4
A.M. Tomkiel et al. / Steroids xxx (2016) xxx–xxx
ꢁ
1
1
0
0
(
(
AcOEt); IR,
m
max (cm ): 3572, 3436, 1133, 1052; H NMR, d: 5.37
H-4 ), 3.42 (m, 4H, –OCH
3
and H-2 ), 3.37 (m, 1H, H-3a), 3.21 (m,
0
m, 1H, H-6), 4.76 (d, 1H, J = 2.7 Hz, H-1 ), 3.88 (dd, 1H, J = 11.7 Hz,
J = 3.6 Hz, H-6a ), 3.81 (dd, 1H, J = 11.7 Hz, J = 4.6 Hz, H-6b ), 3.67
m, 1H, H-5 ), 3.59 (m, 1H, H-3
.45 (m, 1H, H-4 ), 3.44 (s, 3H, –OCH
1H, H-3 ), 1.00 (s, 9H, H-19), 0.92 (d, 9H, J = 6.5 Hz, H-21), 0.877
(d, 9H, J = 6.6 Hz, H-26 or H-27), 0.872 (d, 9H, J = 6.6 Hz, H-26 or
H-27), 0.683 (s, 6H, H-18), 0.677 (s, 3H, H-18); C NMR (ppm),
d: 140.9 (C), 140.6 (C), 140.2 (C), 122.1 (CH), 122.0 (CH), 121.7
a
0
0
0
0
0
13
(
3
a
), 3.54 (m, 2H, H-2 and H-3 ),
0
3
), 1.01 (s, 3H, H-19), 0.92
(
0
(
d, 3H, J = 6.5 Hz, H-21), 0.88 (d, 3H, J = 6.6 Hz, H-26 or H-27),
(CH), 99.6 (CH), 80.5 (CH), 80.1 (CH), 80.0 (CH), 78.6 (CH), 78.2
.87 (d, 3H, J = 6.6 Hz, H-26 or H-27), 0.68 (s, 3H, H-18); 1 C NMR
3
(CH), 70.7 (CH), 69.9 (CH), 67.7 (CH
(CH), 55.2 (CH ), 50.22 (CH), 50.17 (CH), 50.14 (CH), 42.3 (C),
39.8 (CH ), 39.7 (CH ), 39.6 (CH ), 39.5 (CH ), 39.1 (CH ), 37.6
(CH ), 37.3 (CH ), 37.2 (CH ), 37.1 (CH ), 36.9 (C), 36.74 (C),
36.72 (C), 36.2 (CH ), 35.8 (CH), 31.93 (CH ), 31.89 (CH), 30.0
(CH ), 29.4 (CH ), 28.9 (CH ), 28.2 (CH ), 28.0 (CH), 27.1 (CH ),
24.3 (CH ), 23.8 (CH ), 22.8 (CH ), 22.6 (CH ), 21.1 (CH ), 19.42
(CH ), 19.38 (CH ), 19.34 (CH ), 18.7 (CH ), 11.9 (CH ); ESI MS,
m/z: 1322 [(M+Na) , 22%]; HRMS (ESI): m/z calcd for C88
322.1017, found 1322.1004.
2
), 56.8 (CH), 56.7 (CH), 56.2
ppm), d: 140.6 (C), 122.0 (CH), 99.7 (CH), 80.7 (CH), 80.4 (CH),
3
7
5
3
2
2.8 (CH), 71.0 (CH), 70.2 (CH), 62.7 (CH
5.3 (CH ), 50.2 (CH), 42.3 (C), 39.9 (CH ), 39.7 (CH
7.3 (CH ), 36.7 (C), 36.2 (CH ), 35.8 (CH), 31.9 (CH
9.4 (CH ), 28.2 (CH ), 28.0 (CH), 24.3 (CH
), 22.5 (CH ), 21.0 (CH ), 19.4 (CH
ESI MS, m/z: 369 [(cholest-5-en-3-yl) , 8%], 585 [(M+Na) , 100%];
2
), 56.7 (CH), 56.1 (CH),
), 39.5 (CH ),
), 31.8 (CH),
2
2
2
2
2
3
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
), 23.8 (CH
2
), 22.8
);
2
2
2
2
2
(
CH
3
3
2
3
), 18.7 (CH
3
), 11.8 (CH
3
2
2
3
3
2
+
+
3
3
3
3
3
+
+
+
HRMS (ESI): m/z calcd for C34
H
58
O
6
Na : 585.4131, found 585.4124.
