Table 2. Structure assignments of WS070117 and its metabolites with 1H, 13C and HMBC NMR data from off-line measurements.
| metabolite | Peak a | 1H NMR date b (δ, ppm; J, Hz) c | 13C NMR date b (δ, ppm) | HMBC 1H to 13C c |
|---|---|---|---|---|
| 2', 3', 5'-tri-O-acetyl-N 6-(3-hydroxylaniline) adenosine | WS070117 | δ 9.83 (s, 1H, 6-NH), 9.33 (s, 1H, 3'-OH), 8.52 (s, 1H, 8-H), 8.42 (s, 1H, 2-H), 7.49 (dd, J = 2.3, 2.3 Hz, 1H, 2''-H), 7.31 (dd, J = 8.1, 2.3 Hz, 1H, 4''-H), 7.08 (dd, J = 8.1, 8.1 Hz, 1H, 5''-H), 6.46 (dd, J = 8.1, 2.3 Hz, 1H, 6''-H), 6.27 (d, J = 5.2 Hz, 1H, 1'-H), 6.06 (dd, J = 5.6, 5.2 Hz, 1H, 2'-H), 5.65 (dd, J = 5.6, 3.2 Hz, 1H, 3'-H), 4.43 (dd, J = 11.7, 3.7 Hz, 1H, 5'-Ha), 4.39 (m, 1H, 4'-H), 4.25 (dd, J = 11.7, 5.3 Hz, 1H, 5'-Hb), 2.12 (s, 3H, 3'-CH3), 2.04 (s, 3H, 2'-CH3), 2.01 (s, 3H, 5'-CH3). | δ170.01 (5'-C = O), 169.43 (3'-C = O), 169.27 (2'-C = O), 157.36 (3''-C), 152.28 (2-C), 152.21 (6-C), 149.12 (4-C), 140.86 (8-C), 140.38 (1''-C), 128.94 (5''-C), 120.27 (5-C), 111.87 (6''-C), 110.03 (4''-C), 108.11 (2''-C), 85.75 (1'-C), 79.43 (4'-C), 71.99 (2'-C), 70.00 (3'-C), 62.74 (5'-C), 20.46 (5'O-CH3), 20.35 (3'O-CH3), 20.19 (2'O-CH3). | (2,4), (2,6), (8,4), (8,5), (8,1'), (1',2'), (1',4), (1',8), (2',1'), (2',4), (2',2'-C = O), (3',1,), (3',5'), (3',3'-C = O), (4',3'), (4',3'), (5',3'),(5',4'), (5',5'-C = O), (2'',1''), (2'',3''), (2'',4''), (2'',6''), (4'',2''), (4'',3''), (4'',6''), (5'',1''), (5'',2''), (5'',3''), (5'',4''), (5'',6''), (6'',1''), (6'',2''), (6'',4''), (6'',5''), (6-NH,1''), (6-NH,5), (6-NH,6), (6-NH,2''), (6-NH,6''), (3-OH,1''), (3-OH,2''), (3-OH,3'') (3-OH,4''), (2'-CO-CH3, 2'-CO), (3'-CO-CH3,3'-CO), (5'-CO-CH3,5'-CO). |
| N 6-(3-β-D-glucuroniyl-laniline) purin | M2 | δ 8.33 (s, 1H, 2-H), 8.16 (s, 1H, 8-H), 7.64 (dd, J = 2.3, 2.3 Hz, 1H, 2''-H), 7.54 (dd, J = 8.2, 2.3 Hz, 1H, 4''-H), 7.21 (dd, J = 8.2, 8.2 Hz, 1H, 5''-H), 6.71 (dd, J = 8.2, 2.3 Hz, 1H, 6''-H), 4.82 (d, J = 7.5 Hz, 1H, β-glu-H), 3.57 (m, 1H, 4'''-H), 3.44 (m, 1H, 5'''-H), 3.285 (m, 1H, 3'''-H), 3.23 (m, 1H, 2'''-H) | δ 172.78 (6'''-C = O), δ157.98 (3''-C), 150.12 (2-C), 143.50 (8-C), 152.86 (4-C), 151.39 (6-C), 141.35 (1''-C), 129.46 (5''-C), 113.90 (4''-C), 108. 68 (6''-C), 118.23 (5-C), 109.78 (2''-C), 100.23 (1'''-C), 75.86 (3'''-C), 74.33 (4'''-C), 65.31 (5'''-C), 73.37 (2'''-C). d | (2,4), (2,6), (8,4), (8,5), (2'',1''), (2'',3''), (2'',4''), (2'',6''), (4'',2''), (4'',3''), (4'',6''), (5'',1''), (5'',3''), (5'',6''), (6'',2''), (6'',4''), (β-glu-H,3''). |
| N 8-hydroxy-N 6-(3- Sulfo-lailine) purin | M3 | δ 8.87 (s, 6-NH or 3''-OH), 8.17 (s, 1H, 2-H), 7.61 (dd, J = 8.2, 2.3 Hz, 1H, 6''-H), 7.49 (dd, J = 2.3, 2.2 Hz, 1H, 2''-H), 7.18 (dd, J = 8.2, 8.2 Hz, 1H, 5''-H), 6.77 (dd, J = 8.2, 2.2 Hz, 1H, 4''-H). | δ 153.76 (3''-C), 154.43 (2-C), 150.20 (8-C), 149.09 (4-C), 142.48 (6-C), 141.35 (1''-C), 128.95 (5''-C), 114.36 (4''-C), 114.05 (6''-C), 111.38 (2''-C), 106.26 (5-C). d | (8,4), (8,5), (8,1'), (2'',1''), (2'',3''), (2'',4''), (2'',6''), (4'',2''), (4'',3''), (4'',6''), (5'',1''), (5'',2''), (5'',3''), (5'',4''), (5'',6''), (6'',1''), (6'',2''), (6'',4''). |
| N 6-(3-β-D-glucuronide) adenosine | M4 | δ 9.90 (s, 6-NH), 8.52 (s, 1H, 8-H), 8.41 (s, 1H, 2-H), 7.75 (dd, J = 2.3, 2.3 Hz, 1H, 2''-H), 7.58 (dd, J = 8.2, 2.3 Hz, 1H, 6''-H), 7.21 (dd, J = 8.2, 8.2 Hz, 1H, 5''-H), 6.76 (dd, J = 8.2 2.3 Hz, 1H, 4''-H), 5.98 (d, J = 5.9 Hz, 1H, 1'-H), 4.83 (d, J = 7.3 Hz, 1H, β-glu-H), 4.62 (dd, J = 5.9, 5.5 Hz, 1H, 2'-H), 4.17 (dd, J = 5.5, 4.1 Hz, 1H, 3'-H), 3.97 (m, 1H, 4'-H), 3.69 (br d, J = 12.4 Hz, 1H, 5'-Ha), 3.57 (br d, J = 12.4 Hz, 1H, 5'-H), 3.42 (m, 1H, 5'''-H), 3.26 (m, 1H, 3'''-H), 3.23 (m, 1H, 2'''-H), 3.18 (m, 1H, 4'''-H). | δ172.95 (6'''-C = O), 158.05 (3''-C), 152.47 (2,6-C), 149.61 (4-C), 141.21 (8-C), 140.76 (1''-C), 129.73 (5''-C), 120.68 (5-C), 114.61 (6''-C), 111.08 (4''-C), 109.28 (2''-C), 100.69 (β-glu-C), 88.42 (1'-C), 86.28 (4'-C), 76.89 (3'''-H), 74.68 (5'''-H), 74.11 (2'-C), 73.44 (2'''-H), 72.41 (6'''-H), 70.95 (3'-C), 61.96 (5'-C). | (2,4), (2,5), (2,6), (8,4), (8,5), (1',2'), (1',4), (1',8), (2',1'), (2',4'), (3',1,), (3',4), (3',5'), (4',1'), (4',2'), (4',3'), (4',5'), (5',3'),(5',4'), (2'',1''), (2'',3''), (2'',4''), (2'',6''), (4'',2''), (4'',3''), (4'',6''), (5'',1''), (5'',3''), (5'',4''), (5'',6''), (6'',1''), (6'',2''), (6'',4''), (β-glu-H,3''), (6-NH,5), (6-NH,6), (6-NH,2''), (6-NH,6''). |
| N 6-(phenol-3-Sulfo-lailine) adenosine | M5 | δ 9.91 (s, 6-NH), 8.52 (s, 1H, 8-H), 8.40 (s, 1H, 2-H), 7.74 (dd, J = 2.3, 2.3 Hz, 1H, 2''-H), 7.63 (dd, J = 8.4, 2.3 Hz, 1H, 6''-H), 7.20 (dd, J = 8.5, 8.5 Hz, 1H,5''-H), 6.97 (dd, J = 8.5, 2.3 Hz, 1H, 4''-H), 5.95 (d, J = 5.9 Hz, 1H, 1'-H), 4.69 (dd, J = 5.9, 5.4 Hz, 1H, 2'-H), 4.17 (dd, J = 5.4, 3.6 Hz, 1H, 3'-H), 3.97 (m, 1H, 4'-H), 3.67 (br d, J = 11.9 Hz, 1H, 5'-Ha), 3.57 (br d, J = 11.9 Hz, 1H, 5'-Hb). | δ 153.62 (3''-C), 152.34 (6-C), 152.25 (2-C), 149.51 (4-C), 140.98 (8-C), 140.15 (1''-C), 128.75 (5''-C), 120.54 (5-C), 116.25 (6''-C), 115.59 (4''-C), 113.78 (2''-C), 88.20 (1'-C), 86.10 (4'-C), 73.90 (2'-C), 70.78 (3'-C), 61.79 (5'-C). | (2,4), (2,6), (8,4), (8,5), (8,1'), (1',2'), (1',4), (1',8), (2',1'), (2',4), (3',1,), (3',5'), (4',3'), (4',3'), (5',3'),(5',4'), (2'',3''), (4'',2''), (4'',3''), (4'',6''), (5'',1''), (5'',3''), (5'',4''), (5'',6''), (6'',1''), (6'',2''), (6'',4''), (6-NH,5), (6-NH,6), (6-NH,2''), (6-NH,6''). |
| N 8-hydroxy-N 6- (3-hydroxylaniline) purin | M6 | δ 9.00 (s, 6-NH or 3''-OH), 8.13 (s, 1H, 2-H), 7.34 (dd, J = 2.0, 2.2 Hz, 1H, 2''-H), 7.11 (dd, J = 8.0, 2.2 Hz, 1H, 6''-H), 7.05 (dd, J = 8.0, 8.0 Hz, 1H, 5''-H), 6.39 (dd, J = 8.0, 2.2 Hz 1H, 4''-H). | δ 157.66 (3''-C), 154.43(2-C), 149.54 (8-C), 149.25 (4-C), 142.60 (6-C), 141.35 (1''-C), 129.29 (5''-C), 110.12 (6''-C), 108.97 (4''-C), 107.29 (5-C), 106.31 (2''-C). | (8,4), (8,5), (8,1'), (2'',1''), (2'',3''), (2'',4''), (2'',6''), (4'',2''), (4'',3''), (4'',6''), (5'',1''), (5'',2''), (5'',3''), (5'',4''), (5'',6''), (6'',2''), (6'',4''). |
| N 6-(3-hydroxylaniline) purin | M7 | δ 9.44 (s, 6-NH or 3''-OH), 8.29 (s, 1H, 2-H), 8.10 (s, 1H, 8-H), 7.57 (dd, J = 2.2, 2.2 Hz, 1H, 2''-H), 7.30 (dd, J = 8.1 2.2 Hz, 1H, 6''-H), 7.05 (dd, J = 8.1, 8.1 Hz, 1H, 5''-H), 6.38 (dd, J = 8.1, 2.2 Hz, 1H, 4''-H). | δ 157.38 (3''-C), 154.94 (4-C), 150.59 (6-C), 150.12 (2-C), 143.50 (8-C), 141.35 (1''-C), 128.98 (5''-C), 118.97 (5-C), 110.81 (6''-C), 108.86 (4''-C), 106.75 (2''-C). d | (2,4), (2,6), (8,4), (8,5), (8,6), (2'',1''), (2'',3''), (2'',4''), (2'',6''), (4'',2''), (4'',3''), (4'',6''), (5'',1''), (5'',3''), (5'',4''), (5'',6''), (6'',2''), (6'',4''). |
| N 6-(3-hydroxylaniline) adenosine | M8 | δ 9.81 (s, 6-NH or 3'-OH), 8.53 (s, 1H, 8-H), 8.40 (s, 1H, 2-H), 7.51 (dd, J = 2.2, 2.2 Hz, 1H, 2''-H), 7.31 (dd, J = 8.1, 2.2 Hz, 1H, 4''-H), 7.09 (dd, J = 8.1, 8.1 Hz, 1H, 5''-H), 6.46 (dd, J = 8.1, 2.2 Hz 1H, 6''-H), 5.95 (d, J = 6.1 Hz, 1H, 1'-H), 4.64 (dd, J = 6.1, 5.5 Hz, 1H, 2'-H), 4.18 (dd, J = 5.5, 3.3 Hz, 1H, 3'-H), 3.99 (m, 1H, 4'-H), 3.70 (dd, J = 12.1, 3.7 Hz, 1H, 5'-Ha), 3.58 (dd, J = 12.1, 3.8 Hz, 1H, 5'-Hb). | δ157.55 (3''-C), 152.26 (6-C), 152.00 (2-C), 149.33 (4-C), 140.73 (8-C), 140.59 (1''-C), 129.10 (5''-C), 120.45 (5-C), 111.84 6 (''-C), 110.10 (4''-C), 108.14 (2''-C), 87.96 (1'-C), 85.96 (4'-C), 73.71 (2'-C), 70.65 (3'-C), 61.66 (5'-C). | (2,4), (2,6), (8,4), (8,5), (8,1'), (1',2'), (1',4), (1',8), (2',1'), (2',4'), (3',1,), (3',5'), (4',3'), (4',3'), (5',3'),(5',4'), (2'',1''), (2'',3''), (2'',4''), (2'',6''), (4'',2''), (4'',3''), (4'',6''), (5'',1''), (5'',2''), (5'',3''), (5'',4''), (5'',6''), (6'',1''), (6'',2''), (6'',4''), (6'',5''). |
a Retention times with method used for HPLC-PDA analysis.
b 1H (500 MHz), 13C (125 MHz) and HMBC NMR spectral data measured in DMSO-d6.
cMultiplicity of signals: s, singlet; d, doublet; t, triplet; m, multiplet; br, broad. Coupling constants (apparent splittings) reported as numerical values in hertz.
d Obtained from HSQC and HMBC spectra.