Synthesis of (3α,7β,17α)-7-methyl-19-norpregn-5(10)-en-20-yne-3,7,17- triol, a metabolite of ORG OD14, and its 7-epimer

Article abstract of DOI:10.1016/S0039-128X(00)00110-0

The syntheses of the 7β-hydroxy metabolite of ORG OD14 (Livial), (3α,7β,17α)-7-methyl-19-norpregn-5(10)-en-20-yne-3,7,17-triol (35), and its 7-epimer, (3α,7α,17α)-7-methyl-19-norpregn-5(10)-en-20-yne-3,7,17- triol (11), are described. (C) 2000 Elsevier Science Inc.

Full text of DOI:10.1016/S0039-128X(00)00110-0

Steroids 65 (2000) 497–504
Synthesis of (3␣,7␤,17␣)-7-methyl-19-norpregn-5(10)-en-20-yne-3,7,17-
triol, a metabolite of ORG OD14, and its 7-epimer
Ralf Plate*, Rob C.A.L. van Wuijtswinkel, Christan G.J.M. Jans, Marinus B. Groen
Medicinal Chemistry Department, N.V. Organon, P.O. Box 20, 5340 BH Oss, The Netherlands
Received 17 February 2000; received in revised form 4 April 2000; accepted 12 April 2000
Abstract
®
The syntheses of the 7␤-hydroxy metabolite of ORG OD14 (Livial ), (3␣,7␤,17␣)-7-methyl-19-norpregn-5(10)-en-20-yne-3,7,17-triol
(35), and its 7-epimer, (3␣,7␣,17␣)-7-methyl-19-norpregn-5(10)-en-20-yne-3,7,17-triol (11), are described. © 2000 Elsevier Science Inc.
All rights reserved.
Keywords: Tibolone; Livial; ORG OD14; 7-Hydroxy steroids
1
. Introduction
2. Experimental
®
Livial is of great interest in hormone replacement ther-
2.1. General methods
apy in menopausal women at risk of osteoporosis [1,2]. The
active principle of Livial is ORG OD14 (generic name:
®
1
13
H and C NMR spectra were recorded on a Bruker
tibolone, scientific name: (7␣,17␣)-17-hydroxy-7-methyl-
spectrometer (200 MHz, 400 and 600 MHz) with deu-
teriochloroform was as the solvent unless stated other-
wise. Chemical shifts are reported as ␦ values (parts per
million) relative to tetramethylsilane as an internal stan-
dard, and coupling constants are expressed in Hertz. Thin
layer chromatography was performed on pre-coated
Merck Silica gel 60 F254 plates and visualized with UV
light and/or with sulfuric acid in ethanol solution or the
Usui reagent. Column chromatography was performed on
Merck Silica gel 60 (230–400 mesh or 400–600 mesh).
Melting points were determined on a B u¨ chi 535 apparatus
and are uncorrected. Fast atom bombardment (FAB) mass
spectra were recorded with a Finnigan MAT 90 mass
spectrometer (Finnigan MAT, Bremen, FRG). Samples
were dissolved in methanol and mixed with the matrix
compounds on standard stainless steel targets. Exact
masses of the protonated molecular ions were determined
with the peak matching technique at a mass resolution
of Ͼ 7500 (10% valley definition) in the positive ion
mode using reference masses 369 and 461 from glycerol.
Average exact masses were calculated from at least 10
computer-controlled measurements using the bracketing
method.
1
9-norpregn-5(10)-en-20-yn-3-one) I [3]. Recently, the me-
tabolism of ORG OD14 in rat and human hepatocytes has
been reported [4]. In both rat and human hepatocytes the
metabolism of this steroid was found to be virtually com-
plete. Among the metabolites detected, the 7-hydroxy de-
rivative III is a result of phase II metabolism (sulfate con-
jugation) of its precursor 35, which is produced by phase I
metabolism (hydroxylation) of II (Scheme 1). Cytochrome
P450 enzymes [5] are known to be involved in the phase I
7
-hydroxylation of endogenous steroids, such as choles-
terol, pregnenolone and dehydroepiandrosterone. Although
not proven, these enzymes are candidates for the conversion
of II into 35.
The 7-hydroxy derivative III has been found in an in
vivo metabolism study of ORG OD14 in rat, but it could not
be detected in human. Structure verification as well as an
understanding of the pharmacology of this drug metabolite
urged us to develop the synthesis of 35 as described below.
*
Corresponding author. Tel.: ϩ31-412-661957; fax: ϩ31-412-
62546.
E-mail address: R.Plate@organon.oss.akzonobel.nl (R. Plate).
6
0
039-128X/00/$ – see front matter © 2000 Elsevier Science Inc. All rights reserved.
PII: S0039-128X(00)00111-2

4
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R. Plate et al. / Steroids 65 (2000) 497–504
2
1
.1.1. 3-Methoxyestra-1,3,5(10),6-tetraen-17-one cyclic
,2-ethanediyl acetal (2)
2.1.4. 3-Methoxyestra-1,3,5(10)-triene-7,17-dione cyclic
17-(1,2-ethanediyl acetal) (5)
Triethyl orthoformate (66 ml) and p-toluenesulphonic
acid (990 mg) were added to a solution of 1 (33.1 g, 117
Chromic acid (3 ml, 8N, 24 mM) was added to a solution
of 4 (2 g, 5.8 mM) in acetone (30 ml) at Ϫ20°C. The
reaction was stirred for 6 h, whereas the temperature raised
to 20°C. Ethanol (10 ml) was added, and the mixture was
poured into a saturated solution of sodium bicarbonate.
