FGFR4 inhibitor and preparation and application thereof

Abstract

The application belongs to the field of pharmaceutical chemistry, and particularly relates to an FGFR4 inhibitor and preparation and application thereof. The FGFR4 inhibitor is shown as general formula I, has good FGFR4 inhibiting activity and FGFR4 high selectivity, and can be used for treating malignant tumors closely related to the kinase.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of pharmaceutical chemistry, and particularly relates to an FGFR4 inhibitor and preparation and application thereof. BACKGROUND

[0002] FGFR (fibroblast growth factor receptor) is a receptor-type protein tyrosine kinase, including FGFR1, FGFR2, FGFR3, FGFR4 and other different subtypes, each of which has the structural characteristics of an extracellular region binding to a ligand, a transmembrane region and an intracellular region for receptor phosphorylation, and is part of a tyrosine kinase signaling pathway responsible for cell proliferation and differentiation. They play a key role in maintaining cell growth, proliferation, apoptosis, migration, etc. by forming a ternary complex with 18 different fibroblast growth factors (FGF). FGFR molecular changes lead to abnormal FGF / FGFR signals, promoting cell proliferation, neovascularization, invasion, metastasis, anti-apoptosis, etc., thereby participating in a wide range of human malignancies.

[0003] Existing research has found (Clin Cancer Res (2015) 21(12):2684-2694) that FGFR mutation subtypes are not the same in different cancers, such as FGFR1 abnormalities closely related to non-small cell lung cancer, FGFR2 abnormalities closely related to gastric cancer, and FGFR3 abnormalities significantly related to bladder cancer. J. Med. Chem. 2019, 62, 2905-2915 reported that the FGF19 / FGFR4 signaling pathway promotes the survival, proliferation, invasion and metastasis of hepatocellular carcinoma cells through multiple pathways. J. Clin. Oncol. 2015, 33(30), 3401-3408 and J. Clin. Oncol. 2017, 35(2), 157-165, respectively, reported non-selective FGFR inhibitors JNJ-42756493 and BGJ398, which lack selectivity for FGFR subtypes, inhibit FGFR1 and FGFR3, leading to interference with normal phosphate metabolism by blocking FGF23-mediated signaling, thereby causing hyperphosphatemia side effects (Nat. Rev. Drug Discovery 2016, 15(1), 51-69). Therefore, it is necessary to design an inhibitor that only shows high inhibitory activity against FGFR4 while not affecting the activity of other FGFR1-3 subtypes, in combination with the specificity of the FGFR subtypes ligand structure, thereby improving tumor drug targeting, reducing drug toxicity, improving drug compliance, and being very necessary. SUMMARY

[0004] An object of the present application is to provide a highly selective FGFR4 inhibitor of general formula I, 3-substituted ethynyl benzamide compounds of general formula I have good FGFR4 inhibitory activity and high FGFR4 selectivity, and can be used for treating malignant tumors closely related to this kinase.

[0005] Another object of the present application is to provide a method for preparing the compound of general formula I of the present application.

[0006] Still another object of the present application is to provide a composition comprising the compound of general formula I of the present application and a pharmaceutically acceptable carrier, and a composition comprising the compound of general formula I of the present application and another or more drugs.

[0007] Still another object of the present application is to provide a method for treating and / or preventing FGFR4 related diseases by the compound of general formula I of the present application, and the use of the compound of general formula I of the present application in the preparation of a medicament for treating and / or preventing FGFR4 related diseases.

[0008] To achieve the above objects, the present application provides the following technical solutions:

[0009] In a first aspect, the present application provides a compound of general formula I,

[0010]

[0011] wherein,

[0012] R1 is selected from hydrogen or C 1-6 alkyl;

[0013] R2 is selected from hydrogen or C 1-6 alkyl.

[0014] In some preferred embodiments, R1 is selected from hydrogen or methyl.

[0015] In some preferred embodiments, R2 is selected from hydrogen or methyl.

[0016] In a second aspect, the present application provides a compound of general formula I,

[0017]

[0018]

[0019]

[0020] In a second aspect, the present application provides a method for preparing a compound of general formula I, the method comprising:

[0021] (1) reacting a compound of formula (1) with a compound of formula (2) to form a compound of formula (3);

[0022] (2) the compound of formula (4) is reacted with a compound of formula (5) to form a compound of formula (6);

[0023] (3) the compound of formula (3) is condensed with the compound of formula (6) to form a compound of formula (7);

[0024] (4) the compound of formula (7) is hydrolyzed to form a compound of formula (8);

[0025] (5) the compound of formula (8) is condensed with a compound of formula (9) to form a compound of formula (10); (6) the compound of formula (10) is reduced to form a compound of formula (11);

[0026] (7) the compound of formula (11) is reacted with an enoyl chloride to form a compound of formula (I), the reaction route is as follows:

[0027]

[0028] In a third aspect, the present application provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of general formula (I) and a pharmaceutically acceptable carrier.

[0029] In a fourth aspect, the present application relates to the use of a compound of general formula (I) in the preparation of a medicament for treating or preventing FGFR4-related diseases, especially for treating or preventing liver cancer.

[0030] Terminology

[0031] The "alkyl" as used herein refers to a straight-chain or branched saturated hydrocarbon group, preferably C 1-6 alkyl, further preferably C 1-3 alkyl, preferably C 1-3 alkyl is methyl, ethyl, propyl or isopropyl.

[0032] The "pharmaceutical composition" as used herein refers to a mixture of any one of the compounds described herein and one or more pharmaceutically acceptable carriers and / or excipients. The purpose of a pharmaceutical composition is to facilitate the administration of the compound to an organism. The composition is typically used for the treatment and / or prevention of diseases mediated by FGFR. DETAILED DESCRIPTION

[0033] A method for preparing the above-mentioned phenylacetylene compound targeting FGFR4, the steps are as follows: Example 1: Preparation of 3-((2-((2-acrylamido-6-methylphenyl)amino)pyrimidin-5-yl)ethynyl)-4-methyl-N-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)benzamide

[0034] Synthesis of 3-ethynyl-4-methylbenzoic acid methyl ester

[0035]

[0036] Methyl 3-iodo-4-methylbenzoate (4 g, 14.5 mmol) was dissolved in N,N- dimethylformamide (15 ml) under ice bath condition, bis(triphenylphosphine)palladium dichloride (0.2 g, 0.28 mmol) and cuprous iodide (0.275 g, 1.44 mmol) were added, triethylamine solution (4.39 g, 43.46 mmol) was added dropwise, the system was replaced with nitrogen for 3 times, ethynyltrimethylsilane was added dropwise slowly under ice bath condition, after stirring for 10 min, the ice bath was removed, the reaction mixture was reacted at room temperature for 2 h, then tetrabutylammonium fluoride (10% tetrahydrofuran solution, 4.6 g, 17.6 mmol) was added. The reaction was stirred at room temperature for 30 min. TLC monitoring showed that the reaction was completed. The reaction was cooled to room temperature, 100 ml of ethyl acetate was slowly added to quench the reaction, the reaction solution was filtered through diatomite to obtain a mother liquor, water was added to the filtrate (100 ml x 3) for extraction, the organic layers were combined, the organic layer was washed with saturated brine, then dried with anhydrous sodium sulfate, the solvent was removed under reduced pressure, the residue was concentrated to obtain a solid, and the solid was purified by silica gel column chromatography to obtain 2.3 g of a white solid compound, with a yield of 91%.

