Réaction #742672

ord-8391641b21ae466d99e9d1053fa5421a

Équation de réaction

OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O
D(+)-glucose
O.O.O.O.O.O.[Cl-].[Cl-].[Mg+2]
magnesium chloride hexahydrate
O=C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO
glucose
[Na+].[OH-]
sodium hydroxide
CC(C)(C)OC(=O)N1CCC2(CC2)C(=O)C1
4-oxo-6-aza-spiro[2.5]octane-6-carboxylic acid tert-butyl ester
NC(=O)c1ccc[n+]([C@@H]2O[C@H](COP(=O)([O-])OP(=O)(O)OC[C@H]3O[C@@H](n4cnc5c(N)ncnc54)[C@H](O)[C@@H]3O)[C@@H](O)[C@H]2O)c1
β-NAD
→
CC(C)(C)OC(=O)N1CCC2(CC2)[C@H](O)C1
title compound
Rendement 93.0%
CC(C)(C)OC(=O)N1CCC2(CC2)[C@H](O)C1
(S)-4-Hydroxy-6-aza-spiro[2.5]octane-6-carboxylic acid tert-butyl ester
Rendement 93.0%

Réactifs

OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O
D(+)-glucose
O.O.O.O.O.O.[Cl-].[Cl-].[Mg+2]
magnesium chloride hexahydrate
—
KRED-NADH-117
O=C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO
glucose
[Na+].[OH-]
sodium hydroxide
CC(C)(C)OC(=O)N1CCC2(CC2)C(=O)C1
4-oxo-6-aza-spiro[2.5]octane-6-carboxylic acid tert-butyl ester
NC(=O)c1ccc[n+]([C@@H]2O[C@H](COP(=O)([O-])OP(=O)(O)OC[C@H]3O[C@@H](n4cnc5c(N)ncnc54)[C@H](O)[C@@H]3O)[C@@H](O)[C@H]2O)c1
β-NAD

Réactifs

Aucun

Solvants

O=S(=O)(O)CCN1CCOCC1
MES
CCCCCCC
heptane

Conditions de réaction

Température
35°CELSIUS
Conditions détaillées
See reaction.notes.procedure_details.

Traitement

  1. 1
    AutreAfter a consumption of 1.307 L (corresponding to 98% conversion; after 17 h) the reaction mixture
  2. 2
    Extractionwas extracted with ethyl acetate (10 L)
  3. 3
    SéchageThe organic phase was dried over sodium sulfate
  4. 4
    Concentrationconcentrated in vacuo (200 mbar/45° C.) until evaporation
  5. 5
    TempératureUpon cooling the oily residue (411 g)
  6. 6
    Autreto crystallize
  7. 7
    workup.STIRRINGwas stirred with heptane (1 L) for 2 h
  8. 8
    FiltrationThe crystals were filtered off
  9. 9
    Autrethe filtrate evaporated to dryness
  10. 10
    workup.DISSOLUTIONredissolved in ethyl acetate (150 ml)
  11. 11
    Concentrationconcentrated in vacuo
  12. 12
    AutreThe crystal suspension formed again
  13. 13
    Températureupon cooling
  14. 14
    Filtrationthe crystals filtered off
  15. 15
    LavageBoth crops of crystals were washed with heptane
  16. 16
    Autredried under high vacuum

Mode opératoire

D(+)-glucose monoydrate (300 g) and magnesium chloride hexahydrate (1.0 g) were dissolved in 10 mM MES buffer pH 6.5 (2.4 L; Sigma M3671). After addition of 4-oxo-6-aza-spiro[2.5]octane-6-carboxylic acid tert-butyl ester (300 g; 1.33 mmol) and β-NAD (3.0 g; free acid; Roche Diagnostics Cat. No. 10 004 626) the pH was re-adjusted and the suspension heated to 35° C. The reaction was started by adding ketoreductase KRED-NADH-117 (3.0 g; former Biocatalytics, now Codexis) and glucose dehydrogenase GDH-102 (300 mg; Biocatalytics). The suspension was vigorously stirred at 35° C. keeping the pH constant at 6.5 by the controlled addition (pH-stat) of 1.0 M aq. sodium hydroxide solution. After a consumption of 1.307 L (corresponding to 98% conversion; after 17 h) the reaction mixture was extracted with ethyl acetate (10 L). The organic phase was dried over sodium sulfate and concentrated in vacuo (200 mbar/45° C.) until evaporation fell off. Upon cooling the oily residue (411 g) started to crystallize and was stirred with heptane (1 L) for 2 h. The crystals were filtered off and the filtrate evaporated to dryness, redissolved in ethyl acetate (150 ml) and concentrated in vacuo as described above. The crystal suspension formed again upon cooling was stirred with heptane (200 ml; 2 h) and the crystals filtered off. Both crops of crystals were washed with heptane and dried under high vacuum to yield the title compound in 93% yield (250.77 g and 34.60 g white crystals), each having a purity of >98.5% GC and 99.8% ee. [α]D=−44.97° (c=1.00, CHCl3).

Source

DOI: 10.6084/m9.figshare.5104873.v1Brevet: US08071586B2uspto-grants-2011_12