Then, the mixture was filtered over a Decalite pad, extracted
with ethyl acetate, and washed successively with sodium
bicarbonate, ammonium chloride, water, and brine. The
organic layer was dried (Na SO ) and evaporated to dry-
mM) in ethylene glycol (330 ml) and CH Cl (50 ml).
2
2
After stirring at room temperature for 6 h, the reaction
mixture was poured into sodium bicarbonate solution and
extracted with ethyl acetate. The extract was washed with
water and brine, dried (Na SO ), and evaporated to dry-
2
4
ness to give 2 (42.5 g, 100% crude). R 0.8 [toluene/ethyl
f
1
2
4
acetate (8:2 v/v)]. H NMR: (CDCl , 200 MHz) ␦ 0.88 (s,
3
ness. Purification by column chromatography [heptane/eth-
3
6
6
1
H, 18CH ), 3.80 (s, 3H, Ar-OCH ), 3.93 (m, 4H, ketal),
3 3
yl acetate (8:2)] gave 5 (1.31 g, 3.8 mM, 66%). R 0.7
f
.01 (dd, J ϭ 2Hz, 1H, 7-H), 6.45 (dd, J ϭ 3 Hz, 1H,
-H), 6.65 (d, 1H, Ar-4H), 6.74 (dd, 1H, Ar-2H), 7.16 (d,
H, Ar-1H).
1
[
toluene/ethyl acetate (1:1 v/v)]. H NMR: (CDCl , 200
3
MHz) ␦ 0.87 (s, 3H, 18CH ), 2.65 (m, 1H, 8␤H), 3.59 (s,
3
2
H, 6-H), 3.80 (s, 3H, Ar-OCH ), 3.93 (m, 4H, ketal), 6.67
3
(
d, 1H, Ar-4H), 6.81 (dd, 1H, Ar-2H), 7.24 (d, 1H, Ar-1H).
2
1
.1.2. (6␣,7␣)-6,7-Epoxy-3-methoxyestra-1,3,5(10) -trien-
7-one cyclic 1,2-ethanediyl acetal (3)
2
.1.5. (7␣)-7-Hydroxy-3-methoxy-7-methylestra-1,3,5(10)-
3
-Chloroperoxybenzoic acid (44.4 g, 260 mM) was
trien-17-one cyclic 1,2-ethanediyl acetal (6)
added to a solution of 2 (42 g, crude) in ethyl acetate (420
ml) under nitrogen was added and stirred at room tem-
perature for 6 h. Then, the reaction mixture was poured
into saturated sodium bicarbonate solution and extracted
with ethyl acetate. The extract was washed with ammo-
nium chloride solution, treated with sodium thiosulfate
solution, filtered, washed with brine, and evaporated to
Cerium(III) chloride heptahydrate (560 mg, 1.5 mM)
was finely ground in a mortar and placed into in a 50 ml
necked flask. The flask was put in an oil bath, evacuated,
and heated to 140°C, whereas the powder was stirred with
a magnetic stirring bar for 16 h. Then, while the flask was
still hot, nitrogen was introduced, the flask was cooled to
0°C, and tetrahydrofuran (5 ml) was added. The flask was
put in an ultrasound bath, and the suspension was stirred for
1 h. After cooling to 0°C, methylmagnesium chloride (0.75
ml of a 2.0-M solution in tetra- hydrofuran, 1.5 mM) was
added to the suspension. After stirring for 1.5 h at 0°C, a
solution of 5 (356 mg, 1 mM) in tetrahydrofuran (3 ml) was
added to the mixture. After 0.5 h of stirring at 0°C, the
reaction mixture was poured into saturated ammonium chlo-
ride solution and extracted with ethyl acetate. The extract
was washed successively sodium bicarbonate, water, and
brine. The organic layer was dried (Na SO ) and evaporated
dryness to give 3 (48.4 g, 100% crude). R 0.5 [toluene/
f
1
ethyl acetate (8:2 v/v)]. H NMR: (CDCl , 200 MHz) ␦
3
0
.89 (s, 3H, 18CH ), 3.58 (d, 1H, 7-H), 3.82 (s, 3H,
3
Ar-OCH ), 3.85 (d, 1H, 6-H), 3.95 (m, 4H, ketal), 6.85
3
(
dd, 1H, Ar-2H), 6.97 (d, 1H, Ar-4H), 7.16 (d, 1H,
Ar-1H).
2
.1.3. (7␣)-7-Hydroxy-3-methoxyestra-1,3,5(10)-trien-17-
one cyclic 1,2-ethanediyl acetal (4)
A solution of 3 (48.4 g, crude) in tetrahydrofuran (480
ml) to a suspension of lithium aluminum hydride (25.7 g,
2
4
to dryness to give the crude product. Purification by column
chromatography [heptane/ethyl acetate (7:3)] gave 6 (190
6
78 mM) in tetrahydrofuran (300 ml), was added dropwise.
mg, 0.5 mM, 53%). R 0.5 [toluene/ethyl acetate (1:1 v/v)].
f
The mixture was heated to reflux. After stirring for 1.5 h, the
mixture was cooled to Ϫ15°C and quenched with ethyl
acetate whereas maintaining the temperature below 10°C.
