Crystallization of pyrimidine compounds

Abstract

The present invention provides a crystal of 7-((3R, 5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (compound (I)), and a crystal thereof with an acid (a salt crystal or a co-crystal). A type II crystal of compound (I) and 1 equivalent of fumaric acid has characteristic peaks at three or more selected from the group consisting of 5.5°, 6.8°, 9.3°, 13.4°, 15.3°, 16.3°, 18.5°, 19.8°, 22.0° and 24.5° in a powder X-ray diffraction pattern. The (free form) type II crystal of compound (I) has characteristic peaks at three or more selected from 8.3°, 14.8°, 17.3°, 18.0°, 19.1°, 20.3°, 21.0°, 22.5°, 23.0° and 26.2° in the powder X-ray diffraction pattern. The (free form) type I crystal of compound (I) has characteristic peaks at three or more selected from 9.9°, 11.7°, 13.2°, 17.7°, 18.1°, 18.8° and 20.8° in the powder X-ray diffraction pattern. A V-type crystalline powder of compound (I) and 1 equivalent of fumaric acid, which has characteristic peaks at 4 or more selected from 6.9°, 9.4°, 10.2°, 13.7°, 21.1°, 23.6° and 26.5° in the X-ray diffraction spectrum. A I-type crystalline powder of compound (I) and 1 equivalent of fumaric acid, which has characteristic peaks at 4 or more selected from 6.4°, 10.3°, 12.8°, 15.0°, 20.7°, 23.4° and 26.6° in the X-ray diffraction spectrum.

Description

Technical Field

[0001] The present invention relates to a crystal of a pyrimidine compound useful as an antitumor agent and a pharmaceutical composition containing the crystal. Background Art

[0002] HER2 (also known as ErbB2) is a receptor-type tyrosine kinase belonging to the ErbB family.

[0003] HER2 is considered to be an oncogene, and HER2 gene amplification, overexpression, mutation, etc. have been reported in various cancers. From non-clinical and clinical research data, it is known that in cancer cells accompanied by these HER2 gene abnormalities, overexpression, etc., HER2 and downstream signal activation play an important role in the survival and proliferation of cancer cells (Non-Patent Document 1).

[0004] Therefore, it is assumed that inhibitors that can control the kinase activity of HER2 exert anti-tumor effects by inhibiting HER2 and downstream signaling in cancer cells with gene amplification, overexpression, or mutation of HER2, and are therefore considered to be useful in the treatment of cancer patients, and in prolonging their lifespan and improving their QOL.

[0005] It is reported that brain metastasis occurs in about 25-40% of lung cancers, about 15-30% of breast cancers, and a certain proportion of other cancers (Non-Patent Documents 2 and 3). In fact, it is reported that brain metastasis occurs in about 20-30% of HER2-positive breast cancers (Non-Patent Document 4).

[0006] As compounds with HER2 inhibitory activity, lapatinib, neratinib, etc. have been approved as therapeutic agents for HER2-positive breast cancer. However, it has been reported that brain metastasis is limited in non-clinical trials because both are substrates of p-gp and Bcrp (Non-patent Document 5). In fact, in clinical trials of lapatinib and neratinib, no sufficient effect on brain metastasis was obtained (Non-patent Documents 6, 7, 8, 9).

[0007] From the viewpoint of controlling the disease state including brain metastases, a HER2 inhibitor that has inhibitory activity against HER2 and also has brain metastasis properties is desired.

[0008] In addition, when a compound is used as an effective active ingredient of a pharmaceutical product, chemical and physical stability of the compound is required in order to stably maintain the quality and / or facilitate storage management. Therefore, a HER2 inhibitor that has chemical and physical stability, inhibitory activity against HER2, and brain metastasis is desired.

[0009] Prior art literature

[0010] Non-patent literature

[0011] Non-patent literature 1: Cancer Treatment Reviews, 40, p.770-780 (2014)

[0012] Non-patent document 2: Current Oncology, 25, p.S103-S114 (2018)

[0013] Non-patent document 3: Breast Cancer Research, 18(1), 8, p.1-9 (2016)

[0014] Non-patent document 4: Journal of Clinical Oncology, 28, p3271-3277 (2010)

[0015] Non-patent document 5: Journal of Medicinal Chemistry, 59, p10030-10066 (2016)

[0016] Non-patent document 6: Journal of Medicinal Chemistry, 26, p2999-3005 (2008)

[0017] Non-patent document 7: Journal of Clinical Oncology, 26, p1993-1999 (2008)

[0018] Non-patent document 8: Journal of Clinical Oncology, 28, p1301-1307 (2010)

[0019] Non-patent document 9: Journal of Clinical Oncology, 34, p945-952 (2016) Summary of the invention

[0020] The present invention aims to provide a HER2 inhibitor that is chemically and physically stable, has inhibitory activity against HER2, and also has brain metastasis. In particular, the present invention aims to provide a HER2 inhibitor crystal that is excellent in stability, has good oral absorbability, and can be obtained with good reproducibility.

[0021] The inventors of the present invention have conducted intensive studies to solve the above-mentioned problems, and as a result, found that 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (hereinafter also referred to as "compound (I)") represented by the following formula (I) has HER2 inhibitory activity and brain metastasis, and can be used as a therapeutic agent for diseases (particularly malignant tumors) involved in HER2 by inhibiting HER2.

[0022]

[0023] Furthermore, the inventors of the present invention have discovered two crystal forms (Type I crystal and Type II crystal) as free forms in Compound (I), and as crystals with acids (crystals of salt forms or co-crystals), crystals with hydrochloric acid (Type I crystal with hydrochloric acid, Type II crystal with hydrochloric acid, Type III crystal with hydrochloric acid), crystals with hydrobromic acid, crystals with 1 equivalent of tartaric acid, crystals with fumaric acid (Type I crystal with 0.5 equivalent of fumaric acid, Type II crystal with 0.5 equivalent of fumaric acid, Type I crystal with 1 equivalent of fumaric acid, Type II crystal with 1 equivalent of fumaric acid, Type III crystal with 1 equivalent of fumaric acid, Type IV crystal with 1 equivalent of fumaric acid, Type V crystal with 1 equivalent of fumaric acid), and crystals with succinic acid.

[0024] It was discovered that among its crystal forms, free form type I crystal, free form type II crystal, type II crystal with 1 equivalent of fumaric acid, and type V crystal with 1 equivalent of fumaric acid respectively have non-hygroscopicity and excellent solid stability, which are beneficial properties in the manufacture of pharmaceuticals. It was also discovered that they have excellent oral absorbability, etc., thereby completing the present invention.

[0025] That is, the present invention includes the following embodiments.

[0026] [1] Type II crystals of 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide and fumaric acid, wherein in a powder X-ray diffraction pattern, the diffraction angle (2θ±0.2°) is selected from 5.5°, 6.8°, 9.3°, 13. There are peaks at three or more of 4°, 15.3°, 16.3°, 18.5°, 19.8°, 22.0° and 24.5°, and the molar ratio of 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide to fumaric acid is 1:1.

[0027] [2] The crystals as described in [1], which have peaks at diffraction angles (2θ±0.2°) of 5.5°, 6.8°, 9.3°, 13.4°, 15.3°, 16.3°, 18.5°, 19.8°, 22.0° and 24.5° in the powder X-ray diffraction pattern.

[0028] [3] The crystal according to [1] or [2], which has Figure 1 Shown is the powder X-ray diffraction pattern of the crystals.

[0029] [4] The crystal according to any one of [1] to [3], wherein the endothermic peak determined by simultaneous differential thermal and thermogravimetric measurement is around 178°C.

[0030] [5] The crystal according to any one of [1] to [4], wherein the purity of the crystal is 90% by mass or more.

[0031] [6] Type II crystals of 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide, which has peaks at three or more diffraction angles (2θ±0.2°) selected from the group consisting of 8.3°, 14.8°, 17.3°, 18.0°, 19.1°, 20.3°, 21.0°, 22.5°, 23.0° and 26.2° in its powder X-ray diffraction pattern.

[0032] [7] The crystals as described in [6], which have peaks at diffraction angles (2θ±0.2°) of 8.3°, 14.8°, 17.3°, 18.0°, 19.1°, 20.3°, 21.0°, 22.5°, 23.0° and 26.2° in the powder X-ray diffraction pattern.

[0033] [8] The crystal according to [6] or [7], which has Figure 3 Shown is the powder X-ray diffraction pattern of the crystals.

[0034] [9] The crystal according to any one of [6] to [8], wherein the endothermic peak determined by simultaneous differential thermal and thermogravimetric measurement is near 182°C.

[0035]

[10] The crystal according to any one of [6] to [9], wherein the purity of the crystal is 90% by mass or more.

[0036]

[11] Type I crystal of 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide, which has peaks at four or more diffraction angles (2θ±0.2°) selected from the group consisting of 9.9°, 11.7°, 13.2°, 17.7°, 18.1°, 18.8° and 20.8° in its powder X-ray diffraction pattern.

[0037]

[12] The crystals according to

[11] , which have peaks at diffraction angles (2θ±0.2°) of 9.9°, 11.7°, 13.2°, 17.7°, 18.1°, 18.8° and 20.8° in a powder X-ray diffraction pattern.

[0038]

[13] The crystal according to

[11] or

[12] , which has Figure 5 Shown is the powder X-ray diffraction pattern of the crystals.

[0039]

[14] The crystal according to any one of

[11] to

[13] , wherein the endothermic peak determined by simultaneous differential thermal and thermogravimetric measurement is near 183°C.

[0040]

[15] The crystal according to any one of

[11] to

[14] , wherein the purity of the crystal is 90% by mass or more.

[0041]

[16] V-type crystals of 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide and fumaric acid, wherein in the powder X-ray diffraction pattern, the diffraction angle (2θ±0.2°) is selected from 6.9°, 9.4° , 10.2°, 13.7°, 21.1°, 23.6° and 26.5°, and the molar ratio of 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide to fumaric acid is 1:1.

[0042]

[17] The crystals described in

[16] , which have peaks at diffraction angles (2θ±0.2°) of 6.9°, 9.4°, 10.2°, 13.7°, 21.1°, 23.6° and 26.5° in the powder X-ray diffraction pattern.

[0043]

[18] The crystal according to

[16] or

[17] , which has Figure 7 Shown is the powder X-ray diffraction pattern of the crystals.

[0044]

[19] The crystal according to any one of

[16] to

[18] , wherein the endothermic peak determined by simultaneous differential thermal and thermogravimetric measurement is near 151°C.

[0045]

[20] The crystal according to any one of

[16] to

[19] , wherein the purity of the crystal is 90% by mass or more.

[0046]

[21] Type I crystals of 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide and fumaric acid, wherein in a powder X-ray diffraction pattern, a diffraction angle (2θ±0.2°) selected from 6.4°, 10.3° , 12.8°, 15.0°, 20.7°, 23.4° and 26.6°, and the molar ratio of 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide to fumaric acid is 1:1.

[0047]

[22] The crystals described in

[21] , which have peaks at diffraction angles (2θ±0.2°) of 6.4°, 10.3°, 12.8°, 15.0°, 20.7°, 23.4° and 26.6° in the powder X-ray diffraction pattern.

[0048]

[23] The crystal according to

[21] or

[22] , which has Fig. 9 Shown is the powder X-ray diffraction pattern of the crystals.

[0049]

[24] The crystal according to any one of

[21] to

[23] , wherein the endothermic peak determined by simultaneous differential thermal and thermogravimetric measurement is near 153°C.

[0050]

[25] The crystal according to any one of

[21] to

[24] , wherein the purity of the crystal is 90% by mass or more.

[0051]

[26] Type I crystals of 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide and fumaric acid, wherein in a powder X-ray diffraction pattern, at a diffraction angle (2θ±0.2°) selected from 6.4°, 7.5°, There were peaks at four or more of 9.7°, 11.7°, 15.1°, 19.6° and 23.9°, and the molar ratio of 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide to fumaric acid was 1:0.5.

[0052]

[27] The crystals described in

[26] , which have peaks at diffraction angles (2θ±0.2°) of 6.4°, 7.5°, 9.7°, 11.7°, 15.1°, 19.6° and 23.9° in the powder X-ray diffraction pattern.

[0053]

[28] The crystal according to

[26] or

[27] , which has Fig.11 Shown is the powder X-ray diffraction pattern of the crystals.

[0054]

[29] The crystal according to any one of

[26] to

[28] , wherein the endothermic peak determined by simultaneous differential thermal and thermogravimetric measurement is near 161°C.

[0055]

[30] The crystal according to any one of

[26] to

[29] , wherein the purity of the crystal is 90% by mass or more.

[0056]

[31] Type II crystals of 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide and fumaric acid, wherein in the powder X-ray diffraction pattern, the diffraction angle (2θ±0.2°) is selected from 5.4°, 6.4° , 7.3°, 12.8°, 13.4°, 14.7° and 15.4°, and the molar ratio of 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide to fumaric acid is 1:0.5.

[0057]

[32] The crystals described in

[31] , which have peaks at diffraction angles (2θ±0.2°) of 5.4°, 6.4°, 7.3°, 12.8°, 13.4°, 14.7° and 15.4° in the powder X-ray diffraction pattern.

[0058]

[33] The crystal according to

[31] or

[32] , which has Fig.13 Shown is the powder X-ray diffraction pattern of the crystals.

[0059]

[34] The crystal according to any one of

[31] to

[33] , wherein the endothermic peak determined by simultaneous differential thermal and thermogravimetric measurement is near 162°C.

[0060]

[35] The crystal according to any one of

[31] to

[34] , wherein the purity of the crystal is 90% by mass or more.

[0061]

[36] A pharmaceutical composition comprising the crystal according to any one of [1] to

[35] .

[0062]

[37] A pharmaceutical composition for oral administration comprising the crystal according to any one of [1] to

[35] .

[0063]

[38] An antitumor agent comprising the crystal according to any one of [1] to

[35] .

[0064]

[39] The crystal according to any one of [1] to

[35] , which is used as a medicine.

[0065]

[40] Use of the crystal according to any one of [1] to

[35] for producing an antitumor agent for oral administration.

[0066]

[41] The crystal according to any one of [1] to

[35] , which is used for treating tumors.

[0067]

[42] The crystal according to any one of [1] to

[35] , which is used for oral administration to treat tumors.

[0068]

[43] A method for treating a tumor, comprising the step of administering an effective amount of the crystal according to any one of [1] to

[35] to a subject in need of treatment.

[0069]

[44] Use of the crystal according to any one of [1] to

[35] for producing a pharmaceutical composition.

[0070]

[45] Use of the crystal according to any one of [1] to

[35] for producing an antitumor agent.

[0071] The free form I crystal, free form II crystal, form II crystal with 1 equivalent of fumaric acid, and form V crystal with 1 equivalent of fumaric acid of the compound (I) 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide of the present invention are superior to other crystal forms from the viewpoints of high stability, handling properties (low hygroscopicity), quality control properties, etc. Furthermore, the free form II crystal and form II crystal with 1 equivalent of fumaric acid are excellent in oral absorption and are useful forms when the compound is used as a raw material for pharmaceuticals. In addition, the type I crystal with 1 equivalent of fumaric acid can be separated from acetone (solvent) as a solid (after separation, it is dried to form type V crystal), and therefore, it can also be used as an intermediate for the production of type V crystal with 1 equivalent of fumaric acid.

[0072] According to the present invention, there are provided novel crystals of Compound (I) or novel crystals of Compound (I) and an acid, pharmaceutical compositions containing these crystals, antitumor agents, or antitumor agents for oral administration.

[0073] In a preferred embodiment, compound (I) has at least one of the properties that are advantageous in pharmaceutical manufacturing, such as non-hygroscopicity, acquisition reproducibility, solid stability, and oral absorbability, as well as type II crystals of compound (I) with 1 equivalent of fumaric acid, free type II crystals, free type I crystals, type V crystals with 1 equivalent of fumaric acid, type I crystals with 0.5 equivalents of fumaric acid, and type II crystals with 0.5 equivalents of fumaric acid. BRIEF DESCRIPTION OF THE DRAWINGS

[0074] Figure 1 The following is a powder X-ray diffraction pattern of type II crystals of compound (I) obtained in Example 1 and 1 equivalent of fumaric acid (the vertical axis represents intensity (counts) and the horizontal axis represents diffraction angle (2θ)).

[0075] Figure 2 The results of simultaneous differential thermal and thermogravimetric measurement (TG-DTA) of compound (I) obtained in Example 1 and type II crystals of 1 equivalent of fumaric acid are shown (the left vertical axis represents the weight (mg) in the TG curve, the right vertical axis represents the heat flux (μV) in the DTA curve, and the horizontal axis represents the temperature (°C)).

[0076] Figure 3 The powder X-ray diffraction pattern of the type II crystal of compound (I) obtained in Example 2 is shown (the vertical axis represents the intensity (counts), and the horizontal axis represents the diffraction angle (2θ)).

[0077] Figure 4The results of differential thermal-thermogravimetric simultaneous measurement (TG-DTA) of type II crystals of compound (I) obtained in Example 2 are shown (the left vertical axis represents the weight (mg) in the TG curve, the right vertical axis represents the heat flux (μV) in the DTA curve, and the horizontal axis represents the temperature (°C)).

[0078] Figure 5 The X-ray powder diffraction pattern of the type I crystal of compound (I) obtained in Example 3 is shown (the vertical axis represents the intensity (counts), and the horizontal axis represents the diffraction angle (2θ)).

[0079] Figure 6 The results of differential thermal-thermogravimetric simultaneous measurement (TG-DTA) of type I crystals of compound (I) obtained in Example 3 are shown (the left vertical axis represents the weight (mg) in the TG curve, the right vertical axis represents the heat flux (μV) in the DTA curve, and the horizontal axis represents the temperature (°C)).

[0080] Figure 7 The X-ray powder diffraction pattern of the V-type crystal of the compound (I) obtained in Example 4 and 1 equivalent of fumaric acid is shown (the vertical axis represents the intensity (counts), and the horizontal axis represents the diffraction angle (2θ)).

[0081] Figure 8 The results of simultaneous differential thermal and thermogravimetric measurement (TG-DTA) of the compound (I) obtained in Example 4 and 1 equivalent of fumaric acid in type V crystals are shown (the left vertical axis represents the weight (mg) in the TG curve, the right vertical axis represents the heat flux (μV) in the DTA curve, and the horizontal axis represents the temperature (°C)).

[0082] Fig. 9 The powder X-ray diffraction pattern of type I crystal of compound (I) obtained in Example 5 and 1 equivalent of fumaric acid is shown (the vertical axis represents intensity (counts), and the horizontal axis represents diffraction angle (2θ)).