146 6
H O Na :
1
0
0
2
.3.11. 2 ,3 -Di-(O-cholest-5-en-3b-yl)-methyl-
18)
Compound 18 was eluted with a hexane-ethyl acetate 7:3 mix-
ture. Colorless crystals; m.p. 102–105 °C (AcOEt-hexane); Rf = 0.47
a-D-glucopyranoside
0
0
0
(
2.3.14. 2 ,4 ,6 -Tri-(O-cholest-5-en-3b-yl)-methyl-
a-D-glucopyra-
noside (21)
Compound 21 was eluted with a hexane-ethyl acetate 92:8
ꢁ
1
1
(
AcOEt-hexane 1:1); IR,
m
max (cm ): 3566, 3481, 1052; H NMR, d:
mixture. Colorless crystals, m.p. 55–57 °C (AcOEt-hexane);
0
ꢁ1
5
.35 (m, 2H, H-6), 4.72 (d, 1H, J = 3.3 Hz, H-1 ), 3.87 (dd, 1H,
Rf = 0.50 (AcOEt-hexane 2:8); IR,
m
max (cm ): 3586, 1067, 1021;
0
1
J = 11.7 Hz, J = 3.4 Hz, H-6a ), 3.81 (dd, 1H, J = 11.7 Hz, J = 4.6 Hz,
H NMR, d: 5.36 (m, 2H, H-6), 5.33 (m, 1H, H-6), 4.80 (d, 1H,
0
0
0
0
0
0
H-6b ), 3.68 (m, 3H, H-3 , H-4 and H-3
a
), 3.43 (s, 3H, –OCH
), 1.00 (s, 6H, H-19), 0.92 (d, 6H,
J = 6.5 Hz, H-21), 0.873 (d, 6H, J = 6.6 Hz, H-26 or H-27), 0.869 (d,
H, J = 6.6 Hz, H-26 or H-27), 0.68 (s, 6H, H-18); 13C NMR (ppm),
d: 140.4 (C), 139.9 (C), 122.3 (CH), 122.1 (CH), 99.6 (CH), 80.5
3
),
J = 3.1 Hz, H-1 ), 3.85 (t, 1H, J = 8.8 Hz, H-3 ), 3.77 (m, 1H, H-6a ),
0
0
0
0
0
0
3
.40 (m, 3H, H-2 , H-5 and H-3
a
3.60 (m, 3H, H-4 , H-5 and H-6b ), 3.42 (m, 2H, H-2 and H-3
a),
3.39 (s, 3H, –OCH ), 3.33 (m, 1H, H-3 ), 3.16 (m, 1H, H-3 ), 1.00
3
a
a
6
(s, 3H, H-19), 0.99 (s, 6H, H-19), 0.92 (d, 9H, J = 6.3 Hz, H-21),
0.871 (d, 9H, J = 6.6 Hz, H-26 or H-27), 0.868 (d, 9H, J = 6.6 Hz, H-
26 or H-27), 0.68 (s, 3H, H-18), 0.67 (s, 6H, H-18); 13C NMR
(ppm), d: 140.9 (C), 140.8 (C), 140.6 (C), 121.9 (CH), 121.7 (CH),
98.6 (CH), 80.3 (CH), 80.2 (CH), 79.9 (CH), 78.3 (CH), 74.9 (CH),
(
(
5
3
(
2
CH), 80.0 (CH), 78.8 (CH), 78.0 (CH), 70.6 (CH), 70.3 (CH), 62.7
CH
2
), 56.71 (CH), 56.66 (CH), 56.12 (CH), 56.08 (CH), 55.2 (CH
0.2 (CH), 50.1 (CH), 42.3 (C), 40.3 (CH ), 39.7 (CH ), 39.5 (CH
7.5 (CH ), 37.2 (CH ), 36.70 (C), 36.66 (C), 36.6 (CH
), 35.8 (CH), 31.9 (CH ), 31.8 (CH), 28.9 (CH ), 28.2 (CH
8.0 (CH), 24.3 (CH ), 23.8 (CH ), 22.8 (CH ), 22.6 (CH ), 21.02
), 21.00 (CH ), 19.4 (CH ), 19.3 (CH ), 18.7 (CH ), 11.8 (CH );
ESI MS, m/z: 953 [(M+Na) , 100%]; HRMS (ESI): m/z calcd for C61
3
),
2
2
2
),
2
2
2
), 36.2
),
72.7 (CH), 70.0 (CH), 66.6 (CH
(CH), 56.2 (CH), 56.09 (CH), 56.06 (CH), 55.1 (CH
50.1 (CH), 50.0 (CH), 42.3 (C), 40.3 (CH ), 39.81 (CH
(CH ), 39.68 (CH ), 39.5 (CH ), 38.7 (CH ), 37.6 (CH ), 37.4 (CH
37.1 (CH ), 36.9 (C), 36.68 (C), 36.66 (C), 36.1 (CH ), 35.8 (CH),
31.9 (CH ), 31.8 (CH), 29.4 (CH ), 29.1 (CH ), 28.7 (CH ), 28.5
CH ), 28.22 (CH ), 28.16 (CH ), 28.0 (CH), 24.3 (CH ), 23.9 (CH
23.8 (CH ), 22.8 (CH ), 22.6 (CH ), 21.04 (CH ), 21.00 (CH
19.40 (CH ), 19.38 (CH ), 19.34 (CH ), 18.7 (CH ), 11.8 (CH
MS, m/z: 1322 [(M+Na) , 100%]; HRMS (ESI): m/z calcd for
2
), 56.78 (CH), 56.75 (CH), 56.68
), 50.3 (CH),
), 39.77
),
CH
2
2
2
2
3
2
2
3
3
2
2
(
CH
2
2
3
3
3
3
2
2
2
2
2
2
+
-
2
2
+
H
102
O
6
Na : 953.7574, found 953.7585.
2
2
2
2
(
2
2
2
2
2
),
),
0
0
2
.3.12. 2 ,6 -Di-(O-cholest-5-en-3b-yl)-methyl-
19)
Compound 19 was eluted with a hexane-ethyl acetate 4:1 mix-
ture. Colorless crystals, m.p. 168–170 °C (AcOEt-hexane); Rf = 0.51
a-
D-glucopyranoside
2
3
3
2
2
(
3
3
3
3
3
); ESI
+
+
C H
88 146
O
6
Na : 1322.1017, found 1299.1002.
ꢁ
1
1
(
AcOEt-hexane 3:7); IR,
mmax (cm ): 3567, 3480, 1054; H NMR, d:
0
0
0
0
0
0
5
.36 (m, 2H, H-6), 4.78 (d, 1H, J = 1.6 Hz, H-1 ), 3.73 (m, 3H, H-3 ,
2.3.15. 2 ,3 ,4 ,6 -Tetra-(O-cholest-5-en-3b-yl)-methyl-
a-D-glucopyra-
0
0
0
0
H-6a and H-6b ), 3.60 (m, 1H, H-3
H-5 ), 3.43 (s, 3H, –OCH
0
a
), 3.55 (m, 3H, H-2 , H-4 and
noside (22)
0
3
), 3.22 (m, 1H, H-3
a
), 1.01 (s, 3H, H-19),
Compound 22 was eluted with a hexane-ethyl acetate 97:3
mixture. Colorless crystals, m.p. 149–152 °C (AcOEt-hexane);
.99 (s, 3H, H-19), 0.92 (d, 6H, J = 6.5 Hz, H-21), 0.871 (d, 6H,
m H
max (cm ): 1073, 1019; 1
ꢁ
1
J = 6.6 Hz, H-26 or H-27), 0.867 (d, 6H, J = 6.6 Hz, H-26 or H-27),
Rf = 0.50 (AcOEt-hexane 1:9); IR,
1
3
0
(
(
.68 (s, 6H, H-18); C NMR (ppm), d: 140.8 (C), 140.6 (C), 121.8
CH), 99.5 (CH), 80.7 (CH), 80.13 (CH), 80.10 (CH), 72.3 (CH), 71.5
CH), 69.8 (CH), 68.2 (CH ), 56.7 (CH), 56.1 (CH), 55.2 (CH ),
0.11 (CH), 50.09 (CH), 42.3 (C), 39.8 (CH ), 39.7 (CH ), 39.5
), 36.8 (C), 36.7 (C), 36.1
), 31.8 (CH), 29.4 (CH ), 29.3 (CH ),
), 28.0 (CH), 24.3 (CH ), 23.8 (CH ), 22.8
), 21.0 (CH ), 19.40 (CH ), 19.36 (CH ), 18.68
); ESI MS, m/z: 953 [(M+Na) , 100%]; HRMS (ESI):
NMR, d: 5.34 (m, 3H, H-6), 5.31 (m, 1H, H-6), 4.73 (d, 1H,
0
0
J = 3.3 Hz, H-1 ), 3.76 (dd, 1H, J = 9.2 Hz, J = 6.3 Hz, H-4 ), 3.74 (t,
0
0
2
3
1H, J = 9.2 Hz, H-3 ), 3.69 (m, 1H, H-3
a
), 3.59 (m, 2H, H-6a and
), 3.40 (dd, 1H, J = 9.2 Hz,
and H-3 ), 3.15 (m, 1H, H-
0
0
5
2
2
H-6b ), 3.52 (m, 2H, H-5 and H-3
a
0
(
(
CH
CH
2
), 39.0 (CH
), 35.8 (CH), 31.9 (CH
), 28.17 (CH
), 22.6 (CH
), 11.8 (CH
2
), 37.3 (CH
2
), 37.1 (CH
2
J = 3.3 Hz, H-2 ), 3.38 (s, 4H, –OCH
3
a
2
2
2
2
3a), 1.002 (s, 3H, H-19), 0.995 (s, 3H, H-19), 0.989 (s, 3H, H-19),
2
(
(
8.21 (CH
2
2
2
2
0.987 (s, 3H, H-19), 0.93 (d, 6H, J = 6.4 Hz, H-21), 0.92 (d, 6H,
J = 6.4 Hz, H-21), 0.879 (d, 12H, J = 6.6 Hz, H-26 or H-27), 0.874
(d, 12H, J = 6.6 Hz, H-26 or H-27), 0.691 (s, 3H, H-18), 0.685 (s,
CH
CH
3
3
2
3
3
+
3
3
+
13
m/z calcd for C61
102
H O
6
Na : 953.7574, found 953.7583.
3H, H-18), 0.681 (s, 6H, H-18); C NMR (ppm), d: 141.2 (C),
1
41.0 (C), 140.8 (C), 140.6 (C), 121.9 (CH), 121.7 (CH), 121.6 (CH),
121.5 (CH), 99.7 (CH), 80.74 (CH), 80.73 (CH), 80.0 (CH), 79.76
(CH), 79.5 (CH), 79.0 (CH), 73.4 (CH), 70.4 (CH), 66.6 (CH ), 56.80
(CH), 56.77 (CH), 56.74 (CH), 56.71 (CH), 56.25 (CH), 56.17 (CH),
56.14 (CH), 56.11 (CH), 54.9 (CH ), 50.4 (CH), 50.22 (CH), 50.20
(CH), 50.1 (CH), 42.33 (C), 42.31 (C), 40.2 (CH
39.81 (CH ), 39.5 (CH ), 39.4 (CH ), 37.49 (CH
37.42 (CH ), 37.40 (CH ), 37.38 (CH ), 37.35 (CH
0
0
0
2
.3.13. 2 ,3 ,6 -Tri-(O-cholest-5-en-3b-yl)-methyl-
a-D-glucopyra-
noside (20)
2
Compound 20 was eluted with a hexane-ethyl acetate 95:5
mixture. Colorless crystals, m.p. 94–97 °C (AcOEt-hexane);
3
ꢁ
1
Rf = 0.44 (AcOEt-hexane 1:9); IR,
m
max (cm ): 3567, 1214, 1060;
2
), 39.79 (CH
), 37.45 (CH
), 37.1 (CH
2
),
2
),
2
),
1
0
H NMR, d: 5.34 (m, 3H, H-6), 4.74 (d, 1H, J = 3.4 Hz, H-1 ), 3.70
2
2
2
2
0
0
0
0
(
m, 5H, H-3 , H-5 , H-6a , H-6b and H-3
a
), 3.53 (t, 1H, J = 9.4 Hz,
2
2
2
2