ethyl 2-aminoacetate;hydrochloride

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: A mixture of 2,4-dimethoxy-benzaldehyde (50 g, 0.30 mole), glycine ethyl ester hydrochloride (44 g, 0.32 mole) and triethylamine (43.9 mL, 0.32 mole) in dichloroethane was stirred for one hour before sodium triacetoxyborohydride (100 g, 0.47 mole) was added in four portions. The mixture was stirred for three days before it was quenched with saturated sodium bicarbonate. The organic layer was extracted with 3N HCl (2×400 mL, 2.4 mole). The aqueous layers were combined, adjusted with solid sodium hydroxide (97.68 g, 2.4 mole) to pH=8 and extracted with ethyl acetate (500 mL). The ethyl acetate extract was washed with brine, dried over sodium sulfate, and concentrated. The residue was purified by flash column chromatography with a gradient of methanol and dichloromethane on silica gel (120 g) to give the title compound as a yellow oil (42.77 g) 1H NMR (CDCl3, 200 MHz): δ=7.11 (m, 1H), 6.41 (m, 2H), 4.15 (q, J=7.2 Hz, 2H), 3.81 (s, 3H), 3.80 (s, 3H), 3.74 (s, 2H), 3.37 (s, 2H), 1.27 (t, J=7.2 Hz, 3H). was added in four portions was quenched with saturated sodium bicarbonate extracted with ethyl acetate (500 mL) The ethyl acetate extract was washed with brine dried over sodium sulfate concentrated The residue was purified by flash column chromatography with a gradient of methanol and dichloromethane on silica gel (120 g)

Conditions: temp:20.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a suspension of glycine ethyl ester hydrochloride (1.2 eq.) in DCE was added TEA (2 eq.) The reaction was stirred for 10 min at 20° C. then 4-chlorobenzaldehyde (1 eq.), acetic acid (4 eq.) and sodium triacetoxyborohydride (2 eq.) were added. The reaction was stirred for 15 h at 20° C. The crude was partitioned between DCM and a saturated aqueous solution of NaHCO3. The aqueous layer was then extracted twice with DCM. The organic layers were combined, washed with brine, dried on MgSO4, filtered and concentrated under reduced pressure to afford (4-chloro-benzylamino)-acetic acid ethyl ester. The reaction was stirred for 15 h at 20° C The crude was partitioned between DCM The aqueous layer was then extracted twice with DCM washed with brine dried on MgSO4 filtered concentrated under reduced pressure

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: In a separable flask, glycine ethyl ester hydrochloride (reagent grade) (279 g, 2 mol) was mixed into acetic anhydride (reagent grade) (800 ml). Then, to the mixture was added dropwise over 3 hours a solution of maleic anhydride (reagent grade) (196 g. 2 mol) in acetic anhydride (600 ml), with stirring at room temperature. After the dropwise addition, stirring was carried out for another 1 hour, and the resulting precipitate was collected by suction filtration, washed with ion-exchanged water, and dried. The product obtained (382 g), triethylamine (reagent grade) (404 g, 4 mol), and toluene (1200 ml) were charged into a separable flask equipped with a Dean-Stark trap. Then, the mixture was subjected to reaction at 120° C. for 2 hours with stirring. During the reaction, the water formed was removed with the Dean-Stark trap. After the reaction, toluene (300 ml) was added and the mixture was washed three times with ion-exchanged water. Then, the organic phase obtained was collected and subjected to removal of the toluene by evaporation using an evaporator and a vacuum dryer to give ethyl maleimidoacetate (165 g). (Yield: ca. 38%. The production of ethyl maleimidoacetate was verified by GPC measurement. The presence of ethyl maleimidoacetate was verified by 1H-NMR measurement using DMSO-d6. This compound is referred to hereinafter as Compound A4). After the dropwise addition stirring the resulting precipitate was collected by suction filtration washed with ion-exchanged water dried were charged into a separable flask equipped with a Dean-Stark trap Then, the mixture was subjected to reaction at 120° C. for 2 hours with stirring During the reaction was removed with the Dean-Stark trap After the reaction, toluene (300 ml) was added the mixture was washed three times with ion-exchanged water Then, the organic phase obtained was collected subjected to removal of the toluene by evaporation an evaporator

