Mutated EGFR-IN-1

Mutated EGFR-IN-1 is a useful intermediate for the inhibitors design for mutated EGFR, such as L858R EGFR, Exonl9 deletion activating mutant and T790M resistance mutant.IC50 value:Target: Mutated EGFR inhibitorMore information can be found in Patent WO 2013014448 A1.2 - (2, 4, 5 - substituted -anilino) pyrimidine derivatives as egfr modulators useful for treating cancer.

In a nitrogen atmosphere, 28.5 (0.060 mol) of the intermediate 2 and 2.5 g of palladium carbon were added to 500 ml of methanol and passed at 25C Hydrogen atmospheric reaction 4h, filtered, the filtrate was concentrated to dryness under reduced pressure to give a brown solid, 26g, a yield of 97.3percent.A 125 ml autoclave was taken, and after replacing nitrogen three times, a compound F (1.50 g, 3.15 mmol) and a Raney nickel catalyst (0.70 g) were added and dissolved in ethyl acetate (30 ml), and the hydrogen was replaced twice and then charged with 2.0 MPa of hydrogen.The temperature was raised to 40 ° C to stir the reaction.Filter off and wash the filter cake twice with ethyl acetate (10 ml x 2).The filtrate was concentrated under reduced pressure at 40 ° C to give a dark green solid: 1.36 g.The molar yield was 96.90percent.Under nitrogen protection, 928 ml of methanol was added to a 2 L four-necked flask, Then, 92.8 g of the compound represented by the formula (III)Stirring, 18.6g was added Raney nickel (water-wet product)8 at atmospheric pressure under hydrogenation, Reaction temperature 25 , The reaction was carried out for 6 hours.Filtered by rotary evaporation to yield 82.6 g of gray solid, yield: 95.0percentN1-(2-(Dimethylamino)ethyl)-5-methoxy-N1-methyl-N4-(4-(1-methyl-1H-indol-3-yl)pyrimidine(Hydroxy-2-yl)-2-nitrobenzene-1, 4-diamine (666 mg, 1.4 mmol)Soluble in ethanol (40mL), Iron powder (470mg, 8.4 mmol) and ammonium chloride (241 mg, 4.5 mmol).The reaction was stirred at reflux for 2 hours.After the reaction is over, The reaction solution was concentrated under reduced pressure.The crude product was isolated and purified by silica gel column chromatography (eluent: dichloromethane:methanol=20:1) to give a brown oil (552 mg, yield: 88.6percent).A mixture of N'-(2-dimethylaminoethyl)-2-methoxy-N’-methyl-N-[4-(1-methylindol-3-yl)pyrimidin-2-yl]-5-nitrobenzene-1, 4-diamine (Intermediate 101, 220 mg, 0.46 mmol), iron (155 mg, 2.78 mmol) and NHSolvent is tetrahydrofuran B, catalyst for A 10percent Pd/C, the pressure of the hydrogen is charged into the is 30bar, in 30 °C stirring under the temperature condition of 16h. The specific process: in 500 ml dry reaction flask, 5gN-(2-dimethyl amino-ethyl) - 2-methoxy-N-methyl-N-[ 4 - (1-methyl -1H-Indol-3-yl)-pyrimidin-2-yl] - 5-nitro-benzene -1, 4-diamine with tetrafuran 140 ml dissolved, add 0.01g10percent Pd/C, access 30bar hydrogen, in the 30 °C stirring under 16h. Filtering, to obtain the product solvent is removed by the spin vaporization N-(2-dimethyl amino-ethyl) - 2-methoxy-N-methyl-N-[ 4 - (1-methyl -1H-Indol-3-yl)-pyrimidin-2-yl] - 5-amino-benzene -1, 4-diamine a kind of white solid powder. The product purity 98.4percent, yield 85.9percent.1) Weigh 200g of the raw material hydrogenation precursor nitro compound, and then add it to a mixture of 4L of methanol and 100g of concentrated hydrochloric acid, stir and dissolve, then add 10g of Pd/C catalyst with Pd content of 10percent, stir well and mix to form Material I, the material I is sent to the preheating module 1 of the microchannel reactor for preheating, and after preheating, enters the reaction module group of the microchannel reactor.2) The reaction module group that delivers hydrogen to the microchannel reactor and the step 1) the preheated material I reacts in the reaction module group, wherein: the flow rate of the slurry pump is adjusted so that the flow rate of the material I is 40.0 g/min. The flow rate of the H2 gas flow meter is adjusted to 500 ml/min, the molar ratio of the raw material hydrogenation precursor nitro compound to hydrogen is 1:3.5, the reaction temperature is 90 ° C, the temperature of the cooling module is 20 ° C, and the residence time of the reaction is 25s, the reaction pressure is 1.5Mpa, and the collection flows out from the outlet of the cooling module.The reaction solution is subjected to post-treatment, which means that the catalyst is recovered by filtration, and the solvent is distilled off under reduced pressure. The residue is added to 1 L of water, stirred and dissolved, and the pH of the system is adjusted to 9.0 by a NaOH solution having a mass fraction of 20percent. 