to form a larger complex, resulting in slower rotation of the fluorescently labeled DNA, thus emitting higher polarization light values. After the addition of NAD + , PARP1 / 2 enzymes undergo autorigosylation and accumulate negative charges, and when the negative charges accumulate to a certain extent, the fluorescently labeled DNA is dissociated, the fluorescently labeled DNA rotates faster, and the polarization light value decreases. The addition of PARP inhibitors affects PARP-DNA trapping, and the degree of its effect can be detected by changes in polarization light intensity. First, 25nL of 1000-fold final concentration of different concentrations of compounds was transferred to a 384-well plate, 25nL of 100% DMSO was added to Min and Max wells, 5μL of enzyme solution containing 10nM PARP1 (BPS, 80501) and 1nM FAM-PARP1-DNA (Generay, customized) or 10nM PARP2 (BPS, 80502) and 1nM FAM-PARP2-DNA (Generay, customized) was added, 1000rpm centrifugation for 1 minute, then incubated at room temperature for 30 minutes, 5μL of 1mM substrate NAD + (MCE, HY-B0445) was added to the Min well, 5μL of 1mM substrate NAD + was added to the Max well, 1000rpm centrifugation for 1 minute, and the reaction was carried out at 25°C. The following different time points were tested:

[0308] For PARP1 capture assay, the plate reader (Envision) was read at 0.25h, 0.5h, 1h, 2h, 4h, 8h, 24h, 30h, 48h and 54h.

[0309] For PARP2 capture assay, the plate reader was read at 0.25h, 0.5h, 1h, 2h, 4h, 8h, 24h and 48h.

[0310] The compound-induced capture ability was calculated using the fluorescence values of Max and Min wells, and the dose-effect curve was fitted using the analysis software GraphPad Prism 5, so as to obtain the EC 50 values of the PARP enzyme DNA capture ability of each compound.

[0311] As shown in Table 8, taking the 2h time point as an example, the ability of the compound of formula (I) to induce PARP1-DNA capture was 48 times more active than the marketed PARPi Olaparib, and was similar to the reference AZD5305. The compound of formula (I) had almost no capture ability for PARP2-DNA, which was similar to the reference AZD5305; and the marketed PARPi Olaparib had similar capture for PARP2-DNA and PARP1-DNA, and had no selectivity.

[0312] It is shown that the compound of formula (I) has a PARP1 selectivity much higher than PARP2. It is expected to significantly reduce the blood toxicity caused by PARP2.

[0313] As shown in Table 8, further comparison of the ability of the compound of formula (I) and the reference AZD5305 to capture PARP1-DNA showed that the activity of the compound of formula (I) could be maintained for more than 54 hours, and the activity was 14 times stronger than the reference AZD5305 at 54 hours. It is suggested that there will be better anti-tumor proliferation activity.

[0314] Table 8 Effect of compounds on PARP1 / 2-DNA capture

[0315] Example 30 Anti-tumor effect of the compound on a human breast cancer cell MDA-MB-436 xenograft mouse model

[0316] The in vivo validation of the compound of formula (I) was further performed by using BRCA1 mutant MDA-MB-436 cell line mouse subcutaneous xenograft model. Human breast cancer MDA-MB-436 cells (ATCC, HTB-130) were cultured in monolayer in vitro, and the culture condition was L-15 medium with 10% fetal bovine serum, 1% penicillin-streptomycin solution, 0.01 mg / mL bovine insulin, and 37°C CO2-free incubator. The cells were routinely digested and passaged by trypsin-EDTA twice a week. When the cell saturation degree was 80%-90%, the cells were collected, counted, and 0.2 mL 1 x 10 7 MDA-MB-436 cells were subcutaneously inoculated in the right back of each mouse (PBS: Matrigel = 1:1). When the average tumor volume reached 152 mm 3 , the mice were grouped and administered, and the day was set as Day 0. The tumor volume and mouse weight were monitored twice a week, and the administration was performed for 28 days.

[0317] The tumor diameter was measured by a vernier caliper. The tumor volume was calculated by the formula: V = 0.5a x b 2 , where a and b represent the long diameter and short diameter of the tumor, respectively. The antitumor effect of the compound was evaluated by the relative tumor shrinkage rate Reg%. Reg% reflects the tumor volume shrinkage rate of the tumor itself after treatment. Reg% = (V0-V t ) / V0 x 100%, where V0 is the tumor volume measured at the time of grouping and administration (i.e., d0), and V t is the tumor volume at a certain time.

[0318] The compound response criteria in the mouse model were revised from mRECIST (modified response evaluation criteria in solid tumors) (Gao et al, 2015), and the specific definitions are as follows:

[0319] CR (complete remission): BestResponse <-95% and BestAvgResponse <-40%;

[0320] PR (partial remission): BestResponse <-50% and BestAvgResponse <-20%;

[0321] SD (stable disease): BestResponse <35% and BestAvgResponse <30%;

[0322] PD (progressive disease): other classification;

[0323] ORR% is equal to the sum of the proportions of complete remission (CR) and partial remission (PR).

[0324] Statistical analysis was performed using Prism software, including the mean and standard error (SEM) of tumor volume at each time point for each group. For statistical analysis of TV, the original data of TV at each measurement were used to compare the differences between groups, and two-way ANOVA was used to analyze the double factors of administration and time. Tukey's multiple comparisons test was used for inspection, and p < 0.05 was considered to be a statistically significant difference.

[0325] The results are shown in Figure 19. At the same dose, both the compound of formula (I) and the reference AZD5305 promoted tumor regression, and the compound of formula (I) promoted tumor regression better than the reference AZD5305, with an ORR of 60%, while the reference ORR was 20%.

[0326] It is shown that in the human breast cancer MDA-MB-436 mouse subcutaneous xenograft model, the anti-tumor growth effect of the compound of formula (I) is better than that of the reference AZD5305.