Water (250 ml) was added. After stirring for 1 h, the
mixture was filtered over a Decalite pad and extracted with
ethyl acetate. The extract was successively washed with
ammonium chloride, sodium bicarbonate solution, and
brine. The organic layer was dried (Na SO ) and evaporated
1
H NMR: (CDCl , 200 MHz) ␦ 0.88 (s, 3H, 18CH ), 1.32
3
3
(
(
(
s, 3H, 7-CH ), 2.33 (m, 1H, 9␣H), 2.48 (m, 1H, 8␤H), 2.72
d, J ϭ 16 Hz, 1H, 6␣H), 2.94 (d, J ϭ 16Hz, 1H, 6␤H), 3.78
3
s, 3H, Ar-OCH ), 3.92 (m, 4H, ketal), 6.64 (d, 1H, Ar-4H),
3
6
.76 (dd, 1H, Ar-2H), 7.22 (d, 1H, Ar-1H).
2
.1.6. (7␣)-7-Hydroxy-3-methoxy-7-methylestra-2,5(10)-
2
4
dien-17-one cyclic 1,2-ethanediyl acetal (7)
to dryness. The residue was purified by column chromatog-
Lithium metal (1.1 g, 158 mM) was cut into small pieces
and dissolved in liquid ammonia (90 ml). After stirring for
0.5 h at Ϫ65°C, a solution of 6 (4.53 g, 11.67 mM) in
tetrahydrofuran (45 ml) was added dropwise. After stirring
at Ϫ35°C for 4.5 h, ethanol (40 ml) was added. After
evaporation of the ammonia, the reaction mixture was
poured into a saturated ammonium chloride solution and
extracted with ethyl acetate. The extract was washed suc-
raphy [heptane/ethyl acetate (8:2)] to give 4 (24.9 g, 72.3
1
mM, 51%). R 0.55 [toluene/ethyl acetate (1:1 v/v)]. H
f
NMR: (CDCl , 200 MHz) ␦ 0.88 (s, 3H, 18CH ), 2.43 (m,
3
3
1
H, 9␣H), 2.63 (m, 1H, 8␤H), 2.91 (m, 1H, 6-H), 3.12 (m,
1
H, 6-H), 3.78 (s, 3H, Ar-OCH ), 3.93 (m, 4H, ketal), 4.15
3
(m, 1H, 7␤H), 6.64 (d, 1H, Ar-4H), 6.75 (dd, 1H, Ar-2H),
7
.25 (d, 1H, Ar-1H).

R. Plate et al. / Steroids 65 (2000) 497–504
499
cessively with sodium bicarbonate, water, and brine. The
organic layer was dried (Na SO ) and evaporated to dryness
CH ), 25.1 (t, C11), 25.4 (t, C15), 26.3 (t, C1), 29.5 (q,
3
7-CH ), 31.8 (t, C2), 31.9 (t, C12), 36.0 (t, C16), 39.3 (t,
2
4
3
to give 7 (4.83 g, 100% crude). R 0.45 [heptane/ethyl
C4), 43.3 (d, C9), 46.6 (d, C8), 46.6 (d, C14), 47.9 (t, C6),
49.4 (s, C13), 67.1 (d, C3), 71.0 (s, C7), 123.4 (s, C5), 129.2
(s, C10), 222.2 (s, C17).
f
1
acetate (1:1 v/v)]. H NMR: (CDCl , 400 MHz) ␦ 0.88 (s,
3
3
3
H, 18CH ), 1.31 (s, 3H, 7-CH ), 3.54 (s, 3H, 3-OCH ),
.90 (m, 4H, ketal), 4.64 (m, 1H, 2-H).
3 3 3
2
.1.10. (3␣,7␣,17␣)-7-Methyl-19-norpregn-5(10)-en-20-
2
.1.7. (7␣)-7-Hydroxy-7-methylestr-5(10)-ene-3,17-dione
yne-3,7,17-diol (11)
cyclic 17-(1,2-ethanediyl acetal) (8)
Acetylene gas was passed through a suspension of po-
tassium tert-butoxide (1.49 g, 13.3 mM) in tetrahydrofuran
(5 ml) at 0°C. After 0.5 h, a suspension of 10 (0.9 g, 2.96
mM) in tetrahydrofuran (10 ml) was added, and the mixture
was stirred for 1 h at 0°C. Then, nitrogen was passed
through the suspension. The reaction mixture was extracted
with ethyl acetate, the extract was washed successively with
sodium bicarbonate and brine, dried (Na SO ) and evapo-
Oxalic acid (1.17 g, 13.0 mM) was added to a solution of
7
(4.83 g, crude) in acetone (120 ml) and water (30 ml),.
After stirring for 6 h, the mixture was poured into a satu-
rated solution of sodium bicarbonate and extracted with
ethyl acetate. The extract was washed successively with
sodium bicarbonate, water, and brine. The organic layer was
dried (Na SO ) and evaporated to dryness to give the crude
2
4
2
4
product. Purification by column chromatography [heptane/
rated to dryness. Purification by column chromatography
[methylene chloride/acetone (8:2)], followed by crystalliza-
ethyl acetate (1:1)] gave 8 (2.21 g, 6.4 mM, 49%). R 0.3
f
1
[
heptane/ethyl acetate (4:6 v/v)]. H NMR: (CDCl , 400
tion from ethyl acetate/acetone gave 11 (580 mg, 1.76 mM,
3
1
MHz) ␦ 0.89 (s, 3H, 18CH ), 1.32 (s, 3H, 7-CH ), 2.65 (d,
59%), m.p. 197°C. H NMR: (CDCl , 400 MHz) ␦ 0.86 (s,
3
3
3
1
H, 4-H), 2.78 (d, 1H, 4-H), 3.92 (m, 4H, ketal).
3H, 18-CH ), 1.23 (m, 1H, 11␤H), 1.27 (s, 3H, 7-CH ), 1.41
3
3
(m, 1H, 8␤H), 1.51 (m, 1H, 2␤H), 1.60 (m, 1H, 12␤H), 1.63
2
.1.8. (3␣,7␣)-3,7-Dihydroxy-7-methylestr-5(10)-en-17-
(m, 1H, 15␤H), 1.77 (m, 1H, 6␣H), 1.78 (m, 1H, 12␣H),
1.83 (m, 1H, 4␤H), 1.87 (m, 1H, 9␣H), 1.93 (m, 1H, 2␣H),
1.96 (m, 1H, 16␤H), 1.97 (m, 1H, 14␣H) 1.98 (m, 1H,
1␤H), 1.98 (m, 1H, 15␣H), 2.05 (m, 1H, 11␣H), 2.17 (m,
1H, 1␣H), 2.19 (m, 1H, 4␣H), 2.19 (m,1H, 6␤H), 2.25 (m,
1H, 16␣H), 2.58 (s, 1H, C ϵ CH), 3.83 (m, 1H, 3␤ H).
13C-NMR (CDCl , 100 MHz) 13.5 (q, 18-CH ), 25.7 (t,
one cyclic 1,2-ethanediyl acetal (9)
A suspension of lithium tri-tert-butoxy aluminum hy-
dride (3.69 g, 14.52 mM) in THF (30 ml) was added drop-
wise to a solution of 8 (2.21 g, 6.3 mM) in THF (20 ml)
under nitrogen and at room temperature. After 3 h, the
reaction mixture was poured into saturated ammonium chlo-
ride solution and extracted with ethyl acetate. The extract
was washed successively with water, brine and dried
3
3
C11), 26.6 (t, C15), 26.9 (t, C1), 29.9 (q, 7-CH ), 32.3 (t,
3
C2), 33.2 (t, C12), 39.1 (t, C16), 39.7 (t, C4), 43.1 (d, C9),
45.4 (d, C14), 47.7 (d, C8), 48.1 (t, C6), 49.0 (s, C13), 67.6
(d, C3), 71.6 (s, C7), 74.4 (d, C-H ϵCH), 78.9 (s, C17),
(Na SO ), and evaporated to dryness to give 9 (2.18 g, 6.3
2 4
1
mM, 98%). R 0.2 [heptane/ethyl acetate (2:8 v/v)]. H
f
NMR: (CDCl , 400 MHz) ␦ 0.88 (s, 3H, 18CH ), 1.29 (s,
88.0 (s, CϵCH), 123.5 (s, C5), 129.8 (s, C10). Exact mass
3
3
ϩ
3
H, 7-CH ), 3.83 (m, 1H, 3␤H), 3.92 (m, 4H, ketal).
calculated for [MϩHϩGlyc.]
23.2745.
is 423.2747, Found:
3
4
2
.1.9. (3␣,7␣)-3,7-Dihydroxy-7-methylestr-5(10)-en-17-
one (10)
Hydrochloric acid (5 ml of a 0.1N HCl solution) was
2.1.11. (17␤ )-17-Hydroxy-3-methoxyestra-1,3,5(10)-trien-
6-one (13)
added to a solution of 9 (2.18 g, 6.3 mM) in acetone (20 ml)
and water (2 ml). After 4 h, the reaction mixture was poured
into a solution of saturated sodium bicarbonate and ex-
tracted with ethyl acetate. The extract was washed succes-
sively with sodium bicarbonate, water, and brine, dried
19-Nortestosterone 12 (46 g, 168 mM) and potassium
acetate (25g, 255 mM) were dissolved in dry DMF (500 ml)
and heated to 120°C. The solution was stirred for 6 h
whereas oxygen gas was passed through. The reaction mix-
ture was cooled to room temperature, potassium carbonate
(138 g, 1 mol) and methyl iodide (80 ml, 1.29 mol) were
added and the solution was stirred for 18 h at room tem-
perature. The reaction mixture was poured into saturated
ammonium chloride solution and extracted with ethyl ace-
tate. The organic layer was dried (Na SO ) and evaporated
(
Na SO ), and evaporated to dryness. The residue was crys-
2 4
tallized (dichloro methane/acetone) to give 10 (1.07 g, 3.5
mM, 56%) as white crystals; m.p. 247°C. R 0.8 [ethanol/
f
1
ethyl acetate (1:1 v/v)]. H NMR: (CDCl , 600 MHz) ␦ 0.90
3
(
s, 3H, 18-CH ), 1.28 (m, 1H, 11␤H), 1.31 (m, 1H, 12␣H),
3
2
4
1
1
1
1
1
1
.36 (s, 3H, 7-CH ), 1.53 (m, 1H, 2␤H), 1.80 (m, 1H,