10.0, then join500ml of ethyl acetate was extracted and separated, the organic phase was discarded, the aqueous phase was stirred and crystallized at 10 ° C for 1 hour, filtered, and the product was washed with a small amount of ethanol, and vacuum dried at 50 ° C for 8 hours to obtain an oxetinib intermediate. Target product 160.65g, yield85.73percent, purity 99.68percent.The above obtainedN-[(4-Dimethylaminoethylamino-2-methoxy-5-nitrophenyl)]-4-(1-methyl-1H-indol-3-yl)-2-pyrimidinamine 3.8kg (7.99mol)It was added to an autoclave containing 38 L of ethanol, and then 380 g of palladium carbon was added to the system, replaced with nitrogen three times, and then hydrogen was replaced twice.The hydrogen pressure was set to 2 MPa, and the reaction was heated to 80 degrees overnight.After the end of the reaction, the autoclave was cooled, the reaction solution was withdrawn, filtered through a celite cake, and the filter cake was rinsed with ethanol.After concentrating to 15 kg, it was cooled, stirred and crystallized, returned to room temperature for 2 hours, filtered, and the filter cake was rinsed with ethanol.Drying gave 3.05 kg of silver-gray powder with a yield of 85.6percent (HPLC purity >99percent).A solution of N1- (2- (dimethylamino) ethyl) -5-methoxy-N1-methyl-N4- (4- (1-methylindol-3-yl) 2-nitrophenyl-1, 4-diamine (30.0 g, 0.063 mol) was added to a 1 L vial, Adding 600 mL of tetrahydrofuran and 10 g of Raney nickel, Hydrogen gas at a pressure of 3 atm was introduced at room temperature, Reaction for 12 h.After the reaction is complete, The reaction solution was filtered through celite, To obtain a brown solution, Concentrated under reduced pressure to give crude brown solid, The crude product was purified by recrystallization from 250 mL of a mixture of ethyl acetate and n-heptane (1: 1 by volume of ethyl acetate and n-heptane in the mixture)To give a light brown solid N1- (2- (dimethylamino) ethyl) -5-methoxy -N1- methyl -N4- (4- (1- methyl-indol-3-yl) pyrimidine-2 - yl) phenyl-1, 2, 4-triamine 21.9g, yield 78percent.A solution of N1- (2- (dimethylamino) ethyl) -5-methoxy-N1-methyl-N4- (4- (1-methylindol-3-yl)Yl) -2-nitrophenyl-1, 4-diamine (30.0 g, 0.063 mol) was added to a 1 L single-Then 600 mL of tetrahydrofuran and 10 g of Raney nickel were added and the pressure was 3 atm of hydrogen and reacted at 25 ° C for 12 h.After completion of the reaction, the reaction solution was filtered through celite and concentrated to give a crude product of brown solid which was recrystallized from 250 mL of a mixture of ethyl acetate and n-heptane (1: 1 by volume of ethyl acetate and n-heptane in the mixture) purification, A light brown solid product was obtainedMethyl-N4- (4- (1-methylindol-3-yl) pyrimidin-2-yl) phenyl -1, 2, 4-triamine 21.9 g, yield 78percent.N′-(2-(dimethylamino)ethyl)-5-methoxy-N′-methyl-N-(4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl)-2-nitrobenzene-1, 4-diamine (32) (5.00 g, 10.53 mmol) and Pd/C (10percent by weight) (250 mg, 2.10 mmol) were suspended in a mixture of MeOH (100 mL) and stirred under a hydrogen atmosphere for 24 h. The reaction mixture was filtered and the filtrate was concentrated in vacuo. The resulting residue was purified by silica gel chromatography (DCM: MeOH: NH3·H2O=25:1:0.1) to afford the title compound (3.51 g, 75.0percent yield) : 1H NMR (400 MHz, DMSO-d6) δ8.43 (d, J = 7.9 Hz, 1H), 8.31 (s, 1H), 8.28 (d, J = 5.3 Hz, 1H), 7.79 (s, 1H), 7.52 (d, J = 8.2 Hz, 1H), 7.49 (s, 1H), 7.28-7.23 (m, 1H), 7.20-7.13 (m, 2H), 4.63 (s, 2H), 3.89 (s, 