[0037] Synthesis of 5-iodo-N-(2-methyl-6-nitrophenyl)pyrimidin-2-amine

[0038]

[0039] Under ice bath condition, 2-amino-5-iodopyrimidine (3 g, 13.57 mmol) was weighed into a flask, DMF was added to stir and disperse uniformly, the reaction was performed for 20 min, then 2-fluoro-3-nitrotoluene (2.74 g, 17.67 mmol) was slowly added dropwise to the reaction flask, stirred for 5 min, and placed at 60°C for 2 h, after the reaction was completed, the reaction system was cooled to room temperature, added to water, extracted with ethyl acetate twice, the organic layers were combined and washed with saturated brine, then dried with anhydrous sodium sulfate, the solvent was removed under reduced pressure, concentrated and purified by silica gel column chromatography to obtain a yellowish solid compound 3.8 g, with a yield of 78%.

[0040] Synthesis of methyl 4-methyl-3-(2-(2-methyl-6-nitrophenyl)amino)pyrimidin-5- yl)ethynyl)benzoate

[0041]

[0042] Into a 250 mL flask, 3-ethynyl-4-methylbenzoic acid methyl ester (2 g, 11.5 mmol), 5-iodo-N-(2-methyl-6-nitrophenyl)pyrimidin-2-amine (2.72 g, 7.66 mmol), bis(triphenylphosphine)palladium(II) chloride (0.12 g, 1.17 mmol) and cuprous iodide (0.33 g, 1.7 mmol) were added and dissolved in 15 mL of dry DMF. Triethylamine (2.67 g, 26.4 mmol) was added dropwise and the system was purged with nitrogen three times. The reaction was then stirred at room temperature under nitrogen for 2 h. TLC monitoring was performed until the reaction was complete. The reaction was quenched by adding 150 mL of ethyl acetate and the solution was filtered through celite to obtain the mother liquor. The filtrate was extracted with water (150 mL x 3), and the organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure and the residue was concentrated to obtain a solid. The solid was purified by silica gel column chromatography to obtain 2.6 g of a yellow solid with a yield of 56%. Synthesis of 4-methyl-3-(2-(2-methyl-6-nitrophenyl)amino)pyrimidin-5-yl)ethynyl)benzoic acid methyl ester

[0043]

[0044] Into a 250 mL flask, 3-ethynyl-4-methylbenzoic acid methyl ester (2 g, 11.5 mmol), 5-iodo-N-(2-methyl-6-nitrophenyl)pyrimidin-2-amine (2.72 g, 7.66 mmol), bis(triphenylphosphine)palladium(II) chloride (0.12 g, 1.17 mmol) and cuprous iodide (0.33 g, 1.7 mmol) were added and dissolved in 15 mL of dry DMF. Triethylamine (2.67 g, 26.4 mmol) was added dropwise and the system was purged with nitrogen three times. The reaction was then stirred at room temperature under nitrogen for 2 h. TLC monitoring was performed until the reaction was complete. The reaction was quenched by adding 150 mL of ethyl acetate and the solution was filtered through celite to obtain the mother liquor. The filtrate was extracted with water (150 mL x 3), and the organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure and the residue was concentrated to obtain a solid. The solid was purified by silica gel column chromatography to obtain 2.6 g of a yellow solid with a yield of 56%. Synthesis of 4-methyl-3-(2-(2-methyl-6-nitrophenyl)amino)pyrimidin-5-yl)ethynyl)benzoic acid methyl ester

[0045]

[0046] To a 250 mL reaction flask were added the compound 4-methyl-3-(2-(2-methyl-6-nitrophenyl)amino)pyrimidin-5-yl)ethynyl)benzoic acid (1.9 g, 4.89 mmol), HATU (2.23 g, 5.87 mmol) and 15 mL of dry DMF. DIEA (1.26 g, 9.76 mmol) was slowly added dropwise and stirred at room temperature for 1 h. 4-(4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline was added and the reaction temperature was raised to 60° C. for 5 h. TLC was monitored until the reaction of the starting material was complete. The reaction was quenched by adding 150 mL of water and extracted with ethyl acetate (150 mL×3). The organic phases were combined, washed with saturated NaCl, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure. The residue was purified by silica gel column chromatography to obtain 2.5 g of a light yellow solid compound in a yield of 79.4%.

[0047] Synthesis of 3-((2-((2-amino-6-methylphenyl)amino)pyrimidin-5-yl)ethynyl)-4-methyl-N-(4-(4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)benzamide

[0048]

[0049] To a 250 mL reaction flask was added the compound 4-methyl-3-(2-(2-methyl-6-nitrophenyl)amino)pyrimidin-5-yl)ethynyl)-N-(4-(4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)benzamide (2.5 g, 3.88 mmol), iron powder (1.09 g, 19.46 mmol), ammonium chloride (1.05 g, 19.44 mmol), 15 mL of ethanol and 3 mL of water. The mixture was heated under reflux at 90° C. for 1 h and monitored by TLC until the reaction was complete. The reaction solution was filtered through celite while hot to obtain a mother liquor. The solvent was removed under reduced pressure, and the product was extracted with saturated sodium carbonate, water, and ethyl acetate (100 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, and the residue was purified by silica gel column chromatography to obtain 600 mg of the compound as a light yellow solid in a yield of 25.2%.