[0327] Apoptosis-promoting effect of the compound of Example 31 on BRCA2-deficient cell lines

[0328] Mechanism analysis of the anti-proliferation effect of the compound on BRCA mutant tumors, using flow cytometry to analyze the apoptosis effect of the compound on BRCA2 mutant cell lines. Human colon cancer DLD-1 cells (ATCC, HTB-130) with BRCA2 deletion were cultured in RPMI1640 medium with 1% fetal bovine serum and 100 μg / mL hygromycin B. When the cells grew to the logarithmic phase, the cells were collected and placed in a 6-well plate at 6 million cells per well. The cells were incubated in a 37°C / 5% CO2 incubator overnight, and different concentrations of the compound were added to each well. After incubation in the incubator for 7 days, the cells were collected and placed in a 96-well plate, and 195 μL of Annexin V-FITC binding buffer was added to each well to resuspend the cells. Then 5 μL of Annexin V-FITC (beyotime, C1052) was added, mixed gently, and incubated at room temperature for 30 minutes. PBS was washed twice, centrifuged at 300g for 5 minutes, and 500 μL of PBS was added to each well to resuspend the cells. Then 5 μL of PI (beyotime, C1052) was added, mixed gently, and incubated at room temperature for 30 minutes. PBS was washed once, centrifuged at 300g for 5 minutes, and 500 μL of PBS was added to resuspend the cells. 300 μL of cells were taken to a flow tube for flow cytometry (BD bioscience, FACSVerse) analysis. FlowJo software was used to analyze the data collected by the flow cytometer.

[0329] Apoptosis analysis: Drag the apoptosis sample into FlowJo, double click the raw data to open the graph window. Select FSC-A for X axis, SSC-A for Y axis. Select the cell population shown in the figure to analyze the apoptosis cells. Cell debris is not analyzed. Double click the "gated cell population" in SSC / FSC graph, select FITC for X axis, which represents Annexin V-FITC. Select PerCP for Y axis, which represents PI. Use the four-gate tool to define the live cells and apoptotic cells. Annexin V positive is the apoptotic cells, Annexin V positive PI negative is the early apoptotic cells, Annexin V positive PI positive is the late apoptotic cells. Use PRISM to make the graph analysis, and use two-way ANOVA to analyze the different compounds and different concentrations, and use Tukey's multiple comparisons test to test the significance, p<0.05 is considered to be statistically significant.

[0330] The results are shown in Figures 20-22, which are the results of three repeated experiments. The apoptosis rates induced by the compound of formula (I) at different concentrations are all higher than the reference AZD5305, and some concentrations show statistically significant differences. It is suggested that the apoptosis-inducing effect of the compound of formula (I) on BRCA2-deficient human colon cancer DLD-1 cells is better than that of the reference AZD5305.

[0331] Apoptosis-inducing effect of the compound of Example 32 on BRCA1 mutant human breast cancer xenografts

[0332] To further verify the pro-apoptotic effect of the compounds in vivo, the changes in cleaved caspase-3 in MDA-MB-436 mouse subcutaneous transplanted tumor tissues were detected by immunoblotting. After 10 days of treatment with different compounds, the animals were euthanized at 0.25 hours and 24 hours after the last administration, and the tumor tissues were analyzed. The quick-frozen tumor tissues were placed on dry ice, 350 μL of complete cell lysis solution (containing 1% protease inhibitor and phosphatase inhibitor) was added, and the tissues were broken up using a Tissue grinder for 5 minutes. The tissue lysate was lysed on ice for 30 minutes. Centrifugation was performed at 12,000 rpm and 4°C for 10 minutes, the supernatant was taken and placed in a new 1.5 mL centrifuge tube, and the protein was quantified using a BCA quantitative kit. According to the quantitative results, the protein concentration of the samples was uniformly adjusted to 2 μg / μL, and LDS loading buffer (4X) and sample reducing agent (10X) were added. The sample was heated at 100°C for 10 minutes. Western blotting was carried out, 10 μL was loaded per well in the SDS-PAGE gel, 80 volts for 30 minutes, and then 120 volts for 90 minutes for electrophoresis. Membrane transfer was performed using an iBlot2 membrane transfer kit and a membrane transfer instrument for 7 minutes. The membrane was cut according to the molecular weight of the protein to be detected, washed with 1xTBST for 3 times, 5 minutes each time, and primary antibody Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb (CST, 9664), Caspase-3 Antibody (CST, 9662), and β-Actin Antibody (CST, 4967) were added for 4°C overnight incubation. The membrane was washed with 1xTBST for 3 times, 10 minutes each time, Goat anti-Rabbit IgG-HRP (Thermo fisher, 31462) was added for 1 hour of incubation at room temperature, the membrane was washed with 1xTBST for 3 times, 10 minutes each time, and HRP substrate in the West Femto ultra-sensitive chemiluminescence kit was added for chemiluminescence. The chemiluminescence was detected on a Tanon5200 Multi machine and photographed and saved. Quantitative analysis was performed using Alpha View software to relatively quantify the density intensity of the immunoblotting luminescence band. β-Actin is a housekeeping protein that detects the consistency of the amount of sample loaded in the immunoblotting detection. The density intensity of the cleaved caspase-3 band was standardized compared with the density intensity of the total caspase-3 band, and the relative density intensity of the cleaved caspase-3 in the vehicle control group was taken as 1 for conversion of the relative expression amount of cleaved caspase-3 in each treatment group.The values were plotted and analysed using Prism software, including the mean and standard error (SEM) of the relative expression of cleaved caspase-3 at each time point for each group, and comparisons between groups were made. Comparisons between multiple groups were analysed using one-way ANOVA, and if the variances were not equal (F value was significantly different), Games-Howell method was used for testing. If the F value was not significantly different, Tukey's multiple comparisons test method was used for analysis. A p<0.05 was considered to be a statistically significant difference.

