2-amino-2-phenylacetamide

Conditions: temp:0.00; pressure:0.00; stir:

Conditions: temp:0.00; pressure:0.00; stir:

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

Procedure: To a solution of D-Phenylglycine amide (850 mg, 7.5 mmol) in tetrahydrofuran (10 mL) and 1,4-dioxane (4 mL) was added 1,3-propanesultone (818 mg, 6.8 mmol). The solution stirred at reflux for 2 hours. The reaction was cooled to room temperature. The solid was collected by filtration and washed with acetone (2×20 mL). The solid was dissolved in 50% water/EtOH mL). Dowex Marathon C ion exchange resin (strongly acidic) was added to the solution. The suspension was stirred for 15 minutes before the resin was removed by filtration. The filtrate was evaporated under reduced pressure. The solid was suspended in acetone (50 mL), filtered and dried in the vacuum oven (50° C.), affording the title compound (720 mg, 34%). 1H NMR (D2O, 500 MHz) δ ppm 7.38 (m, 5H), 4.92, (s, 1H), 3.00 (m, 1H), 2.90 (m, 1H), 2.78 (m, 2H), 1.97 (m, 2H). 13C (D2O, 125 MHz) δ ppm 170.14, 130.95, 130.24, 129.94, 128.74, 63.40, 47.99, 44.92, 21.27. [α]D=+106° (c=0.0016 in water), ES-MS 273 (M+1). at reflux for 2 hours The solid was collected by filtration washed with acetone (2×20 mL) The solid was dissolved in 50% water/EtOH mL) Dowex Marathon C ion exchange resin (strongly acidic) was added to the solution The suspension was stirred for 15 minutes before the resin was removed by filtration The filtrate was evaporated under reduced pressure filtered dried in the vacuum oven (50° C.)

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

Procedure: The reaction is carried out at 25° C. and pH 7.5 in a fixed bed reactor. 10 ml of an aqueous solution of a mixture 0.2 M 7-ADCA and 0.2 M D-phenylglycinamide are passed through a column with 0.5 ml of the immobilized penicillin amidase. During the reaction, the pH is kept constant by adding HCl in an attached stirred reservoir container. Samples are taken and analyzed by HPLC as shown in example 5 at particular time intervals. is carried out at 25° C. During the reaction

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

Procedure: The reaction is carried out at 25° C. and pH 7.5 in a fixed bed reactor. 10 ml of an aqueous solution of a mixture 0.2 M 7-ADCA and 0.2 M D-phenylglycinamide are passed through a column with 0.5 ml of the immobilized penicillin amidase. During the reaction, the pH is kept constant by adding HCl in an attached stirred reservoir container. Samples are taken and analyzed by HPLC as shown in example 5 at particular time intervals. is carried out at 25° C. During the reaction

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

Procedure: To a solution of acid 9 (38 mg, 0.039 mmol) and N-methyl morpholine (4.5 μL, 0.041 mmol) in 1.5 mL of anhydrous THF cooled to 0° C. was added isobutyl chloroformate (5.2 μL, 0.040 mmol) and the mixture was stirred for 1 hour at that temperature. A solution of D-phenylglycinamide (8.6 mg, 0.057 mmol) and Et3N (10 μL) in 1 mL of THF/H2O 4:1 was added and the mixture was stirred for 1 h at 0° C. and 1 h at room temperature. After removal of the THF under vacuum, the residue was suspended in H2O (5 mL) and extracted with EtOAc (5 mL×3). The combined organic layers were dried over Na2SO4 and filtered. The solvent was removed under vacuum and the residue was purified by chromatography on a silica gel column (5-8% MeOH in CH2Cl2) to give conjugate S1 (28 mg, 66%) as a white solid: 1H NMR (600 MHz, DMSO-d6) δ8.41 (d, J=7.9 Hz, 1H), 8.31 (t, J=5.3 Hz, 3H), 7.65 (br. s., 1H), 7.25-7.45 (m, 14H), 7.13 and 7.12 (2×s, 2H), 5.36 (d, J=8.2 Hz, 1H), 3.11-3.17 (m, 6H), 2.35-2.48 (m, 2H), 2.30-2.34 (m, 2H), 2.28 (s, 9H), 2.21 (s, 9H), 1.58-1.67 (m, 6H), 1.37-1.44 (m, 6H); HRMS (ESI) calcd. for C55H63N6O18 (M+H)+: 1095.4193, found 1095.4203. the mixture was stirred for 1 h at 0° C. and 1 h at room temperature After removal of the THF under vacuum extracted with EtOAc (5 mL×3) The combined organic layers were dried over Na2SO4 filtered The solvent was removed under vacuum the residue was purified by chromatography on a silica gel column (5-8% MeOH in CH2Cl2) to give conjugate S1 (28 mg, 66%) as a white solid

