7
20
Journal of Pharmacy and Pharmacology 2011; 63: 718–724
+
at room temperature for 20 min, and after removal of the 56.36, 67.98, 72.24, 75.12, 178.6; HRMS [M-Na] =
acetone, water was added. The aqueous mixture was extracted 655.72304 (C37
with EtOAc (3 ¥ 10 ml) and the extract was worked up to give
H
64
O Si = 632.45623432).
6
an amorphous solid that was fractionated by silica gel flash
3
(
b,16a,23-Trihydroxyolean-12-en-28-oic acid
16a-hydroxyhederagenin, 7)
We obtained compound 7 under the same conditions used to
obtain compound 3. NaBH (1.15 mg, 0.03 mmol) was added
CC (CH
2
Cl
2
-EtOAc 2 : 1) to give 4b (25 mg, 25%), R
F
= 0.81
(
(
CH
CH
2
Cl
Cl
2
-EtOAc 2 : 1), and 4a (10 mg, 10%), R
F
= 0.17
2
2
-EtOAc 1 : 1). 4a: mp 180–190°C; IR (KBr) 3447,
-
1
1
4
2
0
2
925, 2854, 2361, 1712 cm ; H-NMR (CDCl , 500 MHz) d
3
to a solution of 1 (10 mg, 0.02 mmol) in MeOH (1.0 ml) at 0°C.
The reaction mixture was stirred for 2 h at 0°C.Then cold water
.72, 0.90, 0.97, 1.25, 1.34 (3H each, s, Me-26, 30, 29, 27,
5), 2.04 (1H, s, 24-H), 2.71 (1H, d, J = 15.2 Hz, 19-H), 3.35
(
10 ml) was added, and the mixture was worked up and the
solid residue was purified by silica gel flash CC (CH Cl
EtOAc 2 : 1) to give 7 (8 mg), R = 0.50, mp 200–204°C; IR
KBr) 3420, 2947, 2370, 1715, 1654 cm ; H-NMR (DMSO-
, 400 MHz) 5.50 (1H, t, H-12), 4.45 (H-16), 4.03 (1H, dd,
H-3), 3.50 (d, CH ), 3.30 (d, CH J = 11.0 Hz), 3.00 (1H, dd,
H-18), 0.70–1.3 (6 signals, 3H each, s).
(
1H, dd, J = 3.8, 14.4 Hz, 18-H), 3.61 (3H, s, CH ), 4.30 (1H,
3
2
2
-
t, 16-H), 4.51 (1H, s, 3-H), 5.38 (1H, t, J = 3.3 Hz, 12-H);
1
3
F
C-NMR (CDCl
3
, 126 MHz) d 51.92, 72.24, 74.96, 175.5,
-1 1
(
1
2
5
2
77.4, 122.7, 142.7. 4b: mp 240–246°C; IR (KBr) 3411,
956, 2924, 2855, 1719, 1715 cm ; H-NMR (CDCl ,
00 MHz) d 0.87, 0.92, 0.93, 1.17, 1.19 (3H each, s, Me-30,
5, 29, 26, 27), 1.00 (3H, d, J = 6.4 Hz, 24-H), 2.71 (1H, d,
-
1
1
d
6
3
2
2
J = 15.2 Hz, 19-H), 3.33 (1H, dd, J = 3.8; 14.4 Hz, 18-H),
1
3
3
(
.67 (3H, s, CH
CDCl
, 126 MHz) d 52.44, 124.1, 140.5, 173.3, 208.6, 213.6. 16a-dihydroxyolean-12-en-28-oic acid (8)
Compound (100 mg, 0.205 mmol), 1-amino-4-
methylpiperazine (61.2 mg, 0.6 mmol), absolute ethanol
43 ml) and glacial acetic acid (1.7 ml) were heated under
3
), 5.56 (1H, t, J = 3.3 Hz, 12-H); C-NMR 23-(1-Amino-4-methylpiperazinyl) imino-3b,
3
1
Methyl 3b,16a-dihydroxy-23-
t-butyldimethylsilyloxy)-olean-12-en-
8-oate (5)
A mixture of 3 (100 mg), tert-butyldimethysilyl chloride
96 mg, 1.3 mmol) and imidazole (170 mg, 2.5 mmol) in
anhydrous DMF (0.5 ml) was stirred at room temperature
under N for 18 h. Then, hexane (30 ml) was added, and the
solution was washed successively with brine (2 ¥ 10 ml) and
distilled water (20 ml), and dried with Na SO . After evapo-
ration of the solvent, the residue was purified by silica gel CC
(
(
2
reflux for 24 h. The reaction mixture was concentrated to
dryness and the residue was purified by silica gel CC eluting
with CH
2
Cl -EtOAc (1 : 1) to give 8 (35 mg, 35% yield) as a
2
(
yellow powder; mp 200–210°C; IR (KBr) 3424, 2944, 2862,
-
1 1
2
0
2
361, 1610 cm ; H-NMR (CD
.94, 0.95, 0.97, 1.31 (3H each, s, Me-30, 25, 29, 24, 26, 27),
.41 (3H, s, N-CH ), 2.63 (4H, m, -CH -N-CH ), 3.04 (4H, m,
-N-CH -), 3.27 (3H, s, CH ), 3.52 (1H, t, J = 5.1 Hz,
3
OD, 400 MHz) d 0.77, 0.83,
2
3
2
3
2
4
-
CH
2
2
3
H-3), 3.67 (1H, dd, J = 3.8; 14.2 Hz, H-18), 4.40 (1H, s,
(
(
2
0
hexane-EtOAc, 90 : 10) to give 5 (60 mg, 63%), R
CH Cl -EtOAc 2 : 1); mp 290–294°C; IR (KBr) 3542, 3467,
951, 2927, 2856, 1710 cm ; H-NMR (CDCl
.05 (9H, s, Si-(CH ), 0.72, 0.88, 0.90, 0.96, 1.33 (3H each,
F
= 0.80
H-16), 5.27 (1H, t, J = 3.5 Hz, H-12), 6.70 (1H, s, H-23);
2
2
1
3
-
1
1
C-NMR (CD OD, 100 MHz) d 11.09, 14.18, 17.03, 19.49,
3
3
, 400 MHz) d
2
1
2.35, 50.70, 51.76, 53.47, 74.25, 75.32, 121.4, 144.2, 152.6,
79.4. HREIMS calculated for C35 : 583.4303220;
)
3 3
H
57
N
3
O
4
s, Me-30, 25, 29, 26, 27), 3.03 (1H, dd, J = 3.8, 14.4 Hz
8-H), 3.33 (1H, dd, 2H, -CH O), 3.5 (1H, t, 3-H), 3.70 (3H,
s, CH ), 4.50 (1H, s, 16-H), 5.40 (1H, t, J = 3.3, 12-H); C-
NMR (CDCl , 126 MHz) d 11.20, 16.04, 16.98, 23.40, 24.80, Quillaic acid bis-acetate (9)
7.05, 52.91, 56.45, 68.85, 72.79, 75.15, 123.7, 145.2, 178.0. Compound 1 (100 mg, 0,21 mmol) was treated with Ac
found: 583.4300958.
1
2
1
3
3
3
2
2
O
+
HRMS [M-Na] = 639.31305 (C37
H
64
O
5
Si = 616.45230430).
(0.5 ml, 0.41 mmol) in dry pyridine (1.5 ml) in the presence
of 4-dimethylaminopyridine (25 mg) at room temperature for
2 h. The mixture was worked up to give an amorphous solid,
which was purified by silica gel column chromatography
eluting with CH Cl -EtOAc (2 : 1) to give 9 (96 mg, 80%
yield); R = 0.85 (CH Cl -EtOAc 2 : 1); mp 180–186°C; IR
KBr) 3431, 2950, 2861, 1736, 1656 cm ; H-NMR (CDCl
1
Methyl
3
1
b,16a-dihydroxy-23-(t-butyldimethylsilyl)-12,
2
2
3-epoxyolean-28-oate (6)
F
2
2
A mixture of 5 (50 mg, 0.06 mmol), and m-chloroperoxy
benzoic acid (mCPBA) (60%) (43 mg, 0.5 mmol) in CH Cl
3 ml) was stirred at room temperature overnight. Then the
mixture was diluted with CH Cl -Et O (1 : 2) and was worked
-1 1
(
3
,
2
2
Pyr-d , 400 MHz) d 0.70, 0.85, 0.94, 1.00, 1.07, 1.27 (3H
5
(
each, s, Me-30, 25, 29, 24, 26, 27), 2.07 (6H, s, acetoxy), 3.05
1H, dd, J = 3.6; 14.2 Hz, 18-H), 4.51(1H, s, 3-H), 5.39 (1H,
s, 16-H), 5.64 (1H, t, J = 3.4 Hz, 12-H); 9.23 (1H, s, 23-H);
2
2
2
(
up to give a product (65 mg) that was subjected to preparative
TLC (hexanes-EtOAc 1 : 5 : 1) to give epoxide 6 as a crystal-
line solid. An analytically pure, colourless sample was
1
3
C-NMR (CDCl
3
, Pyr-d , 100 MHz) d 9.48, 15.65, 16.93,
5
2
1
0.38, 21.96, 23.24, 73.27, 76.25, 123.1, 142.1, 170.0, 170.4,
80.1, 204.5. HRMS [M-Na] = 593.3448750 (C34
obtained by recrystallization from MeOH, R
EtOAc 2 : 1), mp 295–297°C; IR (KBr) 3411, 2948, 2862,
F
= 0.60 (CH
2
Cl -
2
+
H
50
O =
7
-
1
1
570.367643025).
1
0
(
(
701, 1627 cm ; H-NMR (DMSO-d
.88, 0.92, 0.96, 1.22 (3H each, s, Me-30, 25, 29, 26, 27), 3.42
O), 4.03 (1H, dd, J = 3.8; 14.4 Hz, 18-H),4.34 Quillaic acid bis-hemisuccinate (10)
6
, 400 MHz) d 0.55,
2H, dd, -CH
1H, s, 16-H), 5.21 (1H, brt, 12-H); C-NMR (CDCl
26 MHz) d 11.45, 16.09, 16.18, 23.35, 26.57, 52.90, 55.47, with succinic anhydride (200 mg, 2.03 mmol) and 25 mg of
2
13
3
,
A mixture of 1 (100 mg, 0.21 mmol) and pyridine (138 ml)
1