CN116120323A - Solid form aza-condensed ring amide compound and use thereof - Google Patents

Abstract

The invention provides a compound shown in a solid form, a compound shown in a crystal form shown in the formula (A), a specific crystal form, a pharmaceutical composition containing the compound and application thereof, wherein the compound shown in the crystal form shown in the formula (A) and the specific crystal form have good crystallinity and stability, and the in-vitro and in-vivo efficacy shows that the compound shown in the formula (A) has good crystallinity and stability on wild type and mutant kinase and fine particle sizeThe cell and the in-vivo tumor have good inhibition effect and good potential of patent medicine.

Description

Solid aza-condensed ring amide compounds and uses thereof

This application claims the priority of the prior application filed by the applicant with the State Intellectual Property Office of China on November 15, 2021, with patent application number 202111345026.6 and invention name “Solid-form azo-fused cyclic amide compounds and their uses”. The full text of the prior application is incorporated into this application by reference.

Technical Field

The present application relates to the field of medicine, and specifically to aza-fused cyclic amide compounds in solid form, aza-fused cyclic amide compounds in crystalline form and their specific crystal forms, pharmaceutical compositions containing the same and their use in the preparation of drugs for preventing and/or treating diseases mediated by one or more of TRK, ROS1, and ALK.

Background Art

Tropomyosin-related kinase or tropomyosin receptor kinase (TRK) is a type of nerve growth factor receptor. Its family consists of three highly homologous subtypes, TRKA, TRKB and TRKC, which are encoded by neurotrophic receptor tyrosine kinase 1 (NTRK1), NTRK2 and NTRK3 genes, respectively. When the TRK receptor protein binds to the corresponding ligand, it can achieve different physiological functions by activating downstream signaling pathways, such as the RAS/MAPK pathway, the PLCγ pathway and the PI3K pathway. TRK family proteins are normally expressed in neural tissues, involved in the differentiation and survival of nerve cells, and the formation of axons and dendrites, and play an important role in embryonic development and the maintenance of normal function of the nervous system.

TRK kinase is activated in malignant tumors through a variety of mechanisms, mainly structural rearrangement and expression changes. For example, the gene encoding TRK kinase NTRK rearranges with other genes to produce a fusion oncogene, which causes TRK kinase to change in structure and expression. It is no longer regulated and controlled by nerve growth factor ligands, and is constitutively activated, promoting tumor development. In addition, gene sequencing results also show that TRK kinase is closely related to the occurrence, metastasis and deterioration of various tumors, and is expressed in a variety of tumors, such as non-small cell lung cancer, colorectal cancer, melanoma, gallbladder cancer, thyroid cancer, malignant glioma, etc.

Currently, the first-generation TRK inhibitors Larotrectinib (LOXO-101) and Entrectinib (RXDX-101) were approved for marketing by the U.S. Food and Drug Administration (FDA) in 2018 and 2019, respectively. Larotrectinib is a potent, oral, selective tropomyosin receptor kinase inhibitor. Its efficacy data were announced as early as the ASCO conference in June 2017. In Phase I and Phase II clinical trials, a total of 55 subjects were recruited, of which 46 evaluable patients had an overall response rate (ORR) of 78%. Entrectinib is a potent inhibitor of TRK, ROS1, and ALK proteins, and can pass through the blood-brain barrier. In Phase I clinical trials, the ORR of 24 evaluable patients was 79%.

Similar to other targeted drugs, TRK inhibitors also face the problem of drug resistance. Mutations in the NTRK kinase region can cause conformational changes in the TRK family protein kinase domain or changes in binding affinity with ATP, thereby affecting the binding of TRK inhibitors to targets. Mutation types include G595R, G639R, G667C, etc. In order to solve the drug resistance problem of the first-generation TRK inhibitors, second-generation TRK inhibitors such as LOXO-195 and TPX-005 are already under study.

Summary of the invention

In one aspect, the present application provides a compound represented by formula (A) in solid form.

In some embodiments of the present application, the compound of formula (A) in solid form can be measured by infrared spectroscopy using a tablet compression method. Its infrared spectrum includes characteristic peaks at the following positions (±4 cm ^-1 ): 3429, 1643, 1488, 1453, 1232, 1027.

In some embodiments of the present application, the compound represented by formula (A) in the solid form has an infrared spectrum substantially as shown in FIG. 12 .

In some embodiments of the present application, the compound represented by formula (A) in the solid form can be tested by X-ray powder diffraction.

In some embodiments of the present application, the compound represented by formula (A) in the solid form has a chemical purity of ≥95%; preferably, it has a chemical purity of ≥97%; further preferably, it has a chemical purity of ≥98%.

In some embodiments of the present application, the compound represented by formula (A) in the solid form is amorphous and has an infrared spectrum substantially as shown in FIG. 12 using Cu-Kα radiation.

In another aspect, the present application provides a crystalline compound of formula (A).

On the other hand, the present application provides a crystalline form I of the compound represented by formula (A), which has an X-ray powder diffraction pattern using Cu-Kα radiation and has characteristic diffraction peaks (±0.2°) at the following 2θ angles: 9.2, 17.9, 18.5.

In some schemes of the present application, the above-mentioned Form I, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 8.9, 9.2, 17.9, 18.5, 23.8.

In some schemes of the present application, the above-mentioned Form I, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 8.9, 9.2, 10.1, 17.9, 18.5, 23.8, 28.0.

In some schemes of the present application, the above-mentioned Form I, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 8.9, 9.2, 10.1, 16.1, 17.9, 18.5, 23.8, 27.0, 28.0.

In some schemes of the present application, the above-mentioned Form I, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 8.9, 9.2, 10.1, 16.1, 16.6, 17.9, 18.5, 23.8, 27.0, 28.0.

In some embodiments of the present application, the above-mentioned Form I, using Cu-Kα radiation, has an X-ray powder diffraction pattern substantially as shown in FIG. 1 .

On the other hand, the present application provides a crystalline form II of the compound represented by formula (A), which has an X-ray powder diffraction pattern using Cu-Kα radiation and has characteristic diffraction peaks (±0.2°) at the following 2θ angles: 9.1, 18.3, 20.6.

In some schemes of the present application, the above-mentioned Form II, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 6.4, 9.1, 18.3, 20.6.

In some schemes of the present application, the above-mentioned Form II, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 6.4, 9.1, 18.3, 20.6, 27.7.

In some schemes of the present application, the above-mentioned Form II, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 6.4, 9.1, 18.3, 18.9, 20.6, 27.7.

In some schemes of the present application, the above-mentioned Form II, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 6.4, 9.1, 18.3, 18.9, 20.6, 22.4, 23.9, 27.7.

In some embodiments of the present application, the above-mentioned Form II, using Cu-Kα radiation, has an X-ray powder diffraction pattern substantially as shown in FIG. 2 .

In some schemes of the present application, the above-mentioned Form II has an endothermic peak at 155.27±5°C in its differential scanning calorimetry curve.

In some schemes of the present application, the above-mentioned Form II has endothermic peaks at 58.39±5°C and 155.27±5°C in its differential scanning calorimetry curve.

In some embodiments of the present application, the above-mentioned Form II has a DSC spectrum substantially as shown in FIG. 3 .

In some schemes of the present application, the thermogravimetric analysis curve of the above-mentioned Form II shows a weight loss of 0.1910%±0.2% between room temperature and 75±5°C.

In some embodiments of the present application, the above-mentioned Form II has a TGA spectrum substantially as shown in FIG. 3 .

On the other hand, the present application provides Form III of Formula (A), which has an X-ray powder diffraction pattern using Cu-Kα radiation and has characteristic diffraction peaks (±0.2°) at the following 2θ angles: 5.6, 13.3, 15.8, 18.4.

In some schemes of the present application, the above-mentioned Form III, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 5.6, 13.3, 15.8, 18.4, 23.7.

In some schemes of the present application, the above-mentioned Form III, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 5.6, 10.9, 13.3, 15.8, 18.4, 19.0, 23.7.

In some schemes of the present application, the above-mentioned form III, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 5.6, 10.9, 11.3, 13.3, 15.8, 17.1, 18.4, 19.0, 23.7.

In some schemes of the present application, the above-mentioned form III, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 5.6, 10.9, 11.3, 13.3, 15.8, 17.1, 18.4, 19.0, 19.9, 23.7.

In some schemes of the present application, the above-mentioned form III, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 5.6, 7.4, 10.9, 11.3, 13.3, 15.8, 17.1, 18.4, 19.0, 19.9, 23.7.

In some embodiments of the present application, the above-mentioned Form III, using Cu-Kα radiation, has an X-ray powder diffraction pattern substantially as shown in FIG. 4 .

In some schemes of the present application, the above-mentioned Form III has an endothermic peak at 182.29±5°C in its differential scanning calorimetry curve.

In some embodiments of the present application, the above-mentioned Form III has a DSC spectrum substantially as shown in FIG. 5 .

