Solid forms of mutant idh inhibitor compound and salt thereof

Abstract

The present invention relates to a crystal form and salt form of a compound 6-(5-amino-6-chloro-4-fluoropyridin-2-yl)-N2,N4-bis((R)-1,1,1-trifluoropropyl-2-yl)-1,3,5-triazine-2,4-diamine, and a crystal form of the salt, a composition comprising the crystal form and salt form of the compound and the crystal form of the salt, and the use of the crystal form and salt form of the compound and the crystal form of the salt in the treatment of diseases. The present invention further relates to a method for preparing 6-(5-amino-6-chloro-4-fluoropyridin-2-yl)-N2,N4-bis((R)-1,1,1-trifluoropropyl-2-yl)-1,3,5-triazine-2,4-diamine.

Description

Solid forms of mutant IDH inhibitor compounds and their salts Technical Field

[0001] The present invention relates to 6-(5-amino-6-chloro-4-fluoropyridin-2-yl)-N 2 ,N 4 -bis((R)-1,1,1-trifluoropropyl-2-yl)-1,3,5-triazine-2,4-diamine, a crystalline form, a salt form and a crystalline form of the salt form; and a pharmaceutical composition comprising the crystalline form, the salt form and the crystalline form of the salt form of the compound, and their use in inhibiting IDH1 and / or IDH2 activity. The present invention also relates to 6-(5-amino-6-chloro-4-fluoropyridin-2-yl)-N 2 ,N 4 The present invention also relates to methods for preparing bis((R)-1,1,1-trifluoropropyl-2-yl)-1,3,5-triazine-2,4-diamine, its salts, and its crystalline forms, as well as related intermediate compounds and their preparation methods. The present invention also relates to methods for using the compounds, their salts, and pharmaceutical compositions for treating diseases or conditions associated with IDH1 and / or IDH2 double mutations. Background Art

[0002] In recent years, three metabolic enzymes have been discovered in tumor cells, namely fumarate dehydrogenase, succinate dehydrogenase and isocitrate dehydrogenase (IDH). Gene mutations of these enzymes change cell metabolism and may be related to the occurrence and development of tumors.

[0003] There are three types of human IDH: IDH1, IDH2, and IDH3. IDH1 is located in the cytoplasm and peroxisomes, while IDH2 and IDH3 are located in mitochondria. This type of protease can oxidize isocitrate to oxalosuccinate, which is then converted to α-ketoglutarate (α-KG).

[0004] In 2008, the inadvertent discovery of an IDH1 gene mutation during gene sequencing of human glioblastomas marked the beginning of IDH research in cancer. Subsequent large-scale case-control studies of gliomas have revealed that IDH1 mutations are common in over 75% of low-grade gliomas and 90% of secondary glioblastomas, while IDH2 mutations are common in approximately 20% of acute myeloid leukemias. Furthermore, IDH gene mutations have been reported in cholangiocarcinoma (10%-23%), melanoma (10%), and chondroid tumors (75%). This suggests that IDH mutations are common in a variety of tumors. Common mutations occur at arginine residues located in the catalytic center (IDH1 / R132H, IDH1 / R132C, IDH2 / R140Q, and IDH2 / R172K). Mutated IDHs catalyze the conversion of α-ketoglutarate (α-KG) to 2-hydroxyglutarate (2-HG). Studies have shown that α-KG and 2-HG have similar structures. 2-HG competes with α-KG, thereby reducing the activity of α-KG-dependent enzymes and leading to hypermethylation of chromatin. This hypermethylation is believed to interfere with normal cell differentiation, leading to excessive proliferation of immature cells and thus causing cancer.

[0005] In 2013, Agios Pharmaceuticals published its research results in the journal Science: The company's mutant IDH1 enzyme inhibitors, AGI-5198 (Science, 2013, 340, 626-630) and mutant IDH2 enzyme inhibitors, AGI-6780 (Science, 2013, 340, 622-626), effectively inhibit the production of 2-HG mediated by mutant IDH1 / IDH2 in cells, inducing the differentiation of abnormally proliferating cancer cells. Treatment of glioma cells carrying IDH1 gene mutations with AGI-5198 and leukemia cells carrying IDH2 gene mutations with AGI-6780 both resulted in increased expression of maturation markers in the cells.

[0006] Phase I clinical trials of AG-120, a mutant IDH1 inhibitor developed by Agios Pharmaceuticals, showed that in patients with acute myeloid leukemia (AML) or myelodysplastic syndrome (MDS) with IDH1 gene mutations, a decrease in α-hydroxyglutarate (2-HG) levels was observed in 98% of patients.

[0007] In 2013, Agios Pharmaceuticals reported IDH2 R140Q Inhibitors AGI-6780 and IDH2 R132H The inhibitor AGI-5198 and another IDH2 inhibitor that the company later launched R140QThe inhibitors AG-221, AGI-6780, and AGI-5198 can inhibit 2-HG production in cells carrying common IDH1 and IDH2 mutants, respectively.

[0008] For cancers caused by IDH1 and IDH2 mutations, such as glioma, acute myeloid leukemia, cholangiocarcinoma, and melanoma, the IDH1 single inhibitor AG120 and the IDH2 single inhibitor AG221 are currently on the market, providing clinical drug options. New studies have found that IDH1 and IDH2 mutations may coexist in the same tumor, resulting in limited efficacy of IDH1 or IDH2 single inhibitors and acquired drug resistance. Although studies on drugs that simultaneously inhibit mutant IDH1 and IDH2 for the treatment of cancer have been reported, there is still a need to develop new mutant IDH1 and IDH2 dual inhibitors with strong target inhibition and excellent selectivity for the treatment of related diseases mediated by mutant IDH1 and IDH2, to overcome the problem of acquired drug resistance after long-term use of single inhibitors, and to provide a new drug option for clinical use.

[0009] The present invention discovered 6-(5-amino-6-chloro-4-fluoropyridin-2-yl)-N 2 ,N 4 -bis((R)-1,1,1-trifluoropropyl-2-yl)-1,3,5-triazine-2,4-diamine has excellent cell inhibitory activity, good pharmacokinetic characteristics, and excellent anti-tumor activity. It is an excellent new mutant IDH1 and IDH2 dual inhibitor. The present invention also discovered 6-(5-amino-6-chloro-4-fluoropyridin-2-yl)-N 2 ,N 4 The advantageous crystal form, salt form, and crystal form of bis((R)-1,1,1-trifluoropropyl-2-yl)-1,3,5-triazine-2,4-diamine have drug-forming properties that are beneficial to drug development.

[0010] SUMMARY OF THE INVENTION

[0011] One aspect of the present invention relates to a crystalline form of a compound represented by Formula I (Compound 1),

[0012] In the present invention, the term "compound of formula I", "compound of formula I", "Formula I" or "Compound 1" refers to a compound having the above structure and the chemical name is 6-(5-amino-6-chloro-4-fluoropyridin-2-yl)-N 2 ,N 4 -bis((R)-1,1,1-trifluoropropyl-2-yl)-1,3,5-triazine-2,4-diamine compound.

[0013] Another aspect of the present invention relates to salt forms of the compound represented by formula I, and its crystal form.

[0014] Another aspect of the present invention relates to a pharmaceutical composition comprising 0.5% to 85% by weight of a compound of formula I, or a pharmaceutically acceptable salt thereof, or a mixture thereof.

[0015] Another aspect of the present invention relates to a pharmaceutical composition comprising 0.5% to 85% by weight of a compound of formula I, or its hydrochloride, phosphate, sulfate, methanesulfonate, L-malate, L-tartrate or L-lactate, or a mixture thereof.

[0016] Another aspect of the present invention provides a method for preparing the compound of formula I and its intermediate compounds.

[0017] Another aspect of the present invention provides a crystal form of the compound represented by Formula I or a salt thereof, or use of the pharmaceutical composition in the preparation of a drug.

[0018] Another aspect of the present invention provides a method for treating a disease, comprising administering to a subject a therapeutically effective amount of a crystalline form of the compound of Formula I or a salt thereof, or a pharmaceutical composition comprising the same.

[0019] Detailed Description of the Invention

[0020] The solid forms of compound 1 and its salts provided by the present invention are used as dual inhibitors of IDH1 and IDH2. In addition, the preferred solid forms of compound 1 and its salts according to the present invention have excellent properties such as stability, hygroscopicity, and pharmacokinetics, which are conducive to the development of drugs. Although preferred embodiments of the present invention are shown and described herein, such embodiments are provided only by way of example and are not intended to limit the scope of the present invention. In practicing the present invention, various alternatives to the described embodiments of the present invention may be used.

[0021] Crystalline form of compound 1

[0022] Compound 1 Form B

[0023] In some embodiments, the crystalline form of Compound 1 is Form B, and the X-ray powder diffraction pattern of Form B comprises characteristic peaks with 2θ values ​​of 5.8°±0.2°, 11.7°±0.2°, and 17.6°±0.2°.

[0024] In some embodiments, the crystalline form of Compound 1 is Form B, and the X-ray powder diffraction pattern of Form B comprises characteristic peaks with 2θ values ​​of 5.8°±0.2°, 11.7°±0.2°, 17.6°±0.2°, 19.6°±0.2° and 23.5°±0.2°.

[0025] In some embodiments, the crystalline form of Compound 1 is Form B, and the X-ray powder diffraction pattern of Form B comprises 5 or more (e.g., 6 or more, 7 or more, or 8) characteristic peaks having 2θ values ​​selected from 5.8°±0.2°, 11.7°±0.2°, 14.2°±0.2°, 16.8°±0.2°, 17.6°±0.2°, 18.6°±0.2°, 19.6°±0.2°, and 23.5°±0.2°.

[0026] In some embodiments, the Form B has an X-ray powder diffraction pattern substantially as shown in FIG2 .

[0027] In some embodiments, the Form B has a thermogravimetric analysis (TGA) pattern substantially as shown in FIG3 , with a weight loss of about 0.2% before 200° C.

[0028] In some embodiments, the Form B has a differential scanning calorimetry (DSC) pattern substantially as shown in FIG. 4 .

[0029] Compound 1 Form D

[0030] In some embodiments, the crystalline form of Compound 1 is Form D, and the X-ray powder diffraction pattern of Form D comprises characteristic peaks with 2θ values ​​of 8.6°±0.2°, 10.5°±0.2°, and 21.1°±0.2°.

[0031] In some embodiments, the crystalline form of Compound 1 is Form D, and the X-ray powder diffraction pattern of Form D comprises characteristic peaks with 2θ values ​​of 8.6°±0.2°, 10.5°±0.2°, 11.8°±0.2°, 14.0°±0.2°, 17.3°±0.2° and 21.1°±0.2°.

[0032] In some embodiments, the crystalline form of Compound 1 is Form D, and the X-ray powder diffraction pattern of Form D comprises 5 or more (e.g., 6 or more, 7 or more, 8 or more, or 9) characteristic peaks with 2θ values ​​selected from 8.6°±0.2°, 10.5°±0.2°, 11.8°±0.2°, 14.0°±0.2°, 16.5°±0.2°, 17.3°±0.2°, 18.3°±0.2°, 19.8°±0.2°, and 21.1°±0.2°.

[0033] In some embodiments, the crystalline Form D has an X-ray powder diffraction pattern substantially as shown in FIG5 .

[0034] In some embodiments, the Form D has a thermogravimetric analysis (TGA) pattern substantially as shown in FIG6 , with a weight loss of about 10.9% before 200° C.

[0035] In some embodiments, the Form D has a differential scanning calorimetry (DSC) pattern substantially as shown in FIG. 7 .

[0036] Salt form of compound 1

[0037] The present invention also provides advantageous salt forms of Compound 1. In some embodiments, the salt form is a sulfate salt of Compound 1. In some embodiments, the salt form is a phosphate salt of Compound 1. In some embodiments, the salt form is an L-tartrate salt of Compound 1. In some embodiments, the salt form is an L-malate salt of Compound 1. In some embodiments, the salt form is an L-lactate salt of Compound 1. In some embodiments, the salt form is a methanesulfonate salt of Compound 1. In some embodiments, the salt form is most preferably a hydrochloride salt of Compound 1.

[0038] Compound 1 hydrochloride and its crystalline form

[0039] The present invention provides a hydrochloride salt of compound 1 represented by the following formula II:

[0040] The present invention also provides a crystalline form of the hydrochloride salt of compound 1 represented by formula II.

[0041] In some embodiments, the crystalline form of the hydrochloride of Compound 1 is hydrochloride Form A, and the X-ray powder diffraction pattern of the hydrochloride Form A comprises characteristic peaks with 2θ values ​​of 11.6°±0.2°, 12.7°±0.2°, 13.0°±0.2°, 19.2°±0.2° and 21.1±0.2°.

[0042] In some embodiments, the crystalline form of the hydrochloride salt of Compound 1 is hydrochloride Form A, and the X-ray powder diffraction pattern of the hydrochloride Form A comprises characteristic peaks with 2θ values ​​of 11.6°±0.2°, 12.7°±0.2°, 13.0°±0.2°, 17.3°±0.2°, 18.3°±0.2°, 19.2°±0.2° and 21.1±0.2°.

[0043] In some embodiments, the crystalline form of the hydrochloride salt of Compound 1 is hydrochloride Form A, and the X-ray powder diffraction pattern of the hydrochloride Form A comprises characteristic peaks with 2θ values ​​of 11.6°±0.2°, 12.7°±0.2°, 13.0°±0.2°, 17.3°±0.2°, 18.3°±0.2°, 19.2°±0.2°, 21.1±0.2°, 23.9±0.2° and 25.5±0.2°.

[0044] In some embodiments, the crystalline form of the hydrochloride salt of Compound 1 is hydrochloride Form A, and the X-ray powder diffraction pattern of the hydrochloride Form A contains 5 or more (e.g., 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, or 13 or more, or 14) characteristic peaks with 2θ values ​​selected from 11.6°±0.2°, 12.7°±0.2°, 13.0°±0.2°, 15.4°±0.2°, 16.3°±0.2°, 17.3°±0.2°, 18.3°±0.2°, 19.2°±0.2°, 21.1±0.2°, 22.6±0.2°, 23.9±0.2°, 25.5±0.2°, 28.0±0.2° and 31.8±0.2°.

[0045] In some embodiments, the hydrochloride salt Form A has an X-ray powder diffraction pattern substantially as shown in Figure 8.

[0046] In some embodiments, the Raman spectrum of the hydrochloride salt form A includes a peak at 1572 cm -1 ±2cm -1 , 1421cm -1 ±2cm -1 , 1289cm -1 ±2cm -1 and 1248cm -1 ±2cm -1 The peak shift at .

[0047] In some embodiments, the Raman spectrum of the hydrochloride salt form A includes 5 or more selected from 1572 cm -1 ±2cm -1 , 1421cm -1 ±2cm -1 , 1312cm -1 ±2cm -1 , 1289cm -1 ±2cm -1 , 1269cm -1 ±2cm -1 and 1248cm -1 ±2cm -1 , 1206cm -1 ±2cm -1 , 999cm -1 ±2cm -1 , 841cm -1 ±2cm -1 , 426cm -1 ±2cm -1 , 136cm-1 ±2cm -1 The peak shift at .

[0048] In some embodiments, the hydrochloride salt Form A has a Raman spectrum substantially as shown in FIG. 20 .

[0049] In some embodiments, the infrared spectrum of the hydrochloride salt form A includes a wave number at 3421 cm -1 ±2cm -1 ,3328cm -1 ±2cm -1 , 1655cm -1 ±2cm -1 , 1536cm -1 ±2cm -1 , 1268cm -1 ±2cm -1 , 1177cm -1 ±2cm -1 , 797cm -1 ±2cm -1 The characteristic absorption peak at .

[0050] In some embodiments, the infrared spectrum of the hydrochloride salt form A includes 5 or more wave numbers selected from 3421 cm -1 ±2cm -1 , 3328cm -1 ±2cm -1 , 3048cm -1 ±2cm -1 , 1655cm -1 ±2cm -1 , 1595cm -1 ±2cm -1 , 1536cm -1 ±2cm -1 , 1291cm -1 ±2cm -1 , 1268cm -1 ±2cm -1 , 1177cm -1 ±2cm -1 , 1154cm -1 ±2cm -1 , 797cm -1 ±2cm -1 The characteristic absorption peak at .

[0051] In some embodiments, the hydrochloride salt Form A has an infrared spectrum substantially as shown in Figure 21.

[0052] In some embodiments, the hydrochloride salt form A has a thermogravimetric analysis (TGA) pattern substantially as shown in FIG9 , with almost no weight loss before 200° C.

[0053] In some embodiments, the hydrochloride salt Form A has a differential scanning calorimetry (DSC) pattern substantially as shown in FIG. 10 .

[0054] The hydrochloride crystal form A has been characterized by single crystal analysis. The unit cell stacking projection of the hydrochloride crystal form A single crystal is basically as shown in Figure 14, and the unit cell parameters are as follows:

[0055] The crystal belongs to the orthorhombic system, the space group is P212121, and the unit cell parameters are: α=β=γ=90.00°;

[0056] Unit cell volume

[0057] The number of asymmetric units in the unit cell is Z = 4.

[0058] The final reliability factors R1 = 0.0283, wR2 = 0.0749, S = 1.088. The final stoichiometric formula of the asymmetric unit is C 14 H 13 The molecular weight of a single ClF7N7·HCl molecule is calculated to be 484.22, and the crystal density is calculated to be 1.607 g / cm 3 .

[0059] Crystalline form of compound 1 mesylate

[0060] The present invention provides a mesylate salt of Compound 1, wherein the structure of the mesylate salt of Compound 1 is represented by the following formula III:

[0061] The present invention also provides a crystalline form of the methanesulfonate salt of Compound 1 represented by Formula III.

[0062] In some embodiments, the crystalline form of the mesylate salt of Compound 1 is mesylate salt Form A, and the X-ray powder diffraction pattern of the mesylate salt Form A comprises characteristic peaks with 2θ values ​​of 6.6°±0.2°, 8.2°±0.2°, and 19.3°±0.2°.

[0063] In some embodiments, the crystalline form of the mesylate salt of Compound 1 is mesylate salt Form A, and the X-ray powder diffraction pattern of the mesylate salt Form A comprises characteristic peaks with 2θ values ​​of 6.6°±0.2°, 8.2°±0.2°, 17.3°±0.2° and 19.3°±0.2°.

[0064] In some embodiments, the crystalline form of the mesylate salt of Compound 1 is mesylate salt Form A, and the X-ray powder diffraction pattern of the mesylate salt Form A comprises 3 or more (e.g., 4 or more, 5 or more, 6 or more, or 7) characteristic peaks having 2θ values ​​selected from 6.6°±0.2°, 8.2°±0.2°, 10.5°±0.2°, 14.2°±0.2°, 17.3°±0.2°, 19.3°±0.2°, and 20.2°±0.2°.

[0065] In some embodiments, the mesylate salt Form A has an X-ray powder diffraction pattern substantially as shown in FIG. 11 .

[0066] In some embodiments, the mesylate salt Form A has a thermogravimetric analysis (TGA) pattern substantially as shown in FIG. 12 , with a weight loss of about 1.9% before 200° C.

[0067] In some embodiments, the mesylate salt Form A has a differential scanning calorimetry (DSC) pattern substantially as shown in FIG13 .

[0068] All crystalline forms of the present invention are substantially pure.

[0069] Unless otherwise specified, the X-ray powder diffraction patterns are all measured using the Kα spectrum of a Cu target.

[0070] Unless otherwise specified, the experimental temperature of the present invention is room temperature.

[0071] The term "substantially pure" as used herein means that the content of the crystalline form is not less than 85% by weight, preferably not less than 95% by weight, and more preferably not less than 98% by weight.

[0072] It should be noted that the numerical values ​​and numerical ranges mentioned in the present invention should not be narrowly understood as the numerical values ​​or numerical ranges themselves. Those skilled in the art should understand that they can fluctuate around the specific numerical values ​​based on different specific technical environments without departing from the spirit and principles of the present invention. In the present invention, such floating ranges that can be foreseen by those skilled in the art are mostly expressed by the terms "about" or "substantially".

[0073] It should be noted that the "salt (salt-type)" mentioned in the present invention includes salts in the classical sense formed by significant or complete proton exchange between the API (e.g., the compound of Formula I) and the guest molecule (e.g., hydrochloric acid), and also includes multi-component systems, such as eutectics, formed by the coexistence of the API (e.g., the compound of Formula I) and the guest molecule (e.g., L-malate) through hydrogen bonds and possible other non-covalent bonds. That is, the "salt (salt-type)" mentioned in the present invention includes salts in the classical sense, eutectics, or mixtures thereof, etc., which are well known to those skilled in the art.

[0074] It should be noted that, unless otherwise specified, the salt form of Compound 1 of the present invention may exist in various physical forms, for example, in the form of a solution, a suspension or a solid. When in solid form, the compound may be in an amorphous form, a crystalline form or a mixture thereof.

[0075] As used herein, the term "crystalline form" refers to a crystalline form having the same chemical composition but a different spatial arrangement of the molecules, atoms, and / or ions that form the crystal, including anhydrous crystalline forms, hydrates, and solvates. "Polymorph," "crystalline form," "crystalline form," and "polymorph" are used interchangeably herein to refer to solid forms of the compound represented by Formula I and its salts, which are distinct from the amorphous form of the compound represented by Formula I and its salts. "Amorphous form" refers to a non-crystalline solid form. Polymorphs of a compound may exhibit different chemical and / or physical properties, including, but not limited to, stability, solubility, dissolution rate, optical properties, melting point, mechanical properties, and / or density. These properties may affect the processing and / or manufacturing of the drug substance, as well as the stability, dissolution, and / or bioavailability of the drug. Therefore, polymorphism may affect at least one property of the drug, including, but not limited to, quality, safety, and / or efficacy. Unless otherwise specified, the compound represented by Formula I and its salts encompasses any polymorphs and amorphous forms.

