An fgfr4 inhibitor acid salt and preparation method and application thereof

Abstract

The present application relates to a kind of FGFR4 inhibitor acid salt and its preparation method and application, the FGFR4 inhibitor is the compound N- ((3S, 4S) -3- ((6- (2, 6-difluoro-3, 5-dimethoxyphenyl) -8- (3-methoxy-3-methylazetidin-1-yl) pyridine [3, 4-d] pyrimidine-2-yl) amino) tetrahydro-2H-pyran-4-yl) acrylamide with formula (I) structure, the acid salt is p-toluenesulfonic acid salt, greatly improve the solubility and bioavailability and other physical and chemical properties of free state formula (I) compound, can satisfy the need of clinical drug preparation development.The p-toluenesulfonic acid salt of the present application has very important clinical application value, and is expected to accelerate the development into a new generation of FGFR4 small molecule inhibitor.

Description

Technical Field

[0001] This invention belongs to the field of drug development, specifically relating to an acid salt of an FGFR4 inhibitor, its preparation method, and its application. Background Technology

[0002] Fibroblast growth factor (FGF) is a family of 22 structure-related polypeptides with different biological activities. The corresponding receptors of FGF (FGFRs) belong to the RPTK family of receptor tyrosine kinases. Currently, four receptors—FGFR1, FGFR2, FGFR3, and FGFR4—have been identified. Their interaction with their corresponding FGF ligands leads to receptor dimerization and autophosphorylation, thereby initiating multiple downstream signaling cascades, including MAPK and AKT. Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related deaths in China and one of the fastest-growing cancers in terms of annual incidence. The current first-line treatment is sorafenib, and there are no approved second-line drugs; targeted therapies with antitumor agents are still needed. FGF19 is overexpressed in 5–10% of HCC patients, and FGFR4 is the dominant FGFR in human hepatocytes, and its high expression in hepatocytes is considered to be associated with the invasiveness of HCC tumors. Therefore, FGFR4 plays a very important role in liver cancer. In addition, the interaction between FGF19 and FGFR4 is also believed to be related to the invasiveness of other cancer types, such as stomach cancer, prostate cancer, lung cancer, colorectal cancer, pancreatic cancer, and ovarian cancer.

[0003] Currently, highly selective FGFR4 inhibitors can effectively treat cancers caused by abnormal FGFR4 signaling pathways and avoid related side effects such as hyperphosphatemia caused by FGFR1-3 inhibition. Small molecule inhibitors of highly selective FGFR4 have significant application prospects in the field of tumor targeted therapy. As a good drug candidate, FGFR4 inhibitors can meet the demand for targeted drugs for liver cancer and other tumors at home and abroad, and bring the advantages of good safety and stronger specificity.

[0004] Abbisko Therapeutics Co., Ltd., through long-term research, has invented a novel small molecule compound with highly selective FGFR4 inhibitory effects. The relevant patent is WO2018113584A1 (international publication date: June 28, 2018). Representative compounds are as follows:

[0005]

[0006] The Chinese name is: N-((3S,4S)-3-((6-(2,6-difluoro-3,5-dimethoxyphenyl)-8-(3-methoxy-3-methylacetidin-1-yl)pyrido[3,4-d]pyrimidin-2-yl)amino)tetrahydro-2H-pyran-4-yl)acrylamide (compound of formula (I)). This compound can significantly improve the inhibitory effect on FGFR4 target and the selectivity for other FGFR1-3 kinase receptors, meeting the current domestic and international needs for targeted therapy of tumors such as liver cancer, prostate cancer, pancreatic cancer, esophageal cancer, gastric cancer, lung cancer, breast cancer, ovarian cancer, colon cancer, skin cancer, glioblastoma or rhabdomyosarcoma.