On the other hand, the present application provides a crystalline form IV of the compound represented by formula (A), which has an X-ray powder diffraction pattern using Cu-Kα radiation and has characteristic diffraction peaks (±0.2°) at the following 2θ angles: 8.5, 10.0, 17.2, 17.9.

In some schemes of the present application, the above-mentioned Form IV, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 8.5, 10.0, 13.1, 17.2, 17.9, 25.0.

In some schemes of the present application, the above-mentioned Form IV, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 8.5, 10.0, 13.1, 17.2, 17.9, 20.0, 25.0.

In some schemes of the present application, the above-mentioned form IV, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 8.5, 10.0, 13.1, 15.7, 17.2, 17.9, 20.0, 25.0, 25.9.

In some schemes of the present application, the above-mentioned form IV, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 8.5, 10.0, 13.1, 15.7, 17.2, 17.9, 19.5, 20.0, 25.0, 25.9.

In some schemes of the present application, the above-mentioned form IV, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 5.5, 8.5, 10.0, 13.1, 14.0, 15.7, 17.2, 17.9, 19.5, 20.0, 25.0, 25.9.

In some schemes of the present application, the above-mentioned form IV, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 5.5, 8.5, 10.0, 13.1, 14.0, 15.7, 17.2, 17.9, 19.0, 19.5, 20.0, 25.0, 25.9, 26.4.

In some schemes of the present application, the above-mentioned form IV, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 5.5, 8.5, 10.0, 13.1, 14.0, 15.7, 16.5, 17.2, 17.9, 19.0, 19.5, 20.0, 22.9, 25.0, 25.9, 26.4.

In some schemes of the present application, the above-mentioned form IV, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 5.5, 8.5, 10.0, 13.1, 14.0, 15.7, 16.5, 17.2, 17.9, 19.0, 19.5, 20.0, 22.9, 23.4, 23.7, 25.0, 25.9, 26.4.

In some embodiments of the present application, the above-mentioned Form IV, using Cu-Kα radiation, has an X-ray powder diffraction pattern substantially as shown in FIG. 6 or FIG. 8 .

In some schemes of the present application, the above-mentioned Form IV has an endothermic peak at 192.07±5°C in its differential scanning calorimetry curve.

In some schemes of the present application, the differential scanning calorimetry curve of the above-mentioned Form IV has endothermic peaks at 64.82±5°C and 192.07±5°C.

In some embodiments of the present application, the above-mentioned Form IV has a DSC spectrum substantially as shown in FIG. 7 .

In some schemes of the present application, the thermogravimetric analysis curve of the above-mentioned Form IV shows a weight loss of 1.1526%±0.2% between room temperature and 75±5°C.

In some embodiments of the present application, the above-mentioned Form IV has a TGA spectrum substantially as shown in FIG. 7 .

On the other hand, the present application provides a crystalline form V of the compound represented by formula (A), which has an X-ray powder diffraction pattern using Cu-Kα radiation and has characteristic diffraction peaks (±0.2°) at the following 2θ angles: 6.0, 12.7, 18.2.

In some schemes of the present application, the above-mentioned crystal form V, using Cu-Kα radiation, has an X-ray powder diffraction pattern with characteristic diffraction peaks (±0.2°) at the following 2θ angles: 6.0, 12.7, 18.2, 23.4.

In some schemes of the present application, the above-mentioned crystal form V, using Cu-Kα radiation, has an X-ray powder diffraction pattern with characteristic diffraction peaks (±0.2°) at the following 2θ angles: 6.0, 12.7, 16.7, 18.2, 23.4.

In some schemes of the present application, the above-mentioned form V, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 6.0, 12.7, 16.7, 18.2, 23.4, 25.4, 28.1.

In some schemes of the present application, the above-mentioned form V, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 6.0, 7.0, 12.7, 14.2, 16.7, 18.2, 23.4, 25.4, 28.1.

In some schemes of the present application, the above-mentioned form V, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 6.0, 7.0, 12.7, 14.2, 16.7, 18.2, 23.4, 24.5, 25.4, 28.1.

In some schemes of the present application, the above-mentioned form V, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 6.0, 7.0, 12.1, 12.7, 14.2, 16.7, 17.6, 18.2, 23.4, 24.5, 25.4, 28.1.

In some schemes of the present application, the above-mentioned form V, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 6.0, 7.0, 12.1, 12.7, 14.2, 16.7, 17.6, 18.2, 20.2, 23.4, 24.5, 25.4, 28.1, and 28.8.

In some embodiments of the present application, the above-mentioned Form V, using Cu-Kα radiation, has an X-ray powder diffraction pattern substantially as shown in FIG. 9 .

On the other hand, the present application provides a crystalline form VI of the compound represented by formula (A), which has an X-ray powder diffraction pattern using Cu-Kα radiation and has characteristic diffraction peaks (±0.2°) at the following 2θ angles: 6.1, 16.9, 22.3.

In some schemes of the present application, the above-mentioned Form VI, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 6.1, 16.9, 18.6, 22.3.

In some schemes of the present application, the above-mentioned crystal form VI, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 6.1, 7.4, 11.5, 16.9, 18.6, 22.3.

In some schemes of the present application, the above-mentioned form VI, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 6.1, 7.4, 11.5, 16.9, 18.6, 19.4, 22.3, 25.3.

In some schemes of the present application, the above-mentioned form VI, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 6.1, 7.4, 11.5, 16.9, 18.6, 19.4, 20.1, 22.3, 25.3, 28.6.

In some schemes of the present application, the above-mentioned form VI, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 6.1, 7.4, 11.5, 16.9, 18.6, 19.4, 20.1, 22.3, 22.8, 23.2, 25.3, 28.6.

In some schemes of the present application, the above-mentioned form VI, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks (±0.2°) at the following 2θ angles: 6.1, 7.4, 11.5, 16.9, 18.6, 19.4, 20.1, 22.3, 22.8, 23.2, 25.3, 26.7, 28.6.

In some embodiments of the present application, the above-mentioned Form VI, using Cu-Kα radiation, has an X-ray powder diffraction pattern substantially as shown in FIG. 10 .

In some schemes of the present application, the differential scanning calorimetry curve of the above-mentioned Form VI has an endothermic peak at 184.47±5°C.

In some embodiments of the present application, the above-mentioned Form VI has a DSC spectrum substantially as shown in FIG. 11 .

In some schemes of the present application, the thermogravimetric analysis curve of the above-mentioned Form VI shows a weight loss of 0.253%±0.2% between room temperature and 200°C±5°C.

In some embodiments of the present application, the above-mentioned Form VI has a TGA spectrum substantially as shown in Figure 11.

On the other hand, the present application provides a crystalline composition comprising one or more of the crystalline form I, crystalline form II, crystalline form III, crystalline form IV, crystalline form V, and crystalline form VI of the compound of formula (A).

In some embodiments of the present application, the crystalline form II accounts for more than 50%, more than 60%, more than 70%, more than 80%, more than 90% or more than 95% of the weight of the crystalline composition.

In some embodiments of the present application, the Form III accounts for more than 50%, more than 60%, more than 70%, more than 80%, more than 90% or more than 95% of the weight of the crystalline composition.

In some embodiments of the present application, the Form IV accounts for more than 50%, more than 60%, more than 70%, more than 80%, more than 90% or more than 95% of the weight of the crystalline composition.

In some embodiments of the present application, the Form VI accounts for more than 50%, more than 60%, more than 70%, more than 80%, more than 90% or more than 95% of the weight of the crystalline composition.

On the other hand, the present application provides a pharmaceutical composition comprising a compound of formula (A) in solid form, a compound of formula (A) in crystalline form, or the above-mentioned crystalline composition.

In some embodiments of the present application, the above-mentioned pharmaceutical composition comprises one or more of the crystalline form I, crystalline form II, crystalline form III, crystalline form IV, crystalline form V, and crystalline form VI of the compound represented by formula (A).

On the other hand, the present application also provides a pharmaceutical composition comprising a compound of formula (A) in solid form, a compound of formula (A) in crystalline form, or the above-mentioned crystalline composition, and a pharmaceutically acceptable carrier.

In some embodiments of the present application, the above-mentioned pharmaceutical composition comprises one or more of the crystalline form I, crystalline form II, crystalline form III, crystalline form IV, crystalline form V, and crystalline form VI of the compound of formula (A), and a pharmaceutically acceptable carrier.

On the other hand, the present application provides the use of the compound of formula (A) in solid form, the compound of formula (A) in crystalline form, the above-mentioned crystalline composition, the crystalline form I, crystalline form II, crystalline form III, crystalline form IV, crystalline form V or crystalline form VI of the compound of formula (A) or the above-mentioned pharmaceutical composition as a drug or in the preparation of a drug.

In some embodiments of the present application, the drug is used to treat pain diseases, cell proliferative diseases, inflammatory diseases, neurodegenerative diseases or infectious diseases.

In some embodiments of the present application, the drug is used to prevent and/or treat diseases mediated by one or more of TRK, ROS1 or ALK.