[0333] The results, as shown in Figures 23 and 24-25, the compound of formula (I) promoted the increase of cleaved caspase-3 in MDA-MB-436 mouse subcutaneous transplanted tumor tissues, and the degree of increase was better than the reference AZD5305 at 0.25 hours and 24 hours after the last administration. It is suggested that in the BRCA1 mutant MDA-MB-436 in vivo tumor, the compound of formula (I) has a better effect of promoting tumor cell apoptosis than the reference AZD5305, which is consistent with its better tumor inhibition effect.

[0334] In summary, the compound of formula (I) of the present application is a PARP1 highly selective inhibitor, and its effect of inhibiting tumor growth in the BRCA1 MDA-MB-436 human breast cancer tumor model is better than AZD5305, and this advantage is further confirmed from the comparison of promoting PARP1-DNA capture and inducing apoptosis. Compared with the marketed PAPR inhibitors, the selectivity for PARP2 is significantly improved, and compared with the reference AZD5305, the selectivity for other PARP family is significantly improved. It is suggested that the compound of formula (I) of the present application is expected to become a safer and more effective PARP1 highly selective inhibitor, and can provide a higher quality treatment mode for clinical patients.

[0335] Although the specific embodiments of the present application are described above, those skilled in the art should understand that this is only an example, and the protection scope of the present application is defined by the appended claims. Those skilled in the art can make various changes or modifications to these embodiments without departing from the principles and essence of the present application, and these changes and modifications all fall within the protection scope of the present application.

Claims

1. A salt form of a compound of formula (I), characterized in that, It is a compound of formula (II), formula (III), or formula (IV); 2. A crystal form A of a compound of formula (I); Its features are, The crystal form A, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction peaks at the following positions, as indicated by 2θ: 9.738°±0.2°, 11.350°±0.2°, 12.504°±0.2°, 12.888°±0.2°, 14.438°±0.2°, 15.025°±0.2°, 15.369°±0.2°, and 16.837°±0.2°. Preferably, the X-ray powder diffraction pattern of crystal form A, expressed at an angle of 2θ, has diffraction peaks at the following positions: 9.738°±0.2°, 11.350°±0.2°, 12.504°±0.2°, 12.888°±0.2°, 14.438°±0.2°, 15.025°±0.2°, 15.369°±0.2°, 16.837°±0.2°, 17.813°±0.2°, and 18.285°±0.2°; More preferably, the X-ray powder diffraction pattern of crystal form A, expressed at an angle of 2θ, has diffraction peaks at the following positions: 9.738°±0.2°, 11.350°±0.2°, 12.504°±0.2°, 12.888°±0.2°, 14.438°±0.2°, 15.025°±0.2°, 15.369°±0.2°, 16.837°±0.2°, 17.813°±0.2°, 18.285°±0.2°, and 18.512°±0.2°. 0.2°, 18.734°±0.2°, 19.591°±0.2°, 19.840°±0.2°, 20.162°±0.2°, 20.513°±0.2°, 21.197°±0.2°, 22.152°±0.2°, 22.398°±0.2°, 22.923°±0.2°, 25.274°±0.2°, 25.853°±0.2°, 25.980°±0.2° and 27.316°±0.2°; Furthermore, the X-ray powder diffraction pattern of crystal form A, expressed at a 2θ angle, has the diffraction peaks shown in the table below: Furthermore, the X-ray powder diffraction pattern of crystal form A, expressed at a 2θ angle, has the diffraction peaks shown in the table below: Ideally, the X-ray powder diffraction pattern of the crystal form A, expressed at a 2θ angle, is essentially as shown in Figure 3.

3. A crystal form of a compound of formula (II), characterized in that, It is crystal form I, crystal form II, crystal form III, crystal form IV, crystal form V, crystal form VI, crystal form VII, crystal form VIII, crystal form IX or crystal form X; The crystal form I, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction peaks at the following positions, as indicated by 2θ: 8.487°±0.2°, 9.663°±0.2°, 11.259°±0.2°, 12.684°±0.2°, 13.627°±0.2°, and 13.994°±0.2°. The crystal form II, when subjected to Cu-Kα radiation, exhibits diffraction peaks at the following positions in its 2θ X-ray powder diffraction pattern: 8.262°±0.2°, 10.219°±0.2°, 11.445°±0.2°, 11.887°±0.2°, 12.352°±0.2°, 13.994°±0.2°, 14.536°±0.2°, 15.243°±0.2°, 15.457°±0.2°, 16.462°±0.2°, 17.115°±0.2°, 17.646°±0.2°, 17.831°±0.2°, and 19.041°±0.2°. The crystal form III, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction peaks at the following positions in 2θ: 8.197°±0.2°, 10.619°±0.2°, 11.200°±0.2°, 12.114°±0.2°, 13.285°±0.2°, 13.581°±0.2°, and 14.787°±0.2°. The crystal form IV, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction peaks at the following positions, as indicated by 2θ: 6.648°±0.2°, 7.725°±0.2°, 11.328°±0.2°, 12.101°±0.2°, 13.154°±0.2°, and 13.647°±0.2°. The crystal form V, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction peaks at the following positions in 2θ: 7.935°±0.2°, 9.092°±0.2°, 9.447°±0.2°, and 11.083°±0.2°. The crystal form VI, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction peaks at the following positions, as indicated by 2θ: 8.499°±0.2°, 9.674°±0.2°, 11.290°±0.2°, 13.982°±0.2°, 14.502°±0.2°, 14.910°±0.2°, 17.050°±0.2°, and 17.995°±0.2°. The crystal form VII, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction peaks at the following positions in 2θ: 7.720°±0.2°, 9.747°±0.2°, 11.885°±0.2°, 12.772°±0.2°, 13.392°±0.2°, and 13.606°±0.2°. The crystal form VIII, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction peaks at the following positions, as indicated by 2θ: 9.600°±0.2°, 10.615°±0.2°, 11.319°±0.2°, 12.717°±0.2°, 13.610°±0.2°, and 15.399°±0.2°. The crystal form IX, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction peaks at the following positions, as indicated by 2θ: 10.509°±0.2°, 11.158°±0.2°, 12.607°±0.2°, 13.154°±0.2°, 13.795°±0.2°, and 15.847°±0.2°. The crystal form X, when irradiated with Cu-Kα, exhibits X-ray powder diffraction patterns (denoted as 2θ) with diffraction peaks at the following positions: 7.657°±0.2°, 9.825°±0.2°, 11.917°±0.2°, 13.536°±0.2°, 15.367°±0.2°, and 17.339°±0.2°.