[0076] Polymorphs of a molecule can be obtained by many methods known in the art. Such methods include, but are not limited to, melt recrystallization, melt cooling, solvent recrystallization, desolvation, rapid evaporation, rapid cooling, slow cooling, vapor diffusion, and sublimation. Polymorphs can be detected, identified, classified, and characterized using well-known techniques such as, but not limited to, differential scanning calorimetry (DSC), thermogravimetry (TGA), X-ray powder diffractometry (XRPD), single crystal X-ray diffractometry, solid-state nuclear magnetic resonance (NMR), infrared (IR) spectroscopy, Raman spectroscopy, and hot-stage optical microscopy.

[0077] As used herein, the terms "approximately" and "substantially" in the context of "having an X-ray powder diffraction pattern approximately as shown in FIG. 1," "having an X-ray powder diffraction pattern substantially as shown in FIG. 1," or "having an X-ray powder diffraction pattern substantially as shown in FIG. 1" refer to the precise positions of peaks in the accompanying drawings and should not be interpreted as absolute values. As will be appreciated by those skilled in the art, the 2θ values ​​of an X-ray powder diffraction pattern may vary due to different measurement conditions (e.g., the equipment and instruments used) and different samples (e.g., different batches of samples). The measurement error of the diffraction angle of an X-ray powder diffraction pattern is 5% or less, and generally, a difference of ±0.2° from a given value is considered appropriate. It should also be understood that the relative intensities of the peaks may fluctuate depending on the experimental conditions and sample preparation, such as the preferred orientation of the particles in the sample. The use of an automatic or fixed divergence slit will also affect the calculation of relative intensities. The intensities shown in the XRD curves included herein are exemplary only and should not be used as absolute comparisons, and any crystalline form having a powder diffraction pattern substantially identical to those disclosed herein is within the scope of protection of the present invention.

[0078] Those skilled in the art will understand that due to variations in sample batches, sample purity, sample preparation, and measurement conditions (e.g., heating rate), DSC measured data may vary slightly, and generally, a difference of ±5°C from a given value is acceptable (and still considered characteristic of the specific crystalline form described herein). Therefore, the endotherms cited in this application are not to be taken as absolute values, and such errors should be taken into account when interpreting DSC data.

[0079] In TGA measurements, without being bound by any particular theory, weight loss corresponds to the loss of trace amounts of residual solvent or water. TGA data may vary due to variations in sample batches, sample purity, residual solvent content, sample preparation, and measurement conditions (e.g., heating rate). Therefore, the thermogravimetric curves cited herein are not to be taken as absolute values, and such errors should be taken into account when interpreting TGA data.

[0080] It is also known in the art that IR and Raman spectra can be obtained, which can vary depending on the measurement conditions. Instrumentation, experimental conditions, sampling methods, and instrument calibration can all affect peak positions and intensities. Therefore, the spectra provided herein should not be considered absolute, and any crystalline form having spectra substantially identical to those disclosed herein is within the scope of the present disclosure.

[0081] Pharmaceutical composition

[0082] In some embodiments, the present invention provides a pharmaceutical composition, wherein the pharmaceutical composition contains 0.5 wt % to 85 wt % of Compound 1, or a pharmaceutically acceptable salt thereof, or a mixture thereof.

[0083] In some embodiments, preferably, the pharmaceutical composition contains 0.5 wt% to 85 wt% of Compound 1, or its hydrochloride, phosphate, sulfate, methanesulfonate, L-malate, L-tartrate or L-lactate, or a mixture thereof.

[0084] In some embodiments, preferably, the pharmaceutical composition contains 0.5 wt% to 85 wt% of Compound 1 Form B.

[0085] In some embodiments, preferably, the pharmaceutical composition contains 0.5 wt% to 85 wt% of Compound 1 Form D.

[0086] In some embodiments, preferably, the pharmaceutical composition contains 0.5 wt% to 85 wt% of Compound 1 hydrochloride.

[0087] In some embodiments, preferably, the pharmaceutical composition contains 0.5 wt% to 85 wt% of Compound 1 Hydrochloride Form A.

[0088] In some embodiments, preferably, the pharmaceutical composition contains 0.5 wt% to 85 wt% of Compound 1 mesylate salt Form A.

[0089] In some embodiments, the composition is for oral administration.

[0090] In some embodiments, the oral dosage form includes tablets, capsules, cachets, pills, granules, oral liquids, suspensions, dispersions, emulsions, and powders.

[0091] In some embodiments, the pharmaceutical composition contains 1 wt% to 70 wt% of Compound 1, or a pharmaceutically acceptable salt thereof, or a mixture thereof.

[0092] In some embodiments, the pharmaceutical composition further comprises one or more pharmaceutically acceptable carriers.

[0093] In some embodiments, the pharmaceutically acceptable carrier may comprise one or more of a diluent, a filler, a lubricant, a binder, and a disintegrant.

[0094] In practice, the active ingredient, Compound 1 of the present invention, or a pharmaceutically acceptable salt thereof, or a mixture thereof, is intimately mixed with a pharmaceutical carrier according to conventional pharmaceutical mixing techniques to form a pharmaceutical composition. The pharmaceutical carrier can take a variety of forms, depending on the desired route of administration, for example, oral or parenteral (including intravenous). Thus, the pharmaceutical composition of the present invention can be in the form of individual units suitable for oral administration, such as capsules, cachets, or tablets containing a predetermined dose of the active ingredient. Furthermore, the pharmaceutical composition of the present invention can be in the form of a powder, granules, solution, aqueous suspension, non-aqueous liquid, oil-in-water emulsion, or water-in-oil emulsion. In addition to the common dosage forms mentioned above, the compound represented by Formula I or its pharmaceutically acceptable salt can also be administered via controlled release and / or a delivery device. The pharmaceutical composition of the present invention can be prepared by any pharmaceutical method. Generally, such a method includes the step of combining the active ingredient with a carrier that constitutes one or more essential ingredients. Generally, the pharmaceutical composition is prepared by uniformly and intimately mixing the active ingredient with a liquid carrier or a finely divided solid carrier, or a mixture of the two. In addition, the product can be easily prepared into a desired appearance.

[0095] The pharmaceutical carrier used in the present invention can be, for example, a solid carrier, a liquid carrier, or a gaseous carrier. Solid carriers include lactose, gypsum powder, sucrose, talc, gelatin, agar, pectin, gum arabic, magnesium stearate, and stearic acid. Liquid carriers include syrup, peanut oil, olive oil, and water. Gaseous carriers include carbon dioxide and nitrogen. When preparing oral pharmaceutical preparations, any pharmaceutically convenient medium can be used. For example, water, ethylene glycol, oils, alcohols, flavor enhancers, preservatives, colorants, etc. can be used for oral liquid preparations such as suspensions, elixirs, and solutions; while carriers such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrants, etc. can be used for oral solid preparations such as powders, capsules, and tablets. Considering ease of administration, tablets and capsules are preferred for oral preparations, and solid pharmaceutical carriers are used here. Alternatively, tablet coating can use standard aqueous or non-aqueous formulation technology.

[0096] Tablets containing the compounds or pharmaceutical compositions of the present invention can be formed by compression or molding, optionally with one or more auxiliary ingredients or adjuvants. Compressed tablets can be prepared by mixing the active ingredient in a free-flowing form such as a powder or granules with a binder, lubricant, inert diluent, surfactant, or dispersant and compressing it in a suitable machine. Molded tablets can be prepared by soaking the powdered compound or pharmaceutical composition with an inert liquid diluent and then molding it in a suitable machine. Preferably, each tablet contains about 0.05 mg to 5 g of the active ingredient, and each cachet or capsule contains about 0.05 mg to 5 g of the active ingredient. For example, a formulation intended for oral administration to humans contains about 0.5 mg to about 5 g of the active ingredient, compounded with suitable and conveniently metered auxiliary materials, which may comprise about 5% to 95% of the total pharmaceutical composition. Dosage unit forms generally contain from about 1 mg to about 2 g of active ingredient, typically 2.5 mg, 5 mg, 10 mg, 25 mg, 50 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 800 mg or 1000 mg.

[0097] The pharmaceutical compositions provided herein for parenteral administration can be prepared by adding the active ingredient to water to form an aqueous solution or suspension. Suitable surfactants such as hydroxypropylcellulose can be included. Dispersions can also be prepared using glycerol, liquid polyethylene glycol, or mixtures thereof in oils. Furthermore, preservatives can also be included in the pharmaceutical compositions of the present invention to prevent the growth of harmful microorganisms.

[0098] The present invention provides pharmaceutical compositions suitable for injection, including sterile aqueous solutions or dispersions. Further, the above-mentioned pharmaceutical compositions can be prepared into sterile powder forms for immediate preparation of sterile injections or dispersions. In any case, the final injection form must be sterile and, in order to be easy to inject, must be easy to flow. In addition, the pharmaceutical composition must be stable during preparation and storage. Therefore, preferably, the pharmaceutical composition will be preserved under conditions of antimicrobial such as bacterial and fungal contamination. The carrier can be a solvent or dispersion medium, for example, water, ethanol, polyols (such as glycerol, propylene glycol, liquid polyethylene glycol), vegetable oils and suitable mixtures thereof.

[0099] The pharmaceutical compositions provided herein can be in a form suitable for topical administration, such as an aerosol, cream, ointment, lotion, dusting powder, or other similar dosage forms. Furthermore, the pharmaceutical compositions provided herein can be in a form suitable for use with a transdermal delivery device. These formulations can be prepared using the compound represented by Formula I of the present invention, or a pharmaceutically acceptable salt thereof, by conventional processing methods. As an example, an emulsion or ointment having the desired consistency can be prepared by adding approximately 5% to 10% by weight of a hydrophilic material and water.

[0100] The pharmaceutical composition provided by the present invention can be in a form suitable for rectal administration using a solid as a carrier. Unit-dose suppositories are the most typical and common dosage form. Suitable excipients include cocoa butter and other materials commonly used in the art. Suppositories can be easily prepared by first mixing the pharmaceutical composition with softened or melted excipients, then cooling and molding.

[0101] In addition to the aforementioned excipients, the formulation may also include, where appropriate, one or more additional excipients, such as diluents, buffers, flavorings, binders, surfactants, thickeners, lubricants, and preservatives (including antioxidants). Furthermore, other excipients may include penetration enhancers to adjust the osmotic pressure between the drug and the intended recipient's blood. Pharmaceutical compositions comprising the compound of Formula I, or a pharmaceutically acceptable salt thereof, may be prepared in the form of a powder or concentrate.