[0007] However, at the time of the patent application, suitable raw materials and processes for clinical and industrial applications had not been developed, and no further research was conducted on whether the compound of formula (I) was suitable for drug development. Later research by the inventors revealed that the free base compound disclosed in WO2018113584A1 is poorly soluble in water and physiological solvents, and has relatively poor bioavailability, failing to meet clinical requirements and thus unsuitable for clinical formulation development. Therefore, to meet the needs of clinical research and marketed drug formulations, it is urgent to further study the aggregation state of the drug to improve the physicochemical properties of the compound, meet the needs of pharmaceutical or clinical applications, and develop a salt-form or crystalline compound suitable for drug development to overcome the deficiencies of the existing technology. Summary of the Invention

[0008] To address the problems existing in the prior art, the inventors further investigated the aggregation state of compound (I) N-((3S,4S)-3-((6-(2,6-difluoro-3,5-dimethoxyphenyl)-8-(3-methoxy-3-methylacetidin-1-yl)pyrido[3,4-d]pyrimidin-2-yl)amino)tetrahydro-2H-pyran-4-yl)acrylamide. During long-term research, the inventors unexpectedly discovered that p-toluenesulfonate can significantly improve the solubility and bioavailability of the free form of compound (I). Its physicochemical properties, such as hygroscopicity and chemical stability, also meet the requirements for industrial production and can satisfy the needs of clinical drug formulation development. The p-toluenesulfonate of this invention can be widely used in the preparation of drugs for treating cancers, particularly liver cancer, prostate cancer, pancreatic cancer, esophageal cancer, gastric cancer, lung cancer, breast cancer, ovarian cancer, colon cancer, skin cancer, glioblastoma, or rhabdomyosarcoma. It has significant clinical application value and is expected to accelerate the development into a new generation of FGFR4 small molecule inhibitors.

[0009] The first aspect of the present invention provides a compound of formula (I) p-toluenesulfonate:

[0010]

[0011] As a preferred embodiment, the compound of formula (I) p-toluenesulfonate is a crystalline compound.

[0012] As a further preferred embodiment, the X-ray powder diffraction (XRPD) pattern of the p-toluenesulfonate compound of formula (I) includes peaks at diffraction angles (2θ) of 9.02±0.2°, 17.18±0.2°, 17.68±0.2°, 19.60±0.2° and 22.90±0.2°.

[0013] As a further preferred embodiment, the X-ray powder diffraction (XRPD) pattern of the p-toluenesulfonate compound of formula (I) includes peaks at diffraction angles (2θ) of 9.02±0.2°, 17.18±0.2°, 17.68±0.2°, 19.60±0.2°, 22.90±0.2°, 10.60±0.2°, 12.72±0.2°, 18.56±0.2°, 21.22±0.2°, and 24.90±0.2°.

[0014] As a further preferred embodiment, the X-ray powder diffraction (XRPD) pattern of the p-toluenesulfonate compound of formula (I) includes peaks at diffraction angles (2θ) of 9.02±0.2°, 17.18±0.2°, 17.68±0.2°, 19.60±0.2°, 22.90±0.2°, 10.60±0.2°, 12.72±0.2°, 18.56±0.2°, 21.22±0.2°, 24.90±0.2°, 15.32±0.2°, 23.82±0.2°, 24.12±0.2°, 24.60±0.2°, and 25.58±0.2°.

[0015] As a preferred embodiment, the X-ray powder diffraction pattern of the p-toluenesulfonate compound of formula (I) includes the following θ values: 9.02±0.2°, 10.60±0.2°, 12.04±0.2°, 12.72±0.2°, 15.32±0.2°, 17.18±0.2°, 17.68±0.2°, 18.56±0.2°, 19.60±0.2°, and 20.60±0.2°. Peaks at diffraction angles (2θ) of 21.22±0.2°, 22.90±0.2°, 23.82±0.2°, 24.12±0.2°, 24.60±0.2°, 24.90±0.2°, 25.58±0.2°, 27.78±0.2°, 29.04±0.2°, 31.48±0.2°, 36.60±0.2°, and 38.16±0.2°.

[0016] As the most preferred embodiment, the X-ray diffraction pattern of the p-toluenesulfonate powder of compound (I) is substantially the same as the peak at the diffraction angle (2θ) shown in Table 1, and the diffraction angle (2θ) and related intensity data (±0.2°) are as follows:

[0017] 2θ(°) strength% 2θ(°) strength% 9.02 40.9 27.78 6.8 10.60 19.6 29.04 8.4 12.04 7.5 29.51 1.7 12.72 29.8 30.62 2.8 14.88 4.3 31.48 8.1 15.32 12.4 31.92 2.4 17.18 47 32.38 1.2 17.68 100 32.84 3.7 18.56 23.2 34.04 2.4 19.60 61.3 35.10 2.2 20.60 8.1 36.60 6.9 21.22 29.1 37.10 3 22.90 84.9 38.16 5.1 23.82 18.8 39.72 3.3 24.12 17.7 40.14 2.4 24.60 14.5 41.06 2.1 24.90 31.5 42.38 3.5 25.58 13.8 42.72 2 26.20 4.2 44.24 1.9 26.66 2.9

[0018] As a preferred embodiment, the unit cell of the p-toluenesulfonate compound of formula (I) is P1. The unit cell volume is

[0019] As a preferred embodiment, the compound of formula (I) p-toluenesulfonate is a hydrate.

[0020] As a more preferred embodiment, each molecule of the p-toluenesulfonate hydrate of formula (I) contains 1 to 3 water molecules.

[0021] As a further preferred embodiment, each molecule of the p-toluenesulfonate hydrate of formula (I) contains 1, 2 or 3 water molecules.

[0022] As an invention of this application, the applicant designates the p-toluenesulfonate crystal of compound (I) having any of the aforementioned powder X-ray diffraction data as crystal form A. The obtained crystal, after physical characterization by DSC, TGA, and XRPD, shows that it is a well-crystallized solid. The DSC and TGA results indicate that the acid salt of the crystalline form (I) compound is a monohydrate, such as... Figure 1 , Figure 2 As shown.

[0023] A second aspect of the present invention provides a method for preparing p-toluenesulfonate of the aforementioned compound (I), comprising the following steps:

[0024] 1) Dissolve or disperse the free form (I) compound in an aqueous solvent or a suitable organic solvent, and add p-toluenesulfonic acid or a p-toluenesulfonic acid solution to the above system to carry out a salt formation reaction; or, add the free form (I) compound to a p-toluenesulfonic acid solution to carry out a salt formation reaction;

[0025] 2) Collect the solid product p-toluenesulfonate of formula (I) precipitated during the above salt formation reaction; or obtain the solid product p-toluenesulfonate of formula (I) by creating supersaturation in the salt formation system.

[0026] As a preferred embodiment, the method for creating supersaturation of the salt system in step 2) includes one or more of the following: adding seed crystals, evaporating solvent, adding antisolvent, or cooling.

[0027] As a preferred embodiment, the obtained solid product compound (I) p-toluenesulfonate is a crystalline compound (I) p-toluenesulfonate.

[0028] As a preferred embodiment, the suitable organic solvent used in step 1) of salt formation is selected from alcohols, chlorinated alkanes, ketones, ethers, cyclic ethers, esters, alkanes, cycloalkanes, benzenes, amides, sulfoxides, or mixtures thereof.

[0029] As a preferred embodiment, the suitable organic solvent is selected from methanol, ethanol, n-propanol, isopropanol, dichloromethane, acetonitrile, acetone, 1,4-dioxane, tetrahydrofuran, N,N-dimethylformamide, ethyl acetate, isopropyl acetate, methyl tert-butyl ether, 2-methoxyethyl ether, or mixtures thereof.

[0030] A third aspect of the present invention provides a pharmaceutical composition comprising a therapeutically effective dose of the compound of formula (I) above, p-toluenesulfonate, and a pharmaceutically acceptable carrier.

[0031] The fourth aspect of the present invention provides the use of p-toluenesulfonate of formula (I) in the preparation of FGFR4 inhibitor drugs.

[0032] The fifth aspect of the present invention provides the use of the compound p-toluenesulfonate of formula (I) in the preparation of a drug for treating liver cancer, prostate cancer, pancreatic cancer, esophageal cancer, gastric cancer, lung cancer, breast cancer, ovarian cancer, colon cancer, skin cancer, glioblastoma or rhabdomyosarcoma.

[0033] The sixth aspect of the present invention provides a method for treating liver cancer, prostate cancer, pancreatic cancer, esophageal cancer, gastric cancer, lung cancer, breast cancer, ovarian cancer, colon cancer, skin cancer, glioblastoma, or rhabdomyosarcoma, the method comprising administering to a patient in need of treatment an effective therapeutic amount of the compound of formula (I) p-toluenesulfonate or the pharmaceutical composition thereof. Attached Figure Description

[0034] Figure 1 The differential scanning calorimetry (DSC) plot of compound (I) p-toluenesulfonate (Form A) is shown; the X-axis represents temperature (°C) and the Y-axis represents heat flux (W / G).

[0035] Figure 2 Thermogravimetric analysis plot of compound (I) p-toluenesulfonate (Form A); X-axis represents temperature (°C), Y-axis represents weight loss percentage (%).

[0036] Figure 3 The image shows the powder X-ray diffraction pattern of compound (I) p-toluenesulfonate (Form A); the X-axis represents the diffraction peak angle 2θ (°), and the Y-axis represents the peak intensity.

[0037] Figure 4The simulated powder diffraction pattern of single-crystal p-toluenesulfonate (Form A) of Formula (I) is shown below, and the measured XRPD pattern of p-toluenesulfonate (Form A) of Formula (I) is shown above. The horizontal axis represents the 2θ value (°), and the vertical axis represents the peak intensity.

[0038] Figure 5 This is a single-crystal unit cell diagram of compound (I) p-toluenesulfonate (Form A).

[0039] Figure 6 This is a DVS plot for compound (I) p-toluenesulfonate (Form A). The horizontal axis represents relative humidity (%), and the vertical axis represents weight change (%).

[0040] Figure 7 The time / solubility curves of compound (I) p-toluenesulfonate (Form A) and the free form of compound (I) are shown in 25% PEG300 / 5% Solutol HS15 / 70% water solvent. The horizontal axis represents time (hours), and the vertical axis represents concentration (mg / mL). Detailed Implementation

[0041] Drugs may exhibit different bioavailability, solubility, melting point, and chemical and physical stability depending on their crystal form, thus affecting their safety and efficacy. To develop a suitable salt or crystal form for drug development, the inventors conducted in-depth research on compound (I), N-((3S,4S)-3-((6-(2,6-difluoro-3,5-dimethoxyphenyl)-8-(3-methoxy-3-methylacetidin-1-yl)pyridino[3,4-d]pyrimidin-2-yl)amino)tetrahydro-2H-pyran-4-yl)acrylamide. Extensive screening of salt and crystal forms of compound (I) was performed, ultimately revealing that p-toluenesulfonate can significantly improve the solubility and bioavailability of the free form of compound (I). Its hygroscopicity and chemical stability also meet the requirements for industrial production, satisfying the needs of clinical drug formulation development and potentially accelerating its development into a new generation of FGFR4 small molecule inhibitors.

[0042] Detailed explanation: Unless otherwise stated, the terms used in the specification and claims shall have the following meanings.

[0043] "Pharmaceutical composition" means a mixture containing one or more of the compounds described herein or their physiologically / pharmacologically acceptable salts or prodrugs, along with other chemical components, such as physiologically / pharmacologically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to a living organism, thereby promoting the absorption of the active ingredient and the exertion of its biological activity.

[0044] "Polymorphism" refers to a crystal form with the same chemical composition but different spatial arrangements of the molecules, atoms, and / or ions that constitute the crystal. Although polymorphs have the same chemical composition, their packing and geometric arrangements differ, and they may exhibit different physical properties, such as melting point, shape, color, density, hardness, deformability, stability, solubility, dissolution rate, and similar properties. Based on their temperature-stability relationship, two polymorphs can be either monomorphic or tautomorphic. For monomorphic systems, the relative stability between the two solid phases remains constant with temperature changes. Conversely, in tautomorphic systems, there is a transition temperature at which the stability of the two phases changes. This phenomenon of compounds existing in different crystal structures is called pharmaceutical polymorphism.

[0045] The various crystalline structures of this invention can be distinguished from each other using a variety of analytical techniques known to those skilled in the art. Such techniques include, but are not limited to, X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), and / or thermogravimetric analysis (TGA).