[0083] Fig.10 The results of simultaneous differential thermal-thermogravimetric measurement (TG-DTA) of compound (I) obtained in Example 5 and type I crystals of 1 equivalent of fumaric acid are shown (the left vertical axis represents the weight (mg) in the TG curve, the right vertical axis represents the heat flux (μV) in the DTA curve, and the horizontal axis represents the temperature (°C)).

[0084] Fig.11 The X-ray powder diffraction pattern of type I crystal of compound (I) obtained in Example 6 and 0.5 equivalent of fumaric acid is shown (the vertical axis represents intensity (counts), and the horizontal axis represents diffraction angle (2θ)).

[0085] Fig.12The results of simultaneous differential thermal-thermogravimetric measurement (TG-DTA) of compound (I) obtained in Example 6 and type I crystals of 0.5 equivalent of fumaric acid are shown (the left vertical axis represents the weight (mg) in the TG curve, the right vertical axis represents the heat flux (μV) in the DTA curve, and the horizontal axis represents the temperature (°C)).

[0086] Fig.13 The powder X-ray diffraction pattern of type II crystals of compound (I) obtained in Example 7 and 0.5 equivalent of fumaric acid is shown (the vertical axis represents intensity (counts), and the horizontal axis represents diffraction angle (2θ)).

[0087] Fig.14 The results of simultaneous differential thermal-thermogravimetric measurement (TG-DTA) of compound (I) obtained in Example 7 and type II crystals of 0.5 equivalent of fumaric acid are shown (the left vertical axis represents the weight (mg) in the TG curve, the right vertical axis represents the heat flux (μV) in the DTA curve, and the horizontal axis represents the temperature (°C)).

[0088] Fig.15 The powder X-ray diffraction pattern of the type III crystal of compound (I) obtained in Reference Example 1 and 1 equivalent of fumaric acid is shown (the vertical axis represents the intensity (counts), and the horizontal axis represents the diffraction angle (2θ)).

[0089] Fig.16 The results of simultaneous differential thermal-thermogravimetric measurement (TG-DTA) of compound (I) obtained in Reference Example 1 and type III crystals of 1 equivalent of fumaric acid are shown (the left vertical axis represents the weight (mg) in the TG curve, the right vertical axis represents the heat flux (μV) in the DTA curve, and the horizontal axis represents the temperature (°C)).

[0090] Fig.17 The X-ray powder diffraction pattern of the type IV crystal of compound (I) obtained in Reference Example 2 and 1 equivalent of fumaric acid is shown (the vertical axis represents the intensity (counts), and the horizontal axis represents the diffraction angle (2θ)).

[0091] Fig.18 The results of simultaneous differential thermal and thermogravimetric measurement (TG-DTA) of compound (I) obtained in Reference Example 2 and type IV crystals of 1 equivalent of fumaric acid are shown (the left vertical axis represents the weight (mg) in the TG curve, the right vertical axis represents the heat flux (μV) in the DTA curve, and the horizontal axis represents the temperature (°C)).

[0092] Fig.19 The X-ray powder diffraction pattern of the type I crystal of compound (I) obtained in Reference Example 3 and hydrochloric acid is shown (the vertical axis represents the intensity (counts), and the horizontal axis represents the diffraction angle (2θ)).

[0093] Fig. 20The results of simultaneous differential thermal and thermogravimetric measurement (TG-DTA) of compound (I) obtained in Reference Example 3 and type I crystals of hydrochloric acid are shown (the left vertical axis represents the weight (mg) in the TG curve, the right vertical axis represents the heat flux (μV) in the DTA curve, and the horizontal axis represents the temperature (°C)).

[0094] Fig.21 The powder X-ray diffraction pattern of the type II crystal of compound (I) obtained in Reference Example 4 and hydrochloric acid is shown (the vertical axis represents the intensity (counts), and the horizontal axis represents the diffraction angle (2θ)).

[0095] Fig. 22 The results of simultaneous differential thermal-thermogravimetric measurement (TG-DTA) of compound (I) obtained in Reference Example 4 and type II crystals of hydrochloric acid are shown (the left vertical axis represents the weight (mg) in the TG curve, the right vertical axis represents the heat flux (μV) in the DTA curve, and the horizontal axis represents the temperature (°C)).

[0096] Fig.23 The powder X-ray diffraction pattern of the type III crystal of compound (I) obtained in Reference Example 5 and hydrochloric acid is shown (the vertical axis represents the intensity (counts), and the horizontal axis represents the diffraction angle (2θ)).

[0097] Fig.24 The results of simultaneous differential thermal and thermogravimetric measurement (TG-DTA) of compound (I) obtained in Reference Example 5 and type III crystals of hydrochloric acid are shown (the left vertical axis represents the weight (mg) in the TG curve, the right vertical axis represents the heat flux (μV) in the DTA curve, and the horizontal axis represents the temperature (°C)).

[0098] Fig.25 The powder X-ray diffraction pattern of the crystals of compound (I) obtained in Reference Example 6 and hydrobromic acid is shown (the vertical axis represents the intensity (counts), and the horizontal axis represents the diffraction angle (2θ)).

[0099] Fig.26 The results of differential thermal-thermogravimetric simultaneous measurement (TG-DTA) of the crystals of compound (I) obtained in Reference Example 6 and hydrobromic acid are shown (the left vertical axis represents the weight (mg) in the TG curve, the right vertical axis represents the heat flux (μV) in the DTA curve, and the horizontal axis represents the temperature (°C)).

[0100] Fig. 27 The powder X-ray diffraction pattern of the crystals of Compound (I) obtained in Reference Example 7 and 1 equivalent of L-tartaric acid is shown (the vertical axis represents the intensity (counts), and the horizontal axis represents the diffraction angle (2θ)).

[0101] Fig.28The results of simultaneous differential thermal and thermogravimetric measurement (TG-DTA) of the crystals of compound (I) obtained in Reference Example 7 and 1 equivalent of L-tartaric acid are shown (the left vertical axis represents the weight (mg) in the TG curve, the right vertical axis represents the heat flux (μV) in the DTA curve, and the horizontal axis represents the temperature (°C)).

[0102] Fig.29 The powder X-ray diffraction pattern of the crystals of compound (I) and succinic acid obtained in Reference Example 8 is shown (the vertical axis represents the intensity (counts), and the horizontal axis represents the diffraction angle (2θ)).

[0103] Fig.30 The results of simultaneous differential thermal and thermogravimetric measurement (TG-DTA) of the crystals of compound (I) obtained in Reference Example 8 and succinic acid are shown (the left vertical axis represents the weight (mg) in the TG curve, the right vertical axis represents the heat flux (μV) in the DTA curve, and the horizontal axis represents the temperature (°C)).

[0104] Fig.31 The present invention shows the antitumor effect of compound (I) on the brain direct transplantation model of HER2-expressing cell line (NCI-N87-luc) into which luciferase gene is introduced.

[0105] Fig.32 The results show the effect of compound (I) on the body weight gain and loss rate in the brain direct transplantation model of HER2-expressing cell line (NCI-N87-luc) into which the luciferase gene was introduced. DETAILED DESCRIPTION

[0106] The present invention relates to a crystal of 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide represented by the following formula (I), and a crystal thereof with an acid (a crystal of a salt thereof or a co-crystal).

[0107]

[0108] Specifically, it relates to type II crystals of compound (I) with 1 equivalent of fumaric acid, type II crystals of free form, type I crystals of free form, type V crystals with 1 equivalent of fumaric acid, type I crystals with 1 equivalent of fumaric acid, type I crystals with 0.5 equivalents of fumaric acid, and type II crystals with 0.5 equivalents of fumaric acid.

[0109] In the present specification, the descriptions of Type I, Type II, Type III, Type IV, and Type V are for the convenience of distinguishing the names of crystal forms, and the crystals according to the present invention are not limited to these names.

[0110] In the present specification, the crystal of 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (Compound (I)) and an acid means a crystal of a salt or a co-crystal with an acid.

[0111] The crystal of salt is a crystal formed by the combination of compound (I) and acid molecules through ionic bonds, and the co-crystal is a crystal formed by the combination of compound (I) and acid molecules through non-ionic interactions. In the present invention, the crystal of compound (I) and acid can be a crystal of salt or a co-crystal, including both meanings. For example, if it is a type II crystal of 7-((3R, 5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide and fumaric acid, it means 7-((3R, 5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide and fumaric acid. The term "cocrystal" means a crystal of a fumarate salt of 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide or a cocrystal of 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide and fumaric acid.

[0112] Crystallization refers to a solid in which atoms or molecules are arranged in a regular repeating structure, which is different from an amorphous (non-crystalline) solid that does not have a repeating structure. Crystalline or amorphous solids can be studied by methods such as powder X-ray diffraction measurement (XRD measurement), differential scanning calorimetry (DSC measurement), thermogravimetric-differential thermal analysis (TG-DTA measurement), and single crystal analysis. Polymorphism refers to the same molecule but with different arrangements of atoms or molecules in the crystal. It is known that the peaks obtained by XRD measurement are different depending on the polymorph. In addition, it is known that solubility, oral absorbability, stability, etc. are different between each polymorph.

[0113] In this specification, the terms "crystalline" and "amorphous" are used with ordinary meanings.

[0114] When simply referred to as “Compound (I)” in the present specification, it means 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide, and is used to include both “amorphous” and “crystalline”.

[0115] In the present specification, the term "crystals of Compound (I)" is used to include both crystals of the free form of Compound (I) and crystals of Compound (I) and an acid (crystals of a salt of Compound (I) and a co-crystal of Compound (I)).

[0116] In the present specification, the crystals of molecules other than compound (I) constituting the crystals (salts or other molecules constituting the cocrystal) are not defined as crystals of the free form of 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (compound (I)). When equivalent is used in the present specification, it means molar equivalent.

[0117] In the crystals of compound (I) and an acid (crystals of a salt or a co-crystal), the equivalent of the acid relative to compound (I) can be analyzed by, for example, NMR or ion chromatography.

[0118] In the present specification, the crystal may be a hydrate.

[0119] Furthermore, a labeled form of Compound (I) or a salt thereof, that is, a compound in which one or more atoms of Compound (I) or a salt of Compound (I) are replaced with a radioactive isotope or a non-radioactive isotope, is also encompassed by the present invention.

[0120] The crystal may be a single crystal or a polymorphic mixture of crystals of other compound (I) as long as it contains compound (I). Specifically, it is preferred that the purity of the crystal is 50% by weight or more, i.e., 50% by weight or more is a single crystal, it is preferred that the purity of the crystal is 75% by weight or more, i.e., 75% by weight or more is a single crystal, it is more preferred that the purity of the crystal is 90% by weight or more, i.e., 90% by weight or more is a single crystal, it is further preferred that the purity of the crystal is 95% by weight or more, i.e., 95% by weight or more is a single crystal, it is particularly preferred that the purity of the crystal is 99% by weight or more, i.e., 99% by weight or more is a single crystal.

[0121] In this specification, chemical purity refers to the purity when measured by high performance liquid chromatography (HPLC). When it is recorded as the chemical purity of compound (I), it refers to the purity when compound (I) is measured by high performance liquid chromatography. In this case, the wavelength of the detector used in the purity measurement can be appropriately set. Specifically, the chemical purity of the crystal of compound (I) is preferably 95% or more, more preferably 98% or more, and particularly preferably 99% or more.

[0122] In addition, when determining the identity of crystals based on the properties of the data, the diffraction angle and the overall pattern are important. The relative intensity of the powder X-ray diffraction pattern will vary slightly depending on the direction of crystal growth, particle size, and measurement conditions, so it should not be strictly understood.

[0123] The values ​​obtained from various patterns may have slight errors depending on the direction of crystal growth, particle size, measurement conditions, etc. Therefore, in this specification, the values ​​of the diffraction angle (2θ) in the powder X-ray diffraction pattern may have a measurement error of about ±0.2°.

[0124] In the present specification, "room temperature" generally means about 10°C to about 35°C.

[0125] In addition, the endothermic peak in the differential thermal and thermogravimetric simultaneous measurement (TG-DTA) curve sometimes changes in the measurement temperature depending on the temperature rise amplitude per minute, the chemical purity of the sample, etc., and is usually ±5.0°C. Therefore, when the crystals involved in the present invention are measured by TG-DTA, the error of the peak (peak value) is considered to be ±5.0°C. The term "near" used here means 5.0°C.

[0126] One embodiment of the present invention relates to crystals of a free form of compound (I) (Type I crystal, Type II crystal), crystals of compound (I) and 1 equivalent of fumaric acid (Type I crystal, Type II crystal, Type III crystal, Type IV crystal, Type V crystal), and crystals of compound (I) and 0.5 equivalent of fumaric acid (Type I crystal, Type II crystal).

[0127] (Type II crystal of compound (I) and 1 equivalent of fumaric acid)

[0128] The type II crystal of compound (I) and 1 equivalent of fumaric acid has Figure 1 The powder X-ray diffraction pattern shown in Figure 2 The differential thermal - thermogravimetric measurement (TG-DTA) curve is shown.

[0129] In one embodiment of the present invention, the type II crystal of compound (I) and 1 equivalent of fumaric acid has the diffraction angle (2θ) and intensity (cts) shown in Table 2 in powder X-ray diffraction.

[0130] Among them, the characteristic peaks in the powder X-ray diffraction pattern of type II crystals of compound (I) and 1 equivalent of fumaric acid can be listed as diffraction angles (2θ±0.2°) of 5.5°, 6.8°, 9.3°, 13.4°, 15.3°, 16.3°, 18.5°, 19.8°, 22.0° and 24.5°.

[0131] The type II crystal of compound (I) and 1 equivalent of fumaric acid involved in the present invention is a crystal having 3 or more peaks selected from the above-mentioned characteristic peaks, preferably a crystal having 4 or more peaks selected from the above-mentioned characteristic peaks, more preferably a crystal having 5 or more peaks selected from the above-mentioned characteristic peaks, even more preferably a crystal having 6 or more peaks selected from the above-mentioned characteristic peaks, even more preferably a crystal having 7 or more peaks selected from the above-mentioned characteristic peaks, further preferably a crystal having 8 or more peaks selected from the above-mentioned characteristic peaks, further preferably a crystal having 9 or more peaks selected from the above-mentioned characteristic peaks, and particularly preferably a crystal having all of the above-mentioned peaks.

[0132] The endothermic peak (peak top value) of the type II crystal of compound (I) and 1 equivalent of fumaric acid determined by simultaneous differential thermal-thermogravimetric measurement can be 173 to 183°C, in other words, around 178°C.

[0133] (Free form II crystal of compound (I))

[0134] The type II crystal of compound (I) has Figure 3 The powder X-ray diffraction pattern shown in Figure 4 The differential thermal - thermogravimetric measurement (TG-DTA) curve is shown.

[0135] In one embodiment of the present invention, the type II crystal of the free form of compound (I) has the diffraction angle (2θ) and intensity (cts) shown in Table 3 in powder X-ray diffraction.

[0136] Among them, regarding the characteristic peaks in the powder X-ray diffraction pattern of type II crystal of compound (I), as diffraction angles (2θ±0.2°), 8.3°, 14.8°, 17.3°, 18.0°, 19.1°, 20.3°, 21.0°, 22.5°, 23.0° and 26.2° can be listed.

[0137] The type II crystal of compound (I) involved in the present invention is a crystal having 3 or more peaks selected from the above-mentioned characteristic peaks, preferably a crystal having 4 or more peaks selected from the above-mentioned characteristic peaks, more preferably a crystal having 5 or more peaks selected from the above-mentioned characteristic peaks, more preferably a crystal having 6 or more peaks selected from the above-mentioned characteristic peaks, more preferably a crystal having 7 or more peaks selected from the above-mentioned characteristic peaks, further preferably a crystal having 8 or more peaks selected from the above-mentioned characteristic peaks, further preferably a crystal having 9 or more peaks selected from the above-mentioned characteristic peaks, and particularly preferably a crystal having all of the above-mentioned characteristic peaks.

[0138] The endothermic peak (peak top value) of the type II crystal of compound (I) determined by simultaneous differential thermal and thermogravimetric measurement can be 177 to 187°C, in other words, around 182°C.

[0139] (Free form I crystal of compound (I))

[0140] Type I crystal of compound (I) has Figure 5 The powder X-ray diffraction pattern shown in Figure 6 The differential thermal - thermogravimetric measurement (TG-DTA) curve is shown.

[0141] In one embodiment of the present invention, the type I crystal of the free form of compound (I) has the diffraction angle (2θ) and intensity (cts) shown in Table 4 in powder X-ray diffraction.

[0142] Among them, the characteristic peaks in the powder X-ray diffraction pattern of the type I crystal of compound (I) include diffraction angles (2θ±0.2°) of 9.9°, 11.7°, 13.2°, 17.7°, 18.1°, 18.8° and 20.8°.

[0143] The type I crystal of compound (I) involved in the present invention is a crystal having three or more peaks selected from the above-mentioned characteristic peaks, preferably a crystal having four or more peaks selected from the above-mentioned characteristic peaks, more preferably a crystal having five or more peaks selected from the above-mentioned characteristic peaks, even more preferably a crystal having six or more peaks selected from the above-mentioned characteristic peaks, and particularly preferably a crystal having all the above-mentioned characteristic peaks.

[0144] Among them, the characteristic peaks in the powder X-ray diffraction pattern of the type I crystal of compound (I) include diffraction angles (2θ±0.2°) of 9.9°, 11.7°, 13.2°, 18.8° and 20.8°.

[0145] The type I crystal of compound (I) of the present invention is a crystal having three or more peaks selected from the above characteristic peaks, preferably a crystal having four or more peaks selected from the above characteristic peaks, and particularly preferably a crystal having all the above characteristic peaks.

[0146] The endothermic peak (peak top value) of the type I crystal of compound (I) determined by simultaneous differential thermal and thermogravimetric measurement can be 178 to 188°C, in other words, around 183°C.

[0147] (V-type crystal of compound (I) and 1 equivalent of fumaric acid)

[0148] The V-type crystal of compound (I) and 1 equivalent of fumaric acid has Figure 7The powder X-ray diffraction pattern shown in Figure 8 The differential thermal - thermogravimetric measurement (TG-DTA) curve is shown.

[0149] In one embodiment of the present invention, the V-type crystal of compound (I) and 1 equivalent of fumaric acid has the diffraction angle (2θ) and intensity (cts) shown in Table 5 in powder X-ray diffraction.

[0150] Among them, the characteristic peaks in the powder X-ray diffraction pattern of the V-type crystal of compound (I) and 1 equivalent of fumaric acid can be listed as diffraction angles (2θ±0.2°) of 6.9°, 9.4°, 10.2°, 13.7°, 21.1°, 23.6° and 26.5°.

[0151] The V-type crystals of the compound (I) and 1 equivalent of fumaric acid involved in the present invention are crystals having 3 or more peaks selected from the above-mentioned characteristic peaks, preferably crystals having 4 or more peaks selected from the above-mentioned characteristic peaks, more preferably crystals having 5 or more peaks selected from the above-mentioned characteristic peaks, even more preferably crystals having 6 or more peaks selected from the above-mentioned characteristic peaks, and particularly preferably crystals having all of the above-mentioned characteristic peaks.

[0152] Among them, the characteristic peaks in the powder X-ray diffraction pattern of the V-type crystal of compound (I) and 1 equivalent of fumaric acid include diffraction angles (2θ±0.2°) of 6.9°, 13.7°, 21.1°, 23.6° and 26.5°.

[0153] The V-type crystal of compound (I) and 1 equivalent of fumaric acid involved in the present invention is a crystal having 3 or more peaks selected from the above characteristic peaks, preferably a crystal having 4 or more peaks selected from the above characteristic peaks, and particularly preferably a crystal having all the above characteristic peaks.

[0154] The endothermic peak (peak top value) of the V-type crystal of compound (I) and 1 equivalent of fumaric acid determined by simultaneous differential thermal-thermogravimetric measurement can be 146 to 156°C, in other words, around 151°C.

[0155] (Type I crystal of compound (I) and 1 equivalent of fumaric acid)

[0156] The type I crystal of compound (I) and 1 equivalent of fumaric acid has Fig. 9 The powder X-ray diffraction pattern shown in Fig.10 The differential thermal - thermogravimetric measurement (TG-DTA) curve is shown.

[0157] In one embodiment of the present invention, the type I crystal of compound (I) and 1 equivalent of fumaric acid has the diffraction angle (2θ) and intensity (cts) shown in Table 6 in powder X-ray diffraction.

[0158] Among them, the characteristic peaks in the powder X-ray diffraction pattern of type I crystals of compound (I) and 1 equivalent of fumaric acid can be listed as diffraction angles (2θ±0.2°) of 6.4°, 10.3°, 12.8°, 15.0°, 20.7°, 23.4° and 26.6°.

[0159] The type I crystal of the compound (I) and 1 equivalent of fumaric acid involved in the present invention is a crystal having more than 3 peaks selected from the above-mentioned characteristic peaks, preferably a crystal having more than 4 peaks selected from the above-mentioned characteristic peaks, more preferably a crystal having more than 5 peaks selected from the above-mentioned characteristic peaks, even more preferably a crystal having more than 6 peaks selected from the above-mentioned characteristic peaks, and particularly preferably a crystal having all of the above-mentioned characteristic peaks.

[0160] The endothermic peak (peak top value) of the type I crystal of compound (I) and 1 equivalent of fumaric acid determined by simultaneous differential thermal-thermogravimetric measurement can be 148 to 158°C, in other words, around 153°C.

[0161] Among them, the characteristic peaks in the powder X-ray diffraction pattern of type I crystals of compound (I) and 1 equivalent of fumaric acid can be listed as diffraction angles (2θ±0.2°) of 6.4°, 10.3°, 12.8°, 20.7°, 23.4° and 26.6°.

[0162] The type I crystal of compound (I) and 1 equivalent of fumaric acid involved in the present invention is a crystal having more than 3 peaks selected from the above-mentioned characteristic peaks, preferably a crystal having more than 4 peaks selected from the above-mentioned characteristic peaks, more preferably a crystal having more than 5 peaks selected from the above-mentioned characteristic peaks, and particularly preferably a crystal having all the above-mentioned characteristic peaks.

[0163] The endothermic peak (peak top value) of the type I crystal of compound (I) and 1 equivalent of fumaric acid determined by simultaneous differential thermal-thermogravimetric measurement can be 148 to 158°C, in other words, around 153°C.

[0164] (Type I crystal of compound (I) and 0.5 equivalent of fumaric acid)

[0165] The type I crystal of compound (I) and 0.5 equivalent of fumaric acid has Fig.11 The powder X-ray diffraction pattern shown in Fig.12 The differential thermal - thermogravimetric measurement (TG-DTA) curve is shown.

[0166] Among them, the characteristic peaks in the powder X-ray diffraction pattern of type I crystals of compound (I) and 0.5 equivalent of fumaric acid can be listed as diffraction angles (2θ±0.2°) of 6.4°, 7.5°, 9.7°, 11.7°, 15.1°, 19.6° and 23.9°.

[0167] The type I crystal of compound (I) and 0.5 equivalent of fumaric acid involved in the present invention is a crystal having 4 or more peaks selected from the above-mentioned characteristic peaks, preferably a crystal having 5 or more peaks selected from the above-mentioned characteristic peaks, more preferably a crystal having 6 or more peaks selected from the above-mentioned characteristic peaks, and particularly preferably a crystal having all the above-mentioned peaks.

[0168] The endothermic peak (peak top value) of the type I crystal of compound (I) and 0.5 equivalent of fumaric acid determined by simultaneous differential thermal-thermogravimetric measurement can be 156 to 166°C, in other words, around 161°C.

[0169] (Type II crystal of compound (I) and 0.5 equivalent of fumaric acid)

[0170] The type II crystal of compound (I) and 0.5 equivalent of fumaric acid has Fig.13 The powder X-ray diffraction pattern shown in Fig.14 The differential thermal - thermogravimetric measurement (TG-DTA) curve is shown.

[0171] Among them, the characteristic peaks in the powder X-ray diffraction pattern of type II crystals of compound (I) and 0.5 equivalent of fumaric acid can be listed as diffraction angles (2θ±0.2°) of 5.4°, 6.4°, 7.3°, 12.8°, 13.4°, 14.7° and 15.4°.

[0172] The type II crystal of compound (I) and 0.5 equivalent of fumaric acid involved in the present invention is a crystal having 4 or more peaks selected from the above-mentioned characteristic peaks, preferably a crystal having 5 or more peaks selected from the above-mentioned characteristic peaks, more preferably a crystal having 6 or more peaks selected from the above-mentioned characteristic peaks, and particularly preferably a crystal having all the above-mentioned peaks.

[0173] The endothermic peak (peak top value) of the type II crystal of compound (I) and 0.5 equivalent of fumaric acid determined by simultaneous differential thermal-thermogravimetric measurement can be 157 to 167°C, in other words, around 162°C.

[0174] (Type III crystal of compound (I) and 1 equivalent of fumaric acid)

[0175] The type III crystal of compound (I) and 1 equivalent of fumaric acid has Fig.15The powder X-ray diffraction pattern shown in Fig.16 The differential thermal - thermogravimetric measurement (TG-DTA) curve is shown.

[0176] Among them, the characteristic peaks in the powder X-ray diffraction pattern of the type III crystal of compound (I) and 1 equivalent of fumaric acid can be listed as diffraction angles (2θ±0.2°) of 8.1°, 13.2°, 20.4°, 23.1°, 24.7° and 26.1°.

[0177] The type III crystal of compound (I) and 1 equivalent of fumaric acid involved in the present invention is a crystal having more than 3 peaks selected from the above-mentioned characteristic peaks, preferably a crystal having more than 4 peaks selected from the above-mentioned characteristic peaks, more preferably a crystal having more than 5 peaks selected from the above-mentioned characteristic peaks, and particularly preferably a crystal having all the above-mentioned characteristic peaks.

[0178] The exothermic peak (peak value) of the type III crystal of compound (I) and 1 equivalent of fumaric acid determined by simultaneous differential thermal-thermogravimetric measurement can be 148 to 158°C, in other words, around 153°C.

[0179] (Form IV crystal of compound (I) and 1 equivalent of fumaric acid)

[0180] The IV type crystal of compound (I) and 1 equivalent of fumaric acid has Fig.17 The powder X-ray diffraction pattern shown in Fig.18 The differential thermal - thermogravimetric measurement (TG-DTA) curve is shown.

[0181] Among them, the characteristic peaks in the powder X-ray diffraction pattern of the type IV crystal of compound (I) and 1 equivalent of fumaric acid include 6.0°, 15.7° and 18.8° as diffraction angles (2θ±0.2°).

[0182] The IV-type crystal of the compound (I) and 1 equivalent of fumaric acid according to the present invention is a crystal having all the above-mentioned characteristic peaks.

[0183] The exothermic peak (peak value) of the type IV crystal of compound (I) and 1 equivalent of fumaric acid determined by simultaneous differential thermal-thermogravimetric measurement is 125 to 135°C, in other words, around 130°C.

[0184] (Method for producing type II crystal with 1 equivalent of fumaric acid)

[0185] As one embodiment of the present specification, the type II crystal of compound (I) and 1 equivalent of fumaric acid is obtained by, for example, a method including the following two steps.

[0186] Step 1 is a step of adding fumaric acid and the following solvent to the type I crystal of compound (I), and optionally adding seed crystals.

[0187] Step 2 is a step of stirring the suspension obtained in the above step to obtain crystals of compound (I) and 1 equivalent of fumaric acid.

[0188] The solid is then filtered out to recover the solid, which is preferably washed with water and dried.

[0189] In step 1, the fumaric acid is preferably added in an amount of 1.5 to 10 equivalents (more preferably 2 to 5 equivalents, most preferably 3 equivalents) based on 1 equivalent of the free form of compound (I).

[0190] The solvent in step 1 is preferably added in an amount of 5 to 35 times (v / w) (more preferably 5 to 30 times (v / w), and most preferably 10 times (v / w)).

[0191] Examples of the solvent used in step 1 include alcohols such as methanol, ethanol, isopropanol, and tert-butanol, ethers such as 1,4-dioxane and tetrahydrofuran, ketones such as acetone and methyl ethyl ketone, and aprotic polar solvents such as acetonitrile, etc., either alone or in combination.

[0192] The Type I crystal of Compound (I) can be obtained by the method described in the column “(Method for producing free Type I crystal)” or in Example 3.

[0193] The temperature in the reaction of step 2 is preferably room temperature or higher and below the boiling point of each solvent as described above, but is not particularly limited and can be appropriately set, but is preferably 25 to 70° C. (more preferably 40 to 60° C., most preferably 50° C.).

[0194] Regarding the stirring time in the reaction of step 2, the suspension is preferably stirred for 24 to 124 hours (more preferably 48 to 72 hours, most preferably 60 to 65 hours).

[0195] (Method for producing free-body type II crystal)

[0196] (Method A-1)

[0197] As one embodiment of the present specification, the free-form II crystal of Compound (I) is obtained by, for example, a method (method A-1) comprising the following two steps.

[0198] Step 1 is a step of adding any one of succinic acid, phosphoric acid and adipic acid and a mixed solvent of a good solvent / poor solvent to compound (I), and optionally adding seed crystals.

[0199] Step 2 is a step of stirring the suspension obtained in the above step to obtain solid crystals of Compound (I).

[0200] During the suspension stirring, the additional solvent (for example, 1-propanol as a good solvent) may be added in multiple portions as needed depending on the amount of evaporation of the solvent.

[0201] Then, the solid was collected by filtration and dried.

[0202] The acid in step 1 is preferably added in an amount of 0.5 to 2 equivalents (more preferably 0.75 to 1.5 equivalents, particularly preferably 1 equivalent) based on 1 equivalent of compound (I).

[0203] The good solvent / poor solvent mixed solvent (e.g. 1-propanol / water) in step 1 is preferably added in an amount of 5 to 50 times (v / w) (more preferably 10 to 30 times (v / w), particularly preferably 20 times (v / w)) the amount of the free form of Compound (I).

[0204] The temperature in the reaction of step 2 is preferably 25 to 70°C (more preferably 40 to 60°C, particularly preferably 50°C).

[0205] The stirring time in the reaction of step 2 is preferably 2 to 7 days (more preferably 3 to 5 days, particularly preferably 4 days) of suspension stirring.

[0206] (Method A-2)

[0207] In addition, as another embodiment in the present specification, the free-form type II crystal of compound (I) can be obtained by, for example, a method (method A-2) comprising the following two steps.

[0208] Step 1 is a step of adding a mixed solvent of a good solvent / poor solvent and seed crystals to compound (I), and optionally adding any acid of succinic acid, phosphoric acid and adipic acid.

[0209] Step 2 is a step of stirring the suspension obtained in the above step to obtain solid crystals of Compound (I).

[0210] During the suspension stirring, the additional solvent (for example, 1-propanol as a good solvent) may be added in multiple portions as needed depending on the amount of evaporation of the solvent.

[0211] Then, the solid was collected by filtration and dried.

[0212] The amount of succinic acid added in step 1 is preferably 0.5 to 2 equivalents (more preferably 0.75 to 1.5 equivalents, particularly preferably 1 equivalent) based on 1 equivalent of compound (I).

[0213] Regarding the good solvent / poor solvent mixed solvent (e.g., a 1-propanol / water mixed solvent) of step 1, it is preferred to add 1 to 10 times (v / w) the amount of the free form of compound (I) (more preferably 2 to 5 times (v / w), particularly preferably 3 times (v / w)), and add seed crystals (e.g., seed crystals obtained in either (i) or (ii)).

[0214] The reaction temperature in step 2 is preferably 25 to 70°C (more preferably 40 to 60°C, particularly preferably 50°C).

[0215] The reaction time of step 2 is preferably 6 to 48 hours (more preferably 12 to 24 hours, particularly preferably 17.5 to 24 hours).

[0216] (Method B)

[0217] In addition, as another embodiment in the present specification, the free-form II crystal of Compound (I) is obtained by, for example, a method (method B) comprising the following two steps.

[0218] Step 1 is a step of adding a poor solvent to compound (I) and optionally adding seed crystals.

[0219] Step 2 is a step of adding a good solvent to the solution obtained in the above step, stirring the obtained suspension, and obtaining solid crystals of Compound (I).

[0220] Regarding the amount of the poor solvent in step 1, it is preferred to add a 2- to 10-fold (v / w) amount of the poor solvent (e.g. diisopropyl ether) relative to the free form of compound (I) (more preferably, 3- to 7-fold (v / w), particularly preferably 5-fold (v / w) amount).

[0221] The reaction time of step 1 is preferably 24 to 96 hours (more preferably 36 to 72 hours, particularly preferably 44.5 hours) under suspension stirring.

[0222] The reaction temperature in step 2 is preferably 25 to 70°C (more preferably 40 to 60°C, particularly preferably 50°C).

[0223] The amount of the good solvent in step 2 is preferably 0.5 to 3 times (v / w) (more preferably 0.8 to 2 times (v / w), particularly preferably 1 time (v / w)) the amount of the good solvent (e.g. ethanol) added to the free form of Compound (I).

[0224] The stirring time in step 2 is preferably 6 to 72 hours (more preferably 12 to 48 hours, particularly preferably 21 hours) of suspension stirring, followed by filtration to recover the solid and dry it.

[0225] During the suspension stirring, the additional solvent (for example, diisopropyl ether as a poor solvent and / or ethanol as a good solvent) may be added in multiple portions as necessary depending on the amount of evaporation of the solvent.

[0226] As a combination of a good solvent and a poor solvent used in the production of the free form Type II crystal of Compound (I), methanol (good solvent) and water (poor solvent), methanol (good solvent) and diisopropyl ether (IPE) (poor solvent), methanol (good solvent) and heptane (poor solvent), ethanol (good solvent) and IPE (poor solvent), ethanol (good solvent) and heptane (poor solvent), 1-propanol (good solvent) and water (poor solvent), 1-propanol (good solvent) and IPE (poor solvent), 1-propanol (good solvent) and heptane (poor solvent), 2-propanol (good solvent) and heptane (poor solvent), acetone (good solvent) and IPE (poor solvent), acetone (good solvent) and heptane (poor solvent), dimethyl sulfoxide (DMSO) (good solvent) and water (poor solvent), dimethylacetamide (DMA) (good solvent) and water (poor solvent), tetrahydrofuran (THF) (good solvent) and IPE (poor solvent), or THF (good solvent) and heptane (poor solvent) can be used.

[0227] Among them, the preferred combination of good solvent and poor solvent is a combination of ethyl acetate (good solvent) and n-heptane (poor solvent), ethanol (good solvent) and water (poor solvent), 1-propanol (good solvent) and water (poor solvent), ethanol (good solvent) and diisopropyl ether (poor solvent). The amount of the poor solvent is preferably 1 to 20 times the amount of the good solvent (v / v), and more preferably 1 to 10 times the amount (v / v).

[0228] In the above method, the amount of seed crystals added can be preferably 0.5 to 30 (w / w) % of the amount of compound (I) charged, and more preferably 1 to 5 (w / w) %.

[0229] (Method for producing free-body type I crystals)

[0230] As one embodiment of the present specification, free form I crystals of Compound (I) are obtained by, for example, a method comprising the steps of suspending a crude compound (I) in a solvent and optionally adding seed crystals to obtain solid crystals of Compound (I).

[0231] In order to promote the crystallization of free-body type I crystals, seed crystals may be optionally added. As seed crystals, type I crystals or mixed crystals containing an appropriate amount of type I crystals of compound (I) may be added. Even without adding seed crystals, type I crystals of compound (I) can be obtained by the above method, but by adding seed crystals, the time for obtaining the above compound can be shortened. In addition, in order to shorten the crystallization time and control the particle size, the crystallization may be carried out while stirring.

[0232] The amount of seed crystals added is 0.5 to 30 (w / w) %, preferably 1 to 10 (w / w) % of the amount of compound (I) charged.

[0233] The temperature can be appropriately set, but is preferably 50 to 65°C.

[0234] Compound (I) can be precipitated at the above dissolution temperature. However, if it does not precipitate at the dissolution temperature, free-form I-type crystals can be obtained by cooling to 25°C.

[0235] As the solvent, ethyl acetate, ethanol, acetonitrile, acetone, tert-butyl methyl ether or the like can be used.

[0236] (Method for producing V-type crystals with 1 equivalent of fumaric acid)

[0237] As one embodiment of the present specification, a V-type crystal of compound (I) and 1 equivalent of fumaric acid is obtained, for example, by a method including the following two steps.

[0238] Step 1 is a step of adding 1 to 2 equivalents of fumaric acid to 1 equivalent of the free form of the compound (I), and adding 40 to 60 times (v / w) amount of acetone as a solvent to the free form of the compound (I), and optionally adding seed crystals.

[0239] Step 2 is a step of stirring the suspension obtained in the above step, filtering and recovering the solid.

[0240] Step 3 is a step of drying the solid obtained in the above step under reduced pressure conditions preferably at 20 to 60° C. (preferably a pressure of 2 kPa or less) to obtain a crystal of compound (I) and 1 equivalent of fumaric acid.

[0241] The fumaric acid in step 1 is more preferably added in an amount of 2 equivalents based on 1 equivalent of the free form of compound (I).

[0242] Acetone in step 1 is preferably added in an amount of 40 to 60 times (v / w) (more preferably 50 times (v / w)) the amount of the free form of compound (I).

[0243] The reaction temperature in step 2 is preferably 10 to 30°C (preferably 25°C).

[0244] The reaction time of step 2 is preferably 12 to 96 hours (more preferably 71 hours).

[0245] The reaction temperature of step 3 is such that the obtained solid is 20 to 60° C. (for example, room temperature).

[0246] The drying time under reduced pressure (for example, at a pressure of 2 kPa or less) in step 3 is preferably 6 to 24 hours (more preferably 7.5 hours).

[0247] As another embodiment of the present specification, a V-type crystal of compound (I) and 1 equivalent of fumaric acid is obtained by a method including the following steps.

[0248] Step 1: Dry the type I crystal with 1 equivalent of fumaric acid under reduced pressure (preferably a pressure of 2 kPa or less).

[0249] The reaction temperature in step 1 is 20 to 60°C.

[0250] (Method for producing type I crystal with 1 equivalent of fumaric acid)

[0251] As one embodiment of the present specification, the type I crystal of compound (I) and 1 equivalent of fumaric acid is obtained by, for example, a method including the following two steps.

[0252] Step 1 is a step of adding 1 to 2 equivalents of fumaric acid to 1 equivalent of the free form of Compound (I), adding 40 to 60 times the amount of acetone as a solvent, and optionally adding seed crystals.

[0253] Step 2 is a step of stirring the suspension obtained in the above step at 30°C or less to obtain crystals of fumaric acid with compound (I).

[0254] Then, filtration was performed to recover the solid.

[0255] The amount of fumaric acid in step 1 is preferably 2 equivalents of fumaric acid per 1 equivalent of the free form of compound (I).

[0256] Acetone in step 1 is preferably added in an amount of 40 to 60 times (more preferably 50 times) the amount of the free form of compound (I).

[0257] The reaction temperature in step 2 is preferably 10 to 30°C (more preferably 25°C).

[0258] The reaction time of step 2 is preferably 12 to 24 hours (more preferably 19.5 hours).

[0259] (Method for producing type III crystal with 1 equivalent of fumaric acid)

[0260] As one embodiment of the present specification, the type III crystal of compound (I) and 1 equivalent of fumaric acid is obtained by, for example, a method including the following two steps.

[0261] Step 1 is a step of adding fumaric acid and acetonitrile as a solvent to compound (I), and optionally adding seed crystals.

[0262] Step 2 is a step of stirring the suspension obtained in the above step for a short period of time of 1.5 hours or less to obtain crystals of compound (I) and 1 equivalent of fumaric acid.

[0263] Then, filtration was performed to recover the solid.

[0264] Preferably, 1 to 5 equivalents (more preferably 3 equivalents) of fumaric acid are added based on 1 equivalent of the free form of Compound (I) in Step 1.

[0265] The solvent (eg, acetonitrile) in step 1 is preferably added in an amount of 25 to 35 times (v / w) (more preferably 30 times (v / w)) the amount of the free form of compound (I).

[0266] The reaction temperature in step 2 is preferably 45 to 55°C (particularly preferably 50°C).

[0267] The reaction time in step 2 is preferably 0.5 to 1.5 hours (more preferably 1 hour).

[0268] (Method for producing type IV crystal with 1 equivalent of fumaric acid)

[0269] As one embodiment of the present specification, the type IV crystal of compound (I) and 1 equivalent of fumaric acid is obtained by, for example, a method including the following two steps.

[0270] Step 1 is a step of adding fumaric acid and water as a solvent to compound (I), and optionally adding seed crystals.

[0271] Step 2 is a step of stirring the suspension obtained in the above step for a short period of time of 2 hours or less to obtain crystals of compound (I) and 1 equivalent of fumaric acid.

[0272] Then, filtration was performed to recover the solid.

[0273] In step 1, the fumaric acid is preferably added in an amount of 1 to 5 equivalents (more preferably 3 equivalents) based on 1 equivalent of the free form of compound (I).

[0274] As the solvent in step 1, it is preferred to add water in an amount of 15 to 25 times (more preferably 20 times) the amount of the free form of compound (I).

[0275] The reaction time in step 2 is preferably 45 to 55°C (particularly preferably 50°C).

[0276] The reaction temperature in step 2 is preferably 1 to 2 hours (more preferably 1.5 hours).

[0277] (Method for producing type I crystal with 0.5 equivalent of fumaric acid)

[0278] As one embodiment of the present specification, the type I crystal of compound (I) and 0.5 equivalent of fumaric acid is obtained by, for example, a method including the following two steps.

[0279] Step 1 is a step of adding fumaric acid and water as a solvent to compound (I), and optionally adding seed crystals.

[0280] Step 2 is a step of stirring the suspension obtained in the above step at a temperature preferably above room temperature and below the boiling point of the solvent to obtain crystals.

[0281] Then, the solid was collected by filtration and dried.

[0282] In step 1, the fumaric acid is preferably added in an amount of 0.5 to 1 equivalent (more preferably 0.5 to 0.75 equivalent, most preferably 0.5 equivalent) based on 1 equivalent of the free form of compound (I).

[0283] The water used as the solvent in step 1 is preferably added in an amount of 10 to 40 times (v / w) (more preferably 15 to 25 times (v / w), most preferably 20 times (v / w)) the amount of the free form of compound (I).

[0284] The reaction temperature in step 2 is preferably 40 to 60°C (more preferably 45 to 55°C, most preferably 50°C).

[0285] The reaction time of step 2 is preferably 48 to 120 hours (more preferably 72 to 96 hours, most preferably 93.5 hours).

[0286] (Method for producing type II crystal with 0.5 equivalent of fumaric acid)

[0287] As one embodiment of the present specification, the type II crystal of compound (I) and 0.5 equivalent of fumaric acid is obtained by, for example, a method including the following two steps.

[0288] Step 1 is a step of adding fumaric acid and ethanol to compound (I), and optionally adding seed crystals.

[0289] Step 2 is a step of stirring the suspension obtained in the above step at a temperature preferably above room temperature and below the boiling point of the solvent to obtain crystals.

[0290] Then, the solid was collected by filtration and dried.

[0291] The amount of fumaric acid in step 1 is preferably 0.5 to 1 equivalent (preferably 1 equivalent) per 1 equivalent of the free form of compound (I).

[0292] Ethanol as a solvent in step 1 is preferably added in an amount of 10 to 40 times (v / w) (more preferably 15 to 25 times (v / w), most preferably 20 times (v / w)) the amount of the free form of compound (I).

[0293] The reaction temperature in step 2 is preferably 40 to 60°C (more preferably 45 to 55°C, most preferably 50°C).

[0294] The reaction time of step 2 is preferably 6 to 48 hours (more preferably 12 to 24 hours, most preferably 19.5 hours).

[0295] The type II crystal of compound (I) with 1 equivalent of fumaric acid, the free type II crystal, the free type I crystal, the type V crystal with 1 equivalent of fumaric acid, the type I crystal with 0.5 equivalent of fumaric acid, and the type II crystal with 0.5 equivalent of fumaric acid have at least one of the properties that are advantageous in pharmaceutical manufacturing, such as non-hygroscopicity, acquisition reproducibility, solid stability, and oral absorbability.

[0296] Among them, type II crystals of compound (I) and 1 equivalent of fumaric acid, free type II crystals, free type I crystals, and type V crystals of compound (I) and 1 equivalent of fumaric acid have favorable properties in pharmaceutical manufacturing, such as non-hygroscopicity, acquisition reproducibility, solid stability, and oral absorbability, compared with other crystal forms of compound (I).

[0297] The type II crystal of compound (I) and 1 equivalent of fumaric acid has low hygroscopicity and has stable acquisition. Among candidate compounds for the development of pharmaceuticals, low hygroscopicity and the ability to stably acquire these properties are very important in the industrial production of pharmaceuticals. In addition, it has properties that are easy to handle as a pharmaceutical, such as solid stability, solubility, and excellent oral absorption. Therefore, the type II crystal of compound (I) and 1 equivalent of fumaric acid has excellent properties that are essential as a pharmaceutical or a pharmaceutical original drug.

[0298] The free-body II-type crystal of compound (I) has low hygroscopicity. This is very important for industrial production of stable quality pharmaceuticals among candidate compounds for pharmaceutical development. In addition, it has properties that are easy to handle as a pharmaceutical, such as excellent solid stability, solubility, and oral absorption. Therefore, the free-body II-type crystal of compound (I) has excellent properties that are essential as a pharmaceutical or pharmaceutical original drug.

[0299] The free form I crystal of compound (I) has low hygroscopicity and excellent acquisition stability. It is also important for the industrial production of stable quality pharmaceuticals that candidate compounds for pharmaceutical development have low hygroscopicity and acquisition stability. Therefore, the I crystal of compound (I) has excellent properties essential for pharmaceuticals or pharmaceutical raw materials.

[0300] The V-type crystal of compound (I) and 1 equivalent of fumaric acid has low hygroscopicity and excellent acquisition stability. It is also important for the industrial production of stable quality pharmaceuticals that the candidate compound for pharmaceutical development has low hygroscopicity and acquisition stability. Therefore, the V-type crystal of compound (I) and 1 equivalent of fumaric acid of the present invention has excellent properties essential for pharmaceuticals or pharmaceutical original drugs.

[0301] The I-type crystal of compound (I) and 1 equivalent of fumaric acid has excellent acquisition stability. The acquisition stability of candidate compounds for pharmaceutical development is also important for industrial production of pharmaceuticals with stable quality. Therefore, the I-type crystal of compound (I) and 1 equivalent of fumaric acid has excellent properties essential for pharmaceuticals or pharmaceutical raw materials.

[0302] Since the type I crystal of compound (I) and 1 equivalent of fumaric acid can be recovered as a solid, it is also useful as an intermediate for producing the type V crystal of compound (I) and 1 equivalent of fumaric acid.

[0303] The obtained stability of the type III crystal of compound (I) and 1 equivalent of fumaric acid is excellent. The fact that the candidate compound for the development of pharmaceuticals has the ability to obtain stability is also important for the industrial production of pharmaceuticals with stable quality. Therefore, the type III crystal of compound (I) and 1 equivalent of fumaric acid has excellent properties essential for pharmaceuticals or pharmaceutical raw materials.

[0304] The IV-type crystal of compound (I) and 1 equivalent of fumaric acid has excellent acquisition stability. The acquisition stability of candidate compounds for pharmaceutical development is also important for industrial production of pharmaceuticals with stable quality. Therefore, the IV-type crystal of compound (I) and 1 equivalent of fumaric acid has excellent properties essential for pharmaceuticals or pharmaceutical raw materials.

[0305] The I-type crystal of compound (I) and 0.5 equivalents of fumaric acid has excellent acquisition stability. The acquisition stability of candidate compounds for pharmaceutical development is also important for industrial production of pharmaceuticals with stable quality. Therefore, the I-type crystal of compound (I) and 0.5 equivalents of fumaric acid has excellent properties essential for pharmaceuticals or pharmaceutical raw materials.

[0306] The obtained stability of the type II crystal of compound (I) and 0.5 equivalent of fumaric acid is excellent. The stability of the candidate compound for the development of pharmaceuticals is also important for the industrial production of pharmaceuticals with stable quality. Therefore, the type II crystal of compound (I) and 0.5 equivalent of fumaric acid according to the present invention has excellent properties essential for pharmaceuticals or pharmaceutical original drugs.

[0307] (Crystallization with other acids)

[0308] Other embodiments of the present invention relate to crystals of compound (I) and hydrochloric acid (Type I crystal, Type II crystal, Type III crystal), crystals of compound (I) and hydrobromic acid, crystals of compound (I) and 1 equivalent of L-tartaric acid, or crystals of compound (I) and succinic acid.

[0309] The powder X-ray diffraction patterns and differential thermal-thermogravimetric (TG-DTA) curves of these crystals are shown in Figure 2. Figures 19-30 shown.

[0310] The characteristic peaks in the powder X-ray diffraction patterns of these crystals are described in Reference Examples 3 to 7. These crystals are crystals having three or more (preferably four or more, and if present, more preferably five or more) peaks selected from the characteristic peaks described in the reference examples, and particularly preferably crystals having all the characteristic peaks.

[0311] In addition, the temperatures of the endothermic peaks of these crystals determined by simultaneous differential thermal and thermogravimetric measurement are also described in the reference examples.

[0312] Regarding the crystals of the free form of the above-mentioned compound (I) and the crystals with the acid (crystals of the salt or co-crystals), the precipitated crystals can be separated and purified from a solution or a mixed solution of the above-mentioned crystals by a known separation and purification method such as filtration, washing with water, and drying under reduced pressure.

[0313] The crystallization of the free form of the compound (I) of the present invention or the crystallization with an acid (crystalization or co-crystal of a salt) has excellent HER2 inhibitory activity. In addition, it has excellent selectivity for HER2. Therefore, the crystallization of the free form of the compound (I) of the present invention or the crystallization with an acid (crystalization or co-crystal of a salt) can be used as an anti-tumor agent for diseases or malignant tumors with HER2 overexpression, HER2 gene amplification or HER2 mutation, etc., and has the advantage of few side effects.

[0314] In the present specification, "HER2" includes HER2 of humans or non-human mammals, preferably human HER2. In addition, the term "HER2" includes isoforms.

[0315] The free form crystals or crystals with an acid (salt crystals or co-crystals) of the compound (I) of the present invention are useful as pharmaceuticals for treating diseases involving HER2 due to their excellent HER2 inhibitory activity.

[0316] "Diseases involving HER2" include diseases that can reduce the incidence, alleviate symptoms, relieve and / or cure by deleting, inhibiting and / or hindering the function of HER2. Such diseases include, but are not limited to, malignant tumors. Preferably, malignant tumors with overexpression of HER2, HER2 gene amplification or HER2 mutation are present.

[0317] The free form crystals or crystals with an acid (salt crystals or co-crystals) of compound (I) of one embodiment of the present invention selectively inhibit wild-type HER2 and mutant HER2 having one or more insertion mutations, point mutations or deletion mutations in the HER2 domain, such as exon 20 insertion mutations.

[0318] One embodiment of the present invention provides a free form crystal of compound (I) having inhibitory activity against wild-type HER2 and mutant HER2 including HER2 having a YVMA insertion mutation as one of the exon 20 insertion mutations, or a crystal with an acid (a salt crystal or a co-crystal), or a pharmaceutical or pharmaceutical composition containing the crystal or the crystal with an acid.

[0319] One embodiment of the present invention provides an inhibitor of wild-type HER2 and HER2 variants including HER2 having a YVMA insertion mutation, comprising crystals of the free form of the compound (I) of the present invention or crystals with an acid (crystals or co-crystals of its salt).

[0320] The human HER2 gene is, for example, a gene as shown in SEQ ID NO: 1, SEQ ID NO: 3, or SEQ ID NO: 5, and the wild-type HER2 protein comprises, for example, an amino acid sequence as shown in SEQ ID NO: 2, SEQ ID NO: 4, or SEQ ID NO: 6. The base sequence information of the human HER2 gene and the amino acid sequence information of the wild-type HER2 protein can be obtained, for example, through accession numbers NM_004448, NM_001289936, or NM_001005862.

[0321] In some embodiments, the crystal of the free form of compound (I) of one embodiment of the present invention or the crystal of the free form with acid (crystal of salt or co-crystal) shows inhibitory activity against variant HER2 based on the amino acid sequence shown in SEQ ID NO: 2 and comprising one or more mutations of G309A, S310F, R678Q, L755S, L755_T759del, D769H, A775_G776insYVMA, V777L, V842I, and R896C. In other embodiments, the crystal of the free form of compound (I) of one embodiment of the present invention or the crystal of the free form with acid (crystal of salt or co-crystal) has inhibitory activity against variant HER2 based on the amino acid sequence shown in SEQ ID NO: 2 and comprising A775_G776insYVMA.

[0322] In some embodiments, the crystal of the free form of compound (I) of one embodiment of the present invention or the crystal of the free form with an acid (crystal of a salt or co-crystal) exhibits inhibitory activity against a variant HER2 based on the amino acid sequence shown in SEQ ID NO: 4 and comprising one or more mutations of G294A, S295F, R663Q, L740S, L740_T744del, D754H, A760_G761insYVMA, V762L, V827I, and R881C. In other embodiments, the crystal of the free form of compound (I) of one embodiment of the present invention or the crystal of the free form with an acid (crystal of a salt or co-crystal) exhibits inhibitory activity against a variant HER2 based on the amino acid sequence shown in SEQ ID NO: 4 and comprising A760_G761insYVMA.

[0323] In some embodiments, the compound (I) of one embodiment of the present invention or a crystal with an acid (a crystal of a salt or a co-crystal) exhibits inhibitory activity against a variant HER2 based on the amino acid sequence shown in SEQ ID NO: 6 and comprising one or more mutations of G279A, S280F, R648Q, L725S, L725_T729del, D739H, A745_G746insYVMA, V747L, V812I, and R866C. In other embodiments, a crystal of a free form of the compound (I) of one embodiment of the present invention or a crystal with an acid (a crystal of a salt or a co-crystal) exhibits inhibitory activity against a variant HER2 based on the amino acid sequence shown in SEQ ID NO: 6 and comprising A745_G746insYVMA.

[0324] In addition, in some embodiments, when the position of the amino acid shown in SEQ ID NO. 2 is different due to the deletion or insertion of an amino acid in a variation in a certain HER2 isoform, it can also be understood to be the same as the variation of the position corresponding to the position of the amino acid shown in SEQ ID NO. 2. Therefore, for example, the glycine at position 309 in HER2 shown in SEQ ID NO. 2 is equivalent to the glycine at position 294 in HER2 consisting of the amino acid sequence shown in SEQ ID NO. 4. Therefore, for example, "G309A" means that the glycine at position 309 in HER2 shown in SEQ ID NO. 2 is mutated to alanine, but in HER2 consisting of the amino acid sequence shown in SEQ ID NO. 4, it is the position corresponding to the amino acid at position 294, and therefore, "G294A" in HER2 consisting of the amino acid sequence shown in SEQ ID NO. 4 is equivalent to "G309A" in HER2 shown in SEQ ID NO. 2. In addition, the amino acid at which position of the amino acid shown in SEQ ID NO. 2 is equivalent to a certain amino acid in a certain HER2 isoform can be confirmed by, for example, Multiple Alignment of BLAST.

[0325] Sequence Listing

[0326] [Table A]

[0327]

[0328]

[0329]

[0330]

[0331]

[0332] One embodiment of the present invention provides an antitumor agent containing the crystals of the above-mentioned compound (I) (i.e., crystals of the free form of compound (I) or crystals of compound (I) and acid (crystals of salt or co-crystals)). In addition, one embodiment of the present invention provides a method for treating tumors, which includes the step of administering an effective amount of the crystals of the above-mentioned compound (I) (i.e., crystals of the free form of compound (I) or crystals of compound (I) and acid (crystals of salt or co-crystals)) to a subject in need of treatment. One embodiment of the present invention provides the use of the crystals of compound (I) (i.e., crystals of the free form of compound (I) or crystals of compound (I) and acid (crystals of salt or co-crystals)) in the manufacture of an antitumor agent. In addition, one embodiment of the present invention provides crystals of compound (I) for use in treating tumors (i.e., crystals of the free form of compound (I) or crystals of compound (I) and acid (crystals of salt or co-crystals)).

[0333] The crystal of the present invention can be used for postoperative adjuvant chemotherapy performed after surgical removal of a tumor to prevent recurrence, or can be used for preoperative adjuvant chemotherapy performed in advance for surgical removal of a tumor.

[0334] The tumors that are the objects of the present invention are not particularly limited, and examples thereof include brain tumors, head and neck cancers, digestive system cancers (esophageal cancer, gastric cancer, duodenal cancer, liver cancer, biliary cancer (gallbladder-choledochal cancer, etc.), pancreatic cancer, colorectal cancer (colon cancer, rectal cancer, etc.), lung cancer (non-small cell lung cancer, small cell lung cancer, mesothelioma, etc.), breast cancer, reproductive system cancers (ovarian cancer, uterine cancer (cervical cancer, uterine end cancer, etc.), urinary system cancers (renal cancer, bladder cancer, prostate cancer, testicular tumors, etc.), hematopoietic system tumors (leukemia, malignant lymphoma, multiple myeloma, etc.), bone-soft tissue tumors, skin cancer, etc., preferably lung cancer, breast cancer, gastric cancer, colorectal cancer, bladder cancer, biliary cancer or uterine cancer, more preferably lung cancer, breast cancer, gastric cancer, bladder cancer or biliary cancer.

[0335] In one embodiment, the tumor is a brain tumor. The compounds of the present invention can be used for the treatment of brain symptoms that require passage through the blood-brain barrier. The compound of one embodiment has a preferred permeability of the blood-brain barrier for delivery to the brain, i.e., excellent brain metastasis. As an indicator of the metastasis of the compound to the brain, there are the compound concentration in the brain, the Kp value (ratio of drug concentration in the brain to that in plasma).

[0336] Brain tumors that can be treated with the compounds of the present invention include metastatic brain tumors and primary brain tumors.

[0337] Brain tumors are not particularly limited, and examples thereof include metastatic brain tumors (e.g., brain metastases of lung cancer, breast cancer, gastric cancer, colorectal cancer, bladder cancer, biliary tract cancer, uterine cancer, etc. (preferably lung cancer, breast cancer or gastric cancer)), pilocytic astrocytoma, diffuse astrocytoma, oligodendroglioma-oligoastrocytoma, anaplastic astrocytoma-anaplastic oligodendroglioma, anaplastic oligoastrocytoma, glioblastoma, ependymoma, anaplastic ependymoma, ganglioglioma, central nervous system tumor, medulloblastoma, germ cell tumor (germinoma), malignant lymphoma of the central nervous system, meningioma, schwannoma, GH (growth hormone) type pituitary adenoma, PRL (prolactin) type pituitary adenoma, ACTH (adrenocorticotropic hormone) type pituitary adenoma, non-functional pituitary adenoma, craniopharyngioma, chordoma, hemangioblastoma, epithelioid tumor, etc.

[0338] In this specification, the term "effective amount" of a compound refers to the amount of the compound of the present invention that causes a biological or medical response in a subject, such as a reduction or inhibition of enzyme or protein activity, or improves symptoms, alleviates a condition, slows or delays the progression of a disease, etc. (therapeutically effective amount).

[0339] In this specification, the term "subject" includes mammals and non-mammals. Examples of mammals, without limitation, include humans, chimpanzees, apes, monkeys, horses, cattle, sheep, goats, pigs, rabbits, dogs, cats, rats, mice, guinea pigs, hedgehogs, kangaroos, moles, wild boars, bears, tigers, lions, etc. Examples of non-mammals, without limitation, include birds, fish, insects, etc. In one embodiment, the subject is a human, which may be a person diagnosed as needing treatment for the symptoms, conditions, or diseases disclosed in this specification.

[0340] When using the crystals or co-crystals of compound (I) or its salt as medicine, the crystals may be crushed or not, and various administration forms may be adopted according to the therapeutic purpose, such as oral preparations such as tablets, capsules, granules, fine granules, powders, dry syrups, etc.; any of non-oral preparations such as suppositories, inhalants, nasal drops, ointments, patches, injections, etc. Pharmaceutical compositions suitable for these administration forms can be manufactured by conventional preparation methods known to those skilled in the art using pharmaceutically acceptable carriers.

[0341] One embodiment of the present invention provides an antitumor agent containing crystals of the above-mentioned compound (I) (i.e., crystals of the free form of compound (I) or crystals of compound (I) and acid (crystals of salt or co-crystals)). In addition, one embodiment of the present invention provides a method for treating tumors, which includes the step of administering an effective amount of crystals of the above-mentioned compound (I) (i.e., crystals of the free form of compound (I) or crystals of compound (I) and acid (crystals of salt or co-crystals)) to a subject in need of treatment. In addition, one embodiment of the present invention provides the use of the crystals of the above-mentioned compound (I) (i.e., crystals of the free form of compound (I) or crystals of compound (I) and acid (crystals of salt or co-crystals)) in the manufacture of an antitumor agent. In addition, one embodiment of the present invention provides the crystals of the above-mentioned compound (I) (i.e., crystals of the free form of compound (I) or crystals of compound (I) and acid (crystals of salt or co-crystals)) for treating tumors.

[0342] One embodiment of the present invention provides an anti-tumor agent for oral administration containing crystals of the above-mentioned compound (I) (i.e., crystals of the free form of compound (I) or crystals of compound (I) and acid (crystals of salt or co-crystals)). In addition, one embodiment of the present invention provides a method for treating tumors, which includes the step of orally administering an effective amount of crystals of the above-mentioned compound (I) (i.e., crystals of the free form of compound (I) or crystals of compound (I) and acid (crystals of salt or co-crystals)) to a subject in need of treatment. In addition, one embodiment of the present invention provides the use of the crystals of the above-mentioned compound (I) (i.e., crystals of the free form of compound (I) or crystals of compound (I) and acid (crystals of salt or co-crystals)) for the manufacture of an anti-tumor agent for oral administration. In addition, one embodiment of the present invention provides the crystals of the above-mentioned compound (I) (i.e., crystals of the free form of compound (I) or crystals of compound (I) and acid (crystals of salt or co-crystals)) for oral administration to treat tumors.

[0343] One embodiment of the present invention provides a pharmaceutical composition containing crystals of the above-mentioned compound (I) (i.e., crystals of the free form of compound (I) or crystals of compound (I) and acid (crystals of salt or co-crystals)). A pharmaceutical composition of one embodiment of the present invention contains crystals of the above-mentioned compound (I) (i.e., crystals of the free form of compound (I) or crystals of compound (I) and acid (crystals of salt or co-crystals)) and a pharmaceutically acceptable carrier. In addition, one embodiment of the present invention provides the use of the crystals of the above-mentioned compound (I) (i.e., crystals of the free form of compound (I) or crystals of compound (I) and acid (crystals of salt or co-crystals)) for the manufacture of a pharmaceutical composition. Another embodiment of the present invention provides crystals of the above-mentioned compound (I) for use as a medicine (i.e., crystals of the free form of compound (I) or crystals of compound (I) and acid (crystals of salt or co-crystals)). In one embodiment of the present invention, a medicine kit is provided, which comprises (i) a pharmaceutical composition containing crystals of the above-mentioned compound (I) (i.e., crystals of the free form of compound (I) or crystals of compound (I) and an acid (crystals of a salt or a co-crystal)) and (ii) instructions for use of the composition.

[0344] As pharmaceutically acceptable carriers, various organic or inorganic carrier substances conventionally used as raw materials for preparations can be used, and they can be used as excipients, binders, disintegrants, lubricants, coating agents in solid preparations, solvents, solubilizers, suspending agents, isotonic agents, buffers, analgesics, etc. in liquid preparations. In addition, preparation additives such as preservatives, antioxidants, colorants, sweeteners, stabilizers, etc. can also be used as needed.

[0345] Examples of the excipient include lactose, saccharide, D-mannitol, starch, crystalline cellulose, and calcium silicate.

[0346] Examples of the binder include hydroxypropyl cellulose, methyl cellulose, polyvinyl pyrrolidone, powdered sugar, and hypromellose.

[0347] Examples of the disintegrant include sodium starch glycolate, carboxymethylcellulose calcium, cross-linked carboxymethylcellulose sodium, crospovidone, low-substituted hydroxypropyl cellulose, and partially pregelatinized starch.

[0348] Examples of the lubricant include talc, magnesium stearate, sucrose fatty acid esters, stearic acid, and sodium stearyl fumarate.

[0349] Examples of the coating agent include ethyl cellulose, aminoalkyl methacrylate copolymer RS, hypromellose, and white sugar.

[0350] Examples of the solvent include water, propylene glycol, and physiological saline.

[0351] Examples of the dissolving aid include polyethylene glycol, ethanol, α-cyclodextrin, Macrogol (polyethylene glycol) 400, and polysorbate 80.

[0352] Examples of the suspending agent include carrageenan, crystalline cellulose-carmellose sodium, and polyoxyethylene hydrogenated castor oil.

[0353] Examples of the isotonic agent include sodium chloride, glycerin, potassium chloride and the like.

[0354] Examples of the pH adjuster and buffer include sodium citrate, hydrochloric acid, lactic acid, phosphoric acid, and sodium dihydrogen phosphate.

[0355] Examples of the analgesic include procaine hydrochloride and lidocaine.

[0356] Examples of the preservative include ethyl p-hydroxybenzoate, cresol, and benzalkonium chloride.

[0357] Examples of the antioxidant include sodium sulfite, ascorbic acid, and natural vitamin E.

[0358] Examples of the coloring agent include titanium oxide, ferric oxide, food blue No. 1, and copper chlorophyll.

[0359] Examples of flavoring agents and flavoring agents include aspartame, saccharin, sucralose, l-menthol, and mint flavoring.

[0360] Examples of the stabilizer include sodium pyrosulfite, sodium edetate, erythrosonic acid, magnesium oxide, and butylated hydroxytoluene.

[0361] In the case of preparing a solid preparation for oral administration, an excipient is added to the crystals of compound (I) (i.e., crystals of the free form of compound (I) or crystals of compound (I) and an acid (crystals of a salt or a co-crystal)), and a binder, a disintegrant, a lubricant, a colorant, a flavoring agent and a flavoring agent are added as needed, and then tablets, coated tablets, granules, powders, capsules, etc. can be manufactured by conventional methods.

[0362] In the case of preparing an injection, a pH adjuster, a buffer, a stabilizer, an isotonic agent, a local anesthetic, etc. can be added to the crystals of compound (I) (i.e., crystals of a free form of compound (I) or crystals of compound (I) and an acid (crystals of a salt or a co-crystal)) to produce a subcutaneous, intramuscular or intravenous injection by a conventional method.

[0363] The amount of the crystals of compound (I) to be formulated in each dosage unit form (i.e., crystals of the free form of compound (I) or crystals of compound (I) and an acid (crystals of a salt or co-crystals)) is not fixed depending on the symptoms of the patient to whom the crystals of compound (I) are to be applied, or the dosage form thereof, but is generally, in terms of the free form of compound (I), it is desirable to set the amount to about 0.05 to 1000 mg for oral preparations, about 0.1 to 500 mg for injections, and about 1 to 1000 mg for suppositories or external preparations per dosage unit form.

[0364] The daily dosage of the crystals of compound (I) (i.e., crystals of the free form of compound (I) or crystals of compound (I) and an acid (crystals of a salt or co-crystals)) of the pharmaceutical agent having various dosage forms varies depending on the patient's symptoms, body weight, age, sex, etc. and cannot be generalized. Generally, for an adult (weighing 50 kg), the daily dosage is about 0.05 to 5000 mg, preferably 0.1 to 1000 mg, calculated as the free form of compound (I). It is preferably administered once a day or divided into 2 to 3 doses.

[0365] Example

[0366] Below, enumerate embodiment, more specifically describe the present invention, but the present invention is not limited by these examples in any way. The present invention is fully described by embodiment, but it is understood that various changes and / or modifications can be made by those skilled in the art. Therefore, such changes and / or modifications are all included in the present invention as long as they do not depart from the scope of the present invention.

[0367] In the following examples of compounds, % means weight percentage unless otherwise specified.

[0368] The various reagents used in the examples were all commercially available unless otherwise specified. Silica gel column chromatography used Purif-Pack (registered trademark) SI manufactured by MORITEX, KP-Sil (registered trademark) Silica pre-filled columns manufactured by BIOTAGE, or HP-Sil (registered trademark) Silica pre-filled columns manufactured by BIOTAGE. Alkaline silica gel column chromatography used Purif-Pack (registered trademark) NH manufactured by MORITEX or KP-NH (registered trademark) pre-filled columns manufactured by BIOTAGE. Thin layer chromatography for separation used KieselgelTM60F254, Art.5744 manufactured by Merck, or NH2 SILICAGEL60F254 PLATE manufactured by Wako Pure Chemical Industries, Ltd. The NMR spectrum was measured using an AL400 (400 MHz; JEOL), Mercury400 (400 MHz; Agilent Technologies) spectrometer, or an Inova400 (400 MHz; Agilent Technologies) spectrometer equipped with a 400 M NMR probe (Protasis). When the deuterated solvent contained tetramethylsilane, tetramethylsilane was used as an internal reference. In all other cases, the NMR solvent was used as an internal reference, and all δ values ​​were expressed in ppm.

[0369] The LCMS spectrum was measured using ACQUITY SQD (quadrupole type) manufactured by Waters Corporation under the following conditions.

[0370] Column: ACQUITY UPLC (registered trademark) BEH C18 manufactured by Waters, 2.1×50 mm, 1.7 μm

[0371] MS detection: ESI positive

[0372] UV detection: 254 and 210nm

[0373] Column flow rate: 0.5mL / min

[0374] Mobile phase A: 0.1% formic acid in water

[0375] Mobile phase B: 0.1% formic acid in acetonitrile

[0376] Injection volume: 1 μL

[0377] Gradient (Table 1)

[0378] [Table 1]

[0379] Time (minutes) Mobile Layer A Mobile layer B 0～0.1 95％ 5％ 0.1～2.1 95％→5％ 5％→95％ 2.1～3.1 5％ 95％

[0380] Reverse phase preparative HPLC purification was carried out using a preparative system manufactured by WATERS under the following conditions.

[0381] Column: A column obtained by connecting YMC-Actus Triart C18, 20×50 mm, 5 μm, manufactured by YMC Corporation, and YMC-Actus Triart C18, 20×10 mm, 5 μm, manufactured by YMC Corporation was used.

[0382] UV detection: 254nm

[0383] MS detection: ESI positive

[0384] Column flow rate: 25mL / min

[0385] Mobile phase: water / acetonitrile (0.1% formic acid)

[0386] Injection volume: 0.1-0.5mL

[0387] The meanings of the abbreviations are as follows.

[0388] s: Single peak

[0389] d: Twin Peaks

[0390] t: triplet

[0391] dt: double triplet

[0392] m: multiple peaks

[0393] DMSO-d6: deuterated dimethyl sulfoxide

[0394] CDCl3: deuterated chloroform

[0395] THF: Tetrahydrofuran

[0396] DMA: N,N-dimethylacetamide

[0397] NMP: 1-methyl-2-pyrrolidone

[0398] DMSO: dimethyl sulfoxide

[0399] Powder X-ray diffraction measurement

[0400] Regarding powder X-ray diffraction, an appropriate amount of a test substance was lightly pulverized in an agate mortar as required, and then measured under any of the following test conditions.

[0401] Device: EMPYREAN (Method A) manufactured by PANalytical

[0402] Reflection method (concentration method)

[0403] Target: Cu

[0404] X-ray tube current: 40mA

[0405] X-ray tube voltage: 45kV

[0406] Scanning range: 2θ=5.0~40.0°

[0407] Step size: 2θ = 0.0131°

[0408] Average time / step: 8.670s

[0409] Scanning speed: 0.0015° / s

[0410] Divergence slit: 1°

[0411] Scattering slit: 2.0mm

[0412] Light receiving slit: 8.0mm

[0413] Device: PANalytical EMPYREAN (Method B)

[0414] Transmission method

[0415] Target: Cu

[0416] X-ray tube current: 40mA

[0417] X-ray tube voltage: 45kV

[0418] Scanning range: 2θ = 2.0 ~ 40.0°

[0419] Step size: 2θ = 0.0066°

[0420] Average time / step: 8.670s

[0421] Scanning speed: 0.0008° / s

[0422] Divergence slit: 1 / 2°

[0423] Scattering slit: 2.0mm

[0424] Light receiving slit: None

[0425] The operation of the devices including data processing is performed according to the methods and sequences indicated by each device. The values ​​obtained from various graphs may vary slightly due to the direction of crystal growth, particle size, measurement conditions, etc. Therefore, these values ​​should not be strictly interpreted.

[0426] Regarding the simultaneous differential thermal and thermogravimetric measurement (TG-DTA measurement), 2 to 3 mg of the test substance was tested under the following test conditions.

[0427] Device:TG / DTA7200

[0428] Made by Hitachi High-Tech Science Co., Ltd.

[0429] Sample container: Aluminum

[0430] Heating rate: 10℃ / min to 25~290℃

[0431] Atmosphere gas: air (200 mL / min)

[0432] Control material: Empty plate

[0433] The operations of the devices including data processing are performed according to the methods and sequences indicated by each device.

[0434] Synthesis Example 1 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6- (Cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide ... Synthesis of Compound (I)

[0435] Synthesis Example 1 (1) (2S, 4R)-4-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidine-7- Synthesis of tert-butyl (2-methylpyrrolidine-1-carboxylate)

[0436] (2S, 4S)-4-hydroxy-2-methylpyrrolidine-1-carboxylic acid tert-butyl ester (19.0 g) and 4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (13.1 g) were dissolved in THF (190 mL), cooled to 0°C, triphenylphosphine (37.2 g) and diisopropyl azodicarboxylate (28.1 mL) were added, the temperature was raised to room temperature, and stirred for 1 hour. After the reaction mixture was concentrated under reduced pressure, the obtained residue was purified by silica gel chromatography (hexane: ethyl acetate) to obtain the corresponding coupled product. The obtained compound was used in the next reaction without further purification.

