372
S. Hanashima et al. / Bioorg. Med. Chem. 9 (2001) 367±376
3-O-(6-Deoxy-6-sulfo-ꢀ-D-glucopyranosyl)-1-O-myris-
toyl-glycerol (12). To a solution of 10 (87.2 mg,
0.110 mmol) in ethanol (20 mL) was added 10% Pd±C
(323 mg), and the resulting mixture was hydrogenated
for 16 h with stirring under hydrogen atmosphere. The
catalyst was removed by ®ltration and the ®ltrate
was concentrated to dryness. The residue was puri®ed
by ¯ash chromatography (SiO2, 65:25:4 chloro-
3.75 (1H), 3.68 (1H), 3.51 (dd, 1H, J=9.7 and 3.5 Hz),
3.52 (1H), 3.38 (1H), 3.32 (t, 1H, J=9.2 Hz), 3.04 (dd,
1H, J=13.7 and 7.5 Hz), 2.35 (t, 2H, OCOCH2), 2.32 (s,
3H, SCOCH3), 1.63 (t, 2H, J=6.9 Hz, OCOCCH2), 1.25
(br, 24H, CH2), 0.88 (t, 3H, J=6.5 Hz, CH3); 13C NMR
(75 MHz, CDCl3) d 194.8, 173.8, 138.5, 137.8, 137.8,
128.6±127.6 (overlapped), 98.17, 97.75, 81.67, 80.37,
80.00, 97.92, 77.2, 75.69, 75.26, 73.58, 73.46, 70.23,
69.87, 69.9, 68.95, 68.45, 65.07, 64.88, 34.1, 31.9, 30.8,
30.5, 29.7±29.1 (overlapped), 24.9, 22.7, 14.1.
form:methanol:water) yielded SQMG 12 (50.1mg,
20
0.0946 mmol, 86.8%): ꢀ +57.2 (c 0.50, chloro-
D
form:methanol:water 65:25:4); 1H NMR (300 MHz,
CDCl3, CD3OD, D2O) d 4.47 (m, 1H), 3.91 (m, 1H),
3.77 (m, 3H), 3.30 (m, 2H), 3.09 (m, 2H), 3.01(m), 2.81
(t, 1H, J=9.5 Hz), 2.66 (dd, 1H, J=14.1 and 5.7 Hz),
2.04 (m, 2H, OCOCH2), 1.31 (m, 2H, OCOCCH2), 0.96
(br, 20H, CH2), 0.60 (t, 3H, J=6.4 Hz, CH3);
LRFABMS m/z 527.2 [MÀH]À; HRFABMS calcd
C23H43O11S [MÀH]À 527.2604, found 527.2632.
3-O-(2,3,4-tri-O-Benzyl-6-deoxy-6-sulfo-ꢀ-D-glucopyra-
nosyl)-1,2-di-O-palmitoyl-glycerol sodium salt (15). To a
solution of 13 (133 mg, 0.125 mmol) in glacial acetic acid
(7.0 mL) were added potassium acetate (814 mg) and
OXONE (2KHSO5, KHSO4, K2SO4) (228 mg,
0.371mmol) with stirring. After 16 h at room tempera-
ture, the resulting mixture was diluted with cold water
(20 mL). The aqueous phase was extracted with ethyl
acetate (5Â20 mL), and the combined organic layers
were washed with saturated NaHCO3 solution
(2Â40 mL) and brine (2Â40 mL), dried over anhydrous
Na2SO4, and concentrated under reduced pressure.
Puri®cation by ¯ash chromatography (SiO2, 10:1
3-O-(2,3,4-tri-O-Benzyl-6-deoxy-6-thioacetyl-ꢀ-D-gluco-
pyranosyl)-1,2-di-O-palmitoyl-glycerol (13) and 3-O-
(2,3,4-tri-O-benzyl-6-deoxy-6-thioacetyl-ꢀ-D-glucopyr-
anosyl) - 1-O-palmitoyl-glycerol (14). To a solution of
glycol 6 (20.3 mg, 0.0343 mmol) in dry dichloromethane
(5.0 mL) were added EDCI (19.4 mg, 0.101 mmol),
DMAP (5.7 mg, 0.047 mmol), and palmitic acid (14.1 mg,
0.0549 mmol) with stirring. After 16 h at room tem-
perature, the reaction mixture was diluted with water
(2 mL). The aqueous phase was extracted with di-
chloromethane (3Â5 mL). The combined organic layers
were washed with brine (2Â5 mL), dried over anhydrous
Na2SO4, and concentrated under reduced pressure.
Puri®cation by ¯ash chromatography (SiO2, 7:1±3:1
hexanes:ethyl acetate) yielded 13 and 14 respectively (13
dichloromethane:methanol) yielded 15 (57.9 mg,
20
D
CHCl3); IR (nuj) 1720, 1490, 1180, 1160±1135, 1070±
0.0543 mmol, 43.4%): mp 53±56 ꢀC; ꢀ +65.6 (c 0.34,
1040 cmÀ1 1H NMR (300 MHz, CDCl3) d 7.25±7.21
;
(overlapped, m, 15H, ArH), 5.25 (m, 1H), 4.94 (d, 1H,
J=11.7 Hz, ArCH2a), 4.90 (d, 1H, J=10.9 Hz,
ArCH2b), 4.81(d, 1H, J=11.0 Hz, ArCH2a), 4.70 (d,
1H, J=11.2 Hz, ArCH2b), 4.50 (2H), 4.58 (d, 1H,
J=3.7 Hz), 4.33±3.36 (overlapped, 8H), 3.25 (1H), 3.12
(1H), 2.27±2.18 (overlapped, m, 4H, OCOCH2), 1.53
(br, 4H, OCOCCH2), 1.25 (br, 48H, CH2), 0.88 (t, 6H,
J=6.0 Hz, CH3); LRFABMS m/z 1087.7 [M+H]+.
14.7 mg, 0.0139 mmol, 40.5%, 14 9.1 mg, 0.0111 mmol,
20
D
32.3%): 13 mp 51±56 ꢀC; ꢀ +11.3 (c 0.64, CHCl3),
IR (nuj) 1735, 1700, 1140, 1060 cmÀ1
;
1H NMR
(300 MHz, CDCl3) d 7.33±7.28 (overlapped, m, 15H),
5.2 (m 1H), 4.95 (d, 1H, J=10.8 Hz, ArCH2a), 4.88 (d,
1H, J=10.7 Hz, ArCH2b), 4.82 (d, 1H, J=9.8 Hz,
ArCH2a), 4.78 (d, 1H, J=10.8 Hz, ArCH2b), 4.70 (d,
1H, J=3.0 Hz, anomeric H), 4.60 (d, 2H, J=11.9 Hz,
ArCH2), 4.37 (1H), 4.21 (1H), 3.93 (t, 1H, J=9.3 Hz),
3.79 (1H), 3.76 (1H), 3.73 (1H), 3.50 (1H), 3.40 (1H),
3.30 (t, 1H, J=9.5 Hz), 3.02 (dd, 1H, J=13.8 and
6.4 Hz), 2.33 (s, 3H, SCOCH3), 2.34±2.27 (overlapped,
m, 4H, OCOCH2), 1.60 (m, 4H, OCOCCH2), 1.25 (br
48H CH2), 0.88 (t, 6H, J=6.4 Hz, CH3); 13C NMR
(75 MHz, CDCl3) d 194.8, 179.6, 173.4, 138.6, 138.2,
138.0, 128.6±127.6 (overlapped), 97.17, 96.85, 81.5, 80.3,
80.0, 80.2, 77.2, 75.7, 75.2, 72.98, 73.08, 69.69, 69.79,
66.0, 66.2, 62.4, 62.5, 34.3, 34.2, 34.0, 31.9, 30.8, 30.6,
3-O-(2,3,4-tri-O-Benzyl-6-deoxy-6-sulfo-ꢀ-D-glucopyra-
nosyl)-1-O-palmitoyl-glycerol sodium salt (16). To a
solution of 14 (52.1mg, 0.0635 mmol) in glacial acetic
acid (2.0 mL) were added potassium acetate (102 mg)
and OXONE (2KHSO5, KHSO4, K2SO4) (116 mg,
0.190 mmol) with stirring. After 16 h at room tempera-
ture, the resulting mixture was diluted with cold water
(5.0 mL). The aqueous phase was extracted with ethyl
acetate (5Â5 mL), and the combined layers were washed
with saturated NaHCO3 solution (2Â10 mL) and brine
(2Â10 mL), dried over anhydrous Na2SO4, and con-
centrated under reduced pressure. Puri®cation by ¯ash
chromatography (SiO2, 10:1 dichloromethane:methanol)
20
D
(c 0.30, CHCl3); IR (nuj) 3400, 1720, 1480, 1185, 1155,
yielded 16 (35.1mg, 0.0424 mmol, 66.8%): ꢀ +36.4
29.7±29.0 (overlapped), 24.9, 24.7, 22.7, 14.1;
1060 cmÀ1 1H NMR (300 MHz CDCl3) d 7.26±7.19
;
20
D
1.13, CHCl3); IR (nuj) 3430, 3050, 3000, 1720, 1680,
LRFABMS m/z 1059.7 [M+H]+; 14 ꢀ +47.0 (c
(overlapped, m, 15H, ArH), 4.91±4.54 (overlapped, 6H,
ArCH2), 4.71(1H, anomeric H), 4.28±3.36 (overlapped,
9H), 3.16 (1H), 2.85 (1H), 2.24 (t, 2H, OCOCH2), 1.50
(m, 2H, OCOCCH2), 1.24 (br, 24H, CH2), 0.87 (t, 3H,
CH3).
1580, 1565, 1480, 1140±1030, 915 cmÀ1
;
1H NMR
(300 MHz, CDCl3) d 7.35±7.27 (overlapped, m, 15H,
ArH), 4.95 (d, 1H, J=10.8 Hz, ArCH2a), 4.89 (d, 1H,
J=10.7 Hz, ArCH2b), 4.81(d, H1,
J=11.1 Hz,
ArCH2a), 4.77 (d, 1H, J=11.8 Hz, ArCH2b), 4.70 (d,
1H, J=3.7 Hz, anomeric H), 4.63 (d, 1H, J=10.2 Hz,
ArCH2a), 4.62 (d, 1H, J=10.7 Hz ArCH2b), 4.16 (m,
1H), 4.13 (m, 1H), 3.95 (t, 1H, J=9.2 Hz), 3.8 (m, 1H),
3-O-(6-Deoxy-6-sulfo-ꢀ-D-glucopyranosyl)-1,2-di-O-
palmitoyl-glycerol (17). To a solution of 15 (360 mg,
0.330 mmol) in ethanol (50 mL) was added 10% Pd±C
(1.3 g), and the resulting mixture was hydrogenated for