[0046] Methods and Materials

[0047] 1.1 X-ray powder diffraction

[0048] Those skilled in the art will recognize that X-ray powder diffraction patterns can be obtained with measurement errors that depend on the measurement conditions used. Intensities in X-ray powder diffraction patterns may fluctuate with the material conditions used. Relative intensities may also vary with experimental conditions; accordingly, exact intensities should not be taken into account. Furthermore, the measurement error of conventional powder X-ray diffraction angles is typically about 5% or less, and this degree of measurement error should be considered within the aforementioned diffraction angle range. Therefore, the crystal structure of the present invention is not limited to crystal structures that provide X-ray powder diffraction patterns that are completely identical to those depicted in the accompanying drawings. Any crystal structure that provides substantially the same X-ray powder diffraction patterns as those disclosed in the drawings falls within the scope of the present invention. Those skilled in the art should be able to determine that X-ray powder diffraction patterns are substantially identical. Other suitable standard calibrations known to those skilled in the art are also applicable. However, relative intensities may vary with crystal size and shape.

[0049] The crystal forms of the compounds of the present invention are characterized by their X-ray powder diffraction patterns. Therefore, in the presence of Cu Kα radiation... X-ray powder diffraction patterns of the salt were acquired using a Bruker D8 Discover X-ray powder diffractometer (GADDS CS) operating in reflection mode. The tube voltage and current were set to 40 kV and 40 mA, respectively, for acquisition scanning. The sample was scanned for 60 seconds within the 2θ range of 3.0° to 40°. The diffractometer was calibrated using corundum standards for the peak positions indicated by 2θ. All analyses were performed typically at 20°C to 30°C. Data were acquired and integrated using GADDS software version 4.1.14T of WNT software. The diffraction patterns were analyzed using DiffracPlus software with Eva version 9.0.0.2, released in 2003.

[0050] XRPD sample preparation typically involves placing the sample on a single-crystal silicon wafer and pressing the sample powder with a glass slide or equivalent to ensure a flat surface and appropriate height. The sample holder is then placed in a Bruker XRPD instrument, and X-ray powder diffraction patterns are acquired using the instrument parameters described above. Measurement variations associated with these X-ray powder diffraction analysis results are caused by several factors, including: (a) errors in sample preparation (e.g., sample height), (b) instrument errors, (c) calibration variations, (d) operator errors (including those occurring when determining peak positions), and (e) the properties of the material (e.g., preferred orientation errors). Calibration errors and sample height errors often result in shifts in all peaks in the same direction. Generally, this calibration factor will ensure that the measured peak positions are consistent with the expected peak positions and can be within the expected 2θ value ± 0.2°.

[0051] 1.2 Differential Scanning Calorimetry (DSC)

[0052] Differential scanning calorimetry (DSC) experiments at TA Instruments TM The experiment was conducted in Model Q2000. The sample (approximately 1–6 mg) was weighed in an aluminum pan and recorded precisely to the nearest hundredth of a milligram, then transferred to the DSC apparatus. The instrument was purged with nitrogen at 50 mL / min. Data were collected at a heating rate of 10 °C / min between room temperature and 350 °C. Plots were drawn with the endothermic peak pointing downwards. However, those skilled in the art will note that in DSC measurements, the measured onset and maximum temperatures exhibit a degree of variability depending on the heating rate, crystal shape and purity, and other measurement parameters.

[0053] 1.3 Thermogravimetric Analysis (TGA)

[0054] Thermogravimetric analysis (TGA) experiments at TA Instruments TMPerform in Model Q500. Place the sample (about 10 - 30 mg) in a pre-weighed platinum dish. Accurately measure the sample weight by the instrument and record it to one-thousandth of a milligram. Purge the furnace with nitrogen at 100 mL / min. Collect data at a heating rate of 10 °C / min between room temperature and 300 °C.