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: In a manner similar to the preparation of N2,N4-bis(3-hydroxyphenyl)-5-fluoro-2,4-pyrimidinediamine, with the addition of triethylamine, 2,4-dichloro-6-ethoxycarbonyl-5-nitropyrimidine and glycine ethyl ester hydrochloride were reacted to yield N2,N4-bis(ethoxycarbonylmethyl)-6-ethoxycarbonyl-5-nitro-2,4-pyrimidinediamine. 1H NMR (CDCl3): mixture of rotamers δ 8.99 and 8.80 (2bs, 1H), 6.22 and 6.00 (2bs, 1H), 4.45 (t, 2H, J=7.2 Hz), 4.31-4.21 (m, 6H), 4.14 (d, 2H, J=5.1 Hz), 1.39 (t, 3H, J=7.2 Hz), 1.34-1.28 (m, 6H); LCMS: ret. time: 26.06 min.; purity: 99%; MS (m/e): 400 (MH+). were reacted

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: In a manner similar to the preparation of 2-chloro-N4-(3,4-ethylenedioxyphenyl)-5-fluoro-4-pyrimidineamine, 2,4-dichloro-6-ethoxycarbonyl-5-nitropyrimidine and glycine ethyl ester hydrochloride salt were reacted to yield N-(2-chloro-6-ethoxycarbonyl-5-nitro-4-pyrimidinyl)glycine Ethyl Ester. 1H NMR (CDCl3): δ 8.87 (bs, 1H), 4.48 (q, 2H, J=7.2 Hz), 4.39 (d, 2H, J=5.1 Hz), 1.40 (t, 3H, J=6.9 Hz), 1.33 (t, 3H, J=7.2 Hz); LCMS: ret. time: 28.27 min.; purity: 97%; MS (m/e): 332 (M+). were reacted

Conditions: temp:5.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Glycine ethyl ester hydrochloride (69.1 g, 495 mmol) and toluene (215 g) were mixed, and N-methylpyrrolidone (32.5 g) was poured into the mixture at room temperature. Benzaldehyde (50.0 g, 471 mmol) was added dropwise to the obtained mixture, and then trimethyl orthoformate (52.5 g, 495 mmol) was added dropwise. After completion of the dropwise addition, the temperature of the obtained mixture was adjusted to 7° C. and a mixed solution of triethylamine (52.4 g, 518 mmol) and toluene (26.5 g) was added dropwise over 3 hours. After completion of the dropwise addition, the mixture was stirred at a temperature in a range from 7° C. to room temperature for 6 hours. After completion of the reaction, the reaction mixture was cooled to 5° C. and water (140 mL) was added dropwise thereto. Thereafter, stirring was stopped to perform liquid separation and then the obtained organic layer was washed with 20% brine (95 g). The obtained organic layer was dried over magnesium sulfate and the solvent was distilled off under reduced pressure to obtain a toluene solution (104.5 g) of (E)-N-phenylmethyleneglycine ethyl ester (86.9 g of pure (E)-N-phenylmethyleneglycine ethyl ester). Yield: 96%. After completion of the dropwise addition was adjusted to 7° C. After completion of the dropwise addition After completion of the reaction Thereafter, stirring liquid separation the obtained organic layer was washed with 20% brine (95 g) The obtained organic layer was dried over magnesium sulfate the solvent was distilled off under reduced pressure

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: A 4.7 M aqueous solution of sodium hydroxide (50 mL) was added into a stirred, ice-cooled solution of ethyl glycinate hydrochloride (32.19 g, 0.23 mole) in water (50 mL). Then, ethyl acrylate (23.1 g, 0.23 mole) was added and the mixture was stirred at room temperature until completion of the reaction was ascertained by TLC. After 19 h the solution was partitioned with dichloromethane (3×100 mL). The organic layer was dried with anhydrous sodium sulfate, filtered and evaporated. The residue was distilled to afford AA (28.8 g, 61%). After 19 h the solution was partitioned with dichloromethane (3×100 mL) The organic layer was dried with anhydrous sodium sulfate filtered evaporated The residue was distilled to afford AA (28.8 g, 61%)