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

Procedure: To a solution of acid 9 (38 mg, 0.039 mmol) and N-methyl morpholine (4.5 μL, 0.041 mmol) in 1.5 mL of anhydrous THF cooled to 0° C. was added isobutyl chloroformate (5.2 μL, 0.040 mmol) and the mixture was stirred for 1 hour at that temperature. A solution of D-phenylglycinamide (8.6 mg, 0.057 mmol) and Et3N (10 μL) in 1 mL of THF/H2O 4:1 was added and the mixture was stirred for 1 h at 0° C. and 1 h at room temperature. After removal of the THF under vacuum, the residue was suspended in H2O (5 mL) and extracted with EtOAc (5 mL×3). The combined organic layers were dried over Na2SO4 and filtered. The solvent was removed under vacuum and the residue was purified by chromatography on a silica gel column (5-8% MeOH in CH2Cl2) to give conjugate S1 (28 mg, 66%) as a white solid: 1H NMR (600 MHz, DMSO-d6) δ8.41 (d, J=7.9 Hz, 1H), 8.31 (t, J=5.3 Hz, 3H), 7.65 (br. s., 1H), 7.25-7.45 (m, 14H), 7.13 and 7.12 (2×s, 2H), 5.36 (d, J=8.2 Hz, 1H), 3.11-3.17 (m, 6H), 2.35-2.48 (m, 2H), 2.30-2.34 (m, 2H), 2.28 (s, 9H), 2.21 (s, 9H), 1.58-1.67 (m, 6H), 1.37-1.44 (m, 6H); HRMS (ESI) calcd. for C55H63N6O18 (M+H)+: 1095.4193, found 1095.4203. the mixture was stirred for 1 h at 0° C. and 1 h at room temperature After removal of the THF under vacuum extracted with EtOAc (5 mL×3) The combined organic layers were dried over Na2SO4 filtered The solvent was removed under vacuum the residue was purified by chromatography on a silica gel column (5-8% MeOH in CH2Cl2) to give conjugate S1 (28 mg, 66%) as a white solid

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

Procedure: In accordance with the process described in JP Patent Application Laying Open (Kokai) No. 62-55097, Enterobacter cloacae N-7901 (FERM BP-873) was cultured. This culture solution, 100 mL, was centrifuged, and the wet cells were then suspended in distilled water to prepare a 270 g cell suspension. D, L-phenylglycine amide, 30 g, was dissolved in this suspension, and the mixture was then allowed to react at 40° C. for 18 hours. After the reaction, cells were removed by centrifugation and 295 g of an aqueous solution containing 5.0% by mass each of L-phenylglycine and D-phenylglycine amide was obtained. to prepare a 270 g cell suspension to react at 40° C. for 18 hours After the reaction, cells were removed by centrifugation and 295 g of an aqueous solution containing 5.0% by mass each of L-phenylglycine

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

Procedure: In accordance with the process described in JP Patent Application Laying Open (Kokai) No. 62-55097, Enterobacter cloacae N-7901 (FERM BP-873) was cultured. This culture solution, 100 mL, was centrifuged, and the wet cells were then suspended in distilled water to prepare a 270 g cell suspension. D, L-phenylglycine amide, 30 g, was dissolved in this suspension, and the mixture was then allowed to react at 40° C. for 18 hours. After the reaction, cells were removed by centrifugation and 295 g of an aqueous solution containing 5.0% by mass each of L-phenylglycine and D-phenylglycine amide was obtained. to prepare a 270 g cell suspension to react at 40° C. for 18 hours After the reaction, cells were removed by centrifugation and 295 g of an aqueous solution containing 5.0% by mass each of L-phenylglycine

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

Procedure: To a solution of D-Phenylglycine amide (850 mg, 7.5 mmol) in tetrahydrofuran (10 mL) and 1,4-dioxane (4 mL) was added 1,3-propanesultone (818 mg, 6.8 mmol). The solution stirred at reflux for 2 hours. The reaction was cooled to room temperature. The solid was collected by filtration and washed with acetone (2×20 mL). The solid was dissolved in 50% water/EtOH mL). Dowex Marathon C ion exchange resin (strongly acidic) was added to the solution. The suspension was stirred for 15 minutes before the resin was removed by filtration. The filtrate was evaporated under reduced pressure. The solid was suspended in acetone (50 mL), filtered and dried in the vacuum oven (50° C.), affording the title compound (720 mg, 34%). 1H NMR (D2O, 500 MHz) δ ppm 7.38 (m, 5H), 4.92, (s, 1H), 3.00 (m, 1H), 2.90 (m, 1H), 2.78 (m, 2H), 1.97 (m, 2H). 13C (D2O, 125 MHz) δ ppm 170.14, 130.95, 130.24, 129.94, 128.74, 63.40, 47.99, 44.92, 21.27. [α]D=+106° (c=0.0016 in water), ES-MS 273 (M+1). at reflux for 2 hours The solid was collected by filtration washed with acetone (2×20 mL) The solid was dissolved in 50% water/EtOH mL) Dowex Marathon C ion exchange resin (strongly acidic) was added to the solution The suspension was stirred for 15 minutes before the resin was removed by filtration The filtrate was evaporated under reduced pressure filtered dried in the vacuum oven (50° C.)

Conditions: temp:0.00; pressure:0.00; stir:

Conditions: temp:0.00; pressure:0.00; stir: