Sodium Phosphate, Dibasic → HCID 58332752, compound

Temperature

30 °C

Water (516 kg) was charged to a 1000 L vessel followed by disodium hydrogen orthophosphate (36.3 kg), D-alanine (41.8 kg) and D-glucose (37.6 kg). The mixture was warmed to 30° C. to dissolve the solids. NAD (130.5 g) and PLP (261 g) were added and the pH checked (7.8). ATA-117 transaminase (2.61 kg), lactate dehydrogenase (52.2 g), and finally glucose dehydrogenase (130.5 g) were added and rinsed into the vessel with 2.5 L water. After all enzymes were in solution, A-2 (39.2 kg total, 29 kg assay) was added, followed by a final water rinse (5 kg). The reaction mixture was stirred at 30° C. with the pH being adjusted to pH 7.6 every 2-3 hours by the addition of 5N NaOH for the first 6 hours. The reaction was then left to stir overnight (16 hours) before the pH was again adjusted to pH 7.6. The reaction was allowed to stir for 44 hours. Sodium chloride (90 kg) was added to the reaction mixture and the pH adjusted to pH 3.6 by the addition of 5N HCl (66 L). Dichloromethane (200 kg) was charged to the vessel followed by solka-Flok (15 kg) and the mixture allowed to stir for 10 minutes. The batch was then filtered through more Solka-Flok (5 kg). The Solka-Flok was washed with more dichloromethane (65 kg). The filtrates were returned to the vessel, and the layers allowed to separate. The lower organic layer was run off and the aqueous layer re-extracted with dichloromethane (200 kg). The lower organic layer was run off and the aqueous layer discarded. The organic layers were recombined, returned to the vessel and washed with 8% sodium bicarbonate solution (54 L) for 20 minutes. The layers were allowed to separate and the organic layer run off. The aqueous layer was discarded and the organic layer returned to the vessel. The dichloromethane solution of A-2 (19.3 kg assay) was stored at room temperature until required.

1. dissolutionto dissolve the solids
2. washrinsed into the vessel with 2.5 L water
3. additionwas added
4. washrinse (5 kg)
5. additionbeing adjusted to pH 7.6 every 2-3 hours by the addition of 5N NaOH for the first 6 hours
6. waitThe reaction was then left
7. stirringto stir overnight (16 hours) before the pH
8. stirringto stir for 44 hours
9. additionSodium chloride (90 kg) was added to the reaction mixture
10. additionthe pH adjusted to pH 3.6 by the addition of 5N HCl (66 L)
11. additionDichloromethane (200 kg) was charged to the vessel
12. stirringto stir for 10 minutes
13. filtrationThe batch was then filtered through more Solka-Flok (5 kg)
14. washThe Solka-Flok was washed with more dichloromethane (65 kg)
15. customto separate
16. extractionthe aqueous layer re-extracted with dichloromethane (200 kg)
17. washwashed with 8% sodium bicarbonate solution (54 L) for 20 minutes
18. customto separate
19. customwas stored at room temperature

ORD record

ord-6c9d5ec30a9b4738be4a394a3501167e

Source dataset

uspto-grants-2014_03

DOI

10.6084/m9.figshare.5104873.v1

Patent number

US08674093B2