3H), 3.75 (s, 3H), 2.90 (t, J = 6.7 Hz, 2H), 2.64 (s, 3H), 2.38 (t, J = 6.6 Hz, 2H), 2.19 (s, 6H). MS (ESI) m/z 446.4 [M+H]+.Example 1 (0622) AZD9291 Nitrodiamine (10.0 g), 5percent platinum-on-activated carbon (0.001 molar equivalents, 50percent water wet, 0.2 g, 0.02 relative weight), methanesulfonic acid (3.23 g, 1.6 molar equivalents) and water (100 mL, 10 relative volume) were mixed in a sealed autoclave. The headspace was inerted by 5 cycles of nitrogen pressurisation. The mixture was warmed to 50°C, and the headspace was purged by 3 cycles of pressurisation with hydrogen. The mixture was stirred for 4 hours at 50°C, dosing hydrogen gas to maintain a headspace pressure of 2 barg. The vessel was inerted by nitrogen purge cycles, and the mixture was filtered to remove catalyst particles. The clear filtrates were mixed with 2- methyltetrahydrofuran (60 mL, 6 relative volumes) and 2 M sodium hydroxide solution in water (19 mL, 1.8 mol eq). After a period of mixing, the mixture was settled and the lower aqueous layer was discarded. The organic layer was washed once with water (80 mL, 8 relative volumes) and then diluted with 2-methyltetrahydrofuran (80 mL, 8 relative volumes). The product solution in 2-methyltetrahydrofuran was distilled at 50°C under reduced pressure, to a residual volume of 60 mL. AZD9291 Aniline (seed) was added, and the mixture was cooled to 0°C over 4 hours. The resulting slurry was filtered, and the solids collected were washed with 2-methyltetrahydrofuran (20 mL, 2 relative volumes) to yield AZD9291 Aniline as an off-white powder (7.0 g, 75 percent> yield) after drying under vacuum. No detectable levels of AZD9291 Aniline Hydroxy were detected in the resulting AZD9291 Aniline by UPLC. (0623) UPLC methodology (0624) Apparatus An ultra performance liquid chromatograph fitted with a UV (0625) detector. The system should be capable of delivering a linear gradient. (0626) Column UPLC BEH Phenyl 1.7um, 2.1 x 100mm, or equivalent Phase A 0.06percent v/v trifluoroacetic acid in water (0627) Phase B 0.06percent v/v trifluoroacetic acid in acetonitrile (0628) Gradient profileCompound 7 from the previous step, iron (12.8 g), and ammonium chloride (1.42 g) were heated in ethanol (100 mL) and water (30 mL) at reflux for 1.5 h. The mixture was cooled and filtered. The solids were rinsed with DCM. The filtrate was concentrated to approximately 20 mL and NaOH (1 N, 50 mL) was added. The gray precipitates were filtered off and rinsed with DCM. The mixture was partitioned and the organic layer was washed with NH4OH (50 mL), brine (100 mL) and concentrated to a brown foam (compound 8, 12 g).2-(4-(N-(2-(dimethylamino)ethyl)-N-methylamino)-2-methoxy-5-amino)-4-chloropyrimidine (5.4g, 15 . 4mmol) and 1 - methyl - 1H - indole (2.6g, 20 . 0mmol) dissolved in 1, 2-dichloroethane (100 ml), by adding potassium hydride (1.0g, 24 . 6mmol), the reaction mixture 50 °C stirring reaction for 8 hours, TLC board determine the completion of the reaction, the reaction solution under reduced pressure to dry and concentration, adding ethyl acetate extraction, magnesium sulfate drying, and concentration to dry, ethanol and isopropanol mixed solvent crystallization, 2-(4-(N-(2-(dimethylamino)ethyl)-N-methylamino)-2-methoxy-5-aminophenylamino)-4-(1-methyl-1H-indole-3-yl)pyrimidine, pale yellow solid (6.2g), yield 90.2percent, In a nitrogen atmosphere, 28.5 (0.060 mol) of the intermediate 2 and 2.5 g of palladium carbon were added to 500 ml of methanol and passed at 25C Hydrogen atmospheric reaction 4h, filtered, the filtrate was concentrated to dryness under reduced pressure to give a brown solid, 26g, a yield of 97.3percent.A 125 ml autoclave was taken, and after replacing nitrogen three times, a compound F (1.50 g, 3.15 mmol) and a Raney nickel catalyst (0.70 g) were added and dissolved in ethyl acetate (30 ml), and the hydrogen was replaced twice and then charged with 2.0 MPa of hydrogen.The temperature was raised to 40 ° C to stir the reaction.Filter off and wash the filter cake twice with ethyl acetate (10 ml x 