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: A 4.7 M aqueous solution of sodium hydroxide (50 mL) was added into a stirred, ice-cooled solution of ethyl glycinate hydrochloride (32.19 g, 0.23 mole) in water (50 mL). Then, ethyl acrylate (23.1 g, 0.23 mole) was added and the mixture was stirred at room temperature until completion of the reaction was ascertained by TLC. After 19 h the solution was partitioned with dichloromethane (3×100 mL). The organic layer was dried with anhydrous sodium sulfate, filtered and evaporated. The residue was distilled to afford AA (28.8 g, 61%). After 19 h the solution was partitioned with dichloromethane (3×100 mL) The organic layer was dried with anhydrous sodium sulfate filtered evaporated The residue was distilled to afford AA (28.8 g, 61%)

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To ethyl glycinate hydrochloride (12.9 g, 92 mmol) in water (20 mL) at 0° C. was added cold aqueous NaOH (19.5 mL, 93 mmol, 4.75 M) followed by ethyl acrylate (9.18 g, 92 mmol). The reaction stirred about 18 hours and warmed to room temperature. The mixture was extracted with CH2Cl2 four times and the solution was dried (Na2SO4). The material was distilled at about 110° C. (1.8 torr) to afford the product, 1-a. The mixture was extracted with CH2Cl2 four times the solution was dried (Na2SO4) The material was distilled at about 110° C. (1.8 torr)

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 6-Bromopyridine-2-carbaldehyde (10 g, 53.8 mmol) and glycine ethyl ester hydrochloride (7.50 g, 53.8 mmol) were placed in a flask. Dichloroethane (220 mL) and triethylamine (7.49 mL, 53.8 mmol) were added and the reaction was stirred for 45 minutes. Sodium triacetoxyborohydride (15.95 g, 75 mmol) was added and the reaction was stirred at ambient temperature overnight. The reaction was diluted with ethyl acetate, saturated sodium bicarbonate, and saturated sodium carbonate to make the pH basic. The organic layer was separated and washed with brine. The organic layer was dried over magnesium sulfate, filtered and concentrated. The crude mixture was purified by silica gel chromatography to yield the title compound. the reaction was stirred at ambient temperature overnight The organic layer was separated washed with brine The organic layer was dried over magnesium sulfate filtered concentrated The crude mixture was purified by silica gel chromatography

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Ethyl N-(2-fluoro-4-nitrobenzoyl)glycinate Triethylamine (0.85 ml,6.1 mmol) was added to a solution of 2-fluoro-4-nitrobenzoic acid (T. R. Jones et al., UK Patent GB 2175 903A, 1986) (1.0 g, 5.4 mmol), glycine ethyl ester hydrochloride (0.85 g, 6.1 mmol), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.1 g, 5.7 mmol) in methylene chloride (30m1). After stirring 1 hour at room temperature the solution was passed through a short pad of silica gel eluting with ethyl acetate: methylene chloride (1:3) and the resulting white solid was filtered from hexane and dried under high vacuum to give ethyl N-(2-fluoro-4-nitrobenzoyl)glycinate (1.04 g). 1H NMR (DMSO-d6, 300 MHz) δ: 1.22 (t,J=7 Hz, 3H, CH3), 4.05 (d, J=6 Hz, 2H, glyCH2), 4.15 (q, J=7 Hz, 2H, ester CH2), 7.88 (t, J=8 Hz, 1H, Ar), 8.17 (dd, J=8.5, 1.5 Hz, 1H, Ar), 8.25 (dd, J=10, 2 Hz, 1H), 9.09 (br t, J5.5 Hz, 1H, CONH). Anal. Calculated for C11H11FN2O5: C, 48.89; H, 4.10; N, 10.37. Found: C, 49.00; H, 4.11; N, 10.43. the resulting white solid was filtered from hexane dried under high vacuum

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 36.9 g of cyanuric chloride, 27.9 g of ethyl glycinate hydrochloride and 33.6 g of sodium bicarbonate were heated under reflux in 200 ml of dioxane for 30 minutes. At the end of this time, the dioxane was removed by distillation under reduced pressure, and a mixture of ethyl acetate and water was added to the resulting residue, after which the mixture was shaken. At the end of this time, the dioxane was removed by distillation under reduced pressure a mixture of ethyl acetate and water was added to the resulting residue