[0055] 1.4 Dynamic Vapor Sorption (DVS)

[0056] The experimental method for characterizing the acid salt of the crystalline form (I) compound by Dynamic Vapor Sorption (DVS) is as follows: Take a small amount of the acid salt powder of the crystalline form (I) compound and place it in a precision sample dish compatible with the instrument. After loading the sample, send it into the instrument for detection. In this patent, the instrument model used for all Dynamic Vapor Sorption is DVS Intrinsic, and the experimental parameters are set as follows: Use nitrogen as the carrier gas, set a constant temperature of 25 °C, and the mass percentage change rate per unit time (dm / dt) = 0.01% / min as the criterion for reaching equilibrium. The program humidity change cycle is set as follows: The initial relative humidity is 0%, the relative humidity at the end point is 90%, the cycle is set 2 times, and each 10% R.H. change is a step.

[0057] The reagents in the embodiments of the present invention are known and commercially available. The reagents used are commercially available industrial grade or analytical grade reagents. Or they can be synthesized by methods known in the art or in accordance with such methods. The API raw materials are prepared according to Patent WO2018113584A1.

[0058] Unless otherwise specified, all reactions of the present invention are carried out under continuous magnetic stirring in a dry nitrogen or argon atmosphere. The solvent is a dry solvent, and the temperature unit is degrees Celsius (°C). The term "room temperature" or "RT" used herein refers to an ambient temperature of 20 to 25 °C (68 - 77 °F).

[0059] The following examples further illustrate the present invention in detail and completely. They are only used to illustrate specific embodiments of the present invention and should not be construed as limiting the scope of the present invention in any way.

[0060] Preparation of Specific Examples

[0061] Preparation of the p-toluenesulfonate of the compound of formula (I) in Example 1

[0062] Weigh 5 mg of the free compound of formula (I) and dissolve it in 0.5 mL of ethyl acetate; weigh 1.67 mg of p-toluenesulfonic acid and dissolve it in 0.1 mL of tetrahydrofuran; slowly add the p-toluenesulfonic acid solution to the compound of formula (I) solution under stirring, stir overnight, filter, and dry the filter cake in a vacuum drying oven at 40 °C for DSC, TGA, and XRPD characterization. The DSC and TGA diagrams of the obtained p-toluenesulfonate of the compound of formula (I) are shown in Figure 1 and Figure 2 The XRPD diffraction pattern can be found in [the image]. Figure 3 .

[0063] Example 2 Preparation of p-toluenesulfonate of compound (I)

[0064] Weigh 2g of the free compound (I) and dissolve it in 160mL of ethyl acetate; weigh 668mg of p-toluenesulfonic acid and dissolve it in 4mL of tetrahydrofuran; slowly add the p-toluenesulfonic acid solution to the compound (I) solution while stirring, stir overnight, filter, and dry the filter cake in a vacuum drying oven at 40℃. The XRPD diffraction pattern of the obtained p-toluenesulfonate of compound (I) is compared with... Figure 3 Consistent.

[0065] Example 3: Preparation of p-toluenesulfonate of compound (I)

[0066] Weigh 5g of free compound (I) and dissolve / suspend it in 400mL of acetone; weigh 1670mg of p-toluenesulfonic acid and dissolve it in 10mL of tetrahydrofuran; slowly add the p-toluenesulfonic acid solution to the compound (I) solution while stirring, stir overnight, filter, and dry the filter cake in a vacuum drying oven at 40℃. The XRPD diffraction pattern of the obtained p-toluenesulfonate of compound (I) is compared with... Figure 3 Consistent.

[0067] Example 4 Single Crystal Preparation and Characterization

[0068] Weigh 150 mg of compound (I) p-toluenesulfonate, add 1 mL of solvent (composition: methanol: ethanol = 1:1, plus 2% water), stir at 50 °C for 15 min, filter the suspension through a 0.45 μm filter membrane, and use the filtrate as the mother liquor. Take 340 μL of the mother liquor, add 260 μL of solvent, mix well, cool at room temperature, and recrystallize to obtain single crystals.