4. The crystal form of the compound of formula (II) as described in claim 3, characterized in that, It meets one or more of the following conditions: (1) The crystal form I, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction patterns (denoted as 2θ) with diffraction peaks at the following positions: 8.487°±0.2°, 9.663°±0.2°, 11.259°±0.2°, 12.684°±0.2°, 13.627°±0.2°, 13.994°±0.2°, 16.128°±0.2°, 17.037°±0.2°, 17.989°±0.2°, 19.376°±0.2°, and 20.642°±0.2°. (2) The crystal form II, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction peaks at the following positions in its 2θ X-ray powder diffraction pattern: 8.262°±0.2°, 10.219°±0.2°, 11.445°±0.2°, 11.887°±0.2°, 12.352°±0.2°, 13.994°±0.2°, 14.536°±0.2°, 15.243°±0.2°, 15.457°±0.2°, 16.462°±0.2°, 17.115°±0.2°, 17.646°±0.2°, 17.831°±0.2°, 19.041°±0.2°, 19.522°±0.2°, 19.8 18°±0.2°, 20.295°±0.2°, 20.698°±0.2°, 21.389°±0.2°, 21.821°±0.2°, 22.423°±0.2°, 22.800°±0.2°, 23.485°±0.2°, 23.825°±0.2°, 24.822°±0.2°, 25.332°±0.2°, 25.739°±0.2°, 26.369°±0.2°, 26.936°±0.2°, 27.351°±0.2°, 28.038°±0.2°, 28.394°±0.2°, 29.003°±0.2°, and 29.293°±0.2°; (3) The thermogravimetric analysis of the crystal form II showed a weight loss of 9.7 ± 0.2% when heated to 114 °C ± 2 °C; (4) The differential scanning calorimetry of crystal form II has an endothermic peak at 108℃±2℃; (5) The crystal form II is a hydrate of the compound of formula (II); (6) The crystal form III, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction peaks at the following positions in 2θ: 8.197°±0.2°, 10.619°±0.2°, 11.200°±0.2°, 12.114°±0.2°, 13.285°±0.2°, 13.581°±0.2°, 14.787°±0.2°, 15.730°±0.2°, and 16.943°±0.2°. (7) The crystal form IV, when subjected to Cu-Kα radiation, exhibits diffraction peaks at the following positions in its X-ray powder diffraction pattern expressed as 2θ at Cu-Kα radiation: 6.648°±0.2°, 7.725°±0.2°, 11.328°±0.2°, 12.101°±0.2°, 13.154°±0.2°, 13.647°±0.2°, 16.209°±0.2°, 19.662°±0.2°, and 19.988°±0.2°. (8) The crystal form V, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction patterns at the following positions, expressed as 2θ: 7.935°±0.2°, 9.092°±0.2°, 9.447°±0.2°, 11.083°±0.2°, 14.289°±0.2°, and 20.970°±0.2°. (9) The crystal form VI, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction peaks at the following positions in 2θ: 8.499°±0.2°, 9.674°±0.2°, 11.290°±0.2°, 13.982°±0.2°, 14.502°±0.2°, 14.910°±0.2°, 17.050°±0.2°, 17.995°±0.2°, 19.102°±0.2°, 20.565°±0.2°, and 20.733°±0.2°. (10) The crystal form VII, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction peaks at the following positions in 2θ: 7.720°±0.2°, 9.747°±0.2°, 11.885°±0.2°, 12.772°±0.2°, 13.392°±0.2°, 13.606°±0.2°, 15.221°±0.2°, 15.496°±0.2°, 17.334°±0.2°, 18.018°±0.2°, and 18.884°±0.2°; (11) The crystal form VIII, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction patterns (denoted as 2θ) with diffraction peaks at the following positions: 9.600°±0.2°, 10.615°±0.2°, 11.319°±0.2°, 12.717°±0.2°, 13.610°±0.2°, 15.399°±0.2°, 15.978°±0.2°, 18.161°±0.2°, 19.307°±0.2°, 20.431°±0.2°, 21.367°±0.2°, and 22.253°±0.2°; (12) The crystal form IX, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction patterns at the following positions, expressed as 2θ: 10.509°±0.2°, 11.158°±0.2°, 12.607°±0.2°, 13.154°±0.2°, 13.795°±0.2°, 15.847°±0.2°, 17.362°±0.2°, 19.819°±0.2°, and 21.062°±0.2°. The crystal form X described in (13) is subjected to Cu-Kα radiation, and the X-ray powder diffraction pattern, expressed in 2θ, has diffraction peaks at the following positions: 7.657°±0.2°, 9.825°±0.2°, 11.917°±0.2°, 13.536°±0.2°, 15.367°±0.2°, 17.339°±0.2°, 17.958°±0.2°, 18.896°±0.2°, 19.938°±0.2°, 20.930°±0.2°, and 22.257°±0.2°; Preferably, it satisfies one or more of the following conditions: (1) The crystal form I, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction patterns (denoted by 2θ) with diffraction peaks at the following positions: 8.487°±0.2°, 9.663°±0.2°, 11.259°±0.2°, 12.684°±0.2°, 13.627°±0.2°, 13.994°±0.2°, 16.128°±0.2°, 17.037°±0.2°, 17.989°±0.2°, 19.376°±0.2°, 20.642°±0.2°, 22.187°±0.2°, 22.749°±0.2°, 24.567°±0.2°, 26.171°±0.2°, and 27.445°±0.2°. (2) The crystal form II, when subjected to Cu-Kα radiation, exhibits diffraction peaks at the following positions in its X-ray powder diffraction pattern (denoted as 2θ): 8.262°±0.2°, 10.219°±0.2°, 11.445°±0.2°, 11.887°±0.2°, 12.352°±0.2°, 13.994°±0.2°, 14.536°±0.2°, 15.243°±0.2°, 15.457°±0.2°, 16.462°±0.2°, 17.115°±0.2°. 17.646°±0.2°, 17.831°±0.2°, 19.041°±0.2°, 19.522°±0.2°, 19.818°±0.2°, 20.295°±0.2°, 20.698°±0.2°, 21.389°±0.2°, 21.821°±0.2°, 22.423°±0.2°, 22.800°±0.2°, 23.485°±0.2°, 23.825°±0.2°, 24.822°±0.2°, 25.3 32°±0.2°, 25.739°±0.2°, 26.369°±0.2°, 26.936°±0.2°, 27.351°±0.2°, 28.038°±0.2°, 28.394°±0.2°, 29.003°±0.2°, 29.293°±0.2°, 29.865°±0.2°, 30.078°±0.2°, 30.612°±0.2°, 31.123°±0.2°, 31.440°±0.2°, 31.878°± 0.2°, 32.066°±0.2°, 32.574°±0.2°, 33.187°±0.2°, 33.488°±0.2°, 33.841°±0.2°, 34.489°±0.2°, 35.106°±0.2°, 35.671°±0.2°, 36.108°±0.2°, 37.180°±0.2°, 37.660°±0.2°, 38.180°±0.2°, 38.573°±0.2° and 39.223°±0.2°; (3) In one scheme, the differential scanning calorimetry of the crystal form II has an endothermic peak at 108℃±2℃ and a heat of fusion of 210J / g. (4) The crystal form II is a trihydrate of the compound of formula (II); (5) The crystal form III, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction peaks at the following positions in 2θ: 8.197°±0.2°, 10.619°±0.2°, 11.200°±0.2°, 12.114°±0.2°, 13.285°±0.2°, 13.581°±0.2°, 14.787°±0.2°, 15.730°±0.2°, 16.943°±0.2°, and 17.815°±0.2°. 2°, 20.459°±0.2°, 21.063°±0.2°, 21.371°±0.2°, 21.861°±0.2°, 22.567°±0.2°, 23.589°±0.2°, 24.865°±0.2°, 25.248°±0.2°, 26.057°±0.2°, 27.127°±0.2°, 29.226°±0.2°, 30.399°±0.2°, and 31.800°±0.2°; (6) The crystal form IV, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction peaks at the following positions in 2θ: 6.648°±0.2°, 7.725°±0.2°, 11.328°±0.2°, 12.101°±0.2°, 13.154°±0.2°, 13.647°±0.2°, 16.209°±0.2°, 19.662°±0.2°, 19.988°±0.2°, 22.250°±0.2°, 22.948°±0.2°, and 24.453°±0.2°. (7) Crystal form VI, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction peaks at the following positions, expressed as 2θ: 8.499°±0.2°, 9.674°±0.2°, 11.290°±0.2°, 13.982°±0.2°, 14.502°±0.2°, 14.910°±0.2°, 17.050°±0.2°, 17.995°±0.2°. °±0.2°, 19.102°±0.2°, 20.565°±0.2°, 20.733°±0.2°, 21.158°±0.2°, 22.238°±0.2°, 22.729°±0.2°, 23.370°±0.2°, 24.585°±0.2°, 25.396°±0.2° and 28.831°±0.2°; (8) The crystal form VII, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction peaks at the following positions in 2θ: 7.720°±0.2°, 9.747°±0.2°, 11.885°±0.2°, 12.772°±0.2°, 13.392°±0.2°, 13.606°±0.2°, 15.221°±0.2°, 15.496°±0.2°, 17.334°±0.2°, 18.018°±0.2°, 18.8°±0.2°. 84°±0.2°, 19.866°±0.2°, 20.240°±0.2°, 20.825°±0.2°, 22.174°±0.2°, 22.672°±0.2°, 23.589°±0.2°, 24.748°±0.2°, 25.118°±0.2°, 26.232°±0.2°, 27.328°±0.2°, 28.580°±0.2°, 29.530°±0.2° and 30.857°±0.2°; (9) The crystal form VIII, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction peaks at the following positions in 2θ: 9.600°±0.2°, 10.615°±0.2°, 11.319°±0.2°, 12.717°±0.2°, 13.610°±0.2°, 15.399°±0.2°, 15.978°±0.2°, 18.161°±0.2°, and 19.307°±0.2°. 2°, 20.431°±0.2°, 21.367°±0.2°, 22.253°±0.2°, 22.793°±0.2°, 23.452°±0.2°, 26.059°±0.2°, 26.801°±0.2°, 27.446°±0.2°, 28.235°±0.2°, 29.126°±0.2°, 31.110°±0.2°, and 32.286°±0.2°; (10) The crystal form IX, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction patterns (denoted as 2θ) with diffraction peaks at the following positions: 10.509°±0.2°, 11.158°±0.2°, 12.607°±0.2°, 13.154°±0.2°, 13.795°±0.2°, 15.847°±0.2°, 17.362°±0.2°, 19.819°±0.2°, 21.062°±0.2°, 22.516°±0.2°, 24.987°±0.2°, 25.957°±0.2°, and 26.509°±0.2°; The crystal form X described in (11) is subjected to Cu-Kα radiation, and the X-ray powder diffraction pattern, expressed in 2θ, has diffraction peaks at the following positions: 7.657°±0.2°, 9.825°±0.2°, 11.917°±0.2°, 13.536°±0.2°, 15.367°±0.2°, 17.339°±0.2°, 17.958°±0.2°, 18.896°±0.2°, 19.938°±0.2°, 20.930°±0.2°, 22.257°±0.2°, 22.813°±0.2°, 23.598°±0.2°, 24.087°±0.2°, 24.974°±0.2°, and 27.090°±0.2°.

