steroids 7 1 ( 2 0 0 6 ) 979–983
981
Table 2 – Transformation of I with the pH 7.0 medium
Compound
24 h
48 h
72 h
96 h
HPLC
content (%)
Isolated
yield (%)
HPLC
content (%)
Isolated
yield (%)
HPLC
content (%)
Isolated
yield (%)
HPLC
content (%)
Isolated
yield (%)
I
IV
V
VI
VII
80.6
18.7
0
0
–
78.2
15.4
0
0
0
0
0
99.2
0
0
0
91.3
0
0
0
84.6
14.2
–
0
0
82.3
8.0
0
0
0
82.5
15.9
–
0
0
74.6
8.7
6.0
a
–
0
A 24, 48, 72 and 96 h denote the reaction time.
a
Compound VII could not be detected with the UV detector of the HPLC.
2
5
◦
−1
2
3.6 (C-15), 18.7 (C-19), 12.7 (C-18); R in chloroform/methanol
f
[
3
˛] + 143 (MeOH, c = 1.0); UV ꢀ Mm ea Ox H = 241 nm; IR ꢁmax (cm ):
D
(
12:1): 0.57; Rt: 6.2 min.
430, 1734, 1664, 1609; FAB-MS (3-nitrophenyl methanol,
probe), m/z: 303 [M + H] ; 1H NMR (CDCl3): ı (ppm): 0.95 (3H,
+
s, H-18), 1.35 (3H, s, H-19), 4.08 (1H, ddd, J = 12, 11, 5 Hz, H-11),
2.4.5. 6ˇ,11˛,17ˇ-Trihydroxyandrost-ene-3-one (VI)
5
(
4
(
.76 (1H, s, H-4); 13C NMR (CDCl3): ı (ppm): 218.2 (C-17), 199.8
C-3), 169.9 (C-5), 124.9 (C-4), 68.8 (C-11), 59.3 (C-9), 50.2 (C-14),
8.0 (C-13), 43.1 (C-12), 40.1 (C-10), 37.5 (C-16), 35.7 (C-1), 34.7
C-2), 34.2 (C-8), 33.4 (C-6), 30.4 (C-7), 21.8 (C-15), 18.4 (C-19),
4.7 (C-18); R in chloroform/methanol (12:1): 0.72; Rt: 9.9 min.
Colorless crystals (crystallized from chloroform); mp
◦
25
◦
MeOH
235–236 C; [˛] + 34 (MeOH, c = 1.0); UV ꢀmax = 240 nm;
D
1
−
IR ꢁmax (cm ): 3430, 1668, 1609; FAB-MS (3-nitrophenyl
+
1
methanol, probe), m/z: 321 [M + H] ; H NMR (CD OD): ı (ppm):
3
1
0.83 (3H, s, H-18), 1.45 (3H, s, H-19), 3.60 (1H, t, J = 8 Hz, H-17␣),
f
4
.01 (1H, ddd, J = 12, 11, 5 Hz, H-11), 4.25 (1H, brs, H-6␣), 5.77
1
3
2
.4.2. 6ˇ,11˛-Dihydroxyandrost-4-ene-3,17-dione (III)
(1H, s, H-4); C NMR (CD OD): ı (ppm): 201.5 (C-3), 169.8 (C-5),
3
◦
Colorless crystals (crystallized from ethyl acetate); mp 260 C,
125.2 (C-4), 79.9 (C-17), 71.9 (C-6), 67.5 (C-11), 58.4(C-9), 49.1
(C-14), 48.0 (C-12), 42.8 (C-13), 38.9 (C-10), 38.4 (C-7), 36.7 (C-1),
33.4 (C-2), 28.9 (C-16), 28.2 (C-8), 22.3 (C-15), 18.6 (C-19), 10.8
2
5
◦
◦
25
◦
[
˛] + 79 , literature [18]; mp 259–260 C; [˛] + 72 (MeOH,
D
D
MeOH
max
−1
c = 1.0); UV ꢀ
= 240 nm; IR ꢁmax (cm ): 3423, 1735, 1668,
1
609; FAB-MS (3-nitrophenyl methanol, probe), m/z: 319
f
t
(C-18); R in chloroform/methanol (12:1): 0.32; R : 4.4 min.
+
1
[M + H] ; H NMR (CD3OD): ı (ppm): 0.99 (3H, s, H-18), 1.56 (3H,
s, H-19), 4.12 (1H, ddd, J = 12, 11, 5 Hz, H-11), 4.41 (1H, brs, H-
2
.4.6. 3˛,11˛,17ˇ-Trihydroxy-5˛-androstane (VII)
6
2
␣), 5.84 (1H, s, H-4); 13C NMR (CD3OD): ı (ppm): 220.7 (C-17),
Colorless crystals (crystallized from chloroform); mp
02.4 (C-3), 170.3 (C-5), 126.2 (C-4), 72.4 (C-6), 68.0 (C-11), 59.0
◦
25
◦
−1
2
3
61–263 C; [˛]D − 21 (MeOH, c = 1.0); IR ꢁmax (cm ): 3490,
400; FAB-MS (3-nitrophenyl methanol, probe), m/z: 307
(
7
(
C-9), 50.0 (C-14), 47.6 (C-13), 42.2 (C-12), 39.6 (C-10), 39.0 (C-
), 36.5 (C-16), 35.6 (C-1), 34.1 (C-2), 28.2 (C-8), 21.5 (C-15), 19.4
C-19), 14.0 (C-18); R in chloroform/methanol (12:1): 0.48; Rt:
−
1
[
M − H] ; H NMR (CD3OD): ı (ppm): 0.71 (3H, s, H-18), 1.08 (3H,
f
s, H-19), 3.58 (1H, t, J = 8 Hz, H-17␣), 3.80 (1H, m, H-3), 4.00
5
.7 min.
1H, m, H-11); 13C NMR (CD3OD): ı (ppm): 80.9 (C-17), 68.4
(
(
C-11), 66.9 (C-3), 59.8 (C-9), 50.2 (C-14), 48.2 (C-12), 47.4 (C-5),
2
.4.3. 17ˇ-Hydroxyandrost-4-ene-3-one (IV)
4
(
3.3 (C-13), 38.4 (C-4), 36.4 (C-10), 35.2 (C-8), 33.9 (C-7), 32.6
C-1), 29.5 (C-2), 28.4 (C-6), 27.0 (C-16), 23.6 (C-15), 23.1 (C-19),
11.3 (C-18); R in chloroform/methanol (12:1): 0.43.
◦
−1
Colorless crystals (crystallized from acetone); mp 154–156 C;
2
5
◦
MeOH
[˛] + 106 (MeOH, c = 1.0); UV ꢀ
= 238 nm; IR ꢁmax (cm ):
D
max
f
1
3430, 1664, 1609; H NMR (CDCl3): ı (ppm): 0.80 (3H, s, H-18),
1.20 (3H, s, H-19), 3.66 (1H, t, J = 8 Hz, H-17␣), 5.73 (1H, s, H-
4
); 13C NMR (CDCl3): ı (ppm): 199.9 (C-3), 171.5 (C-5), 124.1 (C-
4), 81.8 (C-17), 54.1 (C-9), 50.7 (C-14), 43.0 (C-13), 38.9 (C-10),
36.6 (C-12), 35.9 (C-8), 35.8 (C-1), 34.2 (C-2), 33.0 (C-6), 31.7 (C-
7), 30.6 (C-16), 23.6 (C-15), 20.8 (C-11), 17.6 (C-19), 11.3 (C-18); Rf
3.
Results
The structures of the transformation products were deter-
mined from FAB-MS, IR, 1H NMR and 13C NMR spectra.
The positions and configurations of the introduced hydroxyl
groups were determined mainly from changes in 1H NMR
spectra compared with the starting material and data in lit-
erature [19].
The FAB-mass spectrum of compound II showed the
[M + 1]+ at m/z 303, indicative of the addition of 16 mass units
to I in agreement with the formula C19H26O3. The IR spec-
trum showed characteristic absorptions at 3430, 1734, 1664,
1609 cm for hydroxyl and saturated ketone and conjugated
ketone groups, respectively. The 1H NMR spectrum showed
a new downfield signal for the oxygen-bearing methine pro-
ton at ı 4.08 ppm (ddd, J = 12, 11, 5 Hz), which indicated intro-
duction of a C-11␣ hydroxyl group in compounds II. The 13C
in chloroform/methanol (12:1): 0.79; Rt: 11.3 min.
2
.4.4. 11˛,17ˇ-Dihydroxyandrost-ene-3-one (V)
◦
◦
−1
Colorless crystals (crystallized from methanol); mp 218–219 C,
2
5
◦
◦
[
[
˛] + 97 (CHCl3, c = 0.8), literature [16,17]; mp 218–219 C;
D
˛] + 91 (MeOH, c = 1.0); UV ꢀ Mm ea Ox H = 240 nm; IR ꢁmax (cm ):
2
D
5
3
430, 1668, 1610; FAB-MS (3-nitrophenyl methanol, probe),
m/z: 305 [M + H] ; 1H NMR (CDCl3): ı (ppm): 0.81 (3H, s, H-18),
+
−
1
1
.31 (3H, s, H-19), 3.68 (1H, t, J = 8 Hz, H-17␣), 4.02 (1H, ddd,
J = 12, 11, 5 Hz, H-11), 5.71 (1H, s, H-4); 13C NMR (CDCl3): ı
(
(
ppm): 200.7 (C-3), 171.9 (C-5), 124.6 (C-4), 81.2 (C-17), 69.0
C-11), 59.4 (C-9), 50.1 (C-14), 48.7 (C-12), 43.9 (C-13), 40.4 (C-10),
3
7.7 (C-1), 35.6 (C-2), 34.5 (C-8), 34.0 (C-6), 31.5 (C-7), 30.8 (C-16),