[0069] Suitable single crystals were selected and analyzed using a Bruker APEX-IICCD single crystal diffractometer. The temperature was maintained at 220 K during data collection. The cell parameters of p-toluenesulfonate of formula (I) are shown in Table 2; the single crystal XRPD simulation results are consistent with the XRPD results of p-toluenesulfonate of formula (I), as shown in Table 2. Figure 4 As shown; the single-crystal unit cell structure is as follows Figure 5 As shown in Table 2:

[0070]

[0071] Example 5: Moisture Absorption Behavior Test

[0072] The hygroscopic weight gain of compound (I) p-toluenesulfonate (Form A) under various relative humidities was determined using the dynamic moisture adsorption method (hygroscopic weight gain / weight before moisture absorption * 100%). The hygroscopicity of compound (I) p-toluenesulfonate (Form A) was evaluated, and the results are as follows: Figure 6 As shown. Experimental results show that the crystal weight of compound (I) of the present invention, p-toluenesulfonate (Form A), changes by no more than 2% with increasing relative humidity (%), maintaining good hygroscopicity, meeting the requirements of industrial production, and satisfying the needs of clinical drug formulation development.

[0073] Example 6 Stability Test

[0074] Stability tests were conducted on compound (I) p-toluenesulfonate (Form A) and free compound (I) under high temperature and accelerated experimental conditions. Chemical purity was used as the evaluation index to evaluate the stability of compound (I) p-toluenesulfonate (Form A) and free compound (I). The experimental results are shown in Table 3.

[0075]

[0076] The above experimental results show that the compound of the present invention remains stable for 14 days under high temperature and accelerated experimental conditions, and the chemical stability of the product meets the requirements of industrial production and can meet the needs of clinical drug formulation development.

[0077] Example 7 Solubility Determination

[0078] 1) Solubility in water

[0079] 2 mg of compound (I) p-toluenesulfonate (Form A) and 2 mg of free compound (I) were weighed and placed in 2 mL glass bottles respectively. 0.4 mL of water was added to each glass bottle. The mixture was stirred on a magnetic stirrer at room temperature for 24 hours, centrifuged, and the content was determined by liquid chromatography. The experimental results are shown in Table 4.

[0080] compound solvent Solubility (μg / mL) Compound (I) p-Toluenesulfonate (Form A) water 1370 Free state of compound (I) water 25.6

[0081] The experimental results above show that the solubility of compound (I) p-toluenesulfonate (Form A) in water is significantly higher than that of free compound (I), which can meet the needs of drug clinical formulation development.

[0082] 2) Solubility in physiological media

[0083] Weigh 80 mg of compound (I) p-toluenesulfonate (Form A) and 80 mg of free compound (I) into separate 4 mL glass bottles. Add 2 mL of solvent (25% PEG300 / 5% Solutol HS15 / 70% water) (v / v / v) to each bottle. Stir on a magnetic stirrer at room temperature. Take samples at different time points, centrifuge, and determine the content using liquid chromatography. The results are as follows. Figure 7 As shown in the figure. The experimental results show that the solubility of compound (I) in toluenesulfonate is significantly higher than that in free compound (I), which meets the needs of drug clinical formulation development.

[0084] Example 8: Animal in vivo pharmacokinetics experiment

[0085] Compound (I) p-toluenesulfonate (Form A) and the free form of compound (I) were subjected to pharmacokinetic (PK) experiments in monkeys at a dose of 200 mpk (content was calculated as free compound). The experimental results are shown in Table 5.

[0086]

[0087]

[0088] The above experimental results show that compound (I) p-toluenesulfonate can significantly increase the exposure of free compound (I) in monkeys, thus indicating that the bioavailability of compound (I) p-toluenesulfonate (Form A) is significantly higher than that of free compound (I).

[0089] All references to this invention are incorporated herein by reference as if each document were individually incorporated herein by reference. Furthermore, it should be understood that after reading this invention, those skilled in the art can make various alterations or modifications to it, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. A compound of formula (I) p-toluenesulfonic acid salt: ； The compound of formula (I) p-toluenesulfonic acid salt is a crystalline compound. The X-ray powder diffraction pattern (XRPD) of the compound of formula (I) p-toluenesulfonic acid salt further comprises peaks at diffraction angles (2 theta) of 10.60 ± 0.2º, 12.72 ± 0.2º, 18.56 ± 0.2º, 21.22 ± 0.2º and 24.90 ± 0.2º.