[0102] Preparation method of the compound represented by formula I

[0103] In some embodiments, the present invention provides a method for preparing a compound of formula I:

[0104] The method comprises performing a deprotection reaction using a compound represented by formula IV.

[0105] Wherein, the R1 is an amino protecting group, and the R2 is H or an amino protecting group.

[0106] Preferably, the amino protecting group can be selected from one or two of -Boc, -Cbz, -Fmoc, -PMB and -Bn, more preferably a -Boc group.

[0107] Preferably, the amino protecting group is a -Boc group, and the compound of formula IV is reacted in a solvent containing TFA.

[0108] In some embodiments, the preparation method further comprises using a compound represented by formula V With compound A07 A coupling reaction is carried out in the presence of a base and a palladium catalyst to obtain a compound represented by formula IV;

[0109] Wherein, the R1 is an amino protecting group, and the R2 is H or an amino protecting group;

[0110] Said R3 is selected from H, C 1-4 Alkyl, or two R3 and the O atom to which they are connected together form a 5- or 6-membered heterocyclic ring containing two oxygen atoms and 1 boron atom, wherein the 5- or 6-membered heterocyclic ring may be optionally replaced by one or more C 1-4 Alkyl substituted.

[0111] In some embodiments, the amino protecting group may be one or two of -Boc, -Cbz, -Fmoc, -PMB and -Bn, more preferably -Boc group.

[0112] In some embodiments, the base used in the reaction of the compound represented by Formula V with Compound A07 can be selected from one or more of K2CO3, AcOK, Na2CO3, AcONa, Cs2CO3, K3PO4, KF or TEA.

[0113] In some embodiments, the palladium catalyst in the reaction of the compound represented by Formula V with Compound A07 is selected from Pd(dppf)Cl2 or Pd(OAc)2.

[0114] In some embodiments, the solvent in the reaction of the compound represented by Formula V with compound A07 can be selected from one or more of toluene, dioxane, DMF, ethanol, acetonitrile, tert-butanol, ethylene glycol dimethyl ether, diethoxymethane, tetrahydrofuran, or a mixture of one or more of the above solvents and water.

[0115] In some embodiments, a ligand may be added during the reaction of the compound represented by Formula V with Compound A07. The ligand may be selected from dppf, PPh3, XantPhos, Xphos, PCy3, SPhos, and P(t-Bu)3.

[0116] In some embodiments, the compound represented by Formula V is reacted with Compound A07 at 70-100°C.

[0117] In some embodiments, the preparation method further comprises using a compound represented by formula VI reacting with diboric acid pinacol ester in the presence of a base and a catalyst to obtain a compound represented by formula V;

[0118] Wherein R1 is an amino protecting group, and R2 is H or an amino protecting group.

[0119] In some embodiments, the amino protecting group may be one or two of -Boc, -Cbz, -Fmoc, -PMB and -Bn, more preferably -Boc group.

[0120] In some embodiments, the base in the reaction of the compound represented by Formula VI with pinacol diboron is AcOK.

[0121] In some embodiments, the palladium catalyst in the reaction of the compound represented by Formula VI with diboronic acid pinacol ester is selected from one or more of Pd(dppf)Cl2, Pd(PPh3)4 and Pd(OAc)2.

[0122] In some embodiments, the solvent used in the reaction of the compound represented by Formula VI with diboronic acid pinacol ester can be selected from one or more of dioxane, DMSO, DMF, toluene, 2-methyltetrahydrofuran, DMA, and NMP.

[0123] In some embodiments, a ligand may be added during the reaction of the compound represented by Formula VI with pinacol diboronate, and the ligand may be selected from dppf or PPh3.

[0124] In some embodiments, the compound represented by Formula VI is reacted with diboronic acid pinacol ester at 70-100°C.

[0125] In some embodiments, the preparation method further comprises using compound A13 and Boc2O under alkaline conditions to obtain a compound of formula VI (e.g., compound A101 or compound A04 ); preferably, the base is selected from LiHMDS or DIEA.

[0126] The present invention also provides a method for preparing compound A13 of formula:

[0127] Method 1

[0128] or

[0129] Method 2

[0130] The present invention also provides a method for preparing the compound represented by formula I.

[0131] The preparation route 1 of the compound represented by formula I is as follows:

[0132] The second preparation route of the compound represented by formula I is as follows:

[0133] The preparation route three of the compound represented by formula I is as follows:

[0134] The present invention also provides an intermediate for preparing the compound of formula I, which is selected from the compounds of the following formula:

[0135] in,

[0136] R1 is an amino protecting group, and R2 is H or an amino protecting group;

[0137] Said R3 is selected from H, C 1-4 Alkyl, or two R3 and the O atom to which they are connected together form a 5- or 6-membered heterocyclic ring containing two oxygen atoms and 1 boron atom, wherein the 5- or 6-membered heterocyclic ring may be optionally replaced by one or more C 1-4 Alkyl substituted.

[0138] In some embodiments, the amino protecting group can be selected from one or two of -Boc, -Cbz, -Fmoc, -PMB and -Bn.

[0139] In some embodiments, the intermediate compound is selected from one or more of the following compounds:

[0140] Crystal forms, salt forms of compound 1, and uses of the crystal forms of the salt forms, and methods for treating solid tumor cancers, hematological malignancies, inflammatory diseases, autoimmune diseases, and other diseases

[0141] The present invention further provides the crystal form, salt form and crystal form of the salt form of the compound 1, or the use of the pharmaceutical composition in the preparation of drugs.

[0142] In some embodiments, preferably, provided is the use of Compound 1 hydrochloride in the preparation of a medicament.

[0143] In some embodiments, preferably, provided is the use of Compound 1 Crystalline Form B in the preparation of a drug.

[0144] In some embodiments, preferably, provided is the use of Compound 1 Crystalline Form D in the preparation of a drug.

[0145] In some embodiments, preferably, provided is the use of Compound 1 hydrochloride Form A in the preparation of a drug.

[0146] In some embodiments, the medicament is used to treat a disease mediated by mutant IDH1 and / or IDH2. Preferably, the disease is cancer.

[0147] Preferably, the cancer is selected from glioma, melanoma, papillary thyroid tumor, bile duct cancer, lung cancer, breast cancer, sarcoma, glioma, glioblastoma multiforme, acute myeloid leukemia, non-Hodgkin's lymphoma, etc. In a specific embodiment, the cancer to be treated is glioma, glioblastoma (glioma), myelodysplastic syndrome (MDS), myeloproliferative neoplasm (MPN), acute myeloid leukemia (AML), sarcoma, melanoma, non-small cell lung cancer, chondrosarcoma, bile duct cancer or angioimmunoblastic lymphoma. In a more specific embodiment, the cancer to be treated is glioma, glioblastoma (glioma), myelodysplastic syndrome (MDS), myeloproliferative neoplasm (MPN), acute myeloid leukemia (AML), melanoma, chondrosarcoma, or angioimmunoblastic non-Hodgkin's lymphoma (NHL).

[0148] The present invention also provides a method for treating and / or preventing diseases mediated by IDH1 and / or IDH2 by administering a therapeutically effective amount of at least any one of the crystalline forms, salt forms, and crystalline forms of the salt forms, or pharmaceutical compositions of Compound 1 to a subject.

[0149] Preferably, in the above method, the disease mediated by IDH1 and / or IDH2 is cancer.

[0150] Preferably, in the above method, a therapeutically effective amount of Form B of Compound 1 is administered to the subject.

[0151] Preferably, in the above method, a therapeutically effective amount of Compound 1 hydrochloride Form A is administered to the subject.

[0152] The present invention also provides a method for treating cancer, comprising administering to a subject a therapeutically effective amount of at least any one of the crystalline forms, salt forms, and crystalline forms of the salt forms of Compound 1, or the pharmaceutical composition. In some embodiments, the present invention relates to a method for treating a cancer characterized by the presence of mutant IDH1 and / or IDH2, comprising administering to a patient in need thereof a therapeutically effective amount of the crystalline form, salt form, and crystalline forms of the salt forms of Compound 1, or the pharmaceutical composition, wherein the cancer is selected from brain glioma, melanoma, papillary thyroid tumor, bile duct cancer, lung cancer, breast cancer, sarcoma, glioma, glioblastoma multiforme, acute myeloid leukemia, non-Hodgkin's lymphoma, and the like.

[0153] Preferably, in the above method, the subject to be treated is a human.

[0154] As used herein, the term "disease" or "disorder" or "condition" refers to any disease, illness, ailment, symptom or indication.

[0155] As used herein, the term "therapeutically effective amount" refers to an amount of a compound that, when administered to a subject, is sufficient to effect treatment of a disease, disorder, or condition, or at least one clinical symptom of a disease or condition. The "therapeutically effective amount" may vary depending on the compound, the disease, disorder, and / or symptoms of the disease or condition, the severity of the disease, disorder, and / or symptoms of the disease or condition, the age of the patient being treated, and / or the weight of the patient being treated. In any particular case, an appropriate amount will be apparent to those skilled in the art or can be determined by routine experimentation. In the context of combination therapy, a "therapeutically effective amount" refers to the total amount of the combined subject effective to treat the disease, disorder, or condition.

[0156] The compounds or crystal forms of the present invention are used alone or in combination as active ingredients and are mixed with a pharmaceutical carrier to form a pharmaceutical composition. The pharmaceutical compositions of the present invention include pharmaceutical compositions suitable for oral, rectal, topical, and parenteral (including subcutaneous administration, intramuscular injection, and intravenous administration) administration, although the most suitable mode of administration of the active ingredient in any given case depends on the specific subject receiving the administration, the subject's properties, and the severity of the condition. The pharmaceutical compositions of the present invention can be conveniently present in unit dosage forms well known in the art and prepared by any preparation method well known in the pharmaceutical field.

[0157] In general, the dosage level for treating the conditions or discomforts indicated above is about 0.01 mg / kg body weight to 150 mg / kg body weight per day, or 0.5 mg to 7 g per patient per day. However, it will be appreciated that lower or higher doses than those described above may be required. The specific dosage level and treatment regimen for any particular patient will depend on a variety of factors, including the activity of the specific compound used, age, body weight, general health, sex, diet, time of administration, route of administration, excretion rate, combination use of the drug, and the severity of the specific disease being treated.

[0158] These and other aspects will become apparent from the following written description of the invention.

[0159] The following examples are provided to better illustrate the present invention. Unless expressly stated otherwise, all parts and percentages are by weight and all temperatures are in degrees Celsius.

[0160] The present invention will be described in more detail by way of specific examples. The following examples are provided for illustrative purposes and are not intended to limit the present invention in any way. Those skilled in the art will readily recognize that various noncritical parameters that can be changed or modified to produce substantially the same results. BRIEF DESCRIPTION OF THE DRAWINGS

[0161] Figure 1: X-ray powder diffraction pattern of Compound 1 Form A.

[0162] Figure 2: X-ray powder diffraction pattern of Compound 1 Form B.