2).The filtrate was concentrated under reduced pressure at 40 ° C to give a dark green solid: 1.36 g.The molar yield was 96.90percent.Under nitrogen protection, 928 ml of methanol was added to a 2 L four-necked flask, Then, 92.8 g of the compound represented by the formula (III)Stirring, 18.6g was added Raney nickel (water-wet product)8 at atmospheric pressure under hydrogenation, Reaction temperature 25 , The reaction was carried out for 6 hours.Filtered by rotary evaporation to yield 82.6 g of gray solid, yield: 95.0percentIntermediate (VII) (15.3 g, 32.19 mmol, 1 eq) was dissolved in 50 mL of EtOH.3 ml of water, ammonium chloride (8 g, 0.15 mol), CoSO4.7H2O (0.9 g, 3.22 mmol, 0.1 eq) and 50 ml of hydrazine hydrate were added separately, and the reaction was stirred at 85 °C for 8 h, followed by TLC until the reaction was completed, and 100 ml of water was added to precipitate. Solid, stirred at room temperature for 1 hour, filtered, washed with water.The intermediate (VIII) was dried to 13.24 g, and the yield was 92.37percent.N1-(2-(Dimethylamino)ethyl)-5-methoxy-N1-methyl-N4-(4-(1-methyl-1H-indol-3-yl)pyrimidine(Hydroxy-2-yl)-2-nitrobenzene-1, 4-diamine (666 mg, 1.4 mmol)Soluble in ethanol (40mL), Iron powder (470mg, 8.4 mmol) and ammonium chloride (241 mg, 4.5 mmol).The reaction was stirred at reflux for 2 hours.After the reaction is over, The reaction solution was concentrated under reduced pressure.The crude product was isolated and purified by silica gel column chromatography (eluent: dichloromethane:methanol=20:1) to give a brown oil (552 mg, yield: 88.6percent).A mixture of N'-(2-dimethylaminoethyl)-2-methoxy-N’-methyl-N-[4-(1-methylindol-3-yl)pyrimidin-2-yl]-5-nitrobenzene-1, 4-diamine (Intermediate 101, 220 mg, 0.46 mmol), iron (155 mg, 2.78 mmol) and NHSolvent is tetrahydrofuran B, catalyst for A 10percent Pd/C, the pressure of the hydrogen is charged into the is 30bar, in 30 °C stirring under the temperature condition of 16h. The specific process: in 500 ml dry reaction flask, 5gN-(2-dimethyl amino-ethyl) - 2-methoxy-N-methyl-N-[ 4 - (1-methyl -1H-Indol-3-yl)-pyrimidin-2-yl] - 5-nitro-benzene -1, 4-diamine with tetrafuran 140 ml dissolved, add 0.01g10percent Pd/C, access 30bar hydrogen, in the 30 °C stirring under 16h. Filtering, to obtain the product solvent is removed by the spin vaporization N-(2-dimethyl amino-ethyl) - 2-methoxy-N-methyl-N-[ 4 - (1-methyl -1H-Indol-3-yl)-pyrimidin-2-yl] - 5-amino-benzene -1, 4-diamine a kind of white solid powder. The product purity 98.4percent, yield 85.9percent.1) Weigh 200g of the raw material hydrogenation precursor nitro compound, and then add it to a mixture of 4L of methanol and 100g of concentrated hydrochloric acid, stir and dissolve, then add 10g of Pd/C catalyst with Pd content of 10percent, stir well and mix to form Material I, the material I is sent to the preheating module 1 of the microchannel reactor for preheating, and after preheating, enters the reaction module group of the microchannel reactor.2) The reaction module group that delivers hydrogen to the microchannel reactor and the step 1) the preheated material I reacts in the reaction module group, wherein: the flow rate of the slurry pump is adjusted so that the flow rate of the material I is 40.0 g/min. The flow rate of the H2 gas flow meter is adjusted to 500 ml/min, the molar ratio of the raw material hydrogenation precursor nitro compound to hydrogen is 1:3.5, the reaction temperature is 90 ° C, the temperature of the cooling module is 20 ° C, and the residence time of the reaction is 25s, the reaction pressure is 1.5Mpa, and the collection flows out from the outlet of the cooling module.The reaction solution is subjected to post-treatment, which means that the catalyst is recovered by filtration, and the solvent is distilled off under reduced pressure. The residue is added to 1 L of water, stirred and dissolved, and the pH of the system is adjusted to 9.0 by a NaOH solution having a mass fraction of 20percent. 10.0, then join500ml of ethyl acetate was extracted and separated, the organic phase was discarded, the aqueous phase was stirred and crystallized at 10 ° C for 1 hour, filtered, and the product was washed with a small amount of ethanol, and vacuum dried at 50 ° C for 8 hours to obtain an oxetinib intermediate. Target product 160.65g, yield85.73percent, purity 99.68percent.The above obtainedN-[(4-Dimethylaminoethylamino-2-methoxy-5-nitrophenyl)]-4-(1-methyl-1H-indol-3-yl)-2-pyrimidinamine 3.8kg (7.99mol)It was added to an autoclave containing 38 L of ethanol, and then 380 g of palladium carbon was added to the system, replaced with nitrogen three times, and then hydrogen was replaced twice.The hydrogen pressure was set to 2 MPa, and the reaction was heated to 80 degrees overnight.After the end of the reaction, the autoclave was cooled, the reaction solution was withdrawn, filtered through a celite cake, and the filter cake was rinsed with ethanol.After concentrating to 15 kg, it was cooled, stirred and crystallized, returned to room temperature for 2 hours, filtered, and the filter cake was rinsed with ethanol.Drying gave 3.05 kg of silver-gray powder with a yield of 85.6percent (HPLC purity >99percent).60 g (126 mmol) of Intermediate 4 was dissolved in 500 mL of ethanol, and 4.7 g (88.2 mmol) of NH4CI, 49 · 4 g (756 mmol) of zinc powder were added to the above mixed solution, respectively.Finally, 150 mL of water was added to the mixed solution at once, and the mixture was heated to 105 ° C and stirred for 3 hours. After the reaction was completed, the mixture was filtered while hot, and the filter cake was washed with 150 mL of anhydrous ethanol. The filtrate was collected, and an appropriate amount of activated carbon was added thereto, heated to reflux at 80 ° C for 30 min, and filtered while hot, and the filter cake was washed with 300 mL of anhydrous ethanol. The filtrate was collected, and most of the solvent was evaporated under reduced pressure, and the residue was recrystallized from tetrahydrofuran.After suction filtration, the filter cake was dried to give 44.8 g of light gray solid.The yield is 79.8percent.A solution of N1- (2- (dimethylamino) ethyl) -5-methoxy-N1-methyl-N4- (4- (1-methylindol-3-yl) 2-nitrophenyl-1, 4-diamine (30.0 g, 0.063 mol) was added to a 1 L vial, Adding 600 mL of tetrahydrofuran and 10 g of Raney nickel, Hydrogen gas at a pressure of 3 atm was introduced at room temperature, Reaction for 12 h.After the reaction is complete, The reaction solution was filtered through celite, To obtain a brown solution, Concentrated under reduced pressure to give crude brown solid, The crude product was purified by recrystallization from 250 mL of a mixture of ethyl acetate and n-heptane (1: 1 by volume of ethyl acetate and n-heptane in the mixture)To give a light brown solid N1- (2- (dimethylamino) ethyl) -5-methoxy -N1- methyl -N4- (4- (1- methyl-indol-3-yl) pyrimidine-2 - yl) phenyl-1, 2, 4-triamine 21.9g, yield 78percent.A solution of N1- (2- (dimethylamino) ethyl) -5-methoxy-N1-methyl-N4- (4- (1-methylindol-3-yl)Yl) -2-nitrophenyl-1, 4-diamine (30.0 g, 0.063 mol) was added to a 1 L single-Then 600 mL of tetrahydrofuran and 10 g of Raney nickel were added and the pressure was 3 atm of hydrogen and reacted at 25 ° C for 12 h.After completion of the reaction, the reaction solution was filtered through celite and concentrated to give a crude product of brown solid which was recrystallized from 250 mL of a mixture of ethyl acetate and n-heptane (1: 1 by volume of ethyl acetate and n-heptane in the mixture) purification, A light brown solid product was obtainedMethyl-N4- (4- (1-methylindol-3-yl) pyrimidin-2-yl) phenyl -1, 2, 4-triamine 21.9 g, yield 78percent.N′-(2-(dimethylamino)ethyl)-5-methoxy-N′-methyl-N-(4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl)-2-nitrobenzene-1, 4-diamine (32) (5.00 g, 10.53 mmol) and Pd/C (10percent by weight) (250 mg, 2.10 mmol) were suspended in a mixture of MeOH (100 mL) and stirred under a hydrogen atmosphere for 24 h. The reaction mixture was filtered and the filtrate was concentrated in vacuo. The resulting residue was purified by silica gel chromatography (DCM: MeOH: NH3·H2O=25:1:0.1) to afford the title compound (3.51 g, 75.0percent yield) : 1H NMR (400 MHz, DMSO-d6) δ8.43 (d, J = 7.9 Hz, 1H), 8.31 (s, 1H), 8.28 (d, J = 5.3 Hz, 1H), 7.79 (s, 1H), 7.52 (d, J = 8.2 Hz, 1H), 7.49 (s, 1H), 7.28-7.23 (m, 1H), 7.20-7.13 (m, 2H), 4.63 (s, 2H), 3.89 (s, 3H), 3.75 (s, 3H), 2.90 (t, J = 6.7 Hz, 2H), 2.64 (s, 3H), 2.38 (t, J = 6.6 Hz, 2H), 2.19 (s, 6H). MS (ESI) m/z 446.4 [M+H]+.Example 1 (0622) AZD9291 Nitrodiamine (10.0 g), 5percent platinum-on-activated carbon (0.001 molar equivalents, 50percent water wet, 0.2 g, 0.02 relative weight), methanesulfonic acid (3.23 g, 1.6 molar equivalents) and water (100 mL, 10 relative volume) were mixed in a sealed autoclave. The headspace was inerted by 5 cycles of nitrogen pressurisation. The mixture was warmed to 50°C, and the headspace was purged by 3 cycles of pressurisation with hydrogen. The mixture was stirred for 4 hours at 50°C, dosing hydrogen gas to maintain a headspace pressure of 2 barg. The vessel was inerted by nitrogen purge cycles, and the mixture was filtered to remove catalyst particles. The clear filtrates were mixed with 2- methyltetrahydrofuran (60 mL, 6 relative volumes) and 2 M sodium hydroxide solution in water (19 mL, 1.8 mol eq). After a period of mixing, the mixture was settled and the lower aqueous layer was discarded. The organic layer was washed once with water (80 mL, 8 relative volumes) and then diluted with 2-methyltetrahydrofuran (80 mL, 8 relative volumes). The product solution in 2-methyltetrahydrofuran was distilled at 50°C under reduced pressure, to a residual volume of 60 mL. AZD9291 Aniline (seed) was added, and the mixture was cooled to 0°C over 4 hours. The resulting slurry was filtered, and the solids collected were washed with 2-methyltetrahydrofuran (20 mL, 2 relative volumes) to yield AZD9291 Aniline as an off-white powder (7.0 g, 75 percent> yield) after drying under vacuum. No detectable levels of AZD9291 Aniline Hydroxy were detected in the resulting AZD9291 Aniline by UPLC. (0623) UPLC methodology (0624) Apparatus An ultra performance liquid chromatograph fitted with a UV (0625) detector. The system should be capable of delivering a linear gradient. (0626) Column UPLC BEH Phenyl 1.7um, 2.1 x 100mm, or equivalent Phase A 0.06percent v/v trifluoroacetic acid in water (0627) Phase B 0.06percent v/v trifluoroacetic acid in acetonitrile (0628) Gradient profileCompound 7 from the previous step, iron (12.8 g), and ammonium chloride (1.42 g) were heated in ethanol (100 mL) and water (30 mL) at reflux for 1.5 h. The mixture was cooled and filtered. The solids were rinsed with DCM. The filtrate was concentrated to approximately 20 mL and NaOH (1 N, 50 mL) was added. The gray precipitates were filtered off and rinsed with DCM. The mixture was partitioned and the organic layer was washed with NH4OH (50 mL), brine (100 mL) and concentrated to a brown foam (compound 8, 12 g).2-(4-(N-(2-(dimethylamino)ethyl)-N-methylamino)-2-methoxy-5-amino)-4-chloropyrimidine (5.4g, 15 . 4mmol) and 1 - methyl - 1H - indole (2.6g, 20 . 0mmol) dissolved in 1, 2-dichloroethane (100 ml), by adding potassium hydride (1.0g, 24 . 6mmol), the reaction mixture 50 °C stirring reaction for 8 hours, TLC board determine the completion of the reaction, the reaction solution under reduced pressure to dry and concentration, adding ethyl acetate extraction, magnesium sulfate drying, and concentration to dry, ethanol and isopropanol mixed solvent crystallization, 2-(4-(N-(2-(dimethylamino)ethyl)-N-methylamino)-2-methoxy-5-aminophenylamino)-4-(1-methyl-1H-indole-3-yl)pyrimidine, pale yellow solid (6.2g), yield 90.2percent, 2-Fluoroacrylic acid (0.5 g) was added dropwise to a stirred solution of compound 8 (2 g) EDC-HCl (1.28 g) and DIPEA (1.15 g) in DMF (10 mL). The mixture was stirred for 16 hours and then diluted with DCM (50 mL) and washed with brine, NH4OH, brine. The organic layer was concentrated and purified by column chromatography. Pure fractions were evaporated to