2. The p-toluenesulfonate compound of formula (I) according to claim 1, characterized in that, The X-ray powder diffraction pattern (XRPD) of the compound of formula (I) p-toluenesulfonic acid salt further comprises peaks at diffraction angles (2 theta) of 10.60 ± 0.2º, 12.72 ± 0.2º, 18.56 ± 0.2º, 21.22 ± 0.2º and 24.90 ± 0.2º.

3. The p-toluenesulfonate compound of formula (I) according to claim 2, characterized in that, The X-ray powder diffraction pattern (XRPD) of the compound of formula (I) p-toluenesulfonic acid salt further comprises peaks at diffraction angles (2 theta) of 10.60 ± 0.2º, 12.72 ± 0.2º, 18.56 ± 0.2º, 21.22 ± 0.2º and 24.90 ± 0.2º.

4. The compound of formula (I) according to claim 1, p-toluenesulfonic acid salt, characterized in that, The X-ray powder diffraction pattern (XRPD) of the compound of formula (I) p-toluenesulfonic acid salt further comprises peaks at diffraction angles (2 theta) of 10.60 ± 0.2º, 12.72 ± 0.2º, 18.56 ± 0.2º, 21.22 ± 0.2º and 24.90 ± 0.2º.

5. The compound of formula (I) according to any one of claims 1 to 4, p-toluenesulfonic acid salt, characterized in that, The compound of formula (I) tosylate crystallizes in the space group PI with a = 9.079 (3) A, b = 10.561 (3) A, c = 10.882 (3) A, and the unit cell volume is 908.1 (4) A3 3 .

6. The compound of formula (I) according to any one of claims 1 to 4, p-toluenesulfonic acid salt, characterized in that, The compound of formula (I) p-toluenesulfonic acid salt is a hydrate.

7. The p-toluenesulfonate compound of formula (I) according to claim 6, characterized in that, The hydrate comprises 1 water molecule per molecule.

8. A process for the preparation of the p-toluenesulfonic acid salt of the compound of formula (I) according to claim 1, characterized in that, The process comprises the following steps: 1) dissolving or dispersing the free form of the compound of formula (I) in an aqueous solvent or a suitable organic solvent, adding p-toluenesulfonic acid or a solution of p-toluenesulfonic acid to the above system to carry out a salt formation reaction; or adding the free form of the compound of formula (I) to a solution of p-toluenesulfonic acid to carry out a salt formation reaction; 2) collecting the solid product of the compound of formula (I) p-toluenesulfonic acid salt precipitated in the above salt formation reaction; or obtaining the solid product of the compound of formula (I) p-toluenesulfonic acid salt by creating supersaturation in the salt formation system.

9. The preparation method according to claim 8, characterized in that, The method for creating supersaturation in the salt formation system in step 2) is selected from one or more of the following: adding seed crystals, volatilizing solvent, adding antisolvent or lowering temperature.

10. The preparation method according to claim 8, characterized in that, The suitable organic solvent used in step 1) is selected from alcohol, chloroalkane, ketone, ether, cyclic ether, ester, alkane, cycloalkane, benzene, amide, sulfoxide organic solvent or a mixture thereof.

11. The preparation method according to claim 10, characterized in that, said suitable organic solvent is selected from methanol, ethanol, n-propanol, isopropanol, dichloromethane, acetone, 1,4-dioxane, tetrahydrofuran, N,N-dimethylformamide, ethyl acetate, isopropyl acetate, methyl-tert-butyl ether, 2-methoxyethyl ether or a mixture thereof.

12. A pharmaceutical composition comprising a therapeutically effective amount of the compound of formula (I) p-toluenesulfonate salt according to any one of claims 1 to 7 and a pharmaceutically acceptable carrier.

13. Use of the compound of formula (I) p-toluenesulfonate salt according to any one of claims 1 to 7 for the manufacture of a medicament for the inhibition of FGFR4.

14. Use according to claim 13, characterized in that, said medicament is for the treatment of liver cancer, prostate cancer, pancreatic cancer, esophageal cancer, gastric cancer, lung cancer, breast cancer, ovarian cancer, colon cancer, skin cancer, glioblastoma or rhabdomyosarcoma.

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