In some schemes of the present application, the drug is used to prevent and/or treat tumors that are positive for NTRK gene rearrangement/fusion and/or drug-resistant mutations, or tumors that are positive for ROS1 gene rearrangement/fusion and/or drug-resistant mutations; preferably, the tumor is a solid tumor or a hematological tumor; further preferably, the tumor is a solid tumor.

In some schemes of the present application, the NTRK resistance mutation is NTRK1-G595R, NTRK1-G667C, NTRK3-G623R or NTRK3-G696A; preferably, the NTRK resistance mutation is NTRK1-G595R, NTRK3-G623R or NTRK1-G667C; further preferably, the NTRK resistance mutation is NTRK1-G595R or NTRK1-G667C.

On the other hand, the present application also provides the above-mentioned solid form of the compound shown in formula (A), the crystalline form of the compound shown in formula (A), the above-mentioned crystalline composition, the crystalline form I, crystalline form II, crystalline form III, crystalline form IV, crystalline form V or crystalline form VI of the compound shown in formula (A) or the above-mentioned pharmaceutical composition, for preventing and/or treating diseases mediated by one or more of TRK, ROS1 or ALK.

On the other hand, the present application also provides a method for preventing and/or treating diseases mediated by one or more of TRK, ROS1 or ALK, comprising: administering to a patient a therapeutically effective amount of the above-mentioned solid form of the compound of formula (A), the crystalline form of the compound of formula (A), the above-mentioned crystalline composition, the crystalline form I, form II, form III, form IV, form V or form VI of the compound of formula (A) or the above-mentioned pharmaceutical composition.

In some schemes of the present application, the above-mentioned disease is a tumor positive for NTRK gene rearrangement/fusion and/or drug-resistant mutation, or a tumor positive for ROS1 gene rearrangement/fusion and/or drug-resistant mutation; preferably, the tumor is a solid tumor or a blood tumor; further preferably, the tumor is a solid tumor.

In some schemes of the present application, the NTRK resistance mutation is NTRK1-G595R, NTRK1-G667C, NTRK3-G623R or NTRK3-G696A; preferably, the NTRK resistance mutation is NTRK1-G595R, NTRK3-G623R or NTRK1-G667C; further preferably, the NTRK resistance mutation is NTRK1-G595R or NTRK1-G667C.

In some embodiments of the present application, the above-mentioned disease is selected from pain diseases, cell proliferative diseases, inflammatory diseases, neurodegenerative diseases or infectious diseases.

In one embodiment, the above-mentioned TRK-mediated disease is selected from diseases mediated by one, two or three of TRKA, TRKB or TRKC.

In one embodiment, the above-mentioned disease involves NTRK gene, TRK protein, or their expression, activity or level disorder; preferably, it involves NTRK gene fusion, amplification, rearrangement, mutation or high expression; further preferably, it involves NTRK gene rearrangement/fusion or mutation.

In some schemes of the present application, the NTRK mutation is NTRK1-G595R, NTRK1-G667C, NTRK3-G623R or NTRK3-G696A; preferably, the NTRK resistance mutation is NTRK1-G595R, NTRK3-G623R or NTRK1-G667C; further preferably, the NTRK resistance mutation is NTRK1-G595R or NTRK1-G667C.

In one embodiment, the above-mentioned disease involves ROS1 gene, ROS1 protein, or disordered expression, activity or level thereof; preferably, involves ROS1 gene fusion, amplification, rearrangement, mutation or high expression; further preferably, involves ROS1 gene rearrangement/fusion or mutation.

In one embodiment, the above-mentioned disease involves one or more genes, proteins, or dysregulation of their expression, activity or level among TRK, ALK, and ROS1; preferably, it involves one or more gene fusion, amplification, rearrangement, mutation or high expression among NTRK, ALK, and ROS1; further preferably, it involves one or more gene fusion or mutation among NTRK, ALK, and ROS1.

In one embodiment, the cell proliferative disease is a tumor or cancer.

In one embodiment, the above-mentioned tumor or cancer is a solid tumor and a blood tumor; preferably a solid tumor; further preferably a solid tumor positive for NTRK gene rearrangement/fusion and/or drug-resistant mutation, or a solid tumor positive for ROS1 gene rearrangement/fusion and/or drug-resistant mutation.

In some schemes of the present application, the NTRK resistance mutation is NTRK1-G595R, NTRK1-G667C, NTRK3-G623R or NTRK3-G696A; preferably, the NTRK resistance mutation is NTRK1-G595R, NTRK3-G623R or NTRK1-G667C; further preferably, the NTRK resistance mutation is NTRK1-G595R or NTRK1-G667C.

In one embodiment, the tumor or cancer is a hematological malignancy, lung cancer, breast cancer, ovarian cancer, prostate cancer, pancreatic cancer, glioma, colorectal cancer, melanoma, cancer of the head and neck, gallbladder cancer, thyroid cancer, malignant glioma, gastric cancer, neuroblastoma or salivary gland cancer; preferably, the lung cancer is non-small cell lung cancer.

Definition and Description

Unless otherwise indicated, the following terms and phrases used herein are intended to contain the following meanings. A specific phrase or term should not be considered as uncertain or unclear in the absence of a particular definition, but should be understood according to common meaning. The "compound shown in the formula (A) of solid form" mentioned in the application refers to the compound shown in the formula (A) of solid form.

The "compound represented by formula (A) in crystalline form" mentioned in the present application refers to the compound represented by formula (A) in crystalline form, including the anhydrous and solvent-free form, hydrate form and solvate form of the compound represented by formula (A).

The term "solvate" or "solvate" refers to an association formed by a stoichiometric or non-stoichiometric ratio of solvent molecules and the compound represented by formula (A) of the present application, including an association containing both water molecules and one or more other solvent molecules, and an association containing only one or more other solvent molecules.

The term "hydrate" refers to an association formed by water molecules in a stoichiometric ratio or a non-stoichiometric ratio and the compound represented by formula (A) of the present application.

The "anhydrous and solvent-free form" refers to the absence of water molecules or solvent molecules, or the water molecules or solvent molecules coexist with the compound represented by formula (A) in a manner that is not bound by intermolecular forces, such as by adsorption.

In the context of the present application, the characteristic diffraction peaks, diffraction peaks and/or 2θ angle values in the X-ray powder diffraction pattern are all in degrees (°).

The term "crystalline composition" refers to a solid form, which contains one, two or more of the crystal form I, crystal form II, crystal form III, crystal form IV, crystal form V, or crystal form VI mentioned in the present application. Moreover, in addition to the crystal form of the present application, the crystalline composition may optionally contain other crystal forms or other amorphous forms of the compound represented by formula (A) or its salt, or impurities other than these substances. It should be understood by those skilled in the art that the sum of the contents of each component in the crystalline composition should be 100%.

The "room temperature" is the room temperature conventionally known in the art, generally 10-30°C, preferably 25°C±5°C.

In the X-ray powder diffraction pattern, the term "substantially" or "substantially as shown" refers to a substantially pure crystalline form, in which at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99% of the peaks in the powder X-ray diffraction pattern appear in the given pattern. Further, when the content of a certain crystalline form in the product gradually decreases, some of the diffraction peaks in its X-ray powder diffraction pattern attributable to the crystalline form may become less due to the detection sensitivity of the instrument. In addition, for any given crystalline form, there may be slight errors in the position of the peak, which is also well known in the field of crystallography. For example, due to changes in temperature, sample movement or instrument calibration during sample analysis, the position of the peak can move, and the measurement error of the 2θ value is sometimes about ±0.3°, usually about ±0.2°. Therefore, when determining each crystalline structure, this error should be taken into account, and the term "substantially" or "substantially as shown in the accompanying drawings" is also intended to cover such differences in the diffraction peak positions, referring to ±0.3°, preferably ±0.2°.

In a DSC spectrum or a TGA spectrum, the term "substantially" or "substantially as shown" means that for the same crystal form of the same compound, in consecutive analyses, the error of the thermal transition onset temperature, the peak temperature of the endothermic peak, the peak temperature of the exothermic peak, the melting point, the starting temperature of weight loss or the end temperature of weight loss is typically within about 5°C, usually within about 3°C. When describing a compound as having a given thermal transition onset temperature, peak temperature of the endothermic peak, peak temperature of the exothermic peak, the melting point, the starting temperature of weight loss or the end temperature of weight loss, it refers to the temperature ±5°C.

As used herein, the term "cell proliferative disorder" refers to a condition in which a population of cells grows at a rate that is slower or faster than expected under given physiological conditions and circumstances.

The term "tumor" includes benign tumors, malignant tumors and borderline tumors, among which malignant tumors are collectively referred to as cancer.

As used herein, the term "prevent" or "prevent" means that when used for a disease or condition (e.g., cancer), the compound or drug (e.g., the combination product claimed herein) can reduce the frequency of symptoms of the medical condition in a subject or delay its onset compared to a subject to which the compound or drug (e.g., the combination product claimed herein) is not administered.

As used herein, the term "treat," "treat," "treat," "treat," or "treat" refers to alleviating, relieving, or ameliorating symptoms of a disease or condition, ameliorating underlying metabolically caused symptoms, inhibiting a disease or symptom, such as arresting the development of a disease or condition, alleviating a disease or condition, causing regression of a disease or condition, alleviating a condition caused by a disease or condition, or arresting symptoms of a disease or condition.

