Asymmetric Synthesis of Optically Pure Pharmacologically Relevant Amines
FULL PAPERS
(2 mg in the cases of ATA-113, ATA-114 and ATA-117, and
20 mL for ATA-103), l- or d-alanine (250 mM), pyruvate re-
ductase mix (=LDH mix) (40 mg, mixture of lactate dehy-
drogenase, glucose dehydrogenase, glucose, NAD+), and
DMSO (150 mL). The reaction mixture contained 50 mM of
the corresponding ketone 1. The reaction was stopped by
addition of aqueous NaOH solution (200 mL, 10 N), fol-
lowed by extraction with ethyl acetate (600 mL, twice), the
organic phase was dried (Na2SO4) and the conversion was
measured by GC.
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Analysis of Optical Purity of Products
The enantiomeric excess of amines 2a–l was analyzed by gas
chromatography on a chiral phase after derivatization to the
acetoamides, which was performed by addition of DMAP
and a 20-fold excess of acetic anhydride. After washing with
water and drying (Na2SO4), the ee of the derivatized com-
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Represenative Example for Amination: Preparation
of (R)-2a
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In a 50-mL screw-topped tube 1a (100 mg, 0.67 mmol) was
suspended in phosphate buffer (17 mL, 100 mM, pH 8.0,
1 mM PLP, 15% v vÀ1 DMSO). d-Alanine (3.35 mmol) and
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crude preparation of lactate dehydrogenase mixture
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(500 mg) were added. The reaction was started by the addi-
tion of w-transaminase ATA-117 (40 mg) and the tube was
shaken at 308C (300 rpm). Conversion and enantiomeric
excess were analyzed as described above. After 24 h, the pH
of the mixture was adjusted to pH 1 with aqueous HCl
(5M), and the ketone 1a was extracted five times with di-
chloromethane (5ꢂ10 mL). After the extraction, 1a was not
detectable in the residual aqueous phase. The pH was now
adjusted to pH 12, and the amine 2a was extracted four
times with dichloromethane (4ꢂ10 mL). The solvent of the
combined organic phases was evaporated under reduced
pressure and 2a was obtained; yield: 63%; [a]D À6.73 (c 0.5,
chloroform), [lit. 6.40 (c 0.47, chloroform) for the (S)-enan-
tiomer, 98% ee];[16] 1H NMR (200 MHz): d=1.10 (d, 3H,
J=6.70 Hz), 1.32 (s, 2H), 1.52–1.75 (m, 2H), 2.52–2.74 (m,
2H), 2.87–2.91 (m, 1H), 7.10–7.52 (m, 5H).
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Adv. Synth. Catal. 2008, 350, 2761 – 2766
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