[0050] Synthesis of 3-((2-((2-acrylamido-6-methylphenyl)amino)pyrimidin-5-yl)ethynyl)-4-methyl-N-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)benzamide

[0051]

[0052] Take compound 3-((2-((2-amino-6-methylphenyl)amino)pyrimidin-5-yl)ethynyl)-4-methyl-N-(4-(4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)benzamide (300 mg, 0.48 mmol) in a 25 mL reaction bottle, add 10 mL of dry DCM, triethylamine (98 mg, 0.97 mmol), nitrogen protection, stirring under ice salt bath for 10 min, slowly drop acryloyl chloride (88 mg, 0.97 mmol) solution in dichloromethane (1 mL), TLC monitor the raw material to complete the reaction, add 2 mL ice water to quench the reaction, extract with ethyl acetate (20 mL x 3), wash with saturated sodium carbonate, saturated NaCl in turn, combine the organic phase, dry over anhydrous sodium sulfate, remove the solvent under reduced pressure, and purify the residue by silica gel column chromatography to obtain 80 mg of white powder with a yield of 14.7%. 1 H NMR (600 MHz, DMSO-d6) δ 10.51 (s, 1H), 9.49 (s, 1H), 8.82 (s, 1H), 8.57 (s, 2H), 8.20 (d, J = 2.2 Hz, 1H), 8.12 (d, J = 2.0 Hz, 1H), 8.06 (m, 1H), 7.90 (m, 1H), 7.70 (d, J = 8.6 Hz, 2H), 7.50 (d, J = 8.1 Hz, 1H), 7.20 (m, 1H), 7.12-7.07 (m, 1H), 6.53 (m, 1H), 6.23 (m, 1H), 5.71 (m, 1H), 3.56 (s, 2H), 2.52 (s, 3H), 2.47-2.24 (m, 8H), 2.16 (s, 3H), 2.14 (s, 3H). 13 CNMR (151 MHz, DMSO-d6) 165.15, 163.91, 160.22, 143.87, 138.64, 137.13, 135.39, 132.61, 132.53, 132.28, 131.67, 130.74, 130.37, 128.47, 127.72, 127.30, 126.97, 126.76, 123.95, 122.75, 117.73, 107.86, 90.94, 89.65, 57.92, 55.15, 53.13, 46.14, 20.87, 18.84.

[0053] Example 2: Preparation of 3-(2-(2-(2-chloroacetamide)-6-methylphenyl)amino)pyrimidin-5-yl)ethynyl)-4-methyl-N-(4-(4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)benzamide;

[0054]

[0055] Operation as example 1, separated by column chromatography to get 83 mg white solid. 1 H NMR (600 MHz, DMSO-d6) δ 10.49 (s, 1H), 9.39 (s, 1H), 8.97 (s, 1H), 8.58 (m, 2H), 8.20 (d, J = 2.2 Hz, 1H), 8.13 (d, J = 2.0 Hz, 1H), 8.06 (m, 1H), 7.90 (m, 1H), 7.71 (m, 2H), 7.49 (d, J = 8.1 Hz, 1H), 7.21 (t, J = 7.9 Hz, 1H), 7.15-7.09 (m, 1H), 4.29 (s, 2H), 3.57 (s, 2H), 2.53 (s, 3H), 2.38-2.36 (m, 8H), 2.20 (s, 3H), 2.16 (s, 3H). 13 C NMR (151 MHz, DMSO-d6) 165.19, 160.74, 160.29, 143.87, 138.67, 137.01, 135.06, 132.64, 132.47, 131.72, 130.79, 130.37, 128.46, 128.02, 127.82, 127.12, 126.92, 123.98, 123.58, 123.18, 122.76, 121.07, 117.82, 116.09, 108.04, 91.01, 89.57, 57.88, 55.05, 52.90, 45.89, 43.75, 20.84, 18.68.

[0056] Example 3: Preparation of (E)-3-(2-((2-(2-(4-(dimethylamino)but-2-enamido)-6- methylphenyl)amino)pyrimidin-5-yl)ethynyl)-4-methyl-N-(4-((4-methylpiperazin-1- yl)methyl)-3-(trifluoromethyl)phenyl)benzamide;

[0057]

[0058] Operation as example 1, separated by column chromatography to get 60 mg white powder. 1H NMR (600 MHz, DMSO-d6) δ 10.60 (s, 1H), 9.60 (s, 1H), 8.91 (s, 1H), 8.57 (s, 2H), 8.24 (d, J = 2.2 Hz, 1H), 8.14 (d, J = 2.0 Hz, 1H), 8.09 (m, 1H), 7.93 (m, 1H), 7.71 (t, J = 8.7 Hz, 2H), 7.49 (d, J = 8.1 Hz, 1H), 7.19 (t, J = 7.8 Hz, 1H), 7.11 - 7.06 (m, 1H), 6.73 (m, 1H), 6.42 (m, 1H), 3.59 (s, 2H), 3.15 - 3.10 (m, 2H), 2.53 (s, 3H), 2.28 (s, 3H), 2.22 (s, 6H), 2.14 (s, 3H). 13 C NMR (151 MHz, DMSO-d6) δ 165.18, 163.86, 160.64, 160.18, 143.84, 140.83, 138.74, 137.12, 135.47, 132.57, 132.27, 131.69, 130.80, 130.35, 129.71, 128.48, 127.98, 127.78, 127.04, 126.82, 126.72, 125.70, 123.99, 123.89, 122.76, 121.48, 117.79, 117.75, 107.85, 90.93, 89.64, 59.81, 57.74, 54.72, 52.41, 45.38, 45.21, 20.86, 18.85.

[0059] Example 4: Preparation of 4-methyl-3-((2-((2-methyl-6-propionamidophenyl)amino)pyrimidin-5-yl)ethynyl)-N-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)benzamide;

[0060]

[0061] Worked as example 1, isolated by column chromatography to get 78 mg white powder. 1H NMR (600 MHz, DMSO-d6) δ 10.51 (s, 1H), 9.21 (s, 1H), 8.75 (s, 1H), 8.57 (s, 2H), 8.20 (d, J = 2.2 Hz, 1H), 8.12 (d, J = 2.0 Hz, 1H), 8.05 (m, 1H), 7.90 (m, 1H), 7.70 (d, J = 8.5 Hz, 1H), 7.58 (d, J = 8.1 Hz, 1H), 7.50 (d, J = 8.1 Hz, 1H), 7.16 (t, J = 7.8 Hz, 1H), 7.06 (d, J = 7.6 Hz, 1H), 3.56 (s, 2H), 2.53 (s, 3H), 2.50-2.32 (m, 8H), 2.29 (m, 2H), 2.16 (s, 3H), 2.14 (s, 3H), 1.01 (t, J = 7.5 Hz, 3H). 13 CNMR (151 MHz, DMSO-d6) δ 174.21, 172.70, 165.14, 160.65, 160.16, 143.83, 138.65, 137.01, 135.54, 132.61, 132.48, 131.66, 130.74, 130.35, 128.45, 127.96, 127.76, 126.67, 126.61, 125.71, 123.93, 122.75, 121.57, 117.73, 107.82, 90.93, 89.64, 57.90, 55.13, 53.05, 46.05, 20.86, 18.84, 17.69, 10.25.

[0062] Example 5: Preparation of 3-((2-((2-acrylamidophenyl)amino)pyrimidin-5-yl)ethynyl)-4-methyl-N-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)benzamide;

[0063]

[0064] Worked as example 1, separated by column chromatography to get 50 mg white powder. 1H NMR (600 MHz, DMSO-d6) δ 10.51 (s, 1H), 9.84 (s, 1H), 9.10 (s, 1H), 8.65 (s, 2H), 8.21 (d, J = 2.2 Hz, 1H), 8.14 (d, J = 2.0 Hz, 1H), 8.06 (m, 1H), 7.91 (m, 1H), 7.74-7.68 (m, 2H), 7.64-7.57 (m, 1H), 7.51 (d, J = 8.1 Hz, 1H), 7.20 (m, 2H), 6.52 (m, 1H), 6.29 (m, 1H), 5.81-5.74 (m, 1H), 3.57 (s, 2H), 2.54 (s, 3H), 2.45-2.22 (m, 8H), 2.16 (s, 3H). 13 C NMR (151 MHz, DMSO-d6) δ 165.13, 164.24, 160.59, 159.31, 143.87, 138.66, 132.65, 132.53, 131.99, 131.68, 131.15, 130.83, 130.36, 128.52, 128.00, 127.80, 127.59, 125.73, 125.59, 125.55, 124.97, 123.95, 123.91, 122.67, 117.79, 117.74, 108.83, 91.28, 89.42, 57.94, 55.18, 53.12, 46.11, 20.86.

[0065] Example 6: Preparation of 3-((2-((2-acrylamido-6-methylphenyl)amino)pyrimidin-5- yl)ethynyl)-4-methoxy-N-(4-((4-methylpiperazin-1-yl)methyl)-3- (trifluoromethyl)phenyl)benzamide;

[0066]

[0067] The operation is the same as Example 1, and 120 mg of white powder is separated by column chromatography. 1H NMR (600 MHz, DMSO-d6) δ 10.42 (s, 1H), 9.51 (s, 1H), 8.80 (s, 1H), 8.52 (s, 2H), 8.17 (m, 2H), 8.04 (m, 2H), 7.70 (t, J = 7.5 Hz, 2H), 7.25 (d, J = 8.8 Hz, 1H), 7.19 (t, J = 7.8 Hz, 1H), 7.10 (d, J = 7.6 Hz, 1H), 6.54 (m, 1H), 6.23 (m, 1H), 5.77-5.69 (m, 1H), 3.94 (s, 3H), 3.56 (s, 2H), 2.36 (s, 8H), 2.16 (s, 3H), 2.14 (s, 3H). 13 C NMR (151 MHz, DMSO-d6) δ 164.76, 163.93, 162.43, 160.55, 160.10, 138.76, 137.12, 135.36, 132.73, 132.33, 131.65, 130.93, 129.84, 127.93, 127.74, 127.54, 127.31, 126.98, 126.88, 126.74, 125.73, 123.88, 121.52, 117.67, 111.74, 111.58, 108.09, 89.08, 88.80, 57.92, 56.69, 55.17, 53.12, 46.13, 18.84.

[0068] Example 7: Preparation of 3-((2-((2-acrylamido-6-methylphenyl)amino)pyrimidin-5- yl)ethynyl)-N-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)benzamide;

[0069]

[0070] Worked as example 1, separated by column chromatography to get 130 mg white powder. 1H NMR (600 MHz, DMSO-d6) δ 10.57 (s, 1H), 9.48 (s, 1H), 8.75-8.82 (m, 1H), 8.55 (s, 2H), 8.21 (d, J = 2.3 Hz, 1H), 8.13 (d, J = 1.8 Hz, 1H), 8.05 (m, 1H), 7.98 (m, 1H), 7.76-7.67 (m, 3H), 7.60 (t, J = 7.8 Hz, 1H), 7.19 (t, J = 7.8 Hz, 1H), 7.10 (d, J = 7.5 Hz, 1H), 6.53 (m, 1H), 6.22 (m, 1H), 5.71 (m, 1H), 3.57 (s, 2H), 2.37 (m, 8H), 2.17 (s, 3H), 2.14 (s, 3H). 13 CNMR (151 MHz, DMSO-d6) δ 165.29, 163.93, 160.76, 160.15, 138.64, 137.12, 135.36, 135.33, 134.63, 132.41, 132.30, 131.74, 130.51, 129.80, 129.58, 128.51, 128.00, 127.81, 127.30, 126.98, 126.75, 123.98, 123.86, 122.96, 121.55, 117.78, 107.62, 92.10, 85.93, 57.73, 54.75, 52.43, 45.40, 18.85.

[0071] Example 8: Preparation of 5-((2-((2-acrylamido-6-methylphenyl)amino)pyrimidin-5-yl)ethynyl)-2-methyl-N-(4-(4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)benzamide;

[0072]

[0073] The operation is the same as Example 1, and 100 mg of white powder is separated by column chromatography. 1H NMR (600 MHz, DMSO-d6) δ 10.61 (s, 1H), 9.47 (s, 1H), 8.70-8.79 (m, 1H), 8.51 (s, 2H), 8.18 (d, J = 2.3 Hz, 1H), 7.95 (m, 1H), 7.74-7.61 (m, 3H), 7.54 (m, 1H), 7.37 (d, J = 8.1 Hz, 1H), 7.19 (t, J = 7.8 Hz, 1H), 7.12-7.04 (m, 1H), 6.52 (m, 1H), 6.22 (m, 1H), 5.76-5.64 (m, 1H), 3.57 (s, 2H), 2.41 (s, 3H), 2.40-2.31 (m, 8H), 2.21 (s, 3H), 2.13 (s, 3H). 13 CNMR (151 MHz, DMSO-d6) δ 167.47, 163.92, 160.65, 160.09, 138.67, 137.29, 137.12, 136.95, 135.34, 132.72, 132.43, 132.29, 131.82, 131.73, 130.40, 128.07, 127.31, 126.96, 126.74, 125.66, 123.84, 123.45, 121.52, 120.15, 117.15, 113.37, 107.84, 92.15, 85.32, 57.80, 54.95, 52.74, 45.75, 19.84, 18.84.

[0074] Example 9: Preparation of 4-((2-((2-acrylamido-6-methylphenyl)amino)pyrimidin-5-yl)ethynyl)-3-methyl-N-(4-(4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)benzamide;

[0075]

[0076] Worked as example 1, isolated by column chromatography to get 77 mg white powder. 1H NMR (600 MHz, DMSO-d6) δ 10.78 (d, J = 4.2 Hz, 1H), 9.91 (s, 1H), 9.02 (s, 1H), 8.56 (s, 2H), 8.29 (d, J = 2.2 Hz, 1H), 8.14 (m, 1H), 8.03 (d, J = 1.8 Hz, 1H), 7.90 (m, 1H), 7.69 (t, J = 9.7 Hz, 2H), 7.61 (d, J = 8.0 Hz, 1H), 7.19 (t, J = 7.8 Hz, 1H), 7.10 (d, J = 7.6 Hz, 1H), 6.59 (m, 1H), 6.24 (m, 1H), 5.71 (m, 1H), 3.61 (s, 2H), 2.70 (s, 3H), 2.53 (s, 8H), 2.43 (s, 3H), 2.13 (s, 3H).

[0077] Example 10: Preparation of (E)-3-((6-((2-(4-(dimethylamino)but-2-enamido)-6- methylphenyl)amino)pyridin-3-yl)ethynyl)-N-(4-(4-methylpiperazin-1-yl)methyl)-3- (trifluoromethyl)benzamide;

[0078]

[0079] Worked up as example 1 and isolated by column chromatography to give 101 mg of white powder. 1 H NMR (600 MHz, DMSO-d6) δ 10.78 (d, J = 4.2 Hz, 1H), 9.91 (s, 1H), 9.02 (s, 1H), 8.56 (s, 2H), 8.29 (d, J = 2.2 Hz, 1H), 8.14 (m, 1H), 8.03 (d, J = 1.8 Hz, 1H), 7.90 (m, 1H), 7.69 (t, J = 9.7 Hz, 2H), 7.61 (d, J = 8.0 Hz, 1H), 7.19 (t, J = 7.8 Hz, 1H), 7.10 (d, J = 7.6 Hz, 1H), 6.59 (m, 1H), 6.24 (m, 1H), 5.71 (m, 1H), 3.61 (s, 2H), 2.70 (s, 3H), 2.53 (s, 8H), 2.43 (s, 3H), 2.13 (s, 3H). 13C NMR (151 MHz, DMSO-d6) δ 165.24, 163.91, 157.30, 151.37, 143.68, 141.41, 140.12, 138.69, 136.92, 135.70, 132.56, 132.41, 131.69, 130.59, 130.44, 130.30, 128.11, 127.97, 127.78, 126.95, 126.66, 126.50, 125.71, 123.96, 123.90, 123.20, 121.59, 117.77, 117.72, 108.27, 92.96, 88.24, 60.02, 57.85, 55.01, 52.87, 49.06, 45.43, 18.83.

[0080] Example 11: Preparation of (E)-3-((2-(2-(but-2-enamido)-6-methylphenyl)amino)pyrimidin-5-yl)ethynyl)-4-methyl-N-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)benzamide;

[0081]

[0082] Isolated by column chromatography as in Example 1 to give 88 mg of white powder. 1 H NMR (600 MHz, DMSO-d6) δ 10.54 (s, 1H), 9.37 (s, 1H), 8.81 (s, 1H), 8.57 (s, 2H), 8.22 (d, J = 2.2 Hz, 1H), 8.13 (d, J = 2.0 Hz, 1H), 8.08 (m, 1H), 7.91 (m, 1H), 7.69 (m, 2H), 7.50 (d, J = 8.1 Hz, 1H), 7.18 (t, J = 7.8 Hz, 1H), 7.10-7.05 (m, 1H), 6.78 (m, 1H), 6.24 (m, 1H), 3.58 (s, 2H), 2.51-2.53 (s, 3H), 2.45 (s, 8H), 2.29 (s, 3H), 2.13 (s, 3H), 1.84 (m, 3H). 13C NMR (151 MHz, DMSO-d6) δ 172.45, 165.16, 164.24, 160.67, 160.17, 143.86, 140.47, 138.72, 137.12, 135.55, 132.59, 132.28, 131.72, 130.75, 130.37, 129.69, 128.46, 127.99, 127.79, 126.75, 126.35, 125.70, 123.96, 122.75, 121.37, 117.77, 107.86, 90.94, 89.66, 57.74, 54.76, 52.45, 45.41, 20.87, 18.85, 17.92.

[0083] Example 12: Preparation of 3-((2-((2-acrylamido-4-methylphenyl)amino)pyrimidin-5-yl)ethynyl)-4-methyl-N-(4-(4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)benzamide;

[0084]

[0085] Worked up as example 1 and isolated by column chromatography to give 65 mg of white powder. 1 H NMR (600 MHz, DMSO-d6) δ 10.49 (s, 1H), 9.76 (s, 1H), 8.98 (s, 1H), 8.61 (s, 2H), 8.20 (s, 1H), 8.14 (d, J = 2.0 Hz, 1H), 8.06 (m, 1H), 7.91 (m, 1H), 7.70 (d, J = 8.5 Hz, 1H), 7.56 (d, J = 8.2 Hz, 1H), 7.50 (d, J = 8.1 Hz, 1H), 7.43 (s, 1H), 7.03 (m, 1H), 6.51 (m, 1H), 6.27 (m, 1H), 5.76 (m, 1H), 3.57 (s, 2H), 2.54 (s, 3H), 2.40 (s, 8H), 2.31 (s, 3H), 2.18 (s, 3H). 13C NMR (151 MHz, DMSO-d6) δ 165.14, 164.16, 160.57, 159.45, 143.85, 138.67, 134.37, 132.63, 132.48, 132.03, 131.69, 131.14, 130.81, 130.35, 129.32, 128.49, 128.00, 127.80, 127.49, 126.24, 125.65, 125.17, 123.95, 122.71, 117.79, 117.75, 108.54, 91.19, 89.50, 57.91, 55.10, 53.00, 45.98, 21.05, 20.85.

[0086] Example 13: Preparation of 3-((6-((2-acrylamido-6-methylphenyl)amino)pyridin-3-yl)ethynyl)-4-methyl-N-(4-(4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)benzamide;

[0087]

[0088] Worked up as example 1 and isolated by column chromatography to give 110 mg of white powder. 1 H NMR (600 MHz, DMSO-d6) δ 10.49 (s, 1H), 9.53 (s, 1H), 8.35 (s, 1H), 8.27-8.19 (m, 2H), 8.10 (d, J = 2.0 Hz, 1H), 8.06 (m, 1H), 7.90-7.86 (m, 1H), 7.73 (d, J = 8.1 Hz, 1H), 7.69 (d, J = 8.5 Hz, 1H), 7.65 (m, 1H), 7.47 (d, J = 8.0 Hz, 1H), 7.19 (t, J = 7.8 Hz, 1H), 7.10 (d, J = 7.6 Hz, 1H), 6.55-6.41 (m, 2H), 6.20 (m, 1H), 5.69 (m, 1H), 3.56 (s, 2H), 2.51 (s, 3H), 2.43-2.23 (m, 8H), 2.16 (s, 3H), 2.14 (s, 3H). 13C NMR (151 MHz, DMSO-d6) δ 165.22, 163.88, 157.26, 151.34, 143.64, 140.08, 138.69, 136.97, 135.47, 132.58, 132.49, 132.43, 131.65, 130.61, 130.26, 128.12, 127.97, 127.77, 127.04, 126.43, 125.73, 123.98, 123.92, 123.20, 121.68, 117.81, 117.77, 108.34, 92.95, 88.28, 57.95, 55.20, 53.17, 46.16, 20.85, 18.86.

[0089] Example 14: Preparation of (E)-3-((2-(2-(4-(dimethylamino)but-2-enamido)-6- methylphenyl)amino)pyrimidin-5-yl)ethynyl)-4-methyl-N-(4-((4-methylpiperazin-1- yl)methyl)phenyl)benzamide;

[0090]

[0091] Worked up as example 1 and isolated by column chromatography to give 100 mg of white powder. 1 H NMR (600 MHz, DMSO-d6) δ 10.30 - 10.24 (m, 1H), 9.56 (s, 1H), 8.88 (s, 1H), 8.55 (s, 2H), 8.10 (d, J = 2.0 Hz, 1H), 7.94 - 7.87 (m, 1H), 7.77 - 7.64 (m, 3H), 7.46 (d, J = 8.0 Hz, 1H), 7.25 (d, J = 8.2 Hz, 2H), 7.18 (t, J = 7.8 Hz, 1H), 7.07 (d, J = 7.5 Hz, 1H), 6.71 (m, 1H), 6.38 (m, 1H), 3.41 (s, 2H), 3.02 (d, J = 6.1 Hz, 2H), 2.51 (s, 3H), 2.33 (m, 8H), 2.15 (s, 6H), 2.14 (s, 3H), 2.13 (s, 3H). 13C NMR (151 MHz, DMSO-d6) δ 164.81, 164.01, 160.61, 160.18, 143.40, 141.85, 138.36, 137.11, 135.50, 134.05, 133.16, 130.82, 130.22, 129.79, 129.46, 128.40, 126.79, 126.67, 126.41, 122.69, 121.51, 120.72, 107.94, 91.05, 89.53, 62.16, 60.19, 55.23, 52.98, 46.20, 45.60, 20.82, 18.84.

[0092] Example 15: Preparation of (E)-3-((2-(2-(4-(dimethylamino)but-2-enamido)-6- methylphenyl)amino)pyrimidin-5-yl)ethynyl)-4-methyl-N-(4-(morpholinomethyl)phenyl)benzamide;

[0093]

[0094] Worked up as example 1 and isolated by column chromatography to give 90 mg of white powder. 1 H NMR (600 MHz, DMSO-d6) δ 10.31 (t, J = 3.6 Hz, 1H), 9.70 - 9.57 (m, 1H), 8.96 - 8.86 (m, 1H), 8.55 (s, 2H), 8.11 (d, J = 1.9 Hz, 1H), 7.92 (m, 1H), 7.72 (m, 3H), 7.46 (d, J = 8.1 Hz, 1H), 7.27 (d, J = 8.4 Hz, 2H), 7.17 (t, J = 7.8 Hz, 1H), 7.07 (d, J = 7.1 Hz, 1H), 6.71 (m, 1H), 6.39 (m, 1H), 3.57 (t, J = 4.6 Hz, 4H), 3.43 (s, 2H), 3.02 (m, 2H), 2.51 (s, 3H), 2.35 (t, J = 4.7 Hz, 4H), 2.15 (s, 6H), 2.13 (s, 3H). 13C NMR (151 MHz, DMSO-d6) δ 164.83, 164.00, 160.61, 160.15, 143.41, 141.84, 138.47, 137.10, 135.47, 133.50, 133.13, 130.84, 130.22, 129.81, 129.58, 128.42, 126.79, 126.65, 126.41, 122.69, 121.55, 120.74, 107.93, 91.05, 89.53, 66.69, 62.54, 60.19, 53.61, 45.60, 20.81, 18.85.

[0095] Example 16: Preparation of (E)-3-(2-((2-(2-(4-(dimethylamino)but-2-enamido)-6- methylphenyl)amino)pyrimidin-5-yl)ethynyl)-4-methyl-N-(4-(4-methylpiperazin-1- yl)phenyl)benzamide;

[0096]

[0097] Worked up as example 1 and isolated by column chromatography to give 70 mg of white powder. 1 H NMR (600 MHz, DMSO-d6) δ 10.14 (s, 1H), 9.71 (s, 1H), 8.95 (s, 1H), 8.55 (s, 2H), 8.13-8.07 (m, 1H), 7.94-7.88 (m, 1H), 7.68 (d, J = 8.1 Hz, 1H), 7.64 (d, J = 8.7 Hz, 2H), 7.45 (d, J = 8.0 Hz, 1H), 7.17 (t, J = 7.8 Hz, 1H), 7.07 (d, J = 7.6 Hz, 1H), 6.91 (d, J = 8.7 Hz, 2H), 6.71 (m, 1H), 6.39 (m, 1H), 3.32 (s, 3H), 3.10 (t, J = 4.9 Hz, 4H), 3.02 (d, J = 6.0 Hz, 2H), 2.46 (s, 4H), 2.22 (s, 3H), 2.14-2.17 (s, 6H), 2.13 (s, 3H). 13C NMR (151 MHz, DMSO-d6) δ 164.30, 164.00, 160.60, 160.12, 148.01, 143.11, 141.80, 137.10, 135.43, 133.30, 131.43, 130.75, 130.16, 129.92, 129.85, 128.31, 126.80, 126.61, 126.44, 122.63, 122.04, 115.88, 107.95, 91.11, 89.43, 60.19, 55.12, 48.98, 46.24, 45.60, 20.79, 18.86.

[0098] Example 17: Preparation of (E)-N-(4-((lH-l,2,4-triazol-l-yl)methyl)phenyl)-3-((2-(2-(4- (dimethylamino)but-2-enamido)-6-methylphenyl)amino)pyrimidin-5-yl)ethynyl)-4- methylbenzamide;

[0099]

[0100] Worked up as example 1 and isolated by column chromatography to give 86 mg of white powder. 1 H NMR (600 MHz, DMSO-d6) δ 10.36 (d, J = 7.1 Hz, 1H), 9.58 (t, J = 9.2 Hz, 1H), 8.89 (d, J = 9.3 Hz, 1H), 8.65 (s, 1H), 8.55 (s, 2H), 8.11 (d, J = 2.0 Hz, 1H), 7.97 (s, 1H), 7.91 (m, 1H), 7.79 (m, 2H), 7.70 (d, J = 8.1 Hz, 1H), 7.47 (d, J = 8.0 Hz, 1H), 7.29 (d, J = 8.4 Hz, 2H), 7.18 (t, J = 7.8 Hz, 1H), 7.08 (d, J = 7.5 Hz, 1H), 6.71 (m, 1H), 6.38 (m, 1H), 5.39 (s, 2H), 3.02 (d, J = 4.3 Hz, 2H), 2.51 (s, 3H), 2.14 - 2.16 (s, 6H), 2.14 (s, 3H). 13C NMR (151 MHz, DMSO-d6) δ 164.94, 164.02, 160.62, 160.18, 152.11, 144.53, 143.53, 141.86, 139.33, 137.11, 135.50, 132.97, 131.86, 130.83, 130.25, 129.79, 128.76, 128.44, 126.79, 126.67, 126.40, 122.71, 121.52, 121.02, 107.92, 91.01, 89.56, 60.18, 52.28, 45.60, 20.82, 18.84.