The term "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" refers to those carriers or excipients that have no significant irritation to organisms and will not impair the biological activity and properties of the active compound.

The intermediate compounds of the present application can be prepared by a variety of synthesis methods known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitution methods known to those skilled in the art. Preferred embodiments include but are not limited to the examples of the present application.

The chemical reactions of the specific embodiments of the present application are carried out in a suitable solvent, which must be suitable for the chemical changes of the present application and the reagents and materials required. In order to obtain the compounds of the present application, it is sometimes necessary for those skilled in the art to modify or select the synthesis steps or reaction processes based on the existing embodiments.

The present application will be described in detail below through examples, which do not imply any limitation to the present application.

All solvents used in this application were commercially available and used without further purification.

Technical Effects

The solid form of the compound of formula (A), the crystalline form of the compound of formula (A), and the specific crystal form provided in the present application have one or more of the following beneficial effects:

(1) The compound represented by formula (A) in solid form has good properties and is easy to weigh, transfer, separate, purify and store.

(2) the compound of formula (A) and the specific crystalline form have good crystallinity;

(3) The compound of formula (A) and its specific crystal forms in crystalline form are easy to purify, filter and separate, especially the crystal form IV, which is easy to prepare and has a high yield;

(4) The preferred crystal form has good physical and chemical stability and has good medicinal prospects;

(5) In vitro kinase activity inhibition tests showed that the compound represented by formula (A) exhibited excellent inhibitory activity against a variety of kinases (e.g., TRK, ALK, ROS1) and their mutants, especially TRK and its mutant forms;

(5) In vitro cell inhibitory activity tests showed that the compound represented by formula (A) had a strong inhibitory effect on cells with various NTRK mutations, and the inhibitory activity against 6 types of cells had an IC ₅₀ of less than 10 nM, preferably less than 5 nM, and more preferably less than 1 nM;

(6) The results of the in vivo tumor inhibition test showed that compared with the control compound, the compound represented by formula (A) had better in vivo anti-tumor effect, better tolerability, and higher drugability;

(7) In vivo mechanism studies have shown that the compound represented by formula (A) can inhibit TRK phosphorylation in tumor tissues, thereby effectively inhibiting the phosphorylation of PLCγ1 and AKT, and inhibiting tumor tissue growth.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1: X-ray powder diffraction pattern of Form I of Example 1.

Figure 2: X-ray powder diffraction pattern of Form II of Example 2.

Figure 3: DSC-TGA spectrum of Form II of Example 2.

Figure 4: X-ray powder diffraction pattern of Form III of Example 3.

Figure 5: DSC-TGA spectrum of Form III of Example 3.

Figure 6: X-ray powder diffraction pattern of Form IV of Example 4.

Figure 7: DSC-TGA spectrum of Form IV of Example 4.

Figure 8: X-ray powder diffraction pattern of Form IV of Example 5.

Figure 9: X-ray powder diffraction pattern of Form V of Example 7.

Figure 10: X-ray powder diffraction pattern of Form VI of Example 8.

Figure 11: DSC-TGA spectrum of Form VI of Example 8.

Figure 12: IR spectrum of the solid obtained in Example 0.

Figure 13: Result diagram of Experimental Example 4.

DETAILED DESCRIPTION

1. X-ray powder diffractometer (XRPD)

(1) Instrument model: Bruker D8 Advance X-ray powder diffractometer

Test method: About 5-20 mg of sample (Examples 1-4, Examples 6-8) was used for XRPD detection

The detailed XRPD parameters are as follows:

X-ray generator: Cu, Kα,

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

Scanning range: 3°-40°(2θ)

Scanning step: 0.02°

Sample pan: Zero background sample pan.

(2) Instrument model: D2 PHASER desktop X-ray diffractometer

Test method: About 100-200 mg of sample (Example 0, Example 5) was used for XRPD detection

The detailed XRPD parameters are as follows:

X-ray generator: Cu, Kα,

Light tube voltage: 30kV, light tube current: 10mA

Scanning range: 3°-60°(2θ)

Scanning step: 0.02°

Sample pan: Zero background sample pan.

2. Differential Scanning Calorimeter (DSC)

Instrument model: TA Discovery 250 Differential Scanning Calorimeter

Test method: Place the sample in a perforated aluminum crucible, equilibrate the sample at 25°C, and heat it to the final temperature at a heating rate of 10°C/min.

Sample amount: 1~3mg

Air flow type: Nitrogen

Flow rate: 50mL/min

Heating starting temperature: 25°C.

3. Thermogravimetric analysis (TGA)

Instrument model: TA Discovery 55 Thermogravimetric Analyzer (TA, US)

Test method: Place the sample in a pre-peeled aluminum crucible. After the sample mass is automatically weighed in the TGA heating furnace, heat the sample to the final temperature at a rate of 10°C/min.

Sample size: 1-5 mg

Air flow type: Nitrogen

Sample chamber air flow rate: 60mL/min

Heating starting temperature: room temperature

Termination temperature: 250/300℃.

4. Dynamic moisture adsorption and desorption analysis (DVS)

Instrument model: DVS Intrinsic dynamic water vapor sorption instrument (SMS)

Test method: Place a sufficient amount of sample (10-20 mg) in a pre-peeled sample chamber and weigh automatically. Dry the sample at 40°C (only for anhydrous, start at 25°C for hydrate) until dm/dt is less than 0.002%. Cool to 25°C and start the test using the operating parameters in the table below.

Stage time 60 minutes Drying/testing temperature 40℃/25℃ cycle The whole cycle Each RH balance time 1 hour Data storage rate 5 seconds Total airflow 200sccm Total gas flow rate after the experiment 200sccm

5. High Performance Liquid Chromatography (HPLC)

Instrument model: Agilent 1260 series (Waters, US)

Express C18 4.6x 100mm,2.7μmChromatographic column:

Express C18 4.6x 100mm, 2.7μm

Test conditions: wavelength 248nm; column temperature 40℃

Flow rate: 1.0mL/min

Injection volume: 5 μL.

6. Nuclear Magnetic Resonance Spectroscopy (NMRS)

Instrument model: Bruker AVANCE III 400 (Bruker, GER)

Content and test solvent: ¹ H-NMR, the test solvent is DMSO-d6.

7. Infrared Spectroscopy (IR)

Detection instrument: PerkinElmer Spectrum 100FT-IR infrared spectrometer

Test method: Weigh 3 mg of sample, dilute with KBr and press into a pellet, and test at room temperature. Specific parameters are: detection range: 4000-400cm ^-1 wavenumber, resolution: 4cm ^-1 .

Abbreviations: DCM: dichloromethane; DIPEA: diisopropylethylamine; DMF: N,N-dimethylformamide; EA: ethyl acetate; PE: petroleum ether; DMSO: dimethyl sulfoxide; TBTU: O-benzotriazol-N,N,N',N'-tetramethyluronium tetrafluoroborate; BOP: benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate; ATP: adenosine 5'-triphosphate; DTT: 1,4-dithiothreitol; MTT: 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium blue bromide.

In order to better understand the content of this application, the following is further described in conjunction with specific examples, but the specific implementation methods are not intended to limit the content of this application. The test methods for which specific conditions are not specified in the following preparation examples, embodiments, comparative examples, test examples or test examples are selected according to conventional methods and conditions, or according to the product instructions.

Preparation Example 1: Preparation of the compound of formula (A)

Step a: A mixed solution of (2R, 4S)-2-(2,5-difluorophenyl)-4-fluoropyrrolidine (5.826 g, 28.958 mmol), 5-chloropyrazolo[1,5-a]pyrimidine-3-carboxylic acid ethyl ester (6.534 g, 28.958 mmol), n-butanol (50 mL) and diisopropylamine (8.790 g, 86.874 mmol) was reacted at 100°C for 4 h, and concentrated under reduced pressure to obtain a crude product of 5-((2R, 4S)-2-(2,5-difluorophenyl)-4-fluoropyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid ethyl ester (B1). The crude product was used directly in the next step without purification, (ES, m/z): 391.05 [M+H] ⁺ .

Step b: Dissolve the crude product of ethyl 5-((2R,4S)-2-(2,5-difluorophenyl)-4-fluoropyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate in anhydrous ethanol (50 mL), stir at 75°C until the system is clear and transparent, then add LiOH (4.86 g, 115.832 mmol) aqueous solution (50 mL), stir at 75°C for 5 h. After cooling to room temperature, concentrate under reduced pressure to remove anhydrous ethanol. Slowly add 1N HCl aqueous solution to adjust pH 3-4, a large amount of white solid precipitates, stir at room temperature for 30 min and filter, and wash the filter cake with a small amount of purified water. Collect the filter cake, dry it and weigh it to obtain a white powder solid 5-((2R,4S)-2-(2,5-difluorophenyl)-4-fluoropyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid (9.9 g). The filtrate was extracted with EA (2×50 mL), the organic phases were combined, washed with water (2×50 mL) and saturated aqueous NaCl solution (50 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. Column chromatography was performed for purification (PE:EA=4:1-2:1, v/v), the product spots were collected, and the product was concentrated under reduced pressure to obtain a white powdery solid 5-((2R,4S)-2-(2,5-difluorophenyl)-4-fluoropyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid (386 mg). A total of 5-((2R,4S)-2-(2,5-difluorophenyl)-4-fluoropyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid pure product (B2, 10.286 g, 98%) was obtained, (ES, m/z): 363.04[M+H] ⁺ .