[0163] Figure 3: Thermogravimetric analysis (TGA) spectrum of Compound 1 Form B.

[0164] Figure 4: Differential scanning calorimetry (DSC) spectrum of Compound 1 Form B.

[0165] Figure 5: X-ray powder diffraction pattern of Compound 1 Form D.

[0166] Figure 6: Thermogravimetric analysis (TGA) spectrum of Compound 1 Form D.

[0167] Figure 7: Differential scanning calorimetry (DSC) spectrum of Compound 1 Form D.

[0168] Figure 8: X-ray powder diffraction pattern of Compound 1 hydrochloride Form A.

[0169] Figure 9: Thermogravimetric analysis (TGA) spectrum of Compound 1 hydrochloride Form A.

[0170] Figure 10: Differential scanning calorimetry (DSC) spectrum of Compound 1 hydrochloride Form A.

[0171] Figure 11: X-ray powder diffraction pattern of Compound 1 mesylate salt Form A.

[0172] Figure 12: Thermogravimetric analysis (TGA) spectrum of Compound 1 mesylate salt Form A.

[0173] Figure 13: Differential scanning calorimetry (DSC) spectrum of Compound 1 mesylate salt Form A.

[0174] Figure 14: Unit cell stacking projection of a single crystal of Compound 1 hydrochloride Form A along the a direction.

[0175] Figure 15: DVS curve of Compound 1 Form A.

[0176] Figure 16: DVS curve of Compound 1 Form B.

[0177] Figure 17: DVS curve of Compound 1 Form D.

[0178] Figure 18: DVS curve of Compound 1 hydrochloride Form A.

[0179] Figure 19: DVS curve of Compound 1 mesylate salt Form A.

[0180] Figure 20: Raman spectrum of Compound 1 hydrochloride Form A.

[0181] Figure 21: Infrared spectrum of Compound 1 hydrochloride Form A.

[0182] Figure 22: shows the concentration change curve of the compound in the plasma of adult beagle dogs after intravenous administration and oral gavage administration, where the horizontal axis is the time after administration (h) and the vertical axis is the concentration of the compound in dog plasma (ng / ml), including the control group AG-881 (Control Example 1) and the compound 1 group.

[0183] Figure 23 shows the concentration change curve of the compound in the plasma of adult beagle dogs after oral administration or gavage, wherein the abscissa represents the time after administration (h) and the ordinate represents the concentration of the compound in the dog's plasma (ng / ml).

[0184] Unless otherwise specified, the detection instrument information and detection method parameters used in the present invention are as follows:

[0185] Table 1

[0186] Table 2

[0187] Table 3

[0188] Table 4

[0189] Table 5

[0190] Table 6

[0191] Single crystal testing

[0192] Single crystal data were collected at 296 K using a Bruker SMART APEX-II. Cu Kα radiation was used to analyze the crystal structure using the direct method (Shelxs97). The least squares method was used to correct the structural parameters and identify the atomic species. The geometric calculation method was used to obtain the positions of all hydrogen atoms.

[0193] Raman spectroscopy

[0194] Data were collected using a Thermo DXR3xi with a laser power of 1.5 mW, an exposure time of 0.00235 s, 5 scans, a 10x objective lens, and a 25 μm pinhole.

[0195] Infrared spectroscopy

[0196] Potassium bromide pellets were used and data were collected using a Bruker Tensor 27 infrared spectrometer in the range of 400-4000 wavenumbers (cm -1 ). DETAILED DESCRIPTION

[0197] The present invention is further illustrated by the following examples, which do not limit the scope of the present invention. In the specific examples of the present invention, unless otherwise specified, the techniques or methods described are conventional techniques or methods in the art. In the following examples, unless otherwise specified, the raw materials and reagents described were commercially available; the percentages, proportions, ratios, and parts are calculated by weight.

[0198] Abbreviations:

[0199] AcOK / KOAc: potassium acetate;

[0200] AcONa: sodium acetate;

[0201] API: Active pharmaceutical ingredient;

[0202] Bn: benzyl;

[0203] Boc: tert-butyloxycarbonyl;

[0204] (BOC)2O: di-tert-butyl dicarbonate;

[0205] Cbz: benzyloxycarbonyl;

[0206] DCM: dichloromethane;

[0207] DIEA: N,N-diisopropylethylamine;

[0208] DMF: N,N-dimethylformamide;

[0209] DMSO: dimethyl sulfoxide;

[0210] DPPA: diphenylphosphoryl azide;

[0211] DMA: dimethylacetamide;

[0212] DMAP: 4-dimethylaminopyridine;

[0213] NMP: N-methylpyrrolidone;

[0214] dppf: 1,1'-bis(diphenylphosphino)ferrocene;

[0215] EA: ethyl acetate;

[0216] Fmoc: fluorenylmethoxycarbonyl;

[0217] K2CO3: potassium carbonate;

[0218] Na2CO3: sodium carbonate;

[0219] Cs2CO3: cesium carbonate;

[0220] K3PO4: potassium phosphate;

[0221] KF: potassium fluoride; PMB: p-methoxybenzyl;

[0222] PPh3: triphenylphosphine;

[0223] PCy3: tricyclohexylphosphine;

[0224] P(t-Bu)3: tri-tert-butylphosphine;

[0225] PG: protecting group;

[0226] SPhos: 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl; THF: tetrahydrofuran;

[0227] TEA: triethylamine;

[0228] TFA: trifluoroacetic acid

[0229] DIEA: diisopropylethylamine;

[0230] LC-MS or LCMS: liquid chromatography-mass spectrometry;

[0231] LiHMDS: lithium bis(trimethylsilyl)amide;

[0232] NBS: N-bromosuccinimide;

[0233] PE: petroleum ether;

[0234] [PdCl2(dppf)]CH2Cl2 / Pd(dppf)Cl2.DCM: [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride dichloromethane complex;

[0235] Pd(dppf)Cl2: [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride;

[0236] Pd(PPh3)4: tetrakis(triphenylphosphine)palladium;

[0237] Pd(OAc)2: palladium acetate;

[0238] XantPhos: 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene;

[0239] Xphos: 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl;

[0240] Pre-HPLC: preparative high performance liquid chromatography;

[0241] RH: relative humidity;

[0242] RRT: relative retention time;

[0243] hrs / h: hours;

[0244] LC-MS / LCMS: liquid chromatography-mass spectrometry;

[0245] 1 H-NMR / HNMR: hydrogen nuclear magnetic resonance;

[0246] XRD / XRPD: X-ray powder diffraction;

[0247] SCXRD: single crystal X-ray diffraction;

[0248] DSC: differential scanning calorimetry;

[0249] TGA: thermogravimetric analysis;

[0250] DVS: Dynamic moisture sorption.

[0251] Example 1 Synthesis of Intermediate Compound A13

[0252] Step 1: Synthesis of Compound A01

[0253] The compound 2-chloro-4-fluoronicotinic acid (64.66 g), TEA (102.4 ml), and tert-butanol (54.6 g) were dissolved in toluene (600 mL) and heated to 90°C. A solution of DPPA (106.44 g) dissolved in toluene (400 mL) was added. LCMS monitored the reaction for completion and the mixture was cooled to room temperature. Saturated aqueous sodium bicarbonate solution (750 mL) was added to quench the reaction. Extraction was performed with EA (300 mL x 3). The combined organic phases were washed twice with water and once with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure. The residue was added with n-heptane / ethyl acetate (V / V = 15 / 1) and stirred for washing. Filtering afforded 53.41 g of a solid. LCMS [M+H] + :247.06.

[0254] Step 2: Synthesis of Compound A12

[0255] Compound A01 (53.41 g) was dissolved in DCM (530 mL), and TFA (246.88 g) was added. The mixture was stirred at room temperature for 2 h. The reaction was monitored by LCMS until completion. The reaction solution was concentrated under reduced pressure, and the residue was dissolved in DCM. Saturated aqueous sodium carbonate was added dropwise under ice-cooling to adjust the pH to 7-8. The organic phase was separated, and the aqueous phase was extracted with DCM (100 mL x 3). The combined organic phases were washed three times with water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was added to n-heptane (500 mL), heated with stirring, filtered while hot, and the filtrate was cooled to room temperature. The resulting solid was dried under vacuum at 40°C to obtain the target compound A12 (23.61 g, 74.4%). LCMS [M+H] + :147.00.

[0256] Step 3: Synthesis of Compound A13

[0257] 2-Chloro-4-fluoropyridin-3-amine (7.00 g), DMF (100.00 mL), and NBS (9.35 g) were added sequentially to a reaction flask and allowed to react at room temperature. After completion of the reaction, monitored by LCMS, water was added to the reaction system and extracted with ethyl acetate. The organic phase was collected and washed with saturated sodium chloride. The organic phase was then dried by rotary evaporation. The crude product was purified by column chromatography (EA / PE = 0-10%) to obtain the target compound A13 (10.10 g, 93.79%). 1 H NMR (300MHz, CDCl3): δ7.16 (d, J=8.3Hz, 1H), 4.13 (s, 2H). LCMS[M+H] + :224.92.

[0258] Example 2 Synthesis Method 1 of Compound 1

[0259] Step 1: Synthesis of Compound A04

[0260] Compound A13 (29.95 g) was dissolved in THF (300 mL), followed by the addition of DIEA (51.51 g), (Boc)2O (86.98 g), and DMAP (8.12 g). The mixture was stirred at room temperature for 1 h. The reaction was monitored by LCMS until the starting material disappeared. The reaction was quenched by the addition of water, the organic phase was separated, and the aqueous phase was extracted with EA (100 mL x 3). The combined organic phases were washed three times with water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The concentrated residue was added with n-heptane (300 mL), heated with stirring, and filtered while hot. The filtrate was cooled to room temperature and filtered. The resulting solid was dried under vacuum at 40°C to yield the target compound A04 (33.45 g, 59.15%). 1HNMR (400MHz, DMSO-d6): δ8.13 (d, J=8.0Hz, 1H), 1.39 (s, 18H). LCMS:447.2,449.0([M+Na] + ).

[0261] Step 2: Synthesis of Compound A05

[0262] Compound A04 (5.00 g) was dissolved in toluene (100 mL), and diboronic acid pinacol ester (3.88 g), AcOK (3.46 g), and Pd(dppf)Cl2.DCM (285 mg) were added sequentially at room temperature. The reaction system was purged with nitrogen three times and heated at 80°C for 4 h. The reaction was monitored by LCMS until the starting material disappeared. The reaction solution was cooled to room temperature, and EA (50 mL) and water (50 mL) were added to the reaction system, stirred to dissolve, and the layers were separated. The organic phase was washed with saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the target compound A05 (4.73 g). LCMS [M+H-82] + :391.12. 1 H NMR (500MHz, CDCl3): δ7.58 (d, J=8.3Hz, 1H), 1.40 (s, 18H), 1.39 (s, 12H).

