Nadide → HCID 12163515, compound

Temperature

40 °C

To 800 mL of 0.2M, pH 6.0 sodium phosphate buffer in a 2 L flask equipped with an overhead stirrer was added D-glucose (38.6 g, 1.2 eq), β-nicotinamide adenine dinucleotide, free acid (1.6 g, mmol), glucose dehydrogenase (36 mg, 3.2 kU, CODEXIS® GDH-102, 90 U/mg), and enzyme KRED-NADH-110 (200 mg, CODEXIS®, 25 U/mg). The vessels containing the reagents above were rinsed with 200 mL of fresh sodium phosphate buffer and added to the reaction which was stirred to dissolution and then heated to 40° C. To this mixture was added a solution of 2-bromo-4′-cyanoacetophenone (40 g, 178.5 mmol) in 100 mL DMSO through an addition funnel in about 30 min. The container was rinsed with 20 mL DMSO and the rinse was added to the reactor. A pH of 5.5-6.0 was maintained by adding 1 M NaOH through a fresh addition funnel (total volume of 200 mL over 6 h) after which HPLC showed complete consumption of the starting material. The reaction mixture was extracted with 800 mL MTBE×2 and the combined extracts were washed with 300 mL of 25% brine. The crude alcohol was transferred to a 3 L 3-neck flask and treated with solid NaOtBu (34.3 g, 357 mmol) stirring for 1 h and then additional NaOtBu (6.9 g, 357 mmol) and stirring for 30 min. The reaction mixture was filtered and the solution was washed with 300 mL 0.2 M pH 6.0 sodium phosphate buffer, brine, and then the solvent was removed in vacuo and the resulting white solid was dried in a vacuum oven to give (S)-4-(oxiran-2-yl)benzonitrile (23 g, 90% yield, 100% e.e.). 1H NMR (400 MHz, CDCl3) δ ppm 7.62 (2H, d), 7.35 (2H, d), 3.88 (1H, dd), 3.18 (1H, app t), 2.73 (1H, dd) Purity=99%.

1. additionThe vessels containing the reagents
2. washabove were rinsed with 200 mL of fresh sodium phosphate buffer
3. additionadded to the reaction which
4. washThe container was rinsed with 20 mL DMSO
5. additionthe rinse was added to the reactor
6. temperatureA pH of 5.5-6.0 was maintained
7. additionby adding 1 M NaOH through a fresh addition funnel (total volume of 200 mL over 6 h) after which HPLC
8. customconsumption of the starting material
9. extractionThe reaction mixture was extracted with 800 mL MTBE×2
10. washthe combined extracts were washed with 300 mL of 25% brine
11. customThe crude alcohol was transferred to a 3 L 3-neck flask
12. stirringstirring for 1 h
13. filtrationThe reaction mixture was filtered
14. washthe solution was washed with 300 mL 0.2 M pH 6.0 sodium phosphate buffer, brine
15. customthe solvent was removed in vacuo
16. customthe resulting white solid was dried in a vacuum oven

ORD record

ord-92e80fe40493421b8d4359d145ea47c5

Source dataset

uspto-grants-2013_03

DOI

10.6084/m9.figshare.5104873.v1

Patent number

US08399451B2