Step c: Add 1-Boc-4-(4-aminophenyl)piperazine (918 mg, 3.312 mmol) to a solution of 5-((2R,4S)-2-(2,5-difluorophenyl)-4-fluoropyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid (Intermediate B2, 1000 mg, 2.76 mmol) and TBTU (1063 mg, 3.312 mmol) in anhydrous DMF (10 mL), then add DIPEA (1284 mg, 9.936 mmol) dropwise at 0°C and react overnight at room temperature. Water (50 mL) was added to the reaction solution and the mixture was stirred. Solid was precipitated. The filter cake was obtained by vacuum filtration and dried in a vacuum oven to obtain tert-butyl 4-(4-(5-((2R,4S)-2-(2,5-difluorophenyl)-4-fluoropyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamido)phenylpiperazine-1-carboxylate (B3, 1320 mg, 77%). (ES, m/z): 622.09 [M+H] ⁺ .

Step d: To tert-butyl 4-(4-(5-((2R,4S)-2-(2,5-difluorophenyl)-4-fluoropyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamido)phenylpiperazine-1-carboxylate (1.320 g, 2.125 mmol) was added DCM and _CF3 COOH (12 mL, 3/1, v/v), stirred at room temperature for 4 h, the reaction solution was concentrated under reduced pressure, water (80 mL) and EA (10 mL) were added to the residue, the base was adjusted with aqueous ammonia (pH = 9), solid was precipitated during stirring, and the filter cake was obtained by vacuum filtration. The filter cake was rinsed with a small amount of water, dried and weighed to obtain 5-((2R, 4S)-2-(2,5-difluorophenyl)-4-fluoropyrrolidin-1-yl)-N-(4-(piperazin-1-yl)phenyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (B4, 907 mg, 82%), (ES, m/z): 522.09 [M+H] ⁺ .

Step e: Hydroxyacetic acid (306 mg, 4.026 mmol) was added to a solution of 5-((2R,4S)-2-(2,5-difluorophenyl)-4-fluoropyrrolidin-1-yl)-N-(4-(piperazine-4-yl)phenyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (Intermediate B4, 700 mg, 1.342 mmol) and BOP (712 mg, 1.610 mmol) in anhydrous DMF (10 mL), and then DIPEA (520 mg, 4.026 mmol) was added dropwise at 0°C and the reaction was stirred at room temperature for 4 h. Water (80 mL) was added to the reaction solution and mixed. The mixture was extracted with EA (55 mL×2), and the combined organic phase was washed with H ₂ O (80 mL) and brine (80 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. Column chromatography was performed on a silica gel column, first with 1% (v/v) MeOH-DCM and then with 2% (v/v) MeOH-DCM. The product spots were collected and concentrated to obtain 5-((2R,4S)-2-(2,5-difluorophenyl)-4-fluoropyrrolidin-1-yl)-N-(4-(4-(2-hydroxyacetyl)piperazin-1-yl)phenyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (A, 666 mg, 86%), (ES, m/z): 580.14 [M+H] ⁺ . ¹ H NMR (600 MHz, DMSO-d ₆ )δ9.810(s,1H),8.906-8.723(m,1H),8.283-8.229(m,1H),7.623(s,1H),7.343(s,1H),7.210(s,2H),7.061-6.842(m,4H),5.711-5.495(m,2H), 4.631(t,J＝5.4Hz,1H),4.556-4.548(m,1H),4.318-4.225(m,1H),4.150(d,J＝5.4Hz,2H),3.637(s,2H),3.513(s,2H),3.124-3.106(m,4H),2.957 -2.912(m,1H).

Reference substance preparation example 1-5:

In addition: refer to the preparation process routes and operations in WO2019029629A1 and WO2012034095A1 patent documents to prepare compounds D1 to D5.

Example 0: Preparation of a solid form of a compound of formula (A)

Take the sample of Preparation Example 1 (about 50 mg), use an oil pump to pump for 3 hours under heating conditions (50°C), and obtain an off-white solid with a purity of 98.6%. IR (KBr, cm-1): 3429.38, 1643.78, 1487.95, 1453.15, 1232.01, 1026.79. See Figure 12. Take the sample for X-ray powder diffraction, which shows that it is amorphous.

Example 1: Preparation of Form I of Compound of Formula (A)

At room temperature, weigh the sample of Example 0 (about 20 mg) into a sample bottle, add tetrahydrofuran (0.2 mL) to obtain a clear solution, gradually add methanol (1.4 mL) dropwise thereto, stir at room temperature for 1 day, and filter to obtain a solid. The sample was subjected to X-ray powder diffraction, which showed a crystalline solid (form I) with good crystallinity, as shown in Figure 1, and its XRPD diffraction peak data is shown in Table 1.

Table 1 XRPD diffraction peak data table of Example 1 Form I

Note: Peaks with relative peak intensity > 4.0% are selected and listed in the table.

Example 2: Preparation of Crystalline Form II of Compound (A)

An appropriate amount of the crystal form I obtained in Example 1 was weighed and dried at 50°C for 3 hours to obtain a solid. The sample was subjected to X-ray powder diffraction, which showed a crystalline solid (crystal form II) with good crystallinity. The spectrum is shown in Figure 2, and the XRPD diffraction peak data is shown in Table 2. The sample was subjected to DSC-TGA test. The DSC graph showed an endothermic peak at 58.39°C and 155.27°C, respectively. The TGA graph showed that the sample had a weight loss of 0.1910% between room temperature and 75°C, as shown in Figure 3.

Table 2 XRPD diffraction peak data table of Example 2 Form II

Example 3: Preparation of Form III of the Compound of Formula (A)

At room temperature, weigh the sample of Example 0 (about 30 mg) into a sample bottle, add acetonitrile (0.3 mL) to prepare a solution, stir at room temperature for 3 days, filter to obtain a solid, and dry at 50°C for 3 hours. Take the sample for X-ray powder diffraction, which shows a crystalline solid (form III) with good crystallinity. The spectrum is shown in Figure 4, and its XRPD diffraction peak data is shown in Table 3. Take the sample for DSC-TGA test, and the DSC graph shows an endothermic peak at 182.29°C, as shown in Figure 5.

Table 3 XRPD diffraction peak data table of Example 3 Form III

Peak position (2θ)° Relative Strength % Peak position (2θ)° Relative Strength % Peak position (2θ)° Relative Strength % 5.618 100.0 15.802 47.2 19.901 6.7 7.431 5.5 16.272 5.6 23.688 9.9 10.937 8.8 17.053 5.9 23.845 5.4 11.319 8.4 18.428 13.2 13.284 25.5 18.983 9.2

Note: Peaks with relative peak intensity > 5.0% are selected and listed in the table.

Example 4: Preparation of Form IV of Compound of Formula (A)

At room temperature, weigh the sample of Example 0 (about 20 mg) in a sample bottle, add tetrahydrofuran (0.2 mL) to prepare a clear solution, gradually add isopropanol (1.3 mL) dropwise thereto, stir at room temperature for 1 day, filter to obtain a solid, and dry at 50°C for 3 hours. Take the sample for X-ray powder diffraction, which shows a crystalline solid (form IV) with good crystallinity. The spectrum is shown in Figure 6, and its XRPD diffraction peak data is shown in Table 4. Take the sample for DSC-TGA test. The DSC graph shows two endothermic peaks at 64.82°C and 192.07°C. The TGA graph shows that the sample has a weight loss of 1.1526% between room temperature and 75°C, as shown in Figure 7.

Table 4 XRPD diffraction peak data table of Example 4 Form IV

Peak position (2θ)° Relative Strength % Peak position (2θ)° Relative Strength % Peak position (2θ)° Relative Strength % 5.390 11.4 17.184 100.0 23.371 15.2 8.527 97.1 17.897 45.7 24.172 8.7 10.046 59.4 18.994 11.8 25.017 27.2 13.117 16.6 19.463 14.1 25.893 24.3 14.091 3.8 19.958 19.0 26.453 15.6 15.258 7.0 21.259 9.7 29.194 5.4 15.731 20.1 22.052 6.8 16.505 9.0 22.958 10.7

Note: Peaks with relative peak intensity > 3.5% are selected and listed in the table.

Example 5-6: Preparation of Form IV of the Compound of Formula (A)

Using the sample obtained in Example 0, Form IV can also be obtained under the conditions listed in Table 5. In particular, Example 5 refers to the preparation method of Example 4.