5. The crystal form of the compound of formula (II) as shown in claim 4, characterized in that, It meets one or more of the following conditions: (1) The X-ray powder diffraction pattern of crystal form I, expressed in terms of 2θ angle, has the diffraction peaks shown in the table below: (2) The X-ray powder diffraction pattern of crystal form II, expressed in terms of 2θ angle, has the diffraction peaks shown in the table below: (3) The X-ray powder diffraction pattern of crystal form III, expressed at an angle of 2θ, has the diffraction peaks shown in the table below: (4) The X-ray powder diffraction pattern of crystal form IV, expressed at an angle of 2θ, has the diffraction peaks shown in the table below: (5) The X-ray powder diffraction pattern of the crystal form V, expressed in terms of 2θ angle, has the diffraction peaks shown in the table below: (6) The X-ray powder diffraction pattern of crystal form VI, expressed in terms of 2θ angle, has the diffraction peaks shown in the table below: (7) The X-ray powder diffraction pattern of crystal form VII, expressed in terms of 2θ angle, has the diffraction peaks shown in the table below: (8) The X-ray powder diffraction pattern of crystal form VIII, expressed at an angle of 2θ, has the diffraction peaks shown in the table below: (9) The X-ray powder diffraction pattern of the crystal form IX, expressed in terms of 2θ angle, has the diffraction peaks shown in the table below: The X-ray powder diffraction pattern of the crystal form X described in (10) in terms of the 2θ angle has the diffraction peaks shown in the table below: Preferably, the crystal form of the compound of formula (II) satisfies one or more of the following conditions: (1) The X-ray powder diffraction pattern of crystal form I, expressed in terms of 2θ angle, has the diffraction peaks shown in the table below: (2) The X-ray powder diffraction pattern of crystal form II, expressed in terms of 2θ angle, has the diffraction peaks shown in the table below: (3) The X-ray powder diffraction pattern of crystal form III, expressed at an angle of 2θ, has the diffraction peaks shown in the table below: (4) The X-ray powder diffraction pattern of crystal form IV, expressed at an angle of 2θ, has the diffraction peaks shown in the table below: (5) The X-ray powder diffraction pattern of the crystal form V, expressed in terms of 2θ angle, has the diffraction peaks shown in the table below: (6) The X-ray powder diffraction pattern of crystal form VI, expressed in terms of 2θ angle, has the diffraction peaks shown in the table below: (7) The X-ray powder diffraction pattern of crystal form VII, expressed in terms of 2θ angle, has the diffraction peaks shown in the table below: (8) The X-ray powder diffraction pattern of crystal form VIII, expressed at an angle of 2θ, has the diffraction peaks shown in the table below: (9) The X-ray powder diffraction pattern of the crystal form IX, expressed in terms of 2θ angle, has the diffraction peaks shown in the table below: The X-ray powder diffraction pattern of the crystal form X described in (10) in terms of the 2θ angle has the diffraction peaks shown in the table below:

6. The crystal form of the compound of formula (II) as shown in claim 5, characterized in that, It meets one or more of the following conditions: (1) The X-ray powder diffraction pattern of the crystal form I, expressed in terms of 2θ angle, is basically shown in Figure 4; (2) The X-ray powder diffraction pattern of the crystal form II, expressed in terms of 2θ angle, is basically shown in Figure 5; (3) The thermogravimetric analysis diagram of the crystal form II is basically shown in Figure 6; (4) The differential scanning calorimetry diagram of the crystal form II is basically as shown in Figure 6; (5) The X-ray powder diffraction pattern of the crystal form III, expressed in terms of 2θ angle, is basically shown in Figure 7; (6) The X-ray powder diffraction pattern of the crystal form IV, expressed in terms of 2θ angle, is basically shown in Figure 8; (7) The X-ray powder diffraction pattern of the crystal form V, expressed in terms of 2θ angle, is basically shown in Figure 9; (8) The X-ray powder diffraction pattern of the crystal form VI, expressed in terms of 2θ angle, is basically shown in Figure 10; (9) The X-ray powder diffraction pattern of the crystal form VII, expressed in terms of 2θ angle, is basically shown in Figure 11; (10) The X-ray powder diffraction pattern of the crystal form VIII, expressed in terms of 2θ angle, is basically shown in Figure 12; (11) The X-ray powder diffraction pattern of the crystal form IX, expressed in terms of 2θ angle, is basically shown in Figure 13; The X-ray powder diffraction pattern of the crystal form X described in (12) in terms of the 2θ angle is basically shown in Figure 14.

7. A crystal form III-A of a compound of formula (III); Its features are, The crystal form III-A, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction peaks at the following positions in 2θ: 5.356°±0.2°, 7.628°±0.2°, 10.731°±0.2°, 12.019°±0.2°, 12.591°±0.2°, 13.678°±0.2°, 13.912°±0.2°, and 14.237°±0.2°. Preferably, the crystal form III-A, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction peaks at the following positions in its 2θ X-ray powder diffraction pattern: 5.356°±0.2°, 7.628°±0.2°, 10.731°±0.2°, 12.019°±0.2°, 12.591°±0.2°, 13.678°±0.2°, 13.912°±0.2°, 14.237°±0.2°, 15.229°±0.2°, 16.126°±0.2°, 16.938°±0.2°, 17.171°±0.2°, and 17.531°±0.2°. 0.2°, 17.837°±0.2°, 18.223°±0.2°, 18.459°±0.2°, 19.965°±0.2°, 20.997°±0.2°, 21.497°±0.2°, 21.892°±0.2°, 22.237°±0.2°, 22.597°±0.2°, 22.847°±0.2°, 23.182°±0.2°, 23.488°±0.2°, 24.103°±0.2°, 24.883°±0.2°, 25.295°±0.2° and 25.585°±0.2°; More preferably, the crystal form III-A, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction peaks at the following positions in its 2θ X-ray powder diffraction pattern: 5.356°±0.2°, 7.628°±0.2°, 10.731°±0.2°, 12.019°±0.2°, 12.591°±0.2°, 13.678°±0.2°, 13.912°±0.2°, 14.237°±0.2°, and 15.2°. 29°±0.2°, 16.126°±0.2°, 16.938°±0.2°, 17.171°±0.2°, 17.531°±0.2°, 17.837°±0.2°, 18.223°±0.2°, 18.459°±0.2°, 19.965°±0.2°, 20.997°±0.2°, 21.497°±0.2°, 21.892°±0.2°, 22. 237°±0.2°, 22.597°±0.2°, 22.847°±0.2°, 23.182°±0.2°, 23.488°±0.2°, 24.103°±0.2°, 24.883°±0.2°, 25.295°±0.2°, 25.585°±0.2°, 26.091°±0.2°, 26.604°±0.2°, 26.959°±0.2°, 2 7.539°±0.2°, 27.989°±0.2°, 28.637°±0.2°, 29.137°±0.2°, 29.994°±0.2°, 30.348°±0.2°, 30.803°±0.2°, 31.256°±0.2°, 32.501°±0.2°, 33.073°±0.2°, 34.286°±0.2°, and 35.467°±0.2°; Preferably, the X-ray powder diffraction pattern of crystal form III-A, expressed at a 2θ angle, has the diffraction peaks shown in the table below: More preferably, the X-ray powder diffraction pattern of crystal form III-A, expressed at a 2θ angle, has the diffraction peaks shown in the table below: Ideally, the X-ray powder diffraction pattern of crystal type III-A expressed at a 2θ angle is basically shown in Figure 16.

8. A crystal form IV-A of a compound of formula (IV); Its features are, The crystal form IV-A, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction peaks at the following positions in 2θ: 5.937°±0.2°, 9.115°±0.2°, 10.105°±0.2°, 11.734°±0.2°, 11.855°±0.2°, 12.790°±0.2°, 13.841°±0.2°, and 14.955°±0.2°. Preferably, the crystal form IV-A, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction peaks at the following positions in 2θ: 5.937°±0.2°, 9.115°±0.2°, 10.105°±0.2°, 11.734°±0.2°, 11.855°±0.2°, 12.790°±0.2°, 13.841°±0.2°, and 14.955°±0.2°. 2°, 15.313°±0.2°, 15.989°±0.2°, 17.356°±0.2°, 17.796°±0.2°, 18.774°±0.2°, 19.307°±0.2°, 19.515°±0.2°, 20.015°±0.2°, 21.640°±0.2°, 21.942°±0.2° and 22.964°±0.2°; More preferably, the crystal form IV-A, when subjected to Cu-Kα radiation, exhibits X-ray powder diffraction peaks at the following positions in its 2θ X-ray powder diffraction pattern: 5.937°±0.2°, 9.115°±0.2°, 10.105°±0.2°, 11.734°±0.2°, 11.855°±0.2°, 12.790°±0.2°, and 13.84°±0.2°. 1°±0.2°, 14.955°±0.2°, 15.313°±0.2°, 15.989°±0.2°, 17.356°±0.2°, 17.796°±0.2°, 18.774°±0.2°, 19.307°±0.2°, 19.515°±0.2°, 20.015°±0.2°, 21. 640°±0.2°, 21.942°±0.2°, 22.964°±0.2°, 23.228°±0.2°, 23.781°±0.2°, 24.302°±0.2°, 25.697°±0.2°, 26.191°±0.2°, 26.872°±0.2°, 27.979°±0.2°, 2 8.632°±0.2°, 28.960°±0.2°, 29.843°±0.2°, 30.845°±0.2°, 31.716°±0.2°, 32.707°±0.2°, 33.815°±0.2°, 35.594°±0.2°, 36.992°±0.2° and 39.202°±0.2°; Preferably, the X-ray powder diffraction pattern of crystal form IV-A, expressed at a 2θ angle, has the diffraction peaks shown in the table below: More preferably, the X-ray powder diffraction pattern of crystal form IV-A, expressed at a 2θ angle, has the diffraction peaks shown in the table below: Ideally, the X-ray powder diffraction pattern of the crystal form IV-A, expressed at a 2θ angle, is essentially as shown in Figure 18.

9. A method for preparing crystal form A of a compound of formula (I), characterized in that, It includes the following steps: adding isopropyl acetate to a DMSO solution of compound (I) at 50-70°C (e.g., 60°C), and cooling to 15-35°C to crystallize. Preferably, the method for preparing crystal form A of the compound of formula (I) satisfies one or more of the following conditions: (1) The temperature drop to 15-35℃ is the temperature drop to 20-30℃; (2) The mass-to-volume ratio of the compound of formula (I) and the DMSO in the DMSO solution of the compound of formula (I) is 0.8-1.2 g / 10 ml, for example 1 g / 10 ml; The mass-to-volume ratio of the compound of formula (I) and (3) and the isopropyl acetate is 0.8-1.2 g / 15 ml, for example 1 g / 15 ml.