[0263] Step 3: Synthesis of Compound A08

[0264] Compound A07 (4.00 g, 11.8 mmol) and compound A05 (6.01 g) were dissolved in a mixture of toluene (80 mL) and H2O (20 mL). Pd(dppf)Cl2.DCM (0.48 g) and K2CO3 (4.90 g) were added sequentially at room temperature. The reaction was allowed to proceed at 80°C under nitrogen for 4.0 h. LCMS was used to monitor the reaction until the starting material disappeared. The reaction system was cooled to 50°C, filtered, and the filtrate was concentrated under reduced pressure. The residue was added to ethyl acetate / n-heptane (V / V = 1 / 20) and dissolved by heating. The mixture was filtered, and the filtrate was concentrated under reduced pressure to yield the target compound A08 (6.50 g, 85.5%). LCMS [M+H] + :648.19.

[0265] Step 4: Synthesis of compound 1

[0266] Compound A08 (3 g) was dissolved in DCM (30 mL), and TFA (8.13 mL) was added and reacted at room temperature. LCMS monitored the disappearance of the starting material. The reaction solution was dried by rotary evaporation, and the crude product was purified by Pre-HPLC (mobile phase: A: 0.1% TFA in water; B: acetonitrile; flow rate: 40 ml / min; gradient: 37%-62% B in 20 min; wavelength: 254 nm; column: Luna C18 (3), 10 μm, 100 Å, 30*250 mm) to obtain the target compound 1 (270 mg, 11.01%). LCMS [M+H] + :448.08. 1 H-NMR (500MHz, DMSO-d6) δ8.45,8.34,8.04,7.84(d,d,d,d,J=9Hz,J=9Hz,J=9Hz,J=9Hz,2H),8.14 ,7.96(d,d,J=11Hz,J=11Hz,1H),6.30(s,2H),5.20-5.15,4.96-4.88(m,m,2H),1.33(d,J=7.0Hz).

[0267] Example 3 Synthesis Method 2 of Compound 1

[0268] Step 1: Synthesis of compound A101

[0269] A mixture of 6-bromo-2-chloro-4-fluoropyridin-3-amine (10.00 g), THF (50 mL), and (BOC)2O (10.65 g) was cooled to -78°C under nitrogen, and LiHMDS (133.07 mL, 1.00 mol / L) was added dropwise. The reaction mixture was slowly warmed to room temperature for reaction. LCMS monitored the disappearance of the starting material. Ice was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic phase was separated, dried, and spin-dried. The crude product was purified by column chromatography (EA / PE = 0-4%) to obtain the target compound A101 (6 g, 41.55%). LCMS [M+H] + :324.97. 1 H NMR (500 MHz, methanol-d4): δ 7.74 (d, J = 7.7 Hz, 1H), 1.43 (s, 9H).

[0270] Step 2: Synthesis of Compound A102

[0271] Compound A101 (5.70 g), diboronic acid pinacol ester (4.89 g), [PdCl(dppf)]CHCl (1.43 g), AcOK (5.15 g), and 1,4-dioxane (60 mL) were added to a reaction flask. The atmosphere was purged with nitrogen three times and the temperature was raised to 80°C for 3 h. After completion of the reaction, the reaction mixture was cooled to room temperature, followed by addition of ethyl acetate and water, followed by stirring and dissolution. The organic phase was separated, washed with water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to afford crude target compound A102 (5.10 g), which was used directly in the next step without purification.

[0272] Step 3: Synthesis of Compound A103

[0273] To the crude compound A102 (5.10 g) were added compound A07 (4.65 g), [PdCl2(dppf)]CH2Cl2 (1.41 g), K2CO3 (7.14 g), 1,4-dioxane (100 mL), and water (10 mL) in sequence. The atmosphere was replaced with nitrogen three times, and the temperature was raised to 80°C for reaction. After completion of the reaction, monitored by LCMS, the reaction solution was poured into water and extracted with ethyl acetate. The organic phase was separated and washed with saturated aqueous sodium chloride solution and concentrated under reduced pressure. The residue was purified by column chromatography (EA / PE = 0-30%) to obtain the target compound A103 (3 g, 31.81%). LCMS [M+H] + :548.13.

[0274] Step 4: Synthesis of compound 1

[0275] The same preparation method as in step 4 of Example 2 was used to obtain a crude product of compound 1, which was then purified by Prep-HPLC (Column: Luna-C18(3)-10-100; Size: 30*250nm, Conditions: acetonitrile / water (0.1% formic acid) 35%-65%, 20min, UV: 308nm) to obtain the target compound 1.

[0276] The obtained target compound 1 was characterized by XRPD, and it had an XRPD spectrum substantially as shown in FIG1 , and the main data of its X-ray powder diffraction pattern were shown in Table 7, and was named Compound 1 Form A.

[0277] Table 7

[0278] Example 5 Preparation of Compound 1 Crystalline Form B

[0279] 10-20 mg of Compound 1 Form A was heated to 160°C using a TGA at a heating rate of 10°C / min and equilibrated at 160°C for 15 minutes before collecting the Compound 1 solid. The Compound 1 solid was characterized by XRPD, TGA, and DSC, which showed an XRPD pattern substantially as shown in Figure 2, a TGA pattern substantially as shown in Figure 3, and a DSC pattern substantially as shown in Figure 4. The DSC melting endotherm peak temperature was approximately 226°C (Onset value), indicating that the obtained product was Compound 1 Form B. The main data of the X-ray powder diffraction pattern of Compound 1 Form B are shown in Table 8.

[0280] Table 8

[0281] Example 6 Preparation of Compound 1 Crystalline Form D

[0282] Approximately 20 mg of Compound 1 Form A was weighed into a sample vial, and 0.5 ml of ethanol was added and stirred at room temperature to dissolve the mixture. 1.6 ml of purified water was slowly added to the reaction solution, resulting in the precipitation of solids. The suspension was stirred overnight, and after solid-liquid separation, it was dried at room temperature for 3 hours to obtain a product. The product was characterized by XRPD, TGA, and DSC, which showed an XRPD pattern substantially as shown in Figure 5, a TGA pattern substantially as shown in Figure 6, and a DSC pattern substantially as shown in Figure 7, indicating that the obtained product was Compound 1 Form D. The main data of the X-ray powder diffraction pattern of Compound 1 Form D are shown in Table 9.

[0283] Table 9

[0284] Example 7 Salt screening experiment of compound 1

[0285] The sample obtained in Example 4 (20 mg) and the corresponding 2 molar equivalents of acid were added to a sample bottle, and the corresponding solvent (0.5 mL) was added. The mixture was stirred at room temperature for 3 days. If the reaction system was a suspension, solid-liquid separation was performed and the sample was tested by XRPD. If the reaction system was a clear solution, the clear sample was slowly evaporated at room temperature to obtain a solid and then tested by XRPD. The experimental results are shown in Table 10.

[0286] Table 10 Salt type screening test results

[0287] As shown in the salt screening results in Table 10, "salt formation" indicates that the reaction can form a classic salt or eutectic product, and the product can exist as a stable solid. These results indicate that salt formation for Compound 1 is unpredictable, with only a few acids successfully forming stable salts with Compound 1. Among these, hydrochloric acid, sulfuric acid, phosphoric acid, L-tartaric acid, L-malic acid, and L-lactic acid have significant salt-forming advantages.

[0288] Example 8 Preparation of Hydrochloride Form A of the Compound Represented by Formula I

[0289] Method 1

[0290] Compound 1 Form B (500 mg) and 1.1 molar equivalents of hydrochloric acid were added to a sample bottle, and ACN (5 mL) was added. The mixture was suspended and stirred at room temperature for 3 days. The solid was separated by centrifugation and dried in vacuo at 50°C for 3 hours to obtain an off-white solid (300 mg). 1 H-NMR (500MHz, DMSO-d6) δ9.35,9.11,8.97(br,br,br,2H),8.34,7.94(d,d,J=11.0Hz,J=10 .5Hz,1H),6.94(br,3H),5.37-5.31,5.17-5.09,4.91-4.84(m,m,m,2H),1.39-1.35(m,6H).

[0291] The solid was subjected to XRPD, TGA, DSC and Raman characterization, which showed an XRPD spectrum substantially as shown in Figure 8, a TGA spectrum substantially as shown in Figure 9, a DSC spectrum substantially as shown in Figure 10, a Raman spectrum substantially as shown in Figure 20, and an infrared spectrum substantially as shown in Figure 21, indicating that the target compound obtained is Compound 1 hydrochloride salt Form A.

[0292] The main data of the X-ray powder diffraction pattern of Compound 1 hydrochloride Form A are shown in Table 11.

[0293] Table 11

[0294] Method 2

[0295] Methanol (15.0 L) was added to a 100-liter reactor, heated to 50°C, and compound 1 (1880.05 g) was added. The solution dissolved and filtered, and the filtrate was transferred to a 100-liter reactor and heated at 50°C with stirring. Hydrochloric acid (830.07 g (2.0 eq)) was diluted with methanol (10.0 L) and added to the reactor. Methanol (12.5 L) was then added while maintaining the temperature and stirring. Purified water (6.6 L) was slowly added dropwise to the reaction system. After the addition was complete, 18.81 g of seed crystals were added. Purified water (31.0 L) was then added dropwise to the reaction solution. After the addition was complete, the reaction system was slowly cooled to room temperature and stirred for 4 hours. The mixture was then filtered, and the filter cake was rinsed with 4.0 L of a methanol / water (1 / 1, v / v) mixture. The resulting filter cake was dried under vacuum at 50°C. XRPD, TGA, and DSC analysis revealed the hydrochloride salt, Form A (1869.43 g).

[0296] About 20 mg of the hydrochloride crystal form A sample was weighed and added to a sample bottle. 1 mL of methanol was added to dissolve the clear solution. After filtering, the sample bottle was sealed with a sealing film, and a hole was pierced in the sealing film. The sample was placed in a dimethyltetrahydrofuran atmosphere and subjected to gas-liquid diffusion at room temperature to obtain a single crystal sample. The single crystal sample was subjected to SCRXD detection to confirm that the hydrochloride crystal form A of the compound 1 according to the present invention was obtained. Then, single crystal testing and analysis were performed on it. The unit cell stacking projection diagram of the hydrochloride crystal form A single crystal is basically shown in Figure 14, and the unit cell parameters are as follows:

[0297] The crystal belongs to the orthorhombic system, the space group is P212121, and the unit cell parameters are: α=β=γ=90.00°;

[0298] Unit cell volume

[0299] The number of asymmetric units in the unit cell is Z = 4.

[0300] The final reliability factors R1 = 0.0283, wR2 = 0.0749, S = 1.088. The final stoichiometric formula of the asymmetric unit is C 14 H 13 The molecular weight of a single ClF7N7·HCl molecule is calculated to be 484.22, and the crystal density is calculated to be 1.607 g / cm 3 .