Table 5 Experimental conditions and results of Examples 5-6

Table 6 XRPD diffraction peak data of Example 5 Form IV

Peak position (2θ)° Relative Strength % Peak position (2θ)° Relative Strength % Peak position (2θ)° Relative Strength % 5.479 9.6 17.202 74.6 22.932 11.6 8.573 100.0 17.468 18.6 23.369 13.6 10.066 55.4 17.928 32.2 23.737 14.3 11.527 5.8 18.612 5.3 24.219 8.2 13.129 23.9 19.015 11.3 25.023 34.6 13.999 10.0 19.469 15.4 25.358 20.6 14.229 6.5 19.982 24.8 25.841 22.6 15.144 5.1 21.321 5.5 26.421 11.6 15.692 19.8 22.032 7.0 16.457 8.4 22.493 5.7

Note: Peaks with relative peak intensity > 5% are selected and listed in the table.

Example 7: Preparation of Form V of the Compound of Formula (A)

At room temperature, weigh the sample of Example 0 (about 34 mg) into a sample bottle, dissolve it in acetone (0.5 mL), stir it at 5°C for 1 day, and filter to obtain a solid. The sample was subjected to X-ray powder diffraction, which showed a crystalline solid (form V) with good crystallinity. The spectrum is shown in Figure 9, and its XRPD diffraction peak data is shown in Table 7.

Table 7 XRPD diffraction peak data table of Example 7 Form V

Peak position (2θ)° Relative Strength % Peak position (2θ)° Relative Strength % Peak position (2θ)° Relative Strength % 6.003 100.0 17.582 8.1 23.448 52.5 7.048 12.4 18.190 45.0 23.986 4.1 10.203 4.1 18.526 4.3 24.451 9.9 11.571 4.3 20.247 8.7 25.395 18.4 12.113 7.2 20.578 6.6 26.628 4.3 12.729 42.2 20.783 8.9 28.120 19.5 14.225 9.6 20.954 6.1 28.793 7.1 16.727 27.7 23.228 18.3

Note: Peaks with relative peak intensity > 4.0% are selected and listed in the table.

Example 8: Preparation of Crystalline Form VI of Compound of Formula (A)

An appropriate amount of the crystal form V obtained in Example 7 was weighed and dried at 50°C for 3 hours to obtain a solid. The sample was subjected to X-ray powder diffraction, which showed a crystalline solid (crystal form VI) with good crystallinity. The spectrum is shown in Figure 10, and its XRPD diffraction peak data is shown in Table 8. The sample was subjected to DSC-TGA test. The DSC graph showed an endothermic peak at 184.47°C, and the TGA graph showed that the sample had a weight loss of 0.253% between room temperature and 200°C, as shown in Figure 11.

Table 8 XRPD diffraction peak data table of Example 8 Form VI

Note: Peaks with relative peak intensity > 3.0% are selected and listed in the table.

Comparative Example 1-2

Weigh an appropriate amount of the sample of Example 0, dissolve it in tetrahydrofuran to prepare a 10 mg/0.3 mL clear solution, divide the resulting solution evenly into centrifuge tubes, 0.3 mL per tube, then add 0.1 mL of solvent 2 shown in the table below to each tube, cover with a film and pierce holes, and evaporate the solvent at room temperature.

Table 9 Experimental conditions and results of comparative examples 1-2

Comparative Example Solvent 2 result Comparative Example 1 Methanol Sticky Comparative Example 2 acetone Sticky

Test Example 1: Solid Stability Test of Different Crystal Forms of Compound (A)

An appropriate amount of Form III of the compound of formula (A) (Example 3) was weighed into a vial and placed under high temperature (60°C, sealed) and accelerated (40°C/75% RH, open) conditions for 7 days respectively. Samples were taken for purity detection and X-ray powder diffraction, respectively, to investigate the stability of Form III of the compound of formula (A) (Example 3) under different conditions. The results are shown in Table 10.

An appropriate amount of Form IV (Example 4) sample was weighed and placed in a small vial. It was placed under high temperature (60°C, sealed), high humidity (25°C/92.5% RH, open) and accelerated (40°C/75% RH, open) conditions for 7 days respectively. The samples were taken for purity testing and X-ray powder diffraction, respectively, to investigate the stability of Form IV (Example 4) of the compound of formula (A) under different conditions. The results are shown in Table 11.

Table 10 Solid stability test results of Form III

Table 11 Solid stability test results of Form IV

The data show that: the crystal form III of Example 3 can maintain chemical stability and crystal stability under high temperature and accelerated conditions; the crystal form IV of Example 4 can maintain chemical stability and crystal stability under high temperature, high humidity and accelerated conditions.

Test Example 2: DVS test of different crystal forms of the compound of formula (A)

The samples of Form III of Example 3 and Form IV of Example 4 were placed in the DVS sample chamber for testing. The samples after DVS testing were subjected to X-ray powder diffraction, and the results are shown in Table 12.

Table 12 DVS test results of different crystal forms

Examples and initial crystal form Crystalline form after DVS Example 3/Crystal Form III Crystal form unchanged Example 4/Form IV Crystal form unchanged

The data show that the crystal forms III and IV remain unchanged after DVS testing.

Test Example 1: TRK kinase inhibition test

1. Operation steps:

1.1 Kinase reaction:

Add a certain concentration gradient of the test compound and enzyme solution (add kinase buffer (1X kinase buffer (Cisbio, Cat#62EZBFDD), pH 7.5; 5mM MgCl ₂ , 1mM DTT)) to the compound plate in sequence, and centrifuge at 1000rpm for 30 seconds. Seal the plate and incubate it in a constant temperature incubator at 25°C for 30 minutes. Prepare substrate solutions of TK-Sub-biotin (Cisbio, Cat#61TKOBL) and ATP (Sigma, Cat#R0441), add the substrate mixed solution to the 384-well plate, and centrifuge at 1000rpm for 30 seconds. Seal the plate and incubate it in a constant temperature incubator at 25°C for 60 minutes.

1.2 Kinase assay:

Dilute and mix TK antibody and XL665 and add to the assay plate. Centrifuge at 1000rpm for 30 seconds. Seal the plate and incubate it in a 25℃ incubator for 60 minutes. Place the assay plate on the Envision machine for reading. (HTRF 665/615 ratio: 665nm signal value/615nm signal value)

Inhibition rate = (ratio of _{negative control wells} - ratio of _{compound wells} ) / (ratio of _{negative control wells} - ratio of _{no enzyme control wells} ) × 100%

1.3 Data analysis and curve fitting

Data were fitted in XLFit excel add-in version 5.4.0.8 to obtain _IC50 values.

1.4QC parameters

Reference compounds were included in each plate and their _IC50s were within 3-fold each time.

2. Test results: as shown in Table 13

Table 13 Inhibitory activity of different compounds on TRK kinase

Note: RXDX-101, LOXO-195 and LOXO-101 are all publicly known compounds and are commercially available as marketed products (pharmaceutical or chemical grade products); Compound represented by formula (A): Sample of Preparation Example 1.

The results show that the compound represented by formula (A) exhibits high kinase inhibitory activity in multiple kinases, and its activity in TRKA, TRKB, TRKC and TRKC-G696A is better than or equivalent to RXDX-101, LOXO-195 and LOXO-101, and its inhibitory activity in multiple mutant resistant kinases (G595R, G667C, G623R) is significantly better than RXDX-101, LOXO-195 and LOXO-101.

Test Example 2: ALK and ROS1 kinase inhibition test

1. Operation steps:

1.1 Kinase reaction:

The compound was diluted with DMSO to a certain concentration and then diluted 4 times in a gradient. A certain concentration of compound, enzyme solution and DMSO were added to a 384-well plate and incubated at room temperature for 10 minutes; fluorescein-labeled peptide and ATP (sigma, Cat. No.: A7699-1G, Lot No.: 987-65-5) were added and incubated at 28°C for a certain period of time; stop solution was added and the reading was taken.

The inhibition rate formula corresponding to a single concentration is: Inhibition rate = (OD _{negative control well} -OD _{compound well} )/(OD _{negative control well} -OD _{no enzyme control well} ) × 100%

1.2 Data analysis and curve fitting

The data were fitted in XLFit excel add-in version 4.3.1 to obtain _IC50 values, and the results are shown in Table 14.

Table 14 Inhibitory activity of different compounds on ALK and ROS1 kinases

Note: Compound represented by formula (A): Sample of Preparation Example 1.

The results showed that the compound represented by formula (A) exhibited strong inhibitory activity in ROS1 kinase, which was significantly better than RXDX-101 and LOXO-101, and better than LOXO-195; it also had good inhibitory activity against ALK kinase, which was significantly better than LOXO-101 and LOXO-195.

Test Example 3: In vitro cell inhibition test

1. Cell lines

6 experimental cell lines from: Kangyuan Bochuang Biotechnology (Beijing) Co., Ltd.