10. A method for preparing the crystal form of a compound of formula (II), characterized in that, It can be either method one or method two: Method 1 includes the following steps: the compound of formula (II) is slurried in a solvent and crystallized; When the solvent is n-heptane, the crystal form of the compound of formula (II) is crystal form I; When the solvent is acetone / water volume ratio = (8-10):1, the crystal form of the compound of formula (II) is crystal form II; When the solvent is methanol, the crystal form of the compound of formula (II) is crystal form III; When the solvent is 1,4-dioxane, the crystal form of the compound of formula (II) is crystal form V; When the solvent is isopropyl ester, the crystal form of the compound of formula (II) is crystal form VI; When the solvent is isopropyl acetate, the crystal form of the compound of formula (II) is crystal form VII; When the solvent is toluene, the crystal form of the compound of formula (II) is crystal form VIII; When the solvent is DMF / EA (volume ratio 1:1), the crystal form of the compound of formula (II) is crystal form IX; Preferably, the method for preparing the crystal form of the compound of formula (II) satisfies one or more of the following conditions: (1) The pulping temperature is 10-30℃, for example 20-30℃; (2) The stirring speed during pulping is 300-700 rpm, for example 500 rpm; (3) The mass-to-volume ratio of the compound of formula (II) and the solvent is (25-35) g / L; (4) The pulping time is 3-7 days, for example, 5 days; (5) The crystallization process is centrifugation. The preparation of the crystal form of compound (II) described in (6) further includes the following post-processing steps: after crystallization, collect the solid and evaporate the solvent under vacuum at 30-50°C; Method 2 includes the following steps: slurry the acetonitrile solution of compound (II) and crystallize it; When the pulping temperature is 10-30℃, the crystal form of the compound of formula (II) is crystal form IV; When the pulping temperature is 40-60℃, the crystal form of the compound of formula (II) is crystal form X; Preferably, the preparation method of the crystal form of the compound of formula (II) satisfies one or more of the following conditions: (1) The stirring speed during pulping is 300-700 rpm, for example 500 rpm; (2) The mass-to-volume ratio of the compound of formula (II) and the acetonitrile is (25-35) g / L; (3) The pulping time is 3-7 days, for example, 5 days; (4) The crystallization is performed by centrifugation. The preparation described in (5) further includes the following post-processing steps: after crystallization, collect the solid and evaporate the solvent under vacuum at 30-50°C.

11. A method for preparing crystal form III-A of a compound of formula (III), characterized in that, It includes the following steps: In DMSO, after the compound of formula (I) reacts with p-toluenesulfonic acid, isopropyl acetate is added at 50-80℃, and crystals are precipitated by cooling to 15-30℃. Preferably, the method for preparing the crystal form III-A of the compound of formula (III) satisfies one or more of the following conditions: (1) The temperature rise to 50-80℃ is the temperature rise to 60-70℃; (2) The reaction temperature of the compound of formula (I) with the p-toluenesulfonic acid is 15-40°C, for example 20-30°C; (3) The reaction time between the compound of formula (I) and the p-toluenesulfonic acid is 0.2-1 h, for example 0.5 h; (4) The molar ratio of the compound of formula (I) and the p-toluenesulfonic acid is 1:(1-1.5), for example 1:1.05; (5) The mass-to-volume ratio of the compound of formula (I) to the DMSO is 0.8-1.2 g / 20 ml, for example, 1 g / 20 ml; The mass-to-volume ratio of the compound of formula (I) and the isopropyl acetate is 0.8-1.2 g / 20 ml, for example, 1 g / 20 ml.

12. A method for preparing the crystal form IV-A of a compound of formula (IV), characterized in that, It includes the following steps: In DMSO, after the compound of formula (I) reacts with maleic acid, isopropyl acetate is added at 50-80℃, and crystals are precipitated by cooling to 15-30℃. Preferably, the method for preparing the crystal form IV-A of the compound of formula (IV) satisfies one or more of the following conditions: (1) The temperature rise to 50-80℃ is the temperature rise to 60-70℃; (2) The reaction temperature of the compound of formula (I) with the maleic acid is 15-40°C, for example 20-30°C; (3) The reaction time between the compound of formula (I) and the maleic acid is 0.2-1 h, for example 0.5 h; (4) The molar ratio of the compound of formula (I) and the maleic acid is 1:(1-1.5), for example 1:1.05; (5) The mass-to-volume ratio of the compound of formula (I) to the DMSO in the DMSO solution of the compound of formula (I) is 0.8-1.2 g / 20 ml, for example, 1 g / 20 ml; The mass-to-volume ratio of the compound of formula (I) and the isopropyl acetate is 0.8-1.2 g / 20 ml, for example, 1 g / 20 ml.

13. A pharmaceutical composition, characterized in that, It includes substance X and pharmaceutical excipients, wherein substance X is a salt form of the compound of formula (I) as claimed in claim 1, a crystal form of the compound of formula (I) as claimed in claim 2, a crystal form of the compound of formula (II) as claimed in any one of claims 3-6, a crystal form of the compound of formula (III) as claimed in claim 7, or a crystal form of the compound of formula (IV) as claimed in claim 8.

14. The use of the pharmaceutical composition of claim 13 or substance X of claim 13 in the preparation of a medicament for treating and / or preventing PARP-mediated diseases; Preferably, the application satisfies one or more of the following conditions: (1) The PARP is PARP1; (2) The PARP-mediated diseases are cancer, ischemic diseases and neurodegenerative diseases; The cancer may lack the HR-dependent DNA DSB repair pathway; The cancer may have a BRCA1 or BRCA2 defect phenotype; Preferably, the cancers are breast cancer, ovarian cancer, pancreatic cancer, prostate cancer, blood cancer, gastrointestinal cancer, and lung cancer.

15. The use of the pharmaceutical composition of claim 13 or the substance X of claim 13 as a PARP inhibitor, preferably, the PARP inhibitor is a PARP1 inhibitor.

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