J. Rademann, N. W. J. Schrçder et al.
blingen, Germany). The MALDI-TOF mass spectra of the glycolipids
were measured on an AB 4700 MALDI-TOF/TOF (Applied Biosystems,
Carlsbad, CA, USA). After internal calibration with two standards
14), 56.4 (Cho C-17), 50.4 (Cho C-9), 42.5 (Cho C-13), 40.0 (Cho C-12),
39.8 (Cho C-24), 39.4 (Cho C-4), 37.5 (Cho C-1), 36.9 (Cho C-10), 36.5
(Cho C-22), 36.1 (Cho C-20), 32.21 (Cho C-7), 32.09 (Cho C-8), 30.3
(Cho C-2), 28.5 (Cho C-16), 28.3 (Cho C-25), 24.5 (Cho C-15), 24.2 (Cho
C-23), 23.0 (Cho C-27), 22.7 (Cho C-26), 21.3 (Cho C-11), 19.4 (Cho C-
(
717.405, 1479.012 Da) a device accuracy of 10 ppm could be achieved.
As matrices 2,5-dihydroxybenzoic acid and 2,4,6-trihydroxyacetophenone
were used and the given m/z is the mean of both signals.
19), 19.0 (Cho C-21), 12.0 ppm (Cho C-18); MALDI-MS: m/z: calcd for
+
C
33
H
56NaO
6
: 571.398 [M+Na] ; found: 571.385.
The molecular modeling was conducted by the Sybyl 7.0 package (Tripos
Inc., St. Louis, MO, 63144 USA).
Acetone oxime oleate (6): Oleic acid (4.81 mL, 15 mmol), acetone oxime
1.1 g, 15 mmol), EDC (3.59 g, 18.8 mmol), and DMAP (183 mg,
(
The NMR spectroscopic assignments of 1 and 2 and the full experimental
details for compounds 5, 7–12, 14–19, and 21 are supplied in the Support-
ing Information.
1
3
.5 mmol) were dissolved in dry CH
h. Silica gel was added and the solution evaporated. The crude product
2 2
Cl (40 mL) and stirred at RT for
was purified by flash chromatography (hexane/acetone 10:1) to give 6 as
a clear, colorless liquid (4.77 g, 94%). R =0.82 (toluene/acetone 3:1);
, 300 K, TMS): d=5.33 (t, J=4.4 Hz, 2H; FA
H-9, H-10), 2.40 (t, J=7.5 Hz, 2H; FA H-2), 2.04 (s, 3H; oxime H-trans),
[
29]
Cholesteryl 2,3,4,6-tetra-O-acetyl-b-d-galactopyranoside (3): Product 2
0.2 g, 0.4 mmol) and cholesterol (0.13 g, 0.34 mmol) were dissolved in
f
(
1
H NMR (300 MHz, CDCl
3
dry dichloromethane (4 mL) at RT and the reaction was started by the
addition of catalytic TMSOTf (5 mL, 30 mmol). After stirring for 1.5 h,
the reaction was quenched with triethylamine (5 mL). The solution was
evaporated and the crude product purified by column chromatography
2
2
1
3
1
.02 (4H; FA H-8, H-11), 1.99 (s, 3H; oxime H-cis), 1.69 (m, J=7.0 Hz,
H; FA H-3), 1.32 (2H; FA H-4), 1.27 (m, 18H; FA H-5–H-7, H-12–H-
1
3
7), 0.88 ppm (t, J=6.3 Hz, 3H; FA H-18); C NMR (75 MHz, CDCl
00 K, CDCl ): d=170.7 (FA C-1), 163.1 (oxime C-2), 129.6 (FA C-10),
29.3 (FA C-9), 32.6 (FA C-2), 31.6 (FA C-16), 29.42, 29.32, 29.18, 28.97
2C), 28.79, 28.76 (FA C-5–C-7, C-12–C-15), 28.72 (FA C-4), 26.86 (FA
3
,
(
(
hexane/ethyl acetate 1:1) to yield 3 as a colorless, crystal-like solid
1
3
0.185 g, 77%). R
, 300 K, TMS): d=5.37 (Cho H-6), 5.37 (s, 1H; Gal H-4), 5.17 (t,
2,3 =10.2 Hz, 1H; Gal H-2), 5.03 (dd, J3,4 =3.0 Hz, 1H; Gal H-3), 4.56 (d,
1,2 =7.8 Hz, 1H; Gal H-1b), 4.18 (m, J6b,5 =4.5 Hz, 1H; Gal H-6b), 4.12
f
=0.59 (hexane/ethyl acetate 3:1); H NMR (300 MHz,
CDCl
3
(
J
J
(
3
2
(
C-11), 26.80 (FA C-8), 24.6 (FA C-3), 22.3 (FA C-17), 21.6 (oxime C-
trans), 16.5 (oxime C-cis), 14.1 ppm (FA C-18); ESI-MS: m/z: calcd for
m, J6a,6b =11.1 Hz, 1H; Gal H-6a), 3.90 (t, J5,6a =6.6 Hz, 1H; Gal H-5),
.49 (Cho H-3), 2.22 (Cho H-4a), 2.18 (Cho H-4b), 2.15 (s, 3H; Ac4 H-
), 2.06 (s, 3H; Ac2 H-2), 2.04 (s, 3H; Ac6 H-2), 2.02 (Cho H-12b), 1.99
+
C
21
H
40NO
2
: 338.3 [M+H] ; found: 338.3.
[
33]
Cholesteryl 6-O-oleoyl-b-d-galactopyranoside (13, Figure 4):
CGal 4
(
2.5 g, 4.6 mmol), compound 6 (1.38 g, 4.1 mmol), lipase Amano PS (3 g),
s, 3H; Ac3 H-2), 1.97 (Cho H-7b), 1.88 (Cho H-2a), 1.86 (Cho H-1b),
and molecular sieve were suspended under argon in dry pyridine (24 mL)
and agitated at 508C for five days. Extra lipase (2.5 g) was added and the
suspension was stirred for an additional 5 days. The lipase was filtered off
and the solution supplied with silica gel evaporated. The yellow powder
was purified by column chromatography by stepwise elution with chloro-
form, chloroform/methanol 98:2, 95:5, 90:10, and 50:50 to provide prod-
1
1
1
1
1
2
2
.61 (Cho H-2b), 1.56 (Cho H-15a), 1.52 (Cho H-11a), 1.51 (Cho H-7a),
.51 (Cho H-25), 1.47 (Cho H-11b), 1.43 (Cho H-8), 1.36 (Cho H-20),
.32 (Cho H-23R), 1.31 (Cho H-22R), 1.27 (Cho H-16b), 1.17 (Cho H-
2a), 1.12 (Cho H-23S), 1.12 (Cho H-24), 1.08 (Cho H-17), 1.05 (Cho H-
a), 1.04 (Cho H-15b), 0.99 (Cho H-14), 0.99 (Cho H-19), 0.99 (Cho H-
2S), 0.92 (Cho H-9), 0.91 (Cho H-21), 0.88 (Cho H-27), 0.85 (Cho H-
1
3
uct 13 as a colorless, amorphous solid (0.53 g, 16%). R
f
=0.56 (chloro-
, 300 K, TMS): d=5.34
FA H-9, H-10), 5.33 (Cho H-6), 4.38 (Gal H-6b), 4.31 (Gal H-1b), 4.24
Gal H-6a), 3.90 (Gal H-4), 3.64 (Gal H-5), 3.62 (Gal H-2), 3.57 (Gal H-
3 3
6), 0.68 ppm (Cho H-18); C NMR (75 MHz, CDCl , 300 K, CDCl ):
1
form/methanol 85:15); H NMR (300 MHz, CDCl
3
d=170.6 (Ac6 C-1), 170.5 (Ac4 C-1), 170.3 (Ac3 C-1), 169.7 (Ac2 C-1),
39.9 (Cho C-5), 122.0 (Cho C-6), 99.9 (Gal C-1), 80.1 (Cho C-3), 70.9
Gal C-3), 70.3 (Gal C-5), 69.0 (Gal C-2), 67.0 (Gal C-4), 61.1 (Gal C-6),
6.5 (Cho C-14), 55.9 (Cho C-17), 49.9 (Cho C-9), 42.1 (Cho C-13), 39.5
Cho C-12), 39.2 (Cho C-24), 38.7 (Cho C-4), 36.9 (Cho C-1), 36.4 (Cho
C-10), 35.9 (Cho C-22), 35.5 (Cho C-20), 31.67 (Cho C-7), 31.61 (Cho C-
(
(
1
(
5
(
3), 3.53 (Cho H-3), 2.40 (Cho H-4a), 2.28 (FA H-2), 2.25 (Cho H-4b),
2.01 (Cho H-12b), 2.01 (FA H-8, H-11), 1.94 (Cho H-7b), 1.82 (Cho H-
1b), 1.59 (Cho H-2a), 1.59 (FA H-3), 1.52 (Cho H-25), 1.47 (Cho H-7a),
1.47 (Cho H-11b), 1.38 (Cho H-8), 1.37 (Cho H-23R), 1.36 (Cho H-20),
8
2
2
), 29.2 (Cho C-2), 27.9 (Cho C-16), 27.7 (Cho C-25), 24.0 (Cho C-15),
3.5 (Cho C-23), 22.4 (Cho C-27), 22.2 (Cho C-26), 20.8 (Cho C-11),
0.43 (Ac2 C-2), 20.26 (Ac6 C-2), 20.23 (Ac4 C-2), 20.18 (Ac3 C-2), 19.0
(
Cho C-19), 18.4 (Cho C-21), 11.5 ppm (Cho C-18); ESI-MS: m/z: calcd
+
for C41
Cholesteryl b-d-galactopyranoside (4):
0.1 mmol) was dissolved in dry methanol (100 mL). With 0.5m sodium
H64NaO10: 739.4 [M+Na] ; found: 739.4.
[
30]
Compound
3
(7.26 g,
1
methylate in methanol the pH value was adjusted to 9–10 and the solu-
tion was stirred at RT for 3 h. The solution was neutralized with amber-
+
lite IR-120(H ), filtered, and evaporated to give the crude product 4 as a
colorless solid (5.48 g, 99%). After further purification by flash chroma-
tography (chloroform/methanol 88:12), product 4 (2.99 g, 54%) was ob-
1
tained.
R
f
=0.18 (chloroform/methanol 85:15); H NMR (300 MHz,
[
D
5
]pyridine, 300 K, TMS): d=6.39 (s, 4H; Gal OH), 5.35 (d, J6,7a
=
4
3
J
.2 Hz, 1H; Cho H-6), 4.94 (d, J1,2 =7.6 Hz, 1H; Gal H-1b), 4.58 (d, J4,5
.0 Hz, 1H; Gal H-4), 4.46 (m, J6b,5 =4.8 Hz, 1H; Gal H-6b), 4.44 (t,
2,3 =10.6 Hz, 1H; Gal H-2), 4.42 (m, J6a,6b =10.2 Hz, 1H; Gal H-6a), 4.19
=
(dd, J3,4 =3.2 Hz, 1H; Gal H-3), 4.08 (t, J5,6a =5.9 Hz, 1H; Gal H-5), 3.95
(Cho H-3), 2.71 (Cho H-4a), 2.44 (Cho H-4b), 2.14 (Cho H-2a), 1.98
(Cho H-12b), 1.92 (Cho H-7b), 1.81 (Cho H-16a), 1.73 (Cho H-1b), 1.73
(Cho H-2b), 1.56 (Cho H-15a), 1.54 (Cho H-25), 1.54 (Cho H-7a), 1.47
(Cho H-11a), 1.42 (Cho H-20), 1.40 (Cho H-11b), 1.39 (Cho H-23R),
1
1
1
0
.38 (Cho H-22R), 1.37 (Cho H-8), 1.26 (Cho H-16b), 1.19 (Cho H-23S),
.17 (Cho H-24), 1.09 (Cho H-15b), 1.09 (Cho H-12a), 1.08 (Cho H-17),
.07 (Cho H-22S), 0.99 (Cho H-1a), 0.94 (Cho H-14), 0.94 (Cho H-19),
.91 (Cho H-9), 0.91 (Cho H-26), 0.90 (Cho H-27), 0.67 ppm (Cho H-18);
Figure 4. Structural depiction of ACGal C18:1: Representation as
a
capped sticks (top) and a space filled model (bottom). Built and mini-
mized by using Sybyl 7.0. First Steepest descent then a conjugate gradient
1
3
C NMR (75 MHz, [D
5 5
]pyridine, 300 K, [D ]pyridine): d=141.0 (Cho C-
À1
À1
5
), 121.9 (Cho C-6), 103.2 (Gal C-1), 78.0 (Cho C-3), 76.9 (Gal C-5), 75.4
until a gradient of 0.05 kcalmol
ꢄ
) was reached for each routine.
(
Gal C-3), 72.7 (Gal C-2), 70.3 (Gal C-4), 62.5 (Gal C-6), 56.9 (Cho C-
3542
ꢃ 2010 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
Chem. Eur. J. 2010, 16, 3536 – 3544