G.-T. Fan et al. / Tetrahedron 61 (2005) 1855–1862
1861
indexZ(value(test compounds)Kvalue(blank))/(value(control)
K
(t, JZ7.4 Hz, 2H), 3.47 (dd, JZ6.8, 12.8 Hz, 2H), 4.26 (dd,
JZ7.4, 10.6 Hz, 1H), 4.35–4.52 (m, 4H), 4.55 (t, JZ5.8 Hz,
1H), 4.60 (d, JZ3.2 Hz, 1H), 4.68 (dd, JZ3.8, 9.8 Hz, 1H),
5.35 (dt, JZ5.2, 7.4 Hz, 1H), 5.56 (d, JZ3.6 Hz, 1H), 8.85
(t, JZ5.4 Hz, 1H). 13C NMR (100 MHz, pyridine-d5) d
14.7, 23.4, 26.6, 27.80, 30.1, 30.1, 30.1, 30.3, 30.4, 32.6,
37.0, 40.4, 53.8, 63.2, 70.1, 70.7, 71.5, 72.01, 73.7, 102.0,
171.3, 173.8. HRMS (MALDI-TOF, MCHC) calcd for
C38H75N2O8: 687.5523. Found: 687.5527.
value(blank)))!100. The value(control) was the detected
value of the cells cultured with medium containing vehicle,
while the value(blank) was that of medium only.
3.9. Cytokine assay
Spleen cells were suspended in RPMI-1640 medium
containing 10% FCS in cell density of 5!106 cells/mL.
The cell solution was added test compound to a final
concentration of 100 ng/mL or none as control and was
dispensed 200 mL/well into 96-well plate. The culture plates
were incubated at 37 8C incubator containing 5% CO2 for 0,
12, 24 and 48 h. At each time point, the culture supernatant
was drawn for IFN-g and IL-4 detection using standard
sandwich ELISA. All procedures were conducted according
to the standard protocol of the assay kits from Quantikinew
M. (catalog number was MIF00 for INF-g and M4000 for
IL-4).
3.9.5. O-(a-D-Glucopyranosyl)-N-pentadecanoyl-L-serine
tetradecyl amide (50). TLC (MeOH/CHCl3Z1:10) Rf
0.15; 1H NMR (CD3OD, 400 MHz) d 4.86 (1H, H-1,
mixed with water peak), 4.56 (1H, 0dd, JZ6.2, 5.9 Hz, H-2),
3.88 (1H, dd, JZ10.5, 6.2 Hz, H-1 ), 3.85 (1H, dd, JZ0 11.8,
2.4 Hz, H-6), 3.78 (1H, dd, JZ10.5, 5.9 Hz, H-1 ), 3.70
(1H, dd, JZ11.8, 5.6 Hz, H-6), 3.63 (1H, dd, JZ9.7,
9.2 Hz, H-3), 3.58 (1H, ddd, JZ9.9, 5.6, 2.4 Hz, H-5), 3.45
(1H, dd, JZ9.7, 3.8 Hz, H-2), 3.32 (1H, dd, JZ9.9, 9.2 Hz,
H-4), 3.26–3.21 (2H, m, H-30), 2.30 (2H, t, JZ7.5 Hz,
H-40), 1.67–1.64 (2H, m), 1.57–1.54 (2H, m), 1.50–1.10
(44H, br), 0.94 (6H, t, CH3!2); 13C NMR (CD3OD,
100 MHz) d 176.4, 172.2, 101.0, 75.2, 74.2, 73.6, 71.8, 69.1,
62.8, 54.9, 40.8, 37.0, 33.2, 31.0, 30.9, 30.9, 30.8, 30.7,
30.6, 30.6, 30.5, 28.2, 27.0, 23.9, 14.6; FAB-MS m/z 687.5
(MCC1); HRMS calcd. for C38H75N2O8 (MCCH)
687.5523, Found: 687.5531.
3.9.1. (2S,3S,4R)-1-O-(a-D-Galactopyranosyl)-2-(N-tetra-
cosanoylamino)-1,3,4-nonanetriol (2). 1H NMR
(400 MHz, pyridine-d5) d 0.82 (t, JZ7.2 Hz, 3H), 0.89 (t,
JZ6.8 Hz, 3H), 1.10–1.50 (br, 46H), 1.58–1.70 (m, 1H),
1.78–1.94 (m, 3H), 2.46 (t, JZ7.4 Hz, 2H), 4.28–4.34
(m, 2H), 4.36–4.47 (m, 3H), 4.52 (t, JZ5.6 Hz, 1H), 4.56
(d, JZ3.2 Hz, 1H), 4.63–4.72 (m, 2H), 5.27 (m, 1H), 5.58
(d, JZ3.6 Hz, 1H), 8.47 (d, JZ8.4 Hz, 1H). 13C NMR
(100 MHz, pyridine-d5) d 14.7, 23.4, 23.5, 26.6, 26.9, 30.1,
30.2, 30.3, 30.5, 32.6, 32.9, 34.8, 37.3, 51.9, 63.1, 69.1,
70.2, 71.5, 72.1, 72.9, 73.5, 77.2, 102.0, 173.7. HRMS
(MALDI-TOF, MCHC) calcd for C39H78NO9: 704.5677.
Found: 704.5663.
3.9.6. O-(a-D-Glucopyranosyl)-N-pentadecanoyl-L-serine
tetradecyl ester (54). TLC (MeOH/CHCl3Z1/7) Rf 0.33;
1H NMR (CD3OD, 400 MHz) d 4.83 (1H, d, JZ3.8 Hz,
H-1), 4.69 (1H, dd, JZ5.6, 4.4 Hz, H-20), 4.18 (2H, t, JZ
6.6 Hz, H-30), 3.99 (1H, dd, JZ10.8, 4.4 Hz, H-10), 3.95
(1H, dd, JZ10.8, 5.6 Hz, H-10), 3.84 (1H, dd, JZ11.8,
2.3 Hz, H-6), 3.71 (1H, dd, JZ11.8, 5.5 Hz, H-6), 3.64 (1H,
t, JZ9.3 Hz, H-3), 3.59 (1 H, ddd, JZ9.8, 5.3, 2.3 Hz, H-5),
3.43 (1H, dd, JZ9.7, 3.8 Hz, H-2), 3.33 (1H, t, JZ9.4 Hz,
H-4), 2.31 (2H, t, JZ7.2 Hz, H-50), 1.73–1.66 (4H, m, H-40,
60), 1.50–1.20 (44H, br), 0.94 (6H, t, CH3!2); 13C NMR
(C5D5N, 100 MHz) d 174.0, 171.7, 102.3, 75.6, 75.1, 74.2,
72.5, 70.2, 65.9, 63.2, 54.4, 36.7, 32.5, 30.4, 30.3, 30.3,
30.2, 30.2, 30.1, 30.0, 29.9, 29.3, 26.6, 26.5, 23.4, 14.7;
FAB-MS m/z 688.1 (MC C1); HRMS calcd. for
C38H74NO9 (MC C1) 688.5364, Found: 688.5389;
HRMS calcd. for C38H73NNaO9 (MCCNa) 710.5183,
Found: 710.5168.
3.9.2. (2S,3S,4R)-1-O-(a-L-Fucopyranosyl)-2-(N-hexa-
cosanoylamino)-1,3,4-octadecanetriol (27). 1H NMR
(CDCl3/MeOHZ1/1, 400 MHz): d 0.86 (t, 6H, JZ
6.8 Hz), 1.21 (d, 3H, JZ6.4 Hz), 1.25 (m, 72H), 2.19 (t,
2H, JZ7.6 Hz), 3.41 (dd, 1H, JZ3.6, 10.0 Hz), 3.46–3.53
(m, 1H), 3.59–3.62 (m, 1H), 3.65–3.76 (m, 2H), 3.72 (dd,
1H, JZ3.6, 6.8 Hz), 3.95–3.97 (m, 2H), 4.17–4.19 (m, 1H),
4.74 (d, 1H, JZ3.6 Hz). 13C NMR (CDCl3, 400 MHz): d
13.0, 15.1, 21.9, 25.3, 28.7, 28.7, 28.9, 28.9, 29.0, 31.2,
63.5, 65.7, 66.7, 68.2, 69.6, 69.9, 71.5, 71.6, 98.6, 173.8.
HRMS (MALDI-TOF, MCNaC) calcd for C50H99NO8Na:
864.7268, found 864.7252.
3.9.3. (2S,3S,4R)-1-O-(b-L-Fucopyranosyl)-2-(N-hexa-
cosanoylamino)-1,3,4-octadecanetriol (30). 1H NMR
(CDCl3/MeOHZ1/1, 400 MHz) d 0.87 (t, 6H, JZ6.8 Hz),
1.25–1.28 (m, 75H), 2.19 (t, 2H, JZ7.6 Hz), 3.45–3.49 (m,
3H), 3.56–3.63 (m, 2H), 3.67 (dd, 1H, JZ5.2, 7.2 Hz),
3.87–3.88 (m, 2H), 4.07–4.11 (m, 1H), 4.23 (d, 1H, JZ
7.2 Hz). 13C NMR (CDCl3, 500 MHz) d 12.9, 15.1, 21.8,
25.1, 25.2, 28.5, 28.6, 28.6, 28.7, 28.8, 28.9, 28.9, 29.0,
31.1, 35.7, 49.8, 68.4, 70.2, 70.4, 71.0, 71.6, 73.1, 73.3,
102.9, 174.1. HRMS (MALDI-TOF, MCHC) calcd for
C50H100NO8: 842.7449, found 842.7440.
Supporting information available. Synthetic procedure,
characterization and NMR spectra of new compounds 4,
12–17, 19, 21, 24, 26, 29, 36–39, 41, 45–48, and 53.
Acknowledgements
We thank National Science Council for financial support
and a Program Project grant awarded by Academia Sinica.
3.9.4. Pentadecanoic acid [1-tetradecylcarbamoyl-2-(a-
D-galactopyranosyl)-ethylamide (40). 1H NMR
(400 MHz, pyridine-d5) d 0.87 (t, JZ6.6 Hz, 6H), 1.14–
1.48 (br, 44H), 1.56–1.68 (m, 2H), 1.72–1.85 (m, 2H), 2.47
References and notes
1. Porcelli, S. A. Adv. Immunol. 1995, 59, 1–98.