[0101] Example 18: Preparation of 3-(2-((2-(2-(cyclopropanecarboxamido)-6- methylphenyl)amino)pyrimidin-5-yl)ethynyl)-4-methyl-N-(4-((4-methylpiperazin-1- yl)methyl)-3-(trifluoromethyl)phenyl)benzamide;

[0102]

[0103] Worked up as example 1 and isolated by column chromatography to give 102 mg of white powder. 1 H NMR (600 MHz, DMSO-d6) δ 10.58 - 10.50 (m, 1H), 9.73 - 9.54 (m, 1H), 8.80 (d, J = 6.2 Hz, 1H), 8.57 (s, 2H), 8.22 (s, 1H), 8.14 (s, 1H), 8.07 (d, J = 8.7 Hz, 1H), 7.92 (m, 1H), 7.69 (d, J = 8.5 Hz, 1H), 7.59 (d, J = 8.2 Hz, 1H), 7.49 (d, J = 8.0 Hz, 1H), 7.15 (t, J = 7.8 Hz, 1H), 7.06 (d, J = 7.6 Hz, 1H), 3.56 (s, 2H), 2.53 (s, 3H), 2.36 (m, 8H), 2.15 - 2.19 (m, 6H), 1.89 (s, 1H), 0.83 - 0.77 (m, 2H), 0.70 - 0.77 (m, 2H). 13C NMR (151 MHz, DMSO-d6) δ 172.49, 165.17, 160.66, 160.22, 143.82, 138.69, 137.10, 135.61, 132.64, 132.53, 131.66, 130.81, 130.33, 129.67, 128.47, 127.98, 127.79, 126.67, 126.57, 125.74, 123.99, 123.92, 122.79, 121.45, 117.82, 117.78, 107.90, 90.96, 89.66, 57.96, 55.21, 53.17, 46.16, 20.85, 18.84, 14.75, 7.70.

[0104] Example 19: Preparation of (E)-3-(2-((2-(2-(4-(dimethylamino)but-2-enamido)-6- methylphenyl)amino)pyrimidin-5-yl)ethynyl)-4-methyl-N-(3-methyl-4-(4-methylpiperazine- 1-carbonyl)phenyl)benzamide;

[0105]

[0106] Worked up as in example 1 and isolated by column chromatography to give 60 mg of white powder. 1 H NMR (600 MHz, DMSO-d6) δ 10.30 (d, J = 6.5 Hz, 1H), 9.43 (s, 1H), 8.82 (s, 1H), 8.56 (s, 2H), 8.10 (s, 1H), 7.89 (d, J = 8.0 Hz, 1H), 7.69 (m, 3H), 7.48 (d, J = 8.2 Hz, 1H), 7.18 (t, J = 7.9 Hz, 1H), 7.13 (d, J = 8.2 Hz, 1H), 7.08 (d, J = 7.6 Hz, 1H), 6.71 (m, 1H), 6.37 (m, 1H), 3.65 (s, 2H), 3.16 (s, 2H), 3.02 (d, J = 6.0 Hz, 2H), 2.52 (s, 3H), 2.36 (s, 2H), 2.22 (s, 5H), 2.19 (s, 3H), 2.15 (s, 6H), 2.14 (s, 3H). 13C NMR (151 MHz, DMSO-d6) δ 168.87, 165.01, 163.92, 160.62, 160.18, 143.59, 141.38, 139.73, 137.10, 135.49, 134.76, 132.94, 132.10, 130.79, 130.27, 129.66, 128.42, 126.78, 126.71, 126.68, 122.70, 122.10, 121.43, 118.07, 107.86, 90.97, 89.57, 59.98, 55.28, 54.78, 46.02, 45.40, 20.83, 19.40, 18.83.

[0107] Example 20: Preparation of (E)-3-(2-((2-(2-(4-(dimethylamino)but-2-enamido)-6- methylphenyl)amino)pyrimidin-5-yl)ethynyl)-4-methyl-N-(4-(4-methylpiperazine-l- carbonyl)-3-(trifluoromethyl)phenyl)benzamide;

[0108]

[0109] Isolated by column chromatography as in example 1 to give 88 mg of white powder. 1 H NMR (600 MHz, DMSO-d6) δ 10.82 (t, J = 3.0 Hz, 1H), 9.71 - 9.61 (m, 1H), 8.93 (s, 1H), 8.56 (s, 2H), 8.35 (d, J = 2.1 Hz, 1H), 8.20 (m, 1H), 8.17 (d, J = 2.0 Hz, 1H), 7.98 (m, 1H), 7.69 (d, J = 8.1 Hz, 1H), 7.50 (d, J = 8.0 Hz, 1H), 7.43 (d, J = 8.4 Hz, 1H), 7.18 (t, J = 7.8 Hz, 1H), 7.08 (d, J = 7.5 Hz, 1H), 6.71 (m, 1H), 6.39 (m, 1H), 3.63 (m, 2H), 3.19 - 3.12 (m, 1H), 3.10 - 3.04 (m, 1H), 3.02 (m, 2H), 2.53 (s, 3H), 2.40 (m, 1H), 2.34 - 2.22 (m, 2H), 2.19 (s, 3H), 2.15 (s, 7H), 2.13 (s, 3H). 13C NMR (151 MHz, DMSO-d6) δ 166.50, 165.44, 164.00, 160.63, 160.16, 143.99, 141.79, 140.50, 137.10, 135.45, 132.41, 130.97, 130.35, 130.04, 129.82, 128.63, 128.59, 126.80, 126.64, 126.44, 126.29, 126.08, 125.00, 124.08, 123.54, 123.19, 122.82, 118.09, 118.05, 107.87, 90.92, 89.69, 60.18, 54.62, 47.07, 46.04, 45.59, 20.86, 18.85.

[0110] Example 21: Preparation of 3-((2-((2-acrylamido-6-chlorophenyl)amino)pyrimidin-5- yl)ethynyl)-4-methyl-N-(4-((4-methylpiperazin-1-yl)methyl)-3- (trifluoromethyl)phenyl)benzamide;

[0111]

[0112] Worked up as example 1 and isolated by column chromatography to give 75 mg of white powder. 1 H NMR (600 MHz, DMSO-d6) δ 10.48 (s, 1H), 9.59 (s, 1H), 9.03 (s, 1H), 8.59 (s, 2H), 8.19 (d, J = 2.3 Hz, 1H), 8.12 (d, J = 2.0 Hz, 1H), 8.05 (m, 1H), 7.96 (m, 1H), 7.90 (m, 1H), 7.70 (d, J = 8.5 Hz, 1H), 7.50 (d, J = 8.1 Hz, 1H), 7.35 - 7.28 (m, 2H), 6.59 (m, 1H), 6.25 (m, 1H), 5.74 (m, 1H), 3.57 (s, 2H), 2.53 (s, 3H), 2.37 (m, 8H), 2.17 (s, 3H). 13C NMR (151 MHz, DMSO-d6) δ 165.16, 164.13, 160.54, 160.23, 143.91, 138.67, 137.68, 133.53, 132.67, 132.55, 132.14, 131.71, 130.81, 130.36, 128.51, 128.41, 128.01, 127.99, 127.82, 127.77, 125.75, 125.73, 123.97, 123.92, 122.71, 122.32, 117.81, 117.76, 108.56, 91.10, 89.46, 57.94, 55.18, 53.12, 46.11, 20.85.

[0113] Example 22: Preparation of (E)-3-((6-((2-(4-(dimethylamino)but-2-enamido)-6- methylphenyl)amino)pyridin-3-yl)ethynyl)-4-methyl-N-(4-((4-methylpiperazin-1-yl)methyl)-3- (trifluoromethyl)phenyl)benzamide;

[0114]

[0115] Worked up as example 1 and isolated by column chromatography to give 78 mg of white powder. 1 H NMR (600 MHz, DMSO-d6) δ 10.53 (s, 1H), 9.46 (s, 1H), 8.35 (s, 1H), 8.24 (d, J = 2.4 Hz, 1H), 8.23 (s, 1H), 8.11 (d, J = 2.0 Hz, 1H), 8.07 (m, 1H), 7.88 (m, 1H), 7.75 (d, J = 8.0 Hz, 1H), 7.70 (d, J = 8.6 Hz, 1H), 7.65 (m, 1H), 7.48 (d, J = 8.1 Hz, 1H), 7.18 (t, J = 7.8 Hz, 1H), 7.12 - 7.07 (m, 1H), 6.69 (m, 1H), 6.43 (s, 1H), 6.35 (m, 1H), 3.57 (s, 2H), 3.04 (m, 2H), 2.51 (s, 3H), 2.41 (s, 8H), 2.21 (s, 3H), 2.16 (s, 6H), 2.14 (s, 3H). 13C NMR (151 MHz, DMSO-d6) δ 165.24, 163.91, 157.30, 151.37, 143.68, 141.41, 140.12, 138.69, 136.92, 135.70, 132.56, 132.41, 131.69, 130.59, 130.44, 130.30, 128.11, 127.97, 127.78, 126.95, 126.66, 126.50, 125.71, 123.96, 123.90, 123.20, 121.59, 117.77, 117.72, 108.27, 92.96, 88.24, 60.02, 57.85, 55.01, 52.87, 49.06, 45.43, 20.85, 18.83. Experimental Example 1 FGFR4 kinase inhibitory activity test

[0116] In 384-well reaction plates, positive control wells and negative control wells were set, and different concentrations of compounds and FGFR4 kinase were added to the rest of the wells. After centrifugation, incubate at room temperature for 15 min, add fluorescent substrate and ATP, after the reaction is completed, add stop solution to stop the reaction, and use Caliper EZ reader to read the conversion rate. Further calculate the percentage inhibition rate, the calculation formula is as follows: enzyme inhibition rate = Mean (maximum) - sample signal / Mean (maximum) - blank multiplied by 100%. The experimental results are shown in Table 1

[0117] Table 1. Inhibition rate of compounds on FGFR4 and FGFR1 at concentrations of 100 and 10 nM a

[0118]

[0119]

[0120] The results show that: in addition to compounds I-9, I-19, 1-20, the compounds of the present application all show FGFR4 selective tendency, especially compound I-7, the inhibition activity on FGFR4 kinase at a concentration of 100 nM reaches more than 90%, and the inhibition activity on FGFR4 kinase at a concentration of 10 nM is also more than 50%, that is, the FGFR4 inhibition IC 50 value is 10 nM, and even at a concentration of 100 nM, it does not show inhibition activity on FGFR1, indicating that the compound has strong FGFR4 inhibition activity and high selectivity, and has excellent application prospect.

[0121] Experimental Example 2 Cell proliferation inhibitory activity test

[0122] The experiment set solvent control group and drug experimental group, drug experimental group each group set 8 concentrations, each concentration set 3 parallel holes. Each experiment was repeated three times. In 96-well plates, each well was added with cell concentration of 1 x 10 5 4 / mL of cell suspension 100 μL, that is, each well contains 3 x 10 3 4 cells, pay attention to make the cells evenly distributed in each hole when inoculation. To prevent liquid evaporation, a circle of holes around the 96-well plate was not inoculated with cells, and PBS was added for moisturizing. After the cells adhered, different concentrations of target compounds were added to each well in the drug experimental group. The 96-well plate was placed in a 37℃, 5% CO2 incubator for further culture, and the culture was terminated after 72h. After 72h of drug treatment, the 96-well plate was taken out, MTT solution was added to each well, and the cells were incubated in the cell culture box for 4 hours. Then the absorbance value (A) of each well was measured at 570nm on the automatic enzyme label instrument. The inhibition rate was calculated by the formula. The inhibition rate was linearly regressed with the drug concentration, and the IC 50 value was calculated by the linear equation. The results were expressed as mean ± standard deviation, and the experimental results were analyzed by SPSS15.0 software. P<0.05 indicated significant difference. The results in Table 2 showed that the IC 50 values of compounds I-7 and I-21 on Huh-7 cells and Hep3B cells were comparable to Ponatinib (purchased from Macclin Company).

[0123] Table 2. Proliferation inhibition activity of compounds on different tumor cell linesa

[0124]

[0125] The results showed that compound I-7 showed strong inhibition on Hep3B and HuH-7 hepatoma cell lines.

Claims

1. A selective FGFR4 inhibitor represented by formula I, in, R1 is selected from hydrogen or C 1-6 alkyl; R2 is selected from hydrogen, C 1-6 Alkoxy or C 1-6 alkyl.

2. The inhibitor according to claim 1, characterized in that: R1 is selected from hydrogen or methyl.

3. The inhibitor according to claim 1, characterized in that: R2 is selected from hydrogen, methoxy or methyl.

4. The inhibitor according to any one of claims 1 to 3, wherein the inhibitor is a compound selected from the group consisting of:

5. A pharmaceutical composition comprising the inhibitor according to any one of claims 1 to 3 or the compound according to claim 4 and a pharmaceutically acceptable carrier.

6. Use of the inhibitor according to any one of claims 1 to 3, the compound according to claim 4, or the composition according to claim 5 in the preparation of a drug for treating and / or preventing tumors.

Images