[0301] Example 9 Preparation of Compound 1 Methanesulfonate Form A

[0302] Compound 1 Form A (150 mg) and 1 equivalent of methanesulfonic acid were added to EtOAc (2 mL), suspended and stirred at room temperature for 3 days, the solid was separated by centrifugation, and dried in vacuo at room temperature overnight to obtain an off-white solid. XRPD analysis showed that it had an XRPD spectrum substantially as shown in Figure 11, and the DSC melting endothermic peak temperature was approximately 242°C (Onset value), indicating that the target compound obtained was Compound 1 mesylate Form A.

[0303] The main data of the X-ray powder diffraction pattern of Compound 1 mesylate salt Form A are shown in Table 12.

[0304] Table 12

[0305] Example 10 Crystal stability experiment

[0306] Sample and experimental preparation: Take an appropriate amount of Compound 1 hydrochloride Form A and place it in a watch glass, spread it into a thin layer of about 3-5 mm thickness, and place it under the following experimental conditions.

[0307] The experimental conditions are: high temperature (60°C, open), high humidity (RH92.5%, open), light (25°C, 4500lux±500lux, open), long term (25°C / 60%RH, aluminum-aluminum packaging) and accelerated (40°C / 75%RH, aluminum-aluminum packaging).

[0308] Test items: Samples were collected for testing on day 0, day 5, day 10, or day 30, and XRPD and HPLC were compared. The test results are shown in Table 13.

[0309] Table 13 Stability test results

[0310] The results show that the hydrochloride crystal form A has excellent physical and chemical stability, which is beneficial for drug development.

[0311] Example 11 Crystal Hygroscopicity Experiment

[0312] The hygroscopicity of Compound 1 Form A, Compound 1 Form B, Compound 1 Form D, Compound 1 Hydrochloride Form A and Methanesulfonate Form A was evaluated using a dynamic moisture sorption instrument (DVS). The samples were tested for moisture absorption and weight gain changes in the range of 0% RH (relative humidity) to 95% RH. The moisture absorption and weight gain test results of the samples at 25°C / 80% RH are shown in Figures 15-19, and the test results are shown in Table 14.

[0313] Table 14 Weight changes of samples in the range of 0% RH to 80% RH

[0314] From the test results, it can be seen that Compound 1 Hydrochloride Form A and Compound 1 Form B have outstanding advantages in hygroscopicity. Compared with Compound 1 Form A, the hygroscopicity of Compound 1 Hydrochloride Form A and Compound 1 Form B is greatly reduced.

[0315] Example 12 Pharmacological Test of Compound 1

[0316] Example A: Enzyme activity inhibition test

[0317] Compound 1 was tested for its effect on IDH1 R132H and IDH2 R140Q The inhibitory capacity of enzyme activity was measured with the half-maximal inhibitory concentration (IC 50 ) values. AG-881 was used as a positive control compound. In this experiment, fluorescence was used to detect the expression of IDH1 R132H and IDH2 R140Q Compounds were detected on the enzyme with a starting concentration of 10,000 nM, 3-fold dilution, 10 concentrations, and single-well detection.

[0318] (1) Prepare 1× Assay buffer.

[0319] (2) Compound concentration gradient preparation: Compound 1 was tested at a starting concentration of 10,000 nM, diluted 3-fold, and 10 concentrations were added to a 100% DMSO solution in a 384-well plate to a final concentration of 200 times.

[0320] (3) Use Echo 550 to transfer 250 nL of DMSO and 250 nL of diluted compound 1 to the reaction plate.

[0321] (4) IDH2 R140Q Enzyme assay: add 25 μL IDH2 R140Q Enzyme solution was added and incubated for 60 minutes. 25 μL of substrate solution was then added and incubated for 150 minutes. 25 μL of detection buffer was then added, shaken for 1 minute, and incubated for 60 minutes. The final reading was performed using a Synergy 2 with an Ex544 / Em590 cutoff of 590.

[0322] (5) IDH1 R132H In enzyme assay: add 40 μL of 1.25 times the final concentration of IDH1 R132H酶 Mix the solution with NADPH and incubate for 60 minutes. Then add 10 μL of substrate solution at 5x the final concentration and incubate for 90 minutes. Then add 25 μL of detection buffer at 3x the final concentration and shake for 1 minute. Read the result using a Synergy 2 with an Ex544 / Em590 cutoff of 590.

[0323] (8) Inhibition rate calculation: %Inhibition = (RFU_sample–RFU_min) / (RFU_max–RFU_min)*100%. Where: RFU-sample is the fluorescence intensity of the sample; RFU-min is the mean value of the negative control wells, representing the fluorescence intensity with enzyme; RFU-max is the mean value of the positive control wells, representing the fluorescence intensity without enzyme. Graphpad Prism software was used for curve fitting to obtain IC 50 The test results are shown in Table 15.

[0324] Example B: Cell 2-HG Experiment

[0325] The inhibitory effect of compound 1 on the 2-HG production ability in the culture supernatant of U87 cell line stably transfected with IDH1-R132H and TF-1 cell line stably transfected with IDH2-R140Q was detected.

[0326] (1) Collect the cultured cells by centrifugation, then suspend them in cell culture medium. After counting the cells, inoculate the cells into a 96-well cell culture plate, with 160 μL of cell suspension per well (20,000 cells / well for U87-IDH1-R132H and 10,000 cells / well for TF-1-IDH2-R140Q).

[0327] (2) Take a 96-well dilution plate and serially dilute compound 1 with DMSO and cell culture medium. Then, take 40 μL of the serially diluted compound 1 and add it to the 96-well culture plate with cells. The starting concentration for U87-IDH1-R132H cells is 2 μM, and the serial dilution is 3-fold. The starting concentration for TF-1-IDH2-R140Q cells is 10 μM, and the serial dilution is 3-fold (the final DMSO concentration is 0.2%). The final volume per well is 200 μL. Finally, the cells are cultured in a 37°C incubator with 5% CO2.

[0328] (3) After the compound and cells were incubated for 72 hours, the cell culture plate was removed and centrifuged at 1500 rpm for 5 minutes. Then 100 μL of the culture supernatant was aspirated for 2-HG determination.

[0329] (4) 2-HG detection: Dilute the cell supernatant with purified water, take 30 μL of the diluted sample into a 1.5 ml centrifuge tube, add 200 μL of the internal standard solution, vortex to mix, and centrifuge at 4°C, 3200 rpm for 15 minutes. Take 100 μL of the supernatant solution and dilute it 1:1 with water, and detect it by LC-MS. The Analyte Peak Area represents the 2-HG level.

[0330] (5) Inhibition rate calculation: %Inhibition = (1-Analyte Peak Area_sample / Analyte Peak Area_max) * 100%. Where: Analyte Peak Area_sample is the Analyte Peak Area of ​​the sample with compound added; Analyte Peak Area_max is the mean Analyte Peak Area of ​​the positive control well (representing the absence of compound). Curve fitting was performed using Graphpad Prism software to obtain the IC 50 The test results are shown in Table 15.

[0331] Table 15

[0332] This shows that the compounds of the present invention have excellent enzymatic and cellular activities.

[0333] Example C: Canine Pharmacokinetics Test

[0334] Adult beagle dogs (6-12 months old) received a single intravenous (IV) or oral gavage (PO) dose of Compound 1 and AG-881. For the IV single dose, Compound 1 and AG-881 were dissolved in 5% DMSO + 5% Solutol + 90% saline and administered intravenously at a dose of 1 mg / kg (2 females, 2 males). Blood was collected at 5 min, 15 min, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 10 h, and 24 h. For the PO single dose, Compound 1 and AG-881 were dissolved in 5% DMSO + 5% Solutol + 90% purified water and administered orally at a dose of 5 mg / kg (2 females, 2 males). Blood was collected orally at 15 min, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 10 h, and 24 h. The collected blood samples were promptly placed in a test tube containing EDTA anticoagulant, then centrifuged at 4°C and 4000rpm for 10 minutes, and the supernatant was transferred to a centrifuge tube and stored at -20°C. During the test, 30 μL of plasma supernatant sample was taken, 200 μL of internal standard solution was added, and centrifuged at 3000rpm for 10 minutes. Then 100 μL of the supernatant solution was diluted with water in a 1:1 ratio and injected, with an injection volume of 5 μL. Liquid chromatography-tandem mass spectrometry (LC-MS / MS) was used to analyze the concentration of the test compound in the plasma sample. The plasma concentration-time data of individual animals were analyzed using Sciex Analyst software. A non-compartmental model was introduced in the concentration analysis, and the pharmacokinetic parameters (Cl_obs, C max , AUC last ), the test results are shown in Table 16, and the PK curve is shown in Figure 22.

[0335] Table 16

[0336] Note: “ / ” means the corresponding parameter was not detected.

[0337] Example D: Pharmacokinetics of different solid forms in dogs

[0338] Adult beagle dogs (6-12 months old) received a single oral gavage (PO) of Compound 1 Form A and Compound 1 Form A hydrochloride. Compound 1 Form A and Compound 1 Form A hydrochloride were encapsulated and administered orally (1 female, 1 male) at a dose of 10 mg / kg. Blood samples were collected at 15 minutes, 0.5 hours, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours, 24 hours, 36 hours, and 48 hours. Blood samples were promptly placed in tubes containing EDTA anticoagulant and centrifuged at 4000 rpm for 10 minutes at 4°C. The supernatant was transferred to a centrifuge tube and stored at -20°C. For testing, 30 μL of plasma supernatant was sampled, 200 μL of internal standard solution was added, and the samples were centrifuged at 3000 rpm for 10 minutes. Then, 100 μL of the supernatant was diluted 1:1 with water and injected into the tubes, with a total injection volume of 5 μL. Liquid chromatography-tandem mass spectrometry (LC-MS / MS) was used to analyze the concentration of the test compound in plasma samples. The plasma concentration-time data of individual animals were analyzed using Sciex Analyst software. A non-compartmental model was introduced into the concentration analysis, and the pharmacokinetic parameters (Cl_obs, C) of the test compound were calculated using WinNonlin (version 4.1; BYkinginsun) software. max , AUC last ), the test results are shown in Table 17, and the PK curve is shown in Figure 23.

[0339] Table 17

Claims

1. A salt form of the compound represented by formula I and a crystalline form of the salt form, characterized in that: The salt type is hydrochloride, phosphate, sulfate, methanesulfonate, L-malate, L-tartrate or L-lactate.

2. A crystalline form of the compound represented by formula I, characterized in that: The X-ray powder diffraction pattern of the crystalline form comprises characteristic peaks with 2θ values ​​of 5.8°±0.2°, 11.7°±0.2° and 17.6°±0.2°, 3. The crystal form according to claim 2, characterized in that The X-ray powder diffraction pattern of the crystal form comprises characteristic peaks with 2θ values ​​of 5.8°±0.2°, 11.7°±0.2°, 17.6°±0.2°, 19.6°±0.2° and 23.5°±0.2°.

4. The crystal form according to any one of claims 2 to 3, characterized in that: The X-ray powder diffraction pattern of the crystalline form comprises 5 or more characteristic peaks having 2θ values ​​selected from 5.8°±0.2°, 11.7°±0.2°, 14.2°±0.2°, 16.8°±0.2°, 17.6°±0.2°, 18.6°±0.2°, 19.6°±0.2° and 23.5°±0.2°.

5. The crystal form according to any one of claims 2 to 4, characterized in that: The crystalline form has a TGA spectrum substantially as shown in FIG3 .

6. The crystal form according to any one of claims 2 to 5, characterized in that: The crystalline form has a DSC spectrum substantially as shown in FIG4 .

7. A crystalline form of the compound represented by formula I, characterized in that: The X-ray powder diffraction pattern of the crystalline form comprises characteristic peaks with 2θ values ​​of 8.6°±0.2°, 10.5°±0.2° and 21.1°±0.2°, 8. The crystal form according to claim 7, characterized in that The X-ray powder diffraction pattern of the crystal form includes characteristic peaks with 2θ values ​​of 8.6°±0.2°, 10.5°±0.2°, 11.8°±0.2°, 14.0°±0.2°, 17.3°±0.2° and 21.1°±0.2°.

9. The crystal form according to any one of claims 2 to 3, characterized in that: The X-ray powder diffraction pattern of the crystalline form comprises 5 or more characteristic peaks having 2θ values ​​selected from 8.6°±0.2°, 10.5°±0.2°, 11.8°±0.2°, 14.0°±0.2°, 16.5°±0.2°, 17.3°±0.2°, 18.3°±0.2°, 19.8°±0.2° and 21.1°±0.2°.

10. The crystal form according to any one of claims 2 to 4, characterized in that: The crystalline form has a TGA spectrum substantially as shown in FIG6 .

11. The crystal form according to any one of claims 2 to 5, characterized in that: The crystalline form has a DSC spectrum substantially as shown in FIG7 .

12. The salt form and the crystalline form of the salt form according to claim 1, characterized in that: The salt form is a hydrochloride salt as shown in Formula II; 13. The salt form and the crystalline form of the salt form according to claim 12, characterized in that: The crystal form is hydrochloride crystal form A, and the X-ray powder diffraction pattern of the hydrochloride crystal form A includes characteristic peaks with 2θ values ​​of 11.6°±0.2°, 12.7°±0.2°, 13.0°±0.2°, 19.2°±0.2° and 21.1±0.2°.

14. The salt form and the crystalline form of the salt form according to claim 13, characterized in that: The X-ray powder diffraction pattern of the hydrochloride salt form A comprises characteristic peaks with 2θ values ​​of 11.6°±0.2°, 12.7°±0.2°, 13.0°±0.2°, 17.3°±0.2°, 18.3°±0.2°, 19.2°±0.2° and 21.1±0.2°.

15. The salt form and the crystal form of the salt form according to any one of claims 13 to 14, characterized in that: The X-ray powder diffraction pattern of the hydrochloride salt form A includes characteristic peaks with 2θ values ​​of 1.6°±0.2°, 12.7°±0.2°, 13.0°±0.2°, 17.3°±0.2°, 18.3°±0.2°, 19.2°±0.2°, 21.1±0.2°, 23.9±0.2° and 25.5±0.2°.

16. The salt form and the crystalline form of the salt form according to any one of claims 13 to 15, characterized in that: The X-ray powder diffraction pattern of the hydrochloride salt form A comprises 5 or more characteristic peaks having 2θ values ​​selected from 11.6°±0.2°, 12.7°±0.2°, 13.0°±0.2°, 15.4°±0.2°, 16.3°±0.2°, 17.3°±0.2°, 18.3°±0.2°, 19.2°±0.2°, 21.1±0.2°, 22.6±0.2°, 23.9±0.2°, 25.5±0.2°, 28.0±0.2° and 31.8±0.2°.

17. The salt form and the crystalline form of the salt form according to any one of claims 13 to 16, wherein: The hydrochloride salt form A has an X-ray powder diffraction pattern substantially as shown in FIG8 .

18. The salt form and the crystalline form of the salt form according to any one of claims 13 to 17, characterized in that: The hydrochloride salt form A has a TGA spectrum substantially as shown in FIG9 .

19. The salt form and the crystalline form of the salt form according to any one of claims 13 to 18, characterized in that: The hydrochloride salt form A has a DSC spectrum substantially as shown in FIG. 7 .

20. The salt form and the crystalline form of the salt form according to any one of claims 13 to 19, characterized in that: The unit cell parameters of the hydrochloride salt form A are as follows: α=β=γ=90.00°; Unit cell volume The number of asymmetric units in the unit cell is Z = 4.

21. The salt form and the crystalline form of the salt form according to claim 1, characterized in that: The salt form is a mesylate salt as shown in formula III; 22. The salt form and the crystalline form of the salt form according to claim 21, characterized in that: The crystal form is mesylate crystal form A, and the X-ray powder diffraction pattern of the mesylate crystal form A includes characteristic peaks with 2θ values ​​of 6.6°±0.2°, 8.2°±0.2° and 19.3°±0.2°.

23. The salt form and the crystalline form of the salt form according to any one of claims 21 to 22, characterized in that: The X-ray powder diffraction pattern of the mesylate salt form A comprises characteristic peaks with 2θ values ​​of 6.6°±0.2°, 8.2°±0.2°, 17.3°±0.2° and 19.3°±0.2°.

24. The salt form and the crystalline form of the salt form according to any one of claims 21 to 23, characterized in that: The X-ray powder diffraction pattern of the mesylate salt form A comprises three or more characteristic peaks having 2θ values ​​selected from 6.6°±0.2°, 8.2°±0.2°, 10.5°±0.2°, 14.2°±0.2°, 17.3°±0.2°, 19.3°±0.2° and 20.2°±0.2°.

25. The salt form and the crystalline form of the salt form according to any one of claims 21 to 25, characterized in that: The mesylate salt Form A has a TGA spectrum substantially as shown in FIG12 .

26. The salt form and the crystalline form of the salt form according to any one of claims 13 to 18, characterized in that: The mesylate salt Form A has a DSC spectrum substantially as shown in FIG13 .

27. A pharmaceutical composition, characterized in that The pharmaceutical composition contains 0.5% to 85% by weight of the compound of formula I, or the crystal form, salt form or crystal form of the salt form as described in any one of claims 1 to 26, or a mixture thereof, 28. The composition of claim 27, wherein The pharmaceutical composition further contains one or more pharmaceutically acceptable carriers, and the pharmaceutically acceptable carriers include one or more of diluents, fillers, lubricants, binders, and disintegrants.

29. Use of the crystalline form or salt form according to any one of claims 1 to 26, or the composition according to claims 27 to 28 in the preparation of a drug, characterized in that: The medicament is used to treat diseases mediated by mutant IDH1 and / or IDH2.

30. The use according to claim 29, characterized in that The disease is cancer.

31. The use according to claim 30, characterized in that The cancer is selected from the group consisting of brain glioma, melanoma, papillary thyroid tumor, bile duct cancer, lung cancer, breast cancer, sarcoma, glioma, glioblastoma multiforme, acute myeloid leukemia, and non-Hodgkin's lymphoma.

32. A method for treating a disease, characterized in that: The method comprises administering a therapeutically effective amount of the crystalline form or salt form of any one of claims 1 to 26, or the composition of claims 27 to 28 to a subject, wherein the disease is a disease mediated by mutant IDH1 and / or IDH2.

33. The method of claim 32, wherein: The disease is cancer, a proliferative disease or an inflammatory disease, and the cancer is selected from brain glioma, melanoma, papillary thyroid tumor, bile duct cancer, lung cancer, breast cancer, sarcoma, glioma, glioblastoma multiforme, acute myeloid leukemia, and non-Hodgkin's lymphoma.

34. A method for preparing a compound of formula I, characterized in that: The method comprises using the compound represented by formula IV to carry out a deprotection reaction to prepare the compound represented by formula I; Wherein, the R1 is an amino protecting group, and the R2 is H or an amino protecting group.

35. The method of claim 34, wherein: The preparation method also includes using a compound represented by the following formula V With compound A07 Carrying out a coupling reaction to obtain a compound represented by formula IV; Wherein, R1 is an amino protecting group, and R2 is H or an amino protecting group; The R3 is selected from H, C 1-4 alkyl, or two R3 and the O atom to which they are connected together form a 5- or 6-membered heterocyclic ring containing two oxygen atoms and one boron atom, wherein the 5- or 6-membered heterocyclic ring may be optionally replaced by one or more C 1-4 Alkyl substituted.

36. The preparation method according to any one of claims 34 to 35, characterized in that: The preparation method further comprises using the compound shown in the following formula VI to react with biboric acid pinacol ester to obtain the compound shown in formula V, Wherein, the R1 is an amino protecting group, and the R2 is H or an amino protecting group.

37. The preparation method according to any one of claims 34 to 36, characterized in that: The amino protecting group may be selected from one or two of -Boc, -Cbz, -Fmoc, -PMB and -Bn.

38. The preparation method according to claim 34, characterized in that The amino protecting group is a -Boc group, and the compound represented by Formula IV is subjected to a deprotection reaction under acidic conditions. Preferably, the acid is TFA.

39. The preparation method according to claim 35, characterized in that: The compound represented by formula V is coupled with compound A07 under alkaline conditions. Preferably, the base can be selected from one or more of K2CO3, AcOK, Na2CO3, AcONa, Cs2CO3, K3PO4, KF or TEA.

40. The preparation method according to claim 36, characterized in that: The compound represented by formula VI reacts with biboric acid pinacol ester under alkaline conditions, preferably, the base is AcOK.

41. The preparation method according to claim 35 or 36, characterized in that: The reaction is carried out under the catalysis of a palladium catalyst, preferably, the palladium catalyst is selected from Pd(dppf)Cl2 or Pd(OAc)2.

42. The preparation method according to claim 35 or 36, characterized in that: The reaction temperature is 70-100°C.

43. An intermediate compound, characterized in that Selected from the following compounds, in, The R1 is an amino protecting group, and the R2 is H or an amino protecting group; The R3 is selected from H, C 1-4 alkyl, or two R3 and the O atom to which they are connected together form a 5- or 6-membered heterocyclic ring containing two oxygen atoms and one boron atom, wherein the 5- or 6-membered heterocyclic ring may be optionally replaced by one or more C 1-4 Alkyl substituted.

44. The compound according to claim 43, characterized in that The amino protecting group may be selected from one or two of -Boc, -Cbz, -Fmoc, -PMB and -Bn.

45. The compound of claim 44, wherein The amino protecting group is a -Boc group.

46. ​​An intermediate compound, characterized in that Selected from the following compounds, 47. A method for preparing an intermediate compound 6-bromo-2-chloro-4-fluoropyridin-3-amine, characterized in that: Select from the following method 1 or method 2; Method 1: Method 2:

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