Cell type: Murine B cells

Culture medium: RPMI-1640 + 10% FBS

2. Test methods

Cells in the logarithmic growth phase were harvested and counted using a platelet counter. Cell suspensions of a certain density were evenly inoculated in a 96-well plate, 100 μL per well, and shaken to disperse evenly into the wells; 100 μL of a certain concentration gradient of drug solution was added to each well, and three replicate wells were set for each drug concentration; cultured in a 37°C CO ₂ incubator for 72 hours; MTT working solution (5 mg/mL) was added, 20 μL per well; 37°C for 4 hours; centrifuged at 1000 rpm/min on a plate centrifuge for 5 minutes, 180 μL of culture medium was aspirated and 150 μL of DMSO was added, the microporous oscillator was shaken to mix, the bottom of the plate was wiped clean, and the optical density (OD) was detected at 550 nm by an ELISA reader.

3. Data Analysis

Inhibition rate = (control well OD - test well OD) / (control well OD - blank well OD) * 100%. According to the inhibition rate of each concentration, the half inhibition concentration _IC50 value was calculated using SPSS software.

4. Test results: The results are shown in Table 15:

Table 15 Inhibitory activity of different compounds on different cell lines

Note: Compound represented by formula (A): Sample of Preparation Example 1.

Table 16 Inhibitory activity of control compounds on different cell lines

The results show that the compound represented by formula (A) of the present application exhibits good in vitro cell activity in a variety of wild-type and mutant drug-resistant cell lines, which is significantly better than RXDX-101, LOXO-195, LOXO-101 and prior art compounds D1-D5.

Experimental Example 4: Study on the in vivo mechanism of the compound represented by formula (A)

1. Test methods

1.1 Model preparation:

Mutant drug-resistant cells Ba/F3 LMNA-NTRK1-G595R in the logarithmic growth phase were collected and resuspended in serum-free medium to a cell concentration of 6×10 ⁷ -10×10 ⁷ /mL, and an equal volume of Matrigel was added to the cell suspension to a final cell concentration of 3×10 ⁷ -5×10 ⁷ /mL. 0.1 mL of tumor cell suspension was subcutaneously inoculated in the axilla of the forelimb of NuNu mice (Beijing Weitong Lihua, 4-6 weeks, female) at an inoculation volume of 3×10 ⁶ -5×10 ⁶ /mouse to prepare an animal model.

1.2 Experimental Grouping:

The maximum and minimum tumor diameters of the nude mouse transplanted tumors were measured with a vernier caliper to calculate the tumor volume: the calculation formula for tumor volume (TV) is: V = 1/2 × a × b ² , where a and b represent the maximum and minimum diameters of the tumor mass, respectively. Nude mice with appropriate tumor volumes were selected and randomly divided into 7 groups (200-300 mm ³ ) according to the tumor volume, with 3 mice in each group.

1.3 Administration

The drug was administered orally according to the animal body weight, with a dosage volume of 10 ml/kg. The compound represented by formula (A) was prepared into the required dosage concentration using "3% DMSO + 96% HP-β-CD (0.5 g/mL) + 1% HCl".

The control group had three rats, and the tumor tissues were frozen 4 hours after the solvent was administered. The other groups were all given 100 mg/kg of the compound represented by formula (A), and the tumor tissues were frozen at 0.25h, 1h, 4h, 8h, 12h and 24h.

1.4 Protein extraction and quantification

A certain amount of tumor tissue was added with a corresponding volume of protein lysis buffer (RIPA lysis buffer (ThermoFisher, Catalog No. 89900): protease inhibitor (cOmplete, Mini, EDTA-free, EASY pack; Roche, Catalog No. 04693159001): phosphatase inhibitor (PhosStop, EASY pack; Roche, Catalog No. 04906837001) = 8:1:1), homogenized, and lysed on ice for 30 minutes. Centrifuge at low temperature and high speed, and take the supernatant for BCA protein quantification (according to the BCA protein quantification kit (Tiangen, Catalog No.: #PA115-01)). Finally, the protein concentration was adjusted to a uniform concentration with the lysis buffer, and the loading buffer was added, and boiled at 100°C for 10 minutes.

1.5 Western-blot

Use 4-20% 10-well precast gel; load 100 μg; electrophoresis at 140V for 1-1.5h; wet transfer at 300mA for 1.5h-2h; block with 5% BSA for 2-3h; incubate with primary antibody at 4°C overnight (Trk 1:5000, p-Trk, PLCγ1, p-PLCγ1, AKT, p-AKT, actin 1:1000); wash with 0.1% TBST for 4×5min; incubate with secondary antibody at room temperature for 2h (1:5000), ECL luminescence, and expose.

2. Test results: as shown in Figure 13.

The experimental results show that: as time goes by, TRK, p-TRK, p-PLCγ1 and p-AKT in Figure 13 are all significantly reduced, proving that the compound shown in formula (A) can significantly reduce the protein levels of TRK and p-TRK, thereby effectively inhibiting the phosphorylation of p-PLCγ1/PLCγ1 and p-AKT/AKT to regulate cell growth and proliferation.

Experimental Example 5: In vivo efficacy study of compounds on NTRK mutation-resistant tumor models

Test methods

1.1 Model preparation

Cells in the logarithmic growth phase were collected and resuspended in serum-free medium to a cell concentration of 6×10 ⁷ -10×10 ⁷ /mL, and an equal volume of Matrigel was added to the cell suspension to a final cell concentration of 3×10 ⁷ -5×10 ⁷ /mL. 0.1 mL of tumor cell suspension was subcutaneously inoculated in the axilla of the forelimb of NuNu mice (Beijing Weitonglihua, 4-6 weeks, female) at an inoculation volume of 3×10 ⁶ -5×10 ⁶ /mouse to prepare an animal model.

1.2 Trial Grouping

The maximum and minimum tumor diameters of the nude mouse transplanted tumors were measured with a vernier caliper to calculate the tumor volume: the calculation formula for tumor volume (TV) is: V = 1/2 × a × b ² , where a and b represent the maximum and minimum diameters of the tumor mass, respectively. Nude mice with appropriate tumor volumes were selected and randomly divided into 7 groups (100-200 mm ³ ) according to the tumor volume, with 6 mice in each group.

1.3 Observation indicators

On the day of grouping, animals were gavaged and administered according to their body weight. The administration volume was 10 mL/kg. LOXO-195 was prepared into the required administration solution using 0.5% CMC-Na, and the compound represented by formula (A) was prepared into the required administration solution using "3% DMSO + 96% HP-β-CD (0.5 g/mL) + 1% HCl". Tumor diameter was measured twice a week and tumor volume was calculated. Specific indicators are as follows:

Animal body weight: Animals were weighed every morning before administration. A weight loss greater than 20% was defined as a toxic reaction to the drug (observed until the day after the last administration);

Tumor volume (TV) = V = 1/2 × a × b ² , where a and b represent the maximum and minimum diameters of the tumor, respectively (observed until the day after the last administration);

Relative tumor proliferation rate T/C (%): T/C (%) = TRTV/CRTV × 100% (TRTV: RTV of the treatment group, CRTV: RTV of the control group);

Tumor growth inhibition rate (TGI) = [1-(Ti-T0)/(Vi-V0)] × 100%. (Ti represents the average tumor volume of a certain dosing group on a certain day; T0 is the average tumor volume of this dosing group at the beginning of dosing; Vi is the average tumor volume of the vehicle control group on a certain day (the same day as Ti); V0 is the average tumor volume of the vehicle control group at the beginning of dosing);

Tumor inhibition rate: At the end of the experiment, the animals were killed by cervical dislocation, the tumor masses were removed and weighed, and photos were taken to calculate the tumor inhibition rate, tumor inhibition rate = (average tumor weight of the control group - average tumor weight of the drug group) / average tumor weight of the control group × 100%.

Test results

2.1Ba/F3 LMNA-NTRK1-G667C model

2.1.1 Effects of drugs on body weight of tumor-bearing mice

The body weight of each compound and each dose group has an upward trend, and the upward trend is more obvious than that of the control group. The body weight of each compound and each dose group increased significantly, which may be related to the compound, or it may be due to the inhibition of tumor growth, which makes the mice in better condition and has obvious weight gain. The results are shown in Table 17.

2.1.2 Effects of drugs on tumor weight and tumor inhibition rate in tumor-bearing mice

The data results show that: at the same dosage (100 mg/kg), the compound represented by formula (A) has a more significant inhibitory effect on tumor growth compared with LOXO-195; further, compared with the LOXO-195 group with a higher dosage (200 mg/kg), the compound represented by formula (A) (100 mg/kg) also exhibits a better tumor inhibition effect. The results are shown in Table 17.

Table 17 In vivo results of Ba/F3 LMNA-NTRK1-G667C model

2.2Ba/F3 LMNA-NTRK1-G595R model

2.2.1 Effects of drugs on body weight of tumor-bearing mice

The body weight of each compound and each dose group has an upward trend, and the upward trend is more obvious than that of the control group. The body weight of each compound and each dose group increased significantly, which may be related to the compound, or it may be due to the inhibition of tumor growth, which makes the mice in better condition and has obvious weight gain. The results are shown in Table 17.

2.2.2 Effects of drugs on tumor weight and tumor inhibition rate in tumor-bearing mice

The data results show that compared with LOXO-195 (100 mg/kg), the compound represented by formula (A) can achieve significant inhibition of tumor tissue weight at a lower dosage (50 mg/kg), with a tumor weight inhibition rate of >90%. The results are shown in Table 18.

Table 18 In vivo results of Ba/F3 LMNA-NTRK1-G595R model

Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be apparent to one skilled in the art from the teachings of this application that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims.

Claims (11)

1. A compound of formula (A) in solid form,

preferably, the compound represented by the formula (A) in solid form is characterized by comprising characteristic peaks (+ -4 cm) at the following positions by infrared spectrometry using a tabletting method ^-1 )：3429，1643，1488，1453，1232，1027；

Further preferred, the compound of formula (a) in solid form has an infrared spectrum substantially as shown in fig. 12.

2. A compound of formula (a) in crystalline form.

3. The compound of formula (a) in crystalline form according to claim 2, characterized in that it is in crystalline form I of the compound of formula (a), using Cu-ka radiation, having an X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2Θ angles: (±0.2°): 9.2 17.9, 18.5; alternatively, using Cu-ka radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks (±0.2°): 8.9,9.2, 17.9, 18.5, 23.8;

Alternatively, its X-ray powder diffraction pattern using Cu-ka radiation has characteristic diffraction peaks (±0.2°) at the following 2θ angles: 8.9,9.2, 10.1, 17.9, 18.5, 23.8, 28.0;

alternatively, using Cu-ka radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks (±0.2°): 8.9,9.2, 10.1, 16.1, 17.9, 18.5, 23.8, 27.0, 28.0;

alternatively, using Cu-ka radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks (±0.2°): 8.9,9.2, 10.1, 16.1, 16.6, 17.9, 18.5, 23.8, 27.0, 28.0;

alternatively, it has an X-ray powder diffraction pattern substantially as shown in figure 1.

4. The compound of formula (a) in crystalline form according to claim 2, characterized in that it is form II of the compound of formula (a), using Cu-ka radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks (±0.2°): 9.1 18.3, 20.6;

alternatively, using Cu-ka radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks (±0.2°): 6.4,9.1, 18.3, 20.6;

alternatively, using Cu-ka radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: (±0.2°): 6.4,9.1, 18.3, 20.6, 27.7;

Alternatively, using Cu-ka radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks (±0.2°): 6.4,9.1, 18.3, 18.9, 20.6, 27.7;

alternatively, using Cu-ka radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks (±0.2°): 6.4,9.1, 18.3, 18.9, 20.6, 22.4, 23.9, 27.7;

alternatively, it has an X-ray powder diffraction pattern substantially as shown in figure 2;

or, the differential scanning calorimetric curve has an endothermic peak at 155.27 +/-5 ℃;

alternatively, it has a DSC profile substantially as shown in figure 3.

5. The compound of formula (a) in crystalline form according to claim 2, characterized in that it is form III of the compound of formula (a), using Cu-ka radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks (±0.2°): 5.6 13.3, 15.8, 18.4;

alternatively, using Cu-ka radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks (±0.2°): 5.6 13.3, 15.8, 18.4, 23.7;

alternatively, using Cu-ka radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks (±0.2°): 5.6 10.9, 13.3, 15.8, 18.4, 19.0, 23.7;

Alternatively, using Cu-ka radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks (±0.2°): 5.6 10.9, 11.3, 13.3, 15.8, 17.1, 18.4, 19.0, 23.7;

alternatively, using Cu-ka radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks (±0.2°): 5.6,7.4, 10.9, 11.3, 13.3, 15.8, 17.1, 18.4, 19.0, 19.9, 23.7;

alternatively, it has an X-ray powder diffraction pattern substantially as shown in figure 4;

or, the differential scanning calorimetric curve has an endothermic peak at 182.29 +/-5 ℃;

alternatively, it has a DSC profile substantially as shown in figure 5.

6. The compound of formula (a) in crystalline form according to claim 2, characterized in that it is form IV of the compound of formula (a), using Cu-ka radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks (±0.2°): 8.5 10.0, 17.2, 17.9;

alternatively, using Cu-ka radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 8.5 10.0, 13.1, 17.2, 17.9, 25.0;

alternatively, using Cu-ka radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks (±0.2°): 8.5 10.0, 13.1, 17.2, 17.9, 20.0, 25.0;

Alternatively, using Cu-ka radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks (±0.2°): 8.5 10.0, 13.1, 15.7, 17.2, 17.9, 20.0, 25.0, 25.9;

alternatively, it has an X-ray powder diffraction pattern substantially as shown in figure 6 or figure 8.

Or, the differential scanning calorimetric curve has an endothermic peak at 192.07 +/-5 ℃;

alternatively, it has a DSC profile substantially as shown in figure 7.

7. The compound of formula (a) in crystalline form according to claim 2, characterized in that it is form V of the compound of formula (a), using Cu-ka radiation, having an X-ray powder diffraction pattern with characteristic diffraction peaks (±0.2°): 6.0 12.7, 18.2;

alternatively, using Cu-ka radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks (±0.2°): 6.0 12.7, 18.2, 23.4;

alternatively, using Cu-ka radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks (±0.2°): 6.0 12.7, 16.7, 18.2, 23.4;

alternatively, using Cu-ka radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks (±0.2°): 6.0 12.7, 16.7, 18.2, 23.4, 25.4, 28.1;

Alternatively, using Cu-ka radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks (±0.2°): 6.0,7.0, 12.7, 14.2, 16.7, 18.2, 23.4, 24.5, 25.4, 28.1;

alternatively, it has an X-ray powder diffraction pattern substantially as shown in figure 9.

8. The compound of formula (a) in crystalline form according to claim 2, characterized in that it is form VI of the compound of formula (a), using Cu-ka radiation, having an X-ray powder diffraction pattern with characteristic diffraction peaks (±0.2°): 6.1 16.9, 22.3;

alternatively, using Cu-ka radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks (±0.2°): 6.1 16.9, 18.6, 22.3;

alternatively, using Cu-ka radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks (±0.2°): 6.1,7.4, 11.5, 16.9, 18.6, 22.3;

alternatively, using Cu-ka radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks (±0.2°): 6.1,7.4, 11.5, 16.9, 18.6, 19.4, 22.3, 25.3;

alternatively, using Cu-ka radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks (±0.2°): 6.1,7.4, 11.5, 16.9, 18.6, 19.4, 20.1, 22.3, 25.3, 28.6;

Alternatively, it has an X-ray powder diffraction pattern substantially as shown in figure 10.

9. A crystalline composition comprising one or more of form I of claim 3, form II of claim 4, form III of claim 5, form IV of claim 6, form V of claim 7, form VI of claim 8; preferably, the form II comprises more than 50% by weight of the crystalline composition; preferably 60% or more; more preferably 70% or more; more preferably 80% or more; more preferably 90% or more; more preferably 95% or more;

or, the crystalline form III comprises more than 50% by weight of the crystalline composition; preferably 60% or more; more preferably 70% or more; more preferably 80% or more; more preferably 90% or more; more preferably 95% or more;

or, the form IV comprises more than 50% by weight of the crystalline composition; preferably 60% or more; more preferably 70% or more; more preferably 80% or more; more preferably 90% or more; more preferably 95% or more;

or the crystalline form VI comprises more than 50% by weight of the crystalline composition; preferably 60% or more; more preferably 70% or more; more preferably 80% or more; more preferably 90% or more; more preferably 95% or more.

10. A pharmaceutical composition comprising a compound of formula (a) according to claim 1 in solid form, or a compound of formula (a) according to claim 2 in crystalline form, or a crystalline composition according to claim 9;

preferably, the pharmaceutical composition comprises one or more of form I of claim 3, form II of claim 4, form III of claim 5, form IV of claim 6, form V of claim 7, form VI of claim 8.

11. Use of a compound of formula (a) according to claim 1 in solid form, a compound of formula (a) according to any one of claims 2-8 in crystalline form, a crystalline composition according to claim 9, or a pharmaceutical composition according to claim 10 as a medicament or in the preparation of a medicament; preferably, the medicament is for the prevention and/or treatment of one or more of TRK, ROS or ALK mediated diseases; further preferably, the disease is selected from pain diseases, cell proliferative diseases, inflammatory diseases, neurodegenerative diseases or infectious diseases; further preferably, the cell proliferative disease is a tumor or cancer; further preferred, the tumor or cancer is a solid tumor or hematological tumor; further preferred, the tumor or cancer is hematological malignancy, lung cancer, breast cancer, ovarian cancer, prostate cancer, pancreatic cancer, glioma, colorectal cancer, melanoma, cancer of the head and neck, gall bladder cancer, thyroid cancer, glioblastoma, gastric cancer, neuroblastoma, or salivary gland cancer; preferably, the lung cancer is non-small cell lung cancer.

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