[0437] The obtained coupler, THF (114 mL) and aqueous ammonia (114 mL) were added to a pressure tube and stirred at 100°C for 14 hours. After the reaction mixture was cooled to room temperature, water (285 mL) was injected and stirred at room temperature for 5 hours. The precipitated solid was filtered, washed with water and dried to obtain the target product (34.5 g).

[0438] 1 HNMR (CDCl3) δ: 8.27 (s, 1H) 7.15 (s, 1H) 5.55-5.73 (m, 2H) 5.12-5.25 (m, 1H) 3.86-4.18 (m , 2H) 3.43-3.57 (m, 1H) 2.59-2.69 (m, 1H) 1.92-2.03 (m, 1H) 1.48 (s, 9H) 1.30-1.40 (m, 3H)

[0439] ESI-MS m / z 444 (MH+ )

[0440] Synthesis Example 1 (2) 4-amino-7-((3R,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-3- Synthesis of 7H-pyrrolo[2,3-d]pyrimidine-5-carboxylic acid

[0441] After adding the compound (28.0 g) of Synthesis Example 1 (1), 10% palladium carbon catalyst (720 mg), NMP (84 mL), methanol (26 mL), and triethylamine (17.6 mL) into a pressure tube, carbon monoxide substitution was performed and the mixture was stirred at 100°C for 2 hours. The reaction mixture was cooled to room temperature, 2M sodium hydroxide aqueous solution (79 mL) was added, and the mixture was stirred at 80°C for 2 hours. The reaction mixture was cooled to room temperature, filtered through diatomaceous earth, washed with methanol, and the methanol in the filtrate was concentrated under reduced pressure. Next, water was added and the aqueous layer was washed with tert-butyl methyl ether. 1M potassium hydrogen sulfate aqueous solution was added to the aqueous layer, the pH was adjusted to about 3, the precipitated solid was filtered, washed with water, and dried to obtain the target product (23.4 g).

[0442] 1 HNMR (400MHz, DMSO-d6) δ: 8.14 (s, 1H) 8.08

[0443] (s, 1H) 5.16-4.93 (m, 1H) 4.07-3.79 (m, 2H) 3.61-3.45 (m, 1H) 2.53 (m, 1H) 2.33-2.02 (m, 1H) 1.42 (s, 9H) 1.29 (d, J = 6.1Hz, 3H)

[0444] ESI-MS m / z 362(MH + )

[0445] Synthesis Example 1(3) (2S,4R)-4-(4-amino-6-bromo-5-(((R)-1-phenylethyl)carbamoyl Synthesis of tert-butyl (7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-methylpyrrolidine-1-carboxylate

[0446] The compound of Synthesis Example 1 (2) (1.00 g), (R)-(+)-1-phenylethylamine (0.503 g), diisopropylethylamine (1.79 g), N,N-dimethylformamide (10 mL) were added, followed by HATU (1.58 g), and the mixture was stirred at room temperature overnight. Ethyl acetate and saturated sodium bicarbonate aqueous solution were added to the reaction mixture, and the mixture was extracted with ethyl acetate. The combined organic layers were washed with water and then with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (hexane: acetone) to obtain an amide (1.53 g). The obtained compound was used in the next reaction without further purification.

[0447] Chloroform (15 mL) was added to the amide (1.53 g), and the mixture was cooled to 0°C, and then N-bromosuccinimide (0.88 g) was added and stirred at 0°C for 1 hour. The reaction mixture was concentrated under reduced pressure, and the obtained residue was purified by silica gel chromatography (hexane:ethyl acetate) to obtain the target product (1.39 g).

[0448] 1 HNMR (CDCl3) δ: 8.21 (s, 1H) 7.42-7.28 (m, 5H) 6.97 (d, J = 7.3Hz, 1H) 5.36-5.29 (m, 1H) 5.20-5.07 (m, 1H) 4.30 (t, J = 1 0.3Hz, 1H) 4.04-3.72 (m, 2H) 3.00-2.86 (m, 1H) 2.38 (dt, J=14.3, 6.0Hz, 1H) 1.63 (d, J=7.0Hz, 3H) 1.53-1.43 (m, 12H)

[0449] ESI-MS m / z 543, 545 (MH + )

[0450] Synthesis Example 1 (4) 7-((3R, 5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino- 6-Bromo-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0451] Chloroform (3 mL) was added to the compound (600 mg) of Synthesis Example 1 (3), and after cooling to 0°C, trifluoroacetic acid (4.44 g) was added and stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure, acetonitrile (5 mL) was added to the residue, and it was concentrated under reduced pressure again to obtain an amine. The obtained compound was used in the next reaction without further purification.

[0452] Acetonitrile (3 mL) was added to the obtained amine, and after cooling to 0°C, acryloyl chloride (99.9 mg) and diisopropylethylamine (713 mg) were added, and stirred at 0°C for 1 hour. The reaction mixture was concentrated under reduced pressure, and the obtained residue was purified by silica gel chromatography (ethyl acetate: methanol) to obtain the target product (281 mg).

[0453] 1HNMR (CDCl3) δ: 8.20 (d, J=7.3Hz, 1H) 7.42-7.36 (m, 4H) 7.32-7.28 (m, 1H) 7.00-6.94 ( m, 1H) 6.57-6.33 (m, 2H) 5.76-5.66 (m, 1H) 5.36-5.29 (m, 1H) 5.14-5.08 (m, 1H) 4, 71 (t , J=9.9Hz, 0.7H)4.42-4.23(m, 1.6H)3.83(t, J=8.6Hz, 0.7H)3.03-2.92(m, 1H)2.60- 2.57 (m, 0.3H) 2.44-2.40 (m, 0.7H) 1.64 (d, J = 6.6Hz, 3H) 1.56 (dd, J = 11.7, 6.2Hz, 3H)

[0454] ESI-MS m / z 497, 499 (MH+)

[0455] Synthesis Example 1 (5) 7-((3R, 5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino- 6-(Cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (Synthesis of Compound (I))

[0456] The compound of Synthesis Example 1 (4) (65 mg), dichlorobis(triphenylphosphine)dipalladium (9.2 mg), copper (I) iodide (5.0 mg), cyclopropylacetylene (13.0 mg), triethylamine (39.7 mg), and N,N-dimethylformamide (1.3 mL) were added, and the system was replaced with nitrogen, followed by stirring at 70°C for 2.5 hours. Ethyl acetate and saturated aqueous ammonium chloride solution were added to the reaction mixture, and the mixture was extracted with ethyl acetate. The combined organic layers were washed with water and then with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (chloroform: methanol) to obtain the target compound (50 mg).

[0457] 1 HNMR (CDCl3) δ: 8.22 (d, J = 5.1Hz, 1H) 7.82 (d, J = 7.3Hz, 1H) 7.43-7.35 (m, 4H) 7.30 (t, J = 6 .8Hz, 1H) 6.58-6.34 (m, 2H) 5.77-5.66 (m, 1H) 5.35-5.20 (m, 2H) 4.54 (t, J = 10.1Hz, 0.7H) 4.35-4.25(m, 1.6H)3.88(t, J=8.8Hz, 0.7H)2.90-2.78(m, 1H)2.65-2.56(m, 0.3H)2.49- 2.40 (m, 0.7H) 1.63 (d, J = 7.0Hz, 3H) 1.56-1.45 (m, 4H) 1.03-0.91 (m, 2H) 0.84-0.69 (m, 2H)

[0458] ESI-MS m / z 483(MH + )

[0459] Example 1 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6- (Cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide ... Preparation of Type II crystals of compound (I)) and 1 equivalent of fumaric acid

[0460] Type II crystals with fumaric acid were prepared by the following two methods described in Preparation Method 1 and Preparation Method 2. Type I crystals of the free form of Compound (I) were synthesized by the same method as in Example 3 described below.

[0461] (i) Manufacturing method 1

[0462] To 30 mg of the free form of type I crystal of compound (I) were added 3 equivalents of fumaric acid and 0.3 mL of tert-butanol. The suspension was stirred at 25°C for about 124 hours and then filtered to recover the solid and dry it to obtain the title crystal.

[0463] (ii) Manufacturing method 2

[0464] 1.06 g of fumaric acid and 22 mL of acetone were added to 2.20 g of the free form of compound (I) Type I crystals, and the suspension was stirred at 50°C for about 62 hours, then naturally cooled for 30 minutes, filtered, and the solid was recovered. The solid was washed with 2-propanol, filtered, and the solid was recovered and dried to obtain 2.27 g of the title crystal.

[0465] Powder X-ray diffraction pattern (method A): shown in Figure 1 The powder X-ray diffraction pattern data are shown in Table 2.

[0466] [Table 2]

[0467] Table 2: Powder X-ray diffraction pattern data of type II crystals of compound (I) and 1 equivalent of fumaric acid

[0468]

[0469] In addition, the characteristic diffraction angle is as follows.

[0470] Characteristic diffraction angle (2θ±0.2°):

[0471] 5.5°, 6.8°, 9.3°, 13.4°, 15.3°, 16.3°, 18.5°, 19.8°, 22.0° and 24.5°

[0472] Differential thermal and thermogravimetric simultaneous measurement curve: shown in Figure 2 .

[0473] Endothermic peak (peak value) in the DTC-TG simultaneous measurement curve: around 178°C

[0474] Example 2 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6- (Cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide ... Preparation of free-body II crystals of compound (I)

[0475] The free-body type II crystal is produced by any of the four methods described in the following production method 1 to production method 4.

[0476] (i) Manufacturing method 1

[0477] To 15 mg of compound (I) obtained by the method of Synthesis Example 1 were added 1 equivalent of succinic acid and 0.3 mL of 1-propanol / water (1:3 v / v), and the mixture was suspended and stirred at 50°C for 4 days, filtered, and the solid was recovered and dried to obtain the title crystal.

[0478] (ii) Manufacturing method 2

[0479] To 300 mg of compound (I) obtained according to the method of Synthesis Example 1, 1 equivalent of succinic acid and 1.5 mL of 1-propanol / water (1:3 v / v) were added, and a small amount of the crystals obtained in (i) was added as seed crystals. After suspending and stirring at 50°C for 5 hours, 0.3 mL of 1-propanol was added. Subsequently, 0.3 mL of 1-propanol was added after suspending and stirring at 50°C for 2 hours. After that, 0.3 mL of 1-propanol was added. After suspending and stirring at 50°C for 17 hours, the mixture was filtered out, the solid was recovered, and dried to obtain 84.5 mg of the title crystals.

[0480] (iii) Manufacturing method 3

[0481] To 1000 mg of compound (I) obtained according to the method of Synthesis Example 1, 1 equivalent of succinic acid and 3 mL of 1-propanol / water (1:1 v / v) were added, and a small amount of the crystals obtained in (ii) was added as seed crystals. After suspension and stirring at 50°C for 17.5 hours, the mixture was filtered out (washed with water during filtration), and the solid was recovered and dried to obtain 487.6 mg of the title crystals.

[0482] (iv) Manufacturing method 4

[0483] To 1000 mg of compound (I) obtained by the method of Synthesis Example 1, 5 mL of diisopropyl ether was added, and a small amount of the crystals obtained in (iii) was added as seed crystals, and the mixture was suspended and stirred at 50°C. After 42.5 hours, due to evaporation of the solvent, 5 mL of diisopropyl ether was added. After 2 hours of suspension and stirring at 50°C, 0.5 mL of ethanol was added. After 2 hours of suspension and stirring at 50°C, 0.5 mL of ethanol was added. After 19 hours of suspension and stirring at 50°C, the mixture was naturally cooled for 30 minutes, filtered, and the solid was recovered and dried to obtain 798.8 mg of the title crystal.

[0484] Powder X-ray diffraction pattern (method A): shown in Figure 3 The powder X-ray diffraction pattern data are shown in Table 3.

[0485] [Table 3]

[0486] Table 3: Powder X-ray diffraction pattern data of free form II crystal of compound (I)

[0487]

[0488] In addition, the characteristic diffraction angle is as follows.

[0489] Characteristic diffraction angle (2θ±0.2°):

[0490] 8.3°, 14.8°, 17.3°, 18.0°, 19.1°, 20.3°, 21.0°, 22.5°, 23.0° and 26.2°

[0491] Differential thermal and thermogravimetric simultaneous measurement curve: shown in Figure 4 .

[0492] Endothermic peak (peak value) in the DTC-TG simultaneous measurement curve: around 182°C

[0493] Example 37 - ((3R, 5S) -1-acryloyl-5-methylpyrrolidin-3-yl) -4-amino-6- (Cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide ... Preparation of free form I crystal of compound (I)

[0494] The crude product of Compound (I) (100 mg) was suspended in ethanol (500 μL) and stirred overnight at 50° C. The solution was cooled to 25° C. and the obtained suspension was filtered to obtain free form I crystals of Compound (I) (31 mg).

[0495] Powder X-ray diffraction pattern (method A): shown in Figure 5 The powder X-ray diffraction pattern data are shown in Table 4.

[0496] [Table 4]

[0497] Table 4: Powder X-ray diffraction pattern data of free form I crystal of compound (I)

[0498]

[0499] In addition, the characteristic diffraction angle is as follows.

[0500] Characteristic diffraction angle (2θ±0.2°):

[0501] 9.9°, 11.7°, 13.2°, 17.7°, 18.1°, 18.8° and 20.8°

[0502] Differential thermal and thermogravimetric simultaneous measurement curve: shown in Figure 6 .

[0503] Endothermic peak (peak value) in the DTC-TG simultaneous measurement curve: around 183°C

[0504] Example 47 - ((3R, 5S) -1-acryloyl-5-methylpyrrolidin-3-yl) -4-amino-6- (Cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide ... Preparation of V-type crystals of compound (I)) and 1 equivalent of fumaric acid

[0505] To 1000 mg of Compound (I) obtained by the method of Synthesis Example 1, 481 mg of fumaric acid and 50 mL of acetone were added, and the mixture was suspended and stirred at 25°C for 71 hours, and then filtered to recover the solid, which was dried under reduced pressure at room temperature for 7.5 hours to obtain 695.0 mg of the title crystal.

[0506] Powder X-ray diffraction pattern (method A): shown in Figure 7 The powder X-ray diffraction pattern data are shown in Table 5.

[0507] [Table 5]

[0508] Table 5: Powder X-ray diffraction pattern data of V-type crystals of compound (I) and 1 equivalent of fumaric acid

[0509]

[0510] In addition, the characteristic diffraction angle is as follows.

[0511] Characteristic diffraction angle (2θ±0.2°):

[0512] 6.9°, 9.4°, 10.2°, 13.7°, 21.1°, 23.6° and 26.5°

[0513] Differential thermal and thermogravimetric simultaneous measurement curve: shown in Figure 8 .

[0514] Endothermic peak (peak value) in the DTC-TG simultaneous measurement curve: around 151°C

[0515] Example 57 - ((3R, 5S) -1-acryloyl-5-methylpyrrolidin-3-yl) -4-amino-6- (Cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide ... Preparation of Type I crystals of compound (I)) and 1 equivalent of fumaric acid

[0516] To 1000 mg of Compound (I) obtained by the method of Synthesis Example 1 were added 481 mg of fumaric acid and 50 mL of acetone, and the mixture was suspended and stirred at 25°C for 19.5 hours, and then filtered to recover the solid to obtain 868.8 mg of the title crystal.

[0517] Powder X-ray diffraction pattern (method A): shown in Fig. 9 The powder X-ray diffraction pattern data are shown in Table 6.

[0518] [Table 6]

[0519] Table 6: Powder X-ray diffraction pattern data of type I crystals of compound (I) and 1 equivalent of fumaric acid

[0520]

[0521] In addition, the characteristic diffraction angle is as follows.

[0522] Characteristic diffraction angle (2θ±0.2°):

[0523] 6.4°, 10.3°, 12.8°, 15.0°, 20.7°, 23.4° and 26.6°

[0524] Differential thermal and thermogravimetric simultaneous measurement curve: shown in Fig.10 .

[0525] Endothermic peak (peak value) in the DTC-TG simultaneous measurement curve: around 153°C

[0526] Example 67 - ((3R, 5S) -1-acryloyl-5-methylpyrrolidin-3-yl) -4-amino-6- (Cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide ... Preparation of Type I crystals of compound (I)) and 0.5 equivalent of fumaric acid

[0527] To 15 mg of compound (I) obtained by the method of Synthesis Example 1 were added 0.5 equivalent of fumaric acid and 0.3 mL of water, and the mixture was suspended and stirred at 50°C for 93.5 hours, and then filtered to recover the solid and dry it to obtain the title crystal.

[0528] Powder X-ray diffraction pattern (B method): shown in Fig.11 .

[0529] Characteristic diffraction angle (2θ±0.2°): 6.4°, 7.5°, 9.7°, 11.7°, 15.1°, 19.6° and 23.9°

[0530] Differential thermal and thermogravimetric simultaneous measurement curve: shown in Fig.12 .

[0531] Endothermic peak (peak value) in the simultaneous differential thermal and thermogravimetric measurement curve: around 161°C

[0532] Example 7 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6- (Cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide ... Preparation of Type II crystals of compound (I)) and 0.5 equivalent of fumaric acid

[0533] To 300 mg of compound (I) obtained by the method of Synthesis Example 1, 1 equivalent of fumaric acid and 6 mL of ethanol were added, and the mixture was suspended and stirred at 50° C. for 19.5 hours, and then naturally cooled for 3 hours, filtered, and the solid was recovered. The solid was washed with water, filtered, and the solid was recovered and dried to obtain 224.6 mg of the title crystal.

[0534] Powder X-ray diffraction pattern: shown in Fig.13 .

[0535] Characteristic diffraction angle (2θ±0.2°): 5.4°, 6.4°, 7.3°, 12.8°, 13.4°, 14.7° and 15.4°

[0536] Differential thermal and thermogravimetric simultaneous measurement curve: shown in Fig.14 .

[0537] Endothermic peak (peak value) in the DTC-TG simultaneous measurement curve: around 162°C

[0538] Example 8 Analysis of the molar ratio of compound (I) to fumaric acid

[0539] The molar ratios of compound (I) to fumaric acid and compound (I) to L-tartaric acid are as follows: 1 H-NMR was used for confirmation. The results of the NMR spectra of the type I crystal of compound (I) with 1 equivalent of fumaric acid, the type II crystal of compound (I) with 1 equivalent of fumaric acid, and the type V crystal of compound (I) with 1 equivalent of fumaric acid are shown below. In addition, the results of the NMR spectra of the free form type I crystal and the free form type II crystal of compound (I) are also shown below.

[0540] (Crystal of the free form of compound (I) Type I)

[0541] 1 HNMR (400MHz, DMSO-d6) δ: 8.32 (brd, J=7.38Hz, 1H) 8.14 (s, 1H) 7.25-7.50 (m, 5H) 6.54-6.72 (m, 1H) 6.13-6.23 (m, 1H) 5.65-5.76 (m, 1H) 5.12-5.39 ( m, 2H) 4.33-4.40 (m, 1H) 4.00-4.22 (m, 2H) 2.44-2.70 (m, 2H) 1.73 (tt, J=4. 99, 8.27Hz, 1H) 1.51 (d, J = 6.88Hz, 3H) 1.36-1.43 (m, 3H) 0.78-1.03 (m, 4H)

[0542] (Crystal of Free Form II of Compound (I))

[0543] 1HNMR (400MHz, DMSO-d6) δ: 8.32 (brd, J=7.38Hz, 1H) 8.14 (s, 1H) 7.27-7.46 (m, 5H) 6.54-6.72 (m, 1H) 6.13-6.23 (m, 1H) 5.65-5.76 (m, 1H) 5.12-5.39 ( m, 2H) 4.32-4.40 (m, 1H) 4.00-4.22 (m, 2H) 2.44-2.69 (m, 2H) 1.73 (tt, J=4. 99, 8.27Hz, 1H) 1.51 (d, J = 6.88Hz, 3H) 1.40-1.43 (m, 3H) 0.78-1.03 (m, 4H)

[0544] (Type I crystal of compound (I) and 1 equivalent of fumaric acid)

[0545] 1 HNMR (400MHz, DMSO-d6) δ: 13.13 (brs, 2H) 8.32 (brd, J = 7.38Hz, 1H) 8.14 (s, 1H )7.24-7.48(m,5H)6.46-6.78(m,3H)6.06-6.30(m,1H)5.63-5.77(m,1H)5.11 -5.38(m,2H)4.27-4.44(m,1H)3.98-4.23(m,2H)2.42-2.73(m,2H)1.73(tt,J =5.02, 8.30Hz, 1H) 1.51 (d, J = 6.88Hz, 3H) 1.32-1.45 (m, 3H) 0.77-1.02 (m, 4H)

[0546] (Type II crystal of compound (I) and 1 equivalent of fumaric acid)

[0547] 1 HNMR (400MHz, DMSO-d6) δ: 13.14 (brs, 2H) 8.32 (brd, J = 7.38Hz, 1H) 8.14 (s, 1H )7.25-7.49(m,5H)6.39-6.89(m,3H)6.07-6.28(m,1H)5.62-5.77(m,1H)5.12 -5.39(m,2H)4.27-4.45(m,1H)3.99-4.24(m,2H)2.42-2.75(m,2H)1.74(tt,J =5.02, 8.30Hz, 1H) 1.51 (d, J = 6.88Hz, 3H) 1.37-1.46 (m, 3H) 0.78-1.03 (m, 4H)

[0548] (V-type crystal of compound (I) and 1 equivalent of fumaric acid)

[0549] 1 HNMR (400MHz, DMSO-d6) δ: 13.13 (brs, 2H) 8.32 (brd, J = 7.38Hz, 1H) 8.14 (s, 1H )7.21-7.49(m,5H)6.47-6.80(m,3H)6.09-6.26(m,1H)5.63-5.77(m,1H)5.11 -5.38(m,2H)4.27-4.44(m,1H)3.98-4.23(m,2H)2.42-2.72(m,2H)1.73(tt,J =4.99, 8.27Hz, 1H) 1.51 (d, J=6.88Hz, 3H) 1.34-1.46 (m, 3H) 0.77-1.02 (m, 4H)

[0550] Reference Example 1 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6- (Cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide ... Preparation of Type III crystals of compound (I)) and 1 equivalent of fumaric acid

[0551] To 300 mg of compound (I) obtained by the method of Synthesis Example 1 were added 3 equivalents of fumaric acid and 9 mL of acetonitrile, and the mixture was suspended and stirred at 50°C for 1 hour, then naturally cooled, filtered, and the solid was recovered and dried to obtain 303.7 mg of the title crystal.

[0552] Powder X-ray diffraction pattern (B method): shown in Fig.15 .

[0553] Characteristic diffraction angle (2θ±0.2°): 8.1°, 13.2°, 20.4°, 23.1°, 24.7° and 26.1°

[0554] Differential thermal and thermogravimetric simultaneous measurement curve: shown in Fig.16 .

[0555] Exothermic peak (peak value) in the simultaneous differential thermal and thermogravimetric measurement curve: around 153°C

[0556] Reference Example 2 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6- (Cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide ... Preparation of IV type crystals of compound (I)) and 1 equivalent of fumaric acid

[0557] To 300 mg of compound (I) obtained by the method of Synthesis Example 1, 3 equivalents of fumaric acid and 6 mL of water were added, and the mixture was suspended and stirred at 50° C. for 1.5 hours, then naturally cooled, filtered, and the solid was recovered. The solid was washed with water, filtered, and the solid was recovered and dried to obtain 311.2 mg of the title crystal.

[0558] Powder X-ray diffraction pattern (B method): shown in Fig.17 .

[0559] Characteristic diffraction angle (2θ±0.2°): 6.0°, 15.7° and 18.8°

[0560] Differential thermal and thermogravimetric simultaneous measurement curve: shown in Fig.18 .

[0561] Exothermic peak (peak value) in the DTC-TG simultaneous measurement curve: around 130°C

[0562] Reference Example 3 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6- (Cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide ... Preparation of Type I crystals of compound (I)) and hydrochloric acid

[0563] To 15 mg of compound (I) obtained by the method of Synthesis Example 1, 15 μL of 2M hydrochloric acid / ethanol solution and 0.3 mL of a 2-propanol / heptane mixture (1:3 v / v) were added, and the mixture was suspended and stirred for 4 days, then naturally cooled, filtered, and the solid was recovered and dried to obtain the title crystal.

[0564] Powder X-ray diffraction pattern (B method): shown in Fig.19 .

[0565] Characteristic diffraction angle (2θ±0.2°): 7.0°, 7.4°, 12.6°, 19.0° and 25.4°

[0566] Differential thermal and thermogravimetric simultaneous measurement curve: shown in Fig. 20 .

[0567] Endothermic peaks (peak values) in the DTC-TG simultaneous measurement curve: around 51°C and 179°C

[0568] Reference Example 4 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6- (Cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide ... Preparation of Type II crystals of compound (I)) and hydrochloric acid

[0569] To 15 mg of compound (I) obtained by the method of Synthesis Example 1, 15 μL of 2M hydrochloric acid / ethanol solution and 0.3 mL of acetone / heptane mixture (1:3 v / v) were added, and the mixture was suspended and stirred for 4 days, then naturally cooled, filtered, and the solid was recovered and dried to obtain the title crystal.

[0570] Powder X-ray diffraction pattern (B method): shown in Fig.21 .

[0571] Characteristic diffraction angle (2θ±0.2°): 6.3°, 7.4°, 8.0°, 17.6°, 22.0° and 23.6°

[0572] Differential thermal and thermogravimetric simultaneous measurement curve: shown in Fig. 22 .

[0573] Endothermic peak (peak value) in the DTC-TG simultaneous measurement curve: around 147°C

[0574] Reference Example 5 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6- (Cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide ... Preparation of Type III crystals of compound (I)) and hydrochloric acid

[0575] 15 μL of 2M hydrochloric acid / ethanol solution and 0.3 mL of ethyl acetate were added to 15 mg of compound (I) obtained by the method of Synthesis Example 1, and the mixture was suspended and stirred for 4 days, then naturally cooled, filtered, and the solid was recovered and dried to obtain the title crystal.

[0576] Powder X-ray diffraction pattern (B method): shown in Fig.23 .

[0577] Characteristic diffraction angle (2θ±0.2°): 5.3°, 6.5°, 8.1°, 15.3° and 24.2°

[0578] Differential thermal and thermogravimetric simultaneous measurement curve: shown in Fig.24 .

[0579] Endothermic peak (peak value) in the DTC-TG simultaneous measurement curve: around 156°C

[0580] Reference Example 6 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6- (Cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide ... Preparation of crystals of compound (I)) and hydrobromic acid

[0581] To 15 mg of compound (I) obtained by the method of Synthesis Example 1, 1 equivalent of hydrobromic acid and 0.3 mL of a 2-propanol / heptane mixture (1:3 v / v) were added, and the mixture was suspended and stirred for 5 days, then naturally cooled, filtered, and the solid was recovered and dried to obtain the title crystal.

[0582] Powder X-ray diffraction pattern (B method): shown in Fig.25 .

[0583] Characteristic diffraction angle (2θ±0.2°): 7.0°, 7.4°, 13.4°, 15.6°, 19.2° and 25.6°

[0584] Differential thermal and thermogravimetric simultaneous measurement curve: shown in Fig.26 .

[0585] Endothermic peak (peak value) in the simultaneous differential thermal and thermogravimetric measurement curve: around 204°C

[0586] Reference Example 7 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6- (Cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide ... Preparation of crystals of compound (I)) and 1 equivalent of L-tartaric acid

[0587] To 500 mg of compound (I) obtained by the method of Synthesis Example 1, 466 mg of L-tartaric acid and 10 mL of acetone were added, and the mixture was stirred at 50°C for 40 minutes to dissolve. After that, the mixture was stirred at room temperature for 17 hours, and the obtained suspension was filtered to recover the solid. The solid was dried to obtain 506.3 mg of the title crystal.

[0588] Powder X-ray diffraction pattern (method A): shown in Fig. 27 .

[0589] Characteristic diffraction angle (2θ±0.2°): 7.2°, 12.6°, 17.3° and 23.4°

[0590] Differential thermal and thermogravimetric simultaneous measurement curve: shown in Fig.28 .

[0591] Endothermic peak (peak value) in the simultaneous differential thermal and thermogravimetric measurement curve: around 121°C

[0592] Reference Example 8 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6- (Cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide ... Preparation of crystals of compound (I)) and succinic acid

[0593] To 15 mg of compound (I) obtained by the method of Synthesis Example 1, 3 equivalents of succinic acid and 0.3 mL of acetonitrile were added, and the mixture was suspended and stirred at 50°C for 3 days, then naturally cooled, filtered, and the solid was recovered and dried to obtain the title crystal.

[0594] Powder X-ray diffraction pattern (B method): shown in Fig.29 .

[0595] Characteristic diffraction angle (2θ±0.2°): 6.5°, 10.7°, 13.2°, 14.6°, 18.7° and 23.9°

[0596] Differential thermal and thermogravimetric simultaneous measurement curve: shown in Fig.30 .

[0597] Endothermic peaks (peak values) in the DTC-TG simultaneous measurement curve: around 148°C and 159°C

[0598] Experimental Example 1 HER2 phosphorylation activity inhibition (in in vitro (in vitro) assay

[0599] In the setting of the conditions for the in vitro inhibitory activity assay of the compound on HER2 phosphorylation activity, ProfilerPro Peptide 22 was used as a substrate based on a report on a HER2 kinase reaction using a peptide with the same sequence (5-FAM-EEPLYWSFPAKKK-CONH2) as that of PerkinElmer's ProfilerPro Peptide 22 as a substrate (Xie H et al., PLoS One. 2011; 6(7): e21487). The purified recombinant human HER2 protein used in the experiment was purchased from Carna Biosciences. In the inhibitory activity assay of the compound, first, compound (I) obtained according to the method of Synthesis Example 1 was gradiently diluted with dimethyl sulfoxide (DMSO). Then, HER2 protein, substrate peptide (final concentration of 1 μM), manganese chloride (final concentration of 10 mM), ATP (final concentration of 5 μM) and DMSO solution of compound (I) (final concentration of DMSO of 5%) were added to kinase reaction buffer (13.5 mM Tris (pH 7.5), 2 mM dithiothreitol, 0.009% Tween20), and incubated at 25°C for 30 minutes to perform kinase reaction. EDTA was added to the reaction in such a way that the final concentration reached 30 mM, thereby stopping the reaction. Finally, the non-phosphorylated substrate peptide (S) and the phosphorylated peptide (P) were separated and detected by microfluidic capillary electrophoresis using LabChip (registered trademark) EZ Reader II (PerkinElmer). The amount of phosphorylation reaction was determined from the height of each peak of S and P, and the concentration of the compound that can inhibit the phosphorylation reaction by 50% was defined as the IC50 value (nM), which is shown in Table 7 below.

[0600] Experimental Example 2 Inhibition of phosphorylation activity of exon 20 insertion variant HER2 (HER2ex20insYVMA) In vitro assay

[0601] In the in vitro inhibitory activity assay of the compound on the phosphorylation activity of exon 20 insertion mutant HER2, ProfilerPro Peptide 22 was used as a substrate in the same manner as HER2. The purified recombinant human exon 20 insertion mutant HER2 (A775_G776insYVMA) protein used in the test was purchased from SignalChem. In the inhibitory activity assay of the compound, the compound (I) obtained according to the method of Synthesis Example 1 was first diluted with dimethyl sulfoxide (DMSO) in a gradient manner. Next, the exon 20 insertion mutant HER2 protein and the compound (I) DMSO solution (the final concentration of DMSO was 5%) were added to the kinase reaction buffer (13.5mM Tris (pH 7.5), 2mM dithiothreitol, 0.009% Tween20), and pre-incubated at 25°C for 30 minutes. Then, substrate peptide (final concentration of 1 μM), manganese chloride (final concentration of 25 mM), magnesium chloride (final concentration of 20 mM), and ATP (final concentration of 200 μM) were added, and the kinase reaction was carried out at 25°C for 220 minutes. EDTA was added to a final concentration of 30 mM to stop the reaction. Finally, the non-phosphorylated substrate peptide (S) and the phosphorylated peptide (P) were separated and detected by microfluidic capillary electrophoresis using LabChip (registered trademark) EZReader II (PerkinElmer). The amount of phosphorylation reaction was calculated from the height of each peak of S and P, and the concentration of the compound that can inhibit the phosphorylation reaction by 50% was defined as the IC50 value (nM), which is shown in Table 7 below.

[0602] [Table 7]

[0603]

[0604] From the above results, it can be seen that compound (I) has excellent inhibitory activity against HER2 phosphorylation and exon 20 insertion mutant HER2 phosphorylation.

[0605] Test Example 3 Determination of proliferation inhibitory activity on HER2 expressing cell lines

[0606] SK-BR-3 cells, a human breast cancer cell line overexpressing HER2, were suspended in McCoy's 5a medium (manufactured by Life Technologies) containing 10% fetal bovine serum. The cell suspension was inoculated into each well of a 384-well flat-bottom microplate and cultured at 37°C in an incubator containing 5% carbon dioxide for 1 day. Compound (I) obtained according to the method of Synthesis Example 1 was dissolved in DMSO, and the compound was diluted to a concentration 500 times the final concentration using DMSO. The DMSO solution of the compound was diluted with the culture medium used for cell suspension, added to each well of the cell culture plate in such a way that the final concentration of DMSO was 0.2%, and cultured for another 3 days at 37°C in an incubator containing 5% carbon dioxide. CellTiter-Glo2.0 (manufactured by Promega) was used to measure the number of cells after culturing for 3 days in the presence of the compound, and the proliferation inhibition rate was calculated by the following formula. The concentration of the compound that can inhibit proliferation by 50% is defined as IC50 (nM).

[0607] Proliferation inhibition rate (%) = (C-T) / (C) × 100

[0608] T: Luminescence intensity of the wells to which the test compound was added

[0609] C: Luminescence intensity of the wells without added test compound

[0610] The results are shown in Table 8 below.

[0611] Test Example 4: Determination of the proliferation inhibitory activity of exon 20 insertion mutant HER2 expressing cell lines

[0612] The proliferation inhibitory activity against exon 20 insertion mutant HER2 was tested using mouse B lymphocyte progenitor cell line Ba / F3 cells into which human exon 20 insertion mutant HER2 gene was introduced. Ba / F3 cells were maintained in RPMI-1640 medium (Thermo Fisher Scientific) containing 10% fetal bovine serum (FBS), 100 U / mL penicillin, 100 μg / mL streptomycin (Thermo Fisher Scientific) and 1 ng / mL mouse interleukin-3 (mIL-3) (CST). Human exon 20 insertion mutant HER2 gene (A775_G776insYVMA (HER2ex20insYVMA)), Internal Ribosome

[0613] Binding Sequence (IRES) and Kusabira-Orange gene were introduced into the pCDNA3.1-hyg(+) vector by electroporation using Amaxa(registered trademark) Cell Line Nucleofector(registered trademark) Kit V. Ba / F3 cells expressing exon 20 insertion mutant HER2 (Ba / F3-HER2insYVMA) selected with hygromycin B (NACALAI TESQUE) showed mIL-3-independent proliferation.

[0614] In the evaluation of cell proliferation inhibitory activity, Ba / F3-HER2insYVMA cells were suspended in RPMI-1640 medium containing 10% FBS, 100 U / mL penicillin, and 100 μg / mL streptomycin, and the cell suspension was inoculated in each well of a 96-well flat-bottom microplate, and cultured at 37°C in an incubator containing 5% carbon dioxide for 1 day. Compound (I) obtained according to the method of Synthesis Example 1 was dissolved in DMSO, diluted with DMSO or the medium used for cell suspension, and added to each well of the cell culture plate in such a way that the final concentration of DMSO became 0.2%, and further cultured at 37°C in an incubator containing 5% carbon dioxide for 3 days. CellTiter-Glo (produced by Promega) was used to measure the number of cells after culturing for 3 days in the presence of the compound, and the proliferation inhibition rate was calculated by the following formula. The concentration of the compound that can inhibit proliferation by 50% is defined as IC50 (nM).

[0615] Proliferation inhibition rate (%) = (C-T) / (C) × 100

[0616] T: Luminescence intensity of the wells to which the test compound was added

[0617] C: Luminescence intensity of the wells without added test compound

[0618] The results are shown in Table 8 below.

[0619] [Table 8]

[0620]

[0621] From the above results, it was found that compound (I) also has excellent cell proliferation inhibitory activity in HER2-expressing cell line (SK-BR-3) and exon 20 insertion mutant HER2-expressing cell line (Ba / F3-HER2insYVMA).

[0622] Test Example 5 Determination of proliferation inhibitory activity against HER2-expressing cell line (NCI-N87)

[0623] HER2-overexpressing human gastric cancer cell line NCI-N87 cells (American Type Culture Collection, Cat No. ATCC (registered trademark) CRL-5822) were suspended in RPMI1640 medium (Wako Pure Chemical Industries, Ltd.) containing 10% fetal bovine serum. Then, the cell suspension was inoculated in each well of a 96-well flat-bottom microplate and cultured at 37°C in an incubator containing 5% carbon dioxide for 1 day. Compound (I) obtained according to the method of Synthesis Example 1 was dissolved in DMSO, and the test compound was diluted with DMSO to a concentration 1000 times the final concentration. The DMSO solution of compound (I) was diluted with the culture medium used for cell suspension, and added to each well of the cell culture plate in such a way that the final concentration of DMSO became 0.1%. In the control wells, DMSO was diluted with the culture medium used for cell suspension, and added to each well of the cell culture plate in such a way that the final concentration of DMSO became 0.1%. After adding the drug solution, the cells were further cultured at 37°C in an incubator containing 5% carbon dioxide for 3 days. The number of cells after 3 days of culture in the presence of the compound was measured using CellTiter-Glo 2.0 (manufactured by Promega) according to the instructions recommended by Promega. The proliferation inhibition rate was calculated by the following formula, and the concentration of the test compound that could inhibit proliferation by 50% was defined as the IC50 value (nM).

[0624] Proliferation inhibition rate (%) = (C-T) / C×100

[0625] T: Luminescence intensity of the wells to which the test compound was added

[0626] C: Luminescence intensity of the wells without added test compound

[0627] The results are shown in Table 9 below.

[0628] [Table 9]

[0629]

[0630] From the above results, it can be seen that compound (I) also has excellent cell proliferation inhibitory activity in the HER2 overexpressing cell line (NCI-N87).

[0631] Test Example 6 Evaluation of oral absorbability

[0632] Compound (I) obtained by the method of Synthesis Example 1 was suspended or dissolved in 0.5% HPMC aqueous solution and 0.1N hydrochloric acid, and orally administered to BALB / cA mice (Clea Co., Ltd., Japan) at a dose of 50 mg / kg / day. Blood was collected from the facial vein over time 0.5, 1, 2, 4, and 6 hours after oral administration to obtain plasma. The compound concentration in the obtained plasma was measured by LC-MS / MS to evaluate oral absorbability.

[0633] The results are shown in Table 10 below.

[0634] [Table 10]

[0635] AUC 0-6hr (μM·hr) 31

[0636] From the above results, sufficient plasma concentrations of Compound (I) were observed, indicating good oral absorbability.

[0637] Test Example 7 Evaluation of brain metastasis

[0638] The compound (I) obtained by the method of Synthesis Example 1 was suspended or dissolved in 0.5% HPMC aqueous solution and 0.1N hydrochloric acid, and was orally administered to BALB / cA mice (Japan Clea Co., Ltd.) at a dose of 50 mg / kg / day. After 0.5 hours after oral administration, blood was collected from the facial vein, and the whole brain was removed to obtain plasma and brain samples. After adding 3 times the amount of water to the obtained brain sample, it was homogenized using an ultrasonic homogenizer to obtain a brain homogenate. The compound concentration in the obtained plasma and brain homogenate was measured by LC-MS / MS, and brain metastasis was evaluated based on the compound concentration in the brain / plasma.

[0639] The results are shown in Table 11 below.

[0640] [Table 11]

[0641]

[0642] Based on the above results, compound (I) showed good brain transferability.

[0643] Experimental Example 8: Brain direct transplantation model of luciferase gene-transferred HER2-expressing cell line (NCI-N87-luc) Anti-tumor effect confirmation test (in vivo)

[0644] For the antitumor effect of the test compound using the brain direct transplantation model, NCI-N87-Luc, in which the luciferase gene was introduced into the human gastric cancer tumor cell NCI-N87 purchased from American Type Culture Collection, was used. The NCI-N87-Luc cell line was cultured in RPMI-1640 (containing 4.5 g / L glucose, 10 mM HEPES and 1 mM sodium pyruvate) (Wako Pure Chemical Industries, Ltd.) medium containing 10% fetal bovine serum (FBS) at 37°C in a 5% CO2 incubator.

[0645] NCI-N87-Luc cells were cultured at 6.25×10 7 cells / mL in PBS.

[0646] Nude mice (BALB / cAJc1-nu / nu, CLEA Co., Ltd., Japan) aged about 6 to 7 weeks were fixed to a brain fixation device using mouse ear bars, and the skin above the brain was disinfected with alcohol cotton and then incised with a scalpel.

[0647] A hole was made in the skull using a micro drill, and 4 μL of the cell suspension was transplanted into the brain at a flow rate of 0.8 μL / min using a needle, a manipulator, and a syringe pump.

[0648] As a rough standard of brain tumor volume, about 3 weeks after transplantation, the total flux (Total Flux) (photon / sec) of all surviving cases was measured using IVIS (PerkinElmer, Inc., model: Lumina II). Based on the results, 6 animals were assigned to each group using the grouping program of MiSTAT (Ver. 2.00).

[0649] The test compound was orally administered once a day starting from the second day after grouping for 21 consecutive days (Day 1-21).

[0650] The presence or absence of an effect was determined by logarithmically transforming (Log10) the total flux on the determination day (Day 22). As a test compound, Compound (I) obtained by the method of Synthesis Example 1 was used and administered at a dose of 25 mg / kg / day.

[0651] The vertical axis was set to the value obtained by logarithmic transformation (Log10) of the average total flux, and the horizontal axis was set to the number of days after transplantation (Day). A graph was prepared to observe the change in total flux over time during the drug administration period.

[0652] As a control (Control), a 0.1N HCl and 0.5% HPMC aqueous solution was used.

[0653] The results are shown in the following Fig.31 and Fig.32 . The results of the significant difference test on the logarithmic transformation (Log10) of the total flux on Day 22 of each group showed that it was statistically significantly (5% on both sides of the significance level) lower than that of the control group. In addition, the body weight was measured using an electronic balance for animals. The body weight change rate (Body weight change; BWCn) on day n was calculated from the body weight on day n (BWn) using the following formula.

[0654] BWCn (%) = [(body weight on day n) - (body weight on the day of grouping)] / (body weight on the day of grouping) × 100

[0655] The test results show that compound (I) has an excellent anti-tumor effect on HER2 overexpressing cell lines (NCI-N87-luc) transplanted into the brain of nude mice. In addition, no body weight loss of more than -20% was observed in any individual, indicating that there were no serious side effects.

[0656] Test Example 9 Solid Stability Test

[0657] The solid stability of the free-form Type II crystal of Compound (I) obtained in Examples and Comparative Examples and the Type II crystal in an amount of 1 equivalent of fumaric acid after storage for 4 weeks was evaluated.

[0658] Storage conditions: 60°C (closed system)

[0659] Storage period: 4 weeks

[0660] Storage capacity: about 30mg

[0661] Storage container: glass bottle

[0662] The results are shown in Table 12 below.

[0663] [Table 12]

[0664] Free body type II crystal Type II crystal with 1 equivalent of fumaric acid Chemical purity before storage (%) 99.87 99.85 Chemical purity after storage (%) 99.88 99.85 Amount of similar substances before storage (%) 0.13 0.15 Amount of similar substances after storage (%) 0.12 0.15

[0665] The change in the mass of the analog (the amount of the substance detected other than compound (I)) was determined by weighing about 1 mg of the sample, dissolving it in about 1 mL of a mixture of water and acetonitrile (1:1), accurately measuring 5 μL of the liquid, and analyzing it by HPLC by the following method.

[0666] HPLC assay (stability test)

[0667] The mass of the analog in the sample solution was determined by HPLC analysis. In addition, the operation of the devices including data processing followed the method and sequence indicated in each device.

[0668] Column: XSelect CSH C18 manufactured by Waters (4.6×150 mm, 3.5 μm)

[0669] UV detection: 246nm

[0670] Column temperature: 40°C

[0671] Flow rate: 1.0mL / min

[0672] Sample temperature: 5°C

[0673] Sample concentration: 1 mg / mL

[0674] Mobile phase A: 5mmol / L ammonium formate buffer (pH 6.5) / acetonitrile (9:1) mixture

[0675] Mobile phase B: acetonitrile

[0676] The gradients are shown in Table 13.

[0677] [Table 13]

[0678] Time (minutes) Mobile phase A (vol%) Mobile phase B (vol%) 0～31 90→30 10→70 31～36 30 70 36～37 30→90 70→10

[0679] As a result, no change in the powder X-ray diffraction pattern was observed for the free form type II crystal of compound (I) and the type II crystal of 1 equivalent fumaric acid, and almost no increase in similar substances was observed, indicating that the crystals were extremely stable.

[0680] Test Example 10 Dynamic moisture adsorption and desorption (DVS) test

[0681] Water adsorption and desorption tests were carried out using type II crystals of compound (I) obtained in the examples and reference examples with 1 equivalent of fumaric acid, free type II crystals, free type I crystals, type V crystals with 1 equivalent of fumaric acid, and crystals with 1 equivalent of L-tartaric acid.

[0682] The moisture adsorption and desorption test was carried out under the following conditions.

[0683] About 10 mg of the sample was placed in a dedicated quartz container (holder), and the weight of the sample at each humidity was continuously measured and recorded under the following conditions: The operation of the devices including the data processing was performed according to the methods and procedures indicated by each device.

[0684] Device: VTI SA+ (manufactured by TA Instruments)

[0685] Drying temperature: 60℃

[0686] Heating rate: 1℃ / min

[0687] Drying balance: within 300 minutes, confirm that the weight does not decrease by 0.01% within 5 minutes

[0688] Measurement temperature: 25℃

[0689] Wet balance: within 120 minutes, confirm that the humidity does not increase by 0.01 wt% within 5 minutes

[0690] Relative humidity program: 5~95%RH rises in 5%RH intervals, 95%RH~5%RH falls in 5%RH intervals

[0691] The weight changes within the measurement condition ranges obtained in these tests are shown in Tables 14 to 18.

[0692] [Table 14]

[0693] Table 14 Water adsorption and desorption test results of type II crystals of compound (I) and 1 equivalent of fumaric acid

[0694] [Table 15]

[0695] Table 15 Water adsorption and desorption test results of free form II crystals of compound (I)

[0696] [Table 16]

[0697] Table 16 Water adsorption and desorption test results of free form I crystal of compound (I)

[0698] [Table 17]

[0699] Table 17 Water adsorption and desorption test results of V-type crystals of compound (I) and 1 equivalent of fumaric acid

[0700]

[0701] [Table 18]

[0702] Table 18 Water adsorption and desorption test results of the crystals of compound (I) and 1 equivalent of L-tartaric acid

[0703]

[0704] As shown in Table 14, the weight increase of type II crystals of compound (I) and 1 equivalent of fumaric acid at a relative humidity of 95% was 0.38%, which was less than 1%, and it was judged that there was basically no hygroscopicity. Similarly, the mass increase of free type II crystals, free type I crystals, and type V crystals of 1 equivalent of fumaric acid of compound (I) at a relative humidity of 95% was less than 1% in the moisture adsorption and desorption test, and it was judged that there was basically no hygroscopicity (Tables 15 to 17). On the other hand, as shown in Table 18, the mass increase of compound (I) and 1 equivalent of L-tartaric acid was confirmed to be as high as 6.41% at a relative humidity of 95% in the moisture adsorption and desorption test, which shows that it has high hygroscopicity.

[0705] Therefore, the type II crystal of compound (I) and 1 equivalent of fumaric acid, the free type II crystal, the free type I crystal, and the type V crystal of compound (I) and 1 equivalent of fumaric acid have lower hygroscopicity than the crystal of compound (I) and 1 equivalent of L-tartaric acid, and can be said to be excellent from the perspective of industrial production of pharmaceuticals with stable quality among candidate compounds for pharmaceutical development. The type II crystal of compound (I) and 1 equivalent of fumaric acid, the free type II crystal, the free type I crystal, and the type V crystal of compound (I) and 1 equivalent of fumaric acid have been confirmed to have excellent properties as pharmaceuticals or pharmaceutical raw materials.

[0706] Test Example 11 Blood Concentration Measurement Test

[0707] The blood concentration test was conducted for the type I crystal of compound (I), the type II crystal of compound (I), and the type II crystal of 1 equivalent fumarate of compound (I) obtained in the examples. Specifically, 0.5% HPMC containing 0.1N hydrochloric acid was used for the type I crystal of compound (I), and 0.5% HPMC was used for the type II crystal of compound (I) and the type II crystal of compound (I) and 1 equivalent fumaric acid, respectively, to prepare suspensions of compound (I) (free body) with molecular weight conversion of 100 mg / 10 mL and 300 mg / 10 mL. These dosing solutions were orally administered to female rats (Crl: CD (SD), Charles River Co., Ltd., Japan) raised under feeding conditions at a volume of 10 mL per 1 kg body weight using an oral administration probe. After the administration, the rats were confirmed to be returned to the cage. In the cage, water and food were freely available. 0.5, 1, 2, 4, and 6 hours after administration, the rats were anesthetized with isoflurane, and about 80 μL of blood was collected from the jugular vein using a heparinized syringe and injection needle (25G).

[0708] The collected blood was cooled and centrifuged to separate the plasma. After the blood collection, the rats were returned to the rat cage to confirm the state of awakening from anesthesia. After the final blood collection, the depth of isoflurane anesthesia was confirmed, and the limbs were fixed in a supine position with a small animal experimental fixator to open the abdomen, and the rats were euthanized by bleeding through the incision of the abdominal arteries and veins.

[0709] The concentration of compound (I) in each plasma was quantified by the standard curve using LC / MS / MS and measured by the MRM method. Phoenix WinNonlin (v6.4.0, Certara USA, Inc.) was used to calculate AUC0-6hr (area under the blood concentration-time curve from 0 to 6 hours after administration calculated by the trapezoidal method), Cmax (maximum blood concentration), and Tmax (time to reach maximum blood concentration).

[0710] The results are shown in Table 19. When the dosage was increased from 100 mg / kg to 300 mg / kg, the C of the type I crystal of compound (I) and the type II crystal of compound (I) to which hydrochloric acid was added was max The AUC 0－6hr The C of the Type II crystal of Compound (I) and 1 equivalent of fumaric acid increased by 1.0 and 1.5 times, respectively. max and AUC0-6hr increased by 1.9 times, respectively. In addition, it was found that the C of the type II crystal of compound (I) and 1 equivalent of fumaric acid at a dose of 300 mg / kg was max Compared with the values ​​of the type I crystal of compound (I) and the type II crystal of compound (I) added with acid at the same dosage, the AUC values ​​were 1.7 times and 1.4 times higher, respectively. 0－6hr The values ​​were 2.4 times higher, respectively. Therefore, it was confirmed that the type II crystal of the compound (I) according to the present invention and 1 equivalent of fumaric acid showed more excellent oral absorbability.

[0711] [Table 19]

[0712]

[0713] In addition, all documents and publications recorded in this specification are incorporated into this specification by reference in their entirety, regardless of their purpose. In addition, this specification includes the scope of the patent request, specification and drawings of Japanese patent application No. 2020-121520 (filed on July 15, 2020) which is the basis for claiming priority of this application.

[0714] Several embodiments of the present invention have been described, but these embodiments are presented as examples and are not intended to limit the scope of the invention. These novel embodiments can be implemented in various other ways, and various omissions, substitutions, and changes can be made without departing from the scope of the subject matter of the invention. These embodiments or their variations are included in the scope and subject matter of the invention, and are also included in the invention described in the scope of the patent claim and its equivalent scope.

Claims

1. Type II crystals of 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide and fumaric acid, characterized in that: In the powder X-ray diffraction pattern, there are peaks at diffraction angles (2θ±0.2°) of 5.5°, 6.8°, 9.3°, 13.4°, 15.3°, 16.3°, 18.5°, 19.8°, 22.0° and 24.5°, and the molar ratio of 7-((3R, 5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide to fumaric acid is 1:

1.

2. The crystal according to claim 1, characterized in that: This is a crystal having a powder X-ray diffraction pattern shown in FIG1 .

3. The crystal according to claim 1 or 2, characterized in that: The endothermic peak determined by simultaneous differential thermal and thermogravimetric measurement is around 178°C.

4. Type II crystals of 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide, characterized by: In the powder X-ray diffraction pattern, there are peaks at diffraction angles (2θ±0.2°) of 8.3°, 14.8°, 17.3°, 18.0°, 19.1°, 20.3°, 21.0°, 22.5°, 23.0°, and 26.2°.

5. The crystal according to claim 4, characterized in that: This is a crystal having a powder X-ray diffraction pattern shown in FIG3 .

6. The crystal according to claim 4 or 5, characterized in that: The endothermic peak determined by simultaneous differential thermal and thermogravimetric measurement is around 182°C.

7. Type I crystals of 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide, characterized by: The powder X-ray diffraction pattern has peaks at diffraction angles (2θ±0.2°) of 9.9°, 11.7°, 13.2°, 17.7°, 18.1°, 18.8°, and 20.8°.

8. The crystal according to claim 7, characterized in that: This is a crystal having a powder X-ray diffraction pattern shown in FIG5 .

9. The crystal according to claim 7 or 8, characterized in that: The endothermic peak determined by simultaneous differential thermal and thermogravimetric measurement is around 183°C.

10. V-type crystals of 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide and fumaric acid, characterized by: In the powder X-ray diffraction pattern, there are peaks at diffraction angles (2θ±0.2°) of 6.9°, 9.4°, 10.2°, 13.7°, 21.1°, 23.6° and 26.5°, and the molar ratio of 7-((3R, 5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide to fumaric acid is 1:

1.

11. The crystal according to claim 10, characterized in that: This is a crystal having a powder X-ray diffraction pattern shown in FIG7 .

12. The crystal according to claim 10 or 11, characterized in that: The endothermic peak determined by simultaneous differential thermal and thermogravimetric measurement is around 151°C.

13. Type I crystals of 7-((3R,5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide and fumaric acid, characterized by: In the powder X-ray diffraction pattern, there are peaks at diffraction angles (2θ±0.2°) of 6.4°, 10.3°, 12.8°, 15.0°, 20.7°, 23.4° and 26.6°, and the molar ratio of 7-((3R, 5S)-1-acryloyl-5-methylpyrrolidin-3-yl)-4-amino-6-(cyclopropylethynyl)-N-((R)-1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide to fumaric acid is 1:

1.

14. The crystal according to claim 13, characterized in that: This is a crystal having a powder X-ray diffraction pattern shown in FIG9 .

15. The crystal according to claim 13 or 14, characterized in that: The endothermic peak determined by simultaneous differential thermal and thermogravimetric measurement is around 153°C.

16. A pharmaceutical composition, characterized in that: Contains the crystal according to any one of claims 1 to 15.

17. A pharmaceutical composition for oral administration, characterized in that: Contains the crystal according to any one of claims 1 to 15.

18. An antitumor agent, characterized in that: Contains the crystal according to any one of claims 1 to 15.

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