N-Acetyl-Neuraminic Acid → HCID 131735083, compound

HRID 928876Notes

Reaction equation

HRID 928876 reaction equation
[O:1]([CH2:13][C@@H:14]([OH:29])[C@@H:15]([OH:28])[C@@H:16]1[O:25][C:20]([OH:24])([C:21](=[O:23])[OH:22])[CH2:19][C@H:18]([OH:26])[C@H:17]1[OH:27])[C@@H:2]1[O:10][C@H:9]([CH2:11][OH:12])[C@H:7]([OH:8])[C@H:5]([OH:6])[C@H:3]1[OH:4].[OH:30][C:31]([C:33]1([O:44][C@@H:43]([C@@H:45]([C@@H:47]([CH2:49][OH:50])[OH:48])[OH:46])[C@H:38]([NH:39][C:40]([CH3:42])=[O:41])[C@@H:36]([OH:37])[CH2:35]1)O)=[O:32].[Mg+2].[Cl-].[Cl-]>C(O)C(N)(CO)CO.Cl>[OH:32][C:31]([C@@:33]1([O:44][C@@H:43]([C@@H:45]([C@@H:47]([CH2:49][OH:50])[OH:48])[OH:46])[C@H:38]([NH:39][C:40]([CH3:42])=[O:41])[C@@H:36]([OH:37])[CH2:35]1)[O:6][C@H:5]1[C@@H:7]([OH:8])[C@@H:9]([CH2:11][OH:12])[O:10][C@@H:2]([O:1][CH2:13][C@@H:14]([OH:29])[C@@H:15]([OH:28])[C@@H:16]2[O:25][C:20]([OH:24])([C:21](=[O:22])[OH:23])[CH2:19][C@H:18]([OH:26])[C@H:17]2[OH:27])[C@@H:3]1[OH:4])=[O:30]

Participants

5

Reactant 3

HCID 439197N-Acetyl-Neuraminic AcidHCID 5360315Magnesium ChlorideHCID 131735084HCID 131735084, compound

Product 1

HCID 131735083HCID 131735083, compound

Reagents, catalysts & solvents 1

HCID 93573Tromethamine hydrochlorideSolvent

Procedure

Neu5Acα2-3Galβ1-9Kdn was prepared first by following the general one-pot two-enzyme α2-3-sialylation system described above from Galβ1-9Kdn (101 mg, 0.22 mmol) in Tris-HCl buffer (100 mM, pH 8.5) containing Neu5Ac (109 mg, 0.33 mmol), CTP (188 mg, 0.33 mmol), MgCl2 (20 mM), NmCSS (0.8 mg), and PmST1 (0.04 mg). The reaction was incubated at 37° C. for 3 h with shaking (120 rpm). The crude product was purified by Bio-Gel P-2 gel filtration to afford the sialoside product Neu5Acα2-3Galβ1-9Kdn (148 mg, 89%). 1H NMR (600 MHz, D2O): δ 4.33 (d, 1H, J=7.8 Hz, H′-1), 4.00 (dd, 1H, J=1.8 and 10.8 Hz), 3.93 (dd, 1H, J=3.6 and 11.2 Hz), 3.82-3.65 (m, 9H), 3.60-3.39 (m, 9H), 2.58 (dd, 1H, J=4.2 and 12.6 Hz, H-3 eq″), 2.01 (dd, 1H, J=4.8 and 13.2 Hz, H-3 eq), 1.85 (s, 3H, CH3), 1.62 (t, 1H, J=12.0 Hz, H-3ax), 1.60 (t, 1H, J=12.0 Hz, H-3ax″). 13C NMR (75 MHz, D2O): δ 176.29, 175.17, 173.95, 103.28, 99.93, 96.35, 75.87, 75.07, 73.02, 71.93, 71.90, 71.86, 71.61, 70.44, 69.64, 69.50, 69.15, 68.48, 68.25, 67.97, 67.67, 62.75, 61.13, 59.50, 51.84, 39.82, 39.14, 22.22. HRMS (ESI) calcd. for C26H41NNaO22 (MNa), 742.2018, found 742.2001.

Workup

  1. filtrationThe crude product was purified by Bio-Gel P-2 gel filtration

Sources & evidence

ORD record
ord-e47c7fd6ea20407bb72f2becd5843381
Source dataset
uspto-grants-2016_03
Patent number
US09290530B2

Reaction notes

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