to dryness to give the crude product that was purified by
column chromatography [silica gel, heptane/ethyl acetate
3
2␤H), 1.81 (m, 1H, 14␣H), 1.82 (m, 1H, 6␣H), 1.82 (m,
H, 15␤H), 1.83 (m, 1H, 4␤H), 1.89 (m, 1H, 9␣H), 1.91 (m,
H, 2␣H), 1.97 (m, 1H, 1␤H), 2.08 (m, 1H, 11␣H), 2.11 (m,
H, 16␣H), 2.16 (m, 1H, 1␣H), 2.21 (m, 1H, 4␣H), 2.24 (m,
H, 6␤H), 2.30 (m, 1H, 15␣H), 2.45 (m, 1H, 16␤H), 3.85
(9:1 v/v)], to give 13 (37.7 g, 125 mM, 75%). R 0.5
f
1
[heptane/ethyl acetate (1:1 v/v)]. H NMR: (CDCl , 600
3
MHz) ␦ 0.79 (s, 3H, 18CH ), 1.34 (m, 1H, 14␣H), 1.34 (m,
3
1H, 15␤H), 1.35 (m, 1H, 12␣H), 1.50 (m, 1H, 16␤H), 1.61
(m, 1H, 11␤H), 1.68 (m, 1H, 15␣H), 1.94 (m, 1H, 8␤H),
1
3
(m, 1H, 3␤H). C-NMR (CDCl , 150 MHz) 14.6 (q, 18-
3

5
00
R. Plate et al. / Steroids 65 (2000) 497–504
2
.02 (m, 1H, 12␤H), 2.14 (m, 1H, 16␣H), 2.21 (dd, J ϭ 13.4
Hz, 1H, 7␣H), 2.39 (m, 1H, 11␣H), 2.48 (m, 1H, 9␣H), 2.75
dd, J ϭ 3.2 Hz, 1H, 7␤H), 3.75 (t, 1H, 17␣H), 3.75 (s, 3H,
Ar-OCH ), 7.10 (dd, 1H, Ar-2H), 7.34 (d, 1H, Ar-1H), 7.56
(d, C1), 132.6 (s, C10), 139.5 (s, C5), 158.5 (s, C3), 202.4
(s, C6). Exact mass calculated for [MϩH] is 429.2825,
Found: 429.2833.
ϩ
(
3
1
3
(d, 1H, Ar-4H). C-NMR (CDCl , 150 MHz) ␦ 11.3 (q,
2.1.14. (6␣,7␣,17␤)-17-[[(1,1-Dimethylethyl)dimethyl-
silyl]oxy]-3-methoxy-7-methylestra-1,3,5(10)-trien-6-ol
(16)
3
C18), 23.2 (t, C15), 25.9 (t, C11), 30.8 (t, C16), 36.7 (t,
C12), 40.5 (d, C8), 43.4 (d, C9), 43.5 (s, C13), 44.4 (t, C7),
5
0.3 (d, C14), 55.9 (q, Ar-OCH ), 81.9 (d, C17), 110.0 (d,
Compound 15 (31 g, 72.4 mM), dissolved in dry THF
(170 ml), was added dropwise over 1 h to a suspension of
lithium aluminumhydride (11 g, 290 mM) in dry THF (120
ml). After 0.5 h of stirring at room temperature, saturated
sodium sulfate solution was added dropwise until the excess
3
C1), 122.0 (d, C4), 127.0 (d, C2), 133.8 (s, C10), 140.0 (s,
C5), 158.6 (s, C3), 198.4 (s, C6).
2
3
.1.12. (17␤)-17-[[(1,1-Dimethylethyl)dimethylsilyl]oxy]-
-methoxyestra-1,3,5(10)-trien-6-one (14)
of LiAlH was quenched. Water was added, and the reaction
4
Imidazole (26.6 g, 391 mM) was added to a solution of
3 (65.3 g, 217 mM) in methylene chloride (500 ml). tert-
mixture was extracted with ethyl acetate, dried (Na SO ),
2
4
1
and evaporated to dryness to give 16 (32 g, 100% crude). R
f
1
Butyldimethylsilylchloride (54 g, 358 mM) was added, and
the orange reaction mixture was stirred for 4 h at room
temperature. The solution was poured into saturated ammo-
nium chloride solution, extracted with ethyl acetate, and
dried (Na SO ). During evaporation of the solvent, the
0.2 [heptane/ethyl acetate (8:2 v/v)]. H NMR: (CDCl , 200
3
MHz) ␦ 0.01 (d, 6H), 0.76 (s, 3H, 18CH ), 0.80 (d, J ϭ 7.2
3
Hz), 7-CH ) 0.90 (s, 9H), 3.66 (t, 1H, 17␣H), 3.81 (s, 3H,
3
Ar-OCH ), 4.87 (m, 1H, 6-H) 6.78 (dd, 1H, Ar-2H), 7.18 (d,
3
1H, Ar-1H), 7.20 (d, 1H, Ar-4H).
2
4
product crystallized. The crystals were collected by filtra-
tion and dried in vacuo to give 14 (46.9 g, 113 mM, 52%).