dryness and triturated with ether to afford compound 10 (0.35 g) as an off-white solidGeneral procedure: We used indole 7 as starting material to synthesize interediates15a-c by the method reported in the literature [32]. Different substituted acrylic acids were chlorinated by POCl3 to obtain the corresponding acyl chloride at 80 C. Under an ice bath, differently substituted acryl chloride was dissolved in dichloromethane and stirred at 0 C for 10-30 min. Another intermediates 15a-c were added to the above acryl chloride solution, and then added NaHCO3 solid powder, stirred at 0 C for 0.5-3 h. After the reaction was completed, the reaction mixture was filtered and solvent was distilled off under reduced pressure. The crude product was purified using flash chromatography with dichloromethane/methanol(v/v, from 50:1 to 20:1) as eluents.General procedure: We used indole 7 as starting material to synthesize interediates15a-c by the method reported in the literature [32]. Different substituted acrylic acids were chlorinated by POCl3 to obtain the corresponding acyl chloride at 80 C. Under an ice bath, differently substituted acryl chloride was dissolved in dichloromethane and stirred at 0 C for 10-30 min. Another intermediates 15a-c were added to the above acryl chloride solution, and then added NaHCO3 solid powder, stirred at 0 C for 0.5-3 h. After the reaction was completed, the reaction mixture was filtered and solvent was distilled off under reduced pressure. The crude product was purified using flash chromatography with dichloromethane/methanol(v/v, from 50:1 to 20:1) as eluents.General procedure: We used indole 7 as starting material to synthesize interediates15a-c by the method reported in the literature [32]. Different substituted acrylic acids were chlorinated by POCl3 to obtain the corresponding acyl chloride at 80 C. Under an ice bath, differently substituted acryl chloride was dissolved in dichloromethane and stirred at 0 C for 10-30 min. Another intermediates 15a-c were added to the above acryl chloride solution, and then added NaHCO3 solid powder, stirred at 0 C for 0.5-3 h. After the reaction was completed, the reaction mixture was filtered and solvent was distilled off under reduced pressure. The crude product was purified using flash chromatography with dichloromethane/methanol(v/v, from 50:1 to 20:1) as eluents.General procedure: We used indole 7 as starting material to synthesize interediates15a-c by the method reported in the literature [32]. Different substituted acrylic acids were chlorinated by POCl3 to obtain the corresponding acyl chloride at 80 C. Under an ice bath, differently substituted acryl chloride was dissolved in dichloromethane and stirred at 0 C for 10-30 min. Another intermediates 15a-c were added to the above acryl chloride solution, and then added NaHCO3 solid powder, stirred at 0 C for 0.5-3 h. After the reaction was completed, the reaction mixture was filtered and solvent was distilled off under reduced pressure. The crude product was purified using flash chromatography with dichloromethane/methanol(v/v, from 50:1 to 20:1) as eluents.General procedure: We used indole 7 as starting material to synthesize interediates15a-c by the method reported in the literature [32]. Different substituted acrylic acids were chlorinated by POCl3 to obtain the corresponding acyl chloride at 80 C. Under an ice bath, differently substituted acryl chloride was dissolved in dichloromethane and stirred at 0 C for 10-30 min. Another intermediates 15a-c were added to the above acryl chloride solution, and then added NaHCO3 solid powder, stirred at 0 C for 0.5-3 h. After the reaction was completed, the reaction mixture was filtered and solvent was distilled off under reduced pressure. The crude product was purified using flash chromatography with dichloromethane/methanol(v/v, from 50:1 to 20:1) as eluents.General procedure: We used indole 7 as starting material to synthesize interediates15a-c by the method reported in the literature [32]. Different substituted acrylic acids were chlorinated by POCl3 to obtain the corresponding acyl chloride at 80 C. Under an ice bath, differently substituted acryl chloride was dissolved in dichloromethane and stirred at 0 C for 10-30 min. Another intermediates 15a-c were added to the above acryl chloride solution, and then added NaHCO3 solid powder, stirred at 0 C for 0.5-3 h. After the reaction was completed, the reaction mixture was filtered and solvent was distilled off under reduced pressure. The crude product was purified using flash chromatography with dichloromethane/methanol(v/v, from 50:1 to 20:1) as eluents.General procedure: We used indole 7 as starting material to synthesize interediates15a-c by the method reported in the literature [32]. Different substituted acrylic acids were chlorinated by POCl3 to obtain the corresponding acyl chloride at 80 C. Under an ice bath, differently substituted acryl chloride was dissolved in dichloromethane and stirred at 0 C for 10-30 min. Another intermediates 15a-c were added to the above acryl chloride solution, and then added NaHCO3 solid powder, stirred at 0 C for 0.5-3 h. After the reaction was completed, the reaction mixture was filtered and solvent was distilled off under reduced pressure. The crude product was purified using flash chromatography with dichloromethane/methanol(v/v, from 50:1 to 20:1) as eluents. 4.1.1.1. N-(2-((2-(dimethylamino)ethyl)(methyl)amino)-4-methoxy-5-((4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl)amin amino)phenyl)-2-fluoroacrylamid (16a). Light yellow solid. 37.9% yield, m.p:190.7-192.1C 1H NMR (400 MHz, DMSO-d6) delta 10.04 (s, 1H), 8.92(s, 1H), 8.54 (s, 1H), 8.31 (dd, J =17.9, 6.6 Hz, 2H), 7.96 (s, 1H), 7.53(d, J 8.1, 1H), 7.29-7.20 (m, 2H), 7.17 (t, J 7.4 Hz, 1H), 7.07 (s, 1H), 5.46-5.41 (m, 1H), 5.08 (d, J 13.4 Hz, 1H), 3.91 (s, 3H), 3.89 (s, 3H), 3.27-3.22 (m, 2H), 2.87 (s, 2H), 2.65 (s, 3H), 2.58 (s, 6H); 13CNMR (400 MHz, DMSO-d6) delta 169.87, 164.85, 161.25, 158.98, 157.65, 144.25, 136.88, 134.21, 129.45, 128.96, 122.54, 122.21, 121.84, 119.98, 114.12, 111.53, 110.11, 109.86, 104.99, 99.32, 95.02, 59.23, 58.45, 56.88, 47.25 x 2, 42.23, 34.58. TOF MS ES (m/z): (M + H)+, calcdfor C28H32FN7O2: 517.2602, found, 518.2680.General procedure: We used indole 7 as starting material to synthesize interediates15a-c by the method reported in the literature [32]. Different substituted acrylic acids were chlorinated by POCl3 to obtain the corresponding acyl chloride at 80 C. Under an ice bath, differently substituted acryl chloride was dissolved in dichloromethane and stirred at 0 C for 10-30 min. Another intermediates 15a-c were added to the above acryl chloride solution, and then added NaHCO3 solid powder, stirred at 0 C for 0.5-3 h. After the reaction was completed, the reaction mixture was filtered and solvent was distilled off under reduced pressure. The crude product was purified using flash chromatography with dichloromethane/methanol(v/v, from 50:1 to 20:1) as eluents.Take N-(2-dimethylaminoethyl)-5-methoxy-N-[4-(1-methylindol-3-yl)pyrimidin-2-yl]benzene-1, 2, 4- triamine (II) (222.5 mg, 0.5 mmol, grind grinding), 7.5 mL of solvent (ethanol) was placed in a 50 mL round bottom flask maintained at a temperature of 60 C.Then o-hydroxybenzaldehyde (III) (160 muL, 1.5 mmol) was added dropwise.Add 2 drops of acetic acid to the reaction after the addition.The reaction was maintained at a temperature of 60 C for 3 hours.After the reaction is completed, the solvent is cooled, and the mixture is separated by silica gel column chromatography.The eluent is ethyl acetate, A mixed solvent of petroleum ether and triethylamine (V ethyl acetate: V petroleum ether: V triethylamine = 7:1:1) was dried to give the purified pale brown product (I).Yield: 80.5%, melting point: 175 to 177 C.

Molecular Weight:445.6
XLogP3:3.3
Hydrogen Bond Donor Count:2
Hydrogen Bond Acceptor Count:7
Rotatable Bond Count:8
Exact Mass:445.25900864
Monoisotopic Mass:445.25900864
Topological Polar Surface Area:84.5
Heavy Atom Count:33
Complexity:607
Covalently-Bonded Unit Count:1
Compound Is Canonicalized:Yes