2.1.15. (17␤)-17-[[(1,1-Dimethylethyl)dimethylsilyl]oxy]-
3-methoxy-7-methylestra-1,3,5(10),6-tetraene (17)
1
R 0.85 [heptane/ethyl acetate (1:1 v/v)]. H NMR: (CDCl ,
f
3
2
2
7
1
00 MHz) ␦ 0.05 (d, 6H), 0.75 (s, 3H, 18CH ), 0.9 (s, 9H),
.20 (dd, J ϭ 13.4 Hz, 1H, 7␣H), 2.74 (dd, J ϭ 3.2 Hz, 1H,
Compound 16 (20 g, 46.4 mM) was dissolved in toluene
(2 liters), and Amberlyst 15 (4 g) was added. The mixture
was heated to reflux for 5 h. The reaction mixture was
cooled to room temperature, filtered and evaporated to dry-
ness. The crude product was dissolved in methylene chlo-
ride (90 ml), and imidazole (6 g, 88 mM) was added. Then,
tert-butyldimethylsilylchloride (54 g, 358 mM) was added,
and the reaction mixture was stirred for 18 h at room
temperature. The solution was poured into water, extracted
with ethyl acetate, and washed with saturated sodium chlo-
ride solution. The organic layer was dried (Na SO ), evap-
3
␤H), 3.66 (t, 1H, 17␣H), 3.85 (s, 3H, Ar-OCH ), 7.10 (dd,
3
H, Ar-2H), 7.34 (d, 1H, Ar-1H), 7.56 (d, 1H, Ar-4H).
2
.1.13. (7␣,17␤)-17-[[(1,1-Dimethylethyl)dimethyl-
silyl]oxy]-3-methoxy-7-methylestra-1,3,5(10)-trien-6-one
15)
Potassium tert-butoxide (10.3 g, 91.5 mM) was added to
(
a solution of 14 (37.5 g, 90.6 mM) in dimethoxyethane (500
ml). Methyl iodide (13 g, 91.5 mM), dissolved in dime-
thoxyethane (50 ml), was added to the red solution over in
2
4
orated to dryness, and purified by column chromatography
3
0-min interval. The reaction mixture was stirred for 1 h at
[silica gel, heptane/ethyl acetate (98:2 v/v)] to give 17 (11.8
room temperature. Then, the solution was poured into 2N
HCl, extracted with ethyl acetate, and washed with saturated
sodium chloride solution. The organic layer was dried
g, 28.6 mM, 67%). R 0.85 [heptane/ethyl acetate (8:2 v/v)].
f
1
H NMR: (CDCl , 400 MHz) ␦ 0.05 (d, 6H), 0.74 (s, 3H,
3
18CH ), 1.00 (s, 9H), 1.95 (s, 3H, 7-CH ), 3. 61 (t, 1H,
3
3
(
Na SO ) and evaporated to dryness. The crude product was
17␣H), 3.78 (s, 3H, Ar-OCH ), 6.20 (s, 1H, H-6), 6.56 (d,
2
4
3
purified by column chromatography [silica gel, heptane/
1H, Ar-4H), 6.69 (dd, 1H, Ar-2H), 7.16 (d, 1H, Ar-1H).
ethyl acetate (9:1 v/v)] to give 15 (24 g, 56.1 mM, 63%). R
f
1
0
.3 [heptane/ethyl acetate (9:1 v/v)]. H NMR: (CDCl , 600
2.1.16. (6␤,7␤,17␤)-17-[[(1,1-Dimethylethyl)dimethyl-
silyl]oxy]-6,7-epoxy-3-methoxy-7-methylestra-1,3,5(10)-
triene (19)
3
MHz) ␦ 0.01 (s, 6H), 0.75 (s, 3H, 18CH ), 0.90 (s, 9H), 1.09
3
(
d, J ϭ 8 Hz, 3H, 7-CH ), 1.25 (m, 1H, 12␣H), 1.31 (m, 1H,
3
1
1
5␤H), 1.42 (m, 1H, 14␣H), 1.49 (m, 1H, 16␤H), 1.56 (m,
H, 11␤H), 1.58 (m, 1H, 15␣H), 1.91 (m, 1H, 12␤H), 1.95
Compound 17 (21g, 51 mM) was dissolved in THF (170
ml). Water (20 ml) was added, and the solution was cooled
to 0°C. N-chlorosuccinimide (6.8 g, 51 mM) was added, and
the reaction mixture was stirred for 18 h at room tempera-
ture. Then, water was added, and the solution was extracted
with ethyl acetate, dried (Na SO ), and evaporated to dry-
(m, 1H, 16␣H), 2.04 (m, 1H, 8␤H), 2.40 (m, 1H, 11␣H),
2
3
.64 (m, 1H, 7␤H), 2.67 (m, 1H, 9␣H), 3.68 (t, 1H, 17␣H),
.84 (s, 3H, Ar-OCH ), 7.10 (dd, 1H, Ar-2H), 7.34 (d, 1H,
3
1
3
Ar-1H), 7.57 (d, 1H, Ar-4H). C-NMR (CDCl , 100 MHz)
3
2
4
␦
-4.9 (q, CH Si), 9.7 (q, 7-CH ), 11.0 (q, C18), 18.5 (s,
ness to give crude (6␤,7␣,17␤)-7-chloro-17-[[(1,1-dimeth-
3
3
C-TBDMS), 21.9 (t, C15), 25.9 (q, CH -TBDMS), 26.1 (t,
ylethyl)dimethylsilyl]oxy]-3-methoxy-7-methylestra-1,3,5
3
C11), 30.6 (t, C16), 36.0 (d, C9), 36.7 (t, C12), 41.8 (d, C8),
(10)-trien-6-ol 18 (R 0. 5 [heptane/ethyl acetate (8:2 v/v)],
f
4
3.1 (d, C7), 43.8 (s, C13), 45.0 (d, C14), 55.0 (q, Ar-
which was used in the next step without further purification.
OCH ), 81.2 (d, C17), 109.3 (d, C4), 121.2 (d, C2), 126.6
Crude 18 (25 g) was dissolved in THF (330 ml), potassium
3

R. Plate et al. / Steroids 65 (2000) 497–504
501
tert-butoxide (5.7 g, 51 mM) was added, and the reaction
mixture was stirred for 1.5 h at room temperature. Then, the
solution was poured into water, extracted with ethyl acetate,
and washed with saturated sodium chloride solution. The
organic layer was dried (Na SO ) and evaporated to dryness
(45 ml). The reaction mixture was heated to reflux for 1.5 h.
Then, saturated sodium sulfate solution was added, drop-
wise until the excess of LiAlH was quenched. Water was
4
added and the reaction mixture was extracted with ethyl
acetate and washed with saturated sodium chloride solution.
The organic portion was dried (Na SO ) and evaporated to
2
4
to give the crude product 19 [R 0.6 (heptane/ethyl acetate
f
2
4
(
8:2 v/v)]. Purification by column chromatography (silica
dryness to give the crude product, which was purified by
column chromatography [silica gel, heptane/ethyl acetate
gel, heptane/ethyl acetate 9/1 (v/v) gave 19 (1 g, 2.3 mM,
5
3
H-6), 3. 65 (t, 1H, 17␣H), 3.81 (s, 3H, Ar-OCH ), 6.84 (dd,
1
1
%) H NMR: (CDCl , 400 MHz) ␦ 0.05 (d, 6H), 0.67 (s,
3
(9:1 v/v)] to give 22 (1.8 g, 4.2 mM, 51%). R 0.35 [heptane/
f
1
H, 18CH ), 0.89 (s, 9H), 1.52 (s, 3H, 7-CH ), 3.4 (s, 1H,
3
3
ethyl acetate (8:2 v/v)] H NMR: (CDCl , 400 MHz) ␦ 0.05
3
3
(d, 6H), 0.79 (s, 3H, 18CH ), 0.90 (s, 9H),1.14 (s, 3H,
3
H, Ar-2H), 7.06 (d, 1H, Ar-2H), 7.17 (d, 1H, Ar-1H).
7
-CH ), 2.77 (d, J ϭ 16.2 Hz, 1H, 6␣H), 2.96 (d, J ϭ 16.2
3
Further elution gave an additional 4 g of a more polar
compound, characterized as (6␣,7␤,17␤)-17-[[(1,1-
dimethylethyl)dimethylsilyl]oxy]-3-methoxy-7-methylestra-
Hz, 1H, 6␤H), 3. 63 (t, 1H, 17␣H), 3.77 (s, 3H, Ar-OCH3),
.57 (d, 1H, Ar-4H), 6.73 (dd, 1H, Ar-2H), 7.20 (d, 1H,
Ar-1H).
6
1
,3,5(10)-triene-6,7-diol 20 (4g, 9.0 mM, 20%). R 0.15 [hep-
f
1
tane/ethyl acetate (8:2 v/v)]. H NMR: (DMSO, 600 MHz) ␦
2
.1.18. (7␤,17␤)-3-Methoxy-7-methylestra-1,3,5(10)-tri-
0
9
1
1
8
1
.01 (s, 6H), 0.71 (s, 3H, 18CH ), 0.83 (s, 3H, 7-CH ), 0.86 (s,
3 3
ene-7,17-diol (23)
H) 1.19 (m, 1H, 12␣H), 1.30 (m, 1H, 16␤H), 1.33 (m, 1H,
4␣H), 1.38 (m, 1H, 11␤H), 1.65 (m, 1H, 15␤H), 1.72(m, 1H,
2␤H), 1.81 (m, 1H, 15␣H), 1.82 (m, 1H, 16␣H), 1.88 (m, 1H,
␤H), 2.11 (m, 1H, 9␣H), 2.28 (m, 1H, 11␣H), 3.61 (t, 1H,
Hydrochloric acid (2N, 3.5 ml) was added to a solution
of 22 (1.75 g, 4.1 mM) in acetone (25 ml), and the reaction
mixture was stirred at room temperature for 5 h. Aqueous
sodium bicarbonate was added, and the solution was ex-
tracted with ethyl acetate. The organic portion was dried
7␣H), 3.70 (s, 3H, Ar-OCH ), 3.90 (d, 1H, H6) 6.76 (d, 1H,
3
13
Ar-4H), 6.78 (dd, 1H, Ar-2H), 7.20 (dd, 1H, Ar-1H). C-NMR
CDCl , 100 MHz) ␦ -4.8 (q, CH Si), 11.7 (q, C18), 18.2 (s,
(
Na SO ) and evaporated to dryness to give the crude prod-
2 4
(
3
3
uct, which was purified by column chromatography [silica
C-TBDMS), 18.8 (q, 7-CH ), 25.9 (q, CH -TBDMS), 26.1 (t,
3
3
gel, heptane/ethyl acetate (9:1–7:3 v/v)] to give 23 (1.15 g,
C15), 27.5 (t, C11), 31.3 (t, C16), 37.6 (t, C12), 41.6 (d, C8), 43.2
1
3
.63 mM, 89%). R 0.2 [heptane/ethyl acetate (1:1 v/v)]. H
f
(
d, C9), 45.0 (s, C13), 45.6 (d, C14), 55.5 (q, Ar-OCH ), 73.6 (s,
3
NMR: (CDCl , 600 MHz) ␦ 0.81 (s, 3H, 18-CH ), 1.14 (s,
3
3
C7), 77.1 (d, C6), 81.6 (d, C17), 115.4 (d, C2), 115.7 (d, C4),
3
H, 7-CH ), 1.28 (m, 1H, 12␣H), 1.40 (m, 1H, 14␣H), 1.48
3
127.7 (d, C1), 131.2 (s, C10), 137.4 (s, C5), 158.5 (s, C3).
(m, 1H, 16␤H), 1.55 (m, 1H, 11␤H), 1.72 (m, 1H, 8␤H),
1.75 (m, 1H, 15␤H), 1.88 (m, 1H, 15␣H), 1.94 (m, 1H,
12␤H), 2.10 (m, 1H, 16␣H), 2.28 (m, 1H, 9␣H), 2.35 (m,
1H, 11␣H), 2.78 (d, J ϭ 16.2 Hz, 1H, 6␣H), 2.97 (d, J ϭ
16.2 Hz, 1H, 6␤H), 3.70 (t, 1H, 17␣H), 3.77 (s, 3H, Ar-
2
.1.17. (7␤,17␤)-17-[[(1,1-Dimethylethyl)dimethyl-
silyl]oxy]-3-methoxy-7-methylestra-1,3,5(10)-trien-7-ol
(22)
OCH ), 6.57 (d, 1H, Ar-4H), 6.74 (dd, 1H, Ar-2H), 7.19 (d,
3
2
.1.17.1. From 19. Compound 19 (320 mg, 0.75 mM),
1
3
1
H, Ar-1H). C-NMR (CDCl , 150 MHz) 11.7 (q, 18-
3
dissolved in dry THF (3 ml), was added to a suspension of
lithium aluminum hydride (136 mg, 3.6 mM) in dry THF (3
ml). The reaction mixture was heated to reflux for 1.5 h.
Then, saturated sodium sulfate solution was added dropwise
until the excess of LiAlH was quenched. Water was added,
and the reaction mixture was extracted with ethyl acetate,
CH ), 21.5 (q, 7-CH ), 26.2 (t, C15), 28.1 (t, C11), 31.1 (t,
3
3
C16), 37.5 (t, C12), 43.8 (d, C9) 44.7 (s, C13), 46.8 (d,
C14), 48.0 (t, C6), 48.5 (d, C8), 55.6 (q, Ar-OCH ), 72.6 (s,
3
C7), 81.7 (d, C17) 112.7 (d, C4), 113.9 (d, C2), 127.4 (d,
C1), 130.5 (s, C10), 136.9 (s, C5), 158.1 (s, C3).
4
dried (Na SO ) and evaporated to dryness to give 22 (300
mg, 0.70 mM, 93%). Data are shown below.
2
4
2
.1.19. (7␤)-7-Hydroxy-3-methoxy-7-methylestra-1,3,5(10)-
trien-17-one (24)
Compound 23 (1.15 g, 3.6 mM) and N-methylmorpho-
line-N-oxide (1.1 g, 9.5 mM) were dissolved in acetone (50
ml). A catalytic amount of tetra-n-propylammonium perru-
thenate(VII) (70 mg, 0.2 mM) was added, and the reaction
mixture was stirred for 1.5 h at room temperature. The
solution was filtered over dicalite, rinsed with ethyl acetate,
evaporated to dryness, and filtered over a short silica gel
column using heptane/ethyl acetate (8:2 v/v) as eluent to
2
.1.17.2. From 20. Mesyl chloride (3.7 g, 32.6 mM) was
added to a cooled (0°C) solution of 20 (3.6 g, 8.1 mM) and
triethylamine (3.3 g, 32.6 mM) in methylene chloride (50
ml). The reaction mixture was stirred for 1.5 h at 0°C and
then for3 h at room temperature. Then, the mixture was
poured into a saturated solution of ammonium chloride,
extracted with methylene chloride, dried (Na SO ), and
2
4
evaporated to dryness to give crude 6␣-O-mesylate 21. Rf
.55 [heptane/ethyl acetate (8:2 v/v)]. Crude 21(5 g), dis-
0
give 24 (1.0 g, 3.2 mM 88%). R 0.3 [heptane/ethyl acetate
f
1
solved in dry THF (45 ml), was added to a suspension of
lithium aluminum hydride (1.48 g, 39.1 mM) in dry THF
(1:1 v/v)]. H NMR: (CDCl , 400 MHz) ␦ 0.96 (s, 3H,
3
18CH ), 1.21 (s, 3H, 7-CH ), 2.82 (d, J ϭ 16 Hz, 1H, 6␣H),
3
3

5
02
R. Plate et al. / Steroids 65 (2000) 497–504
3
.01 (d, J ϭ 16 Hz, 1H, 6␤H), 3.78 (s, 3H, Ar-OCH ), 6.58
dition funnel. Lithium (342 mg, 49 mM) was added, and the
blue solution was stirred for 20 min at Ϫ78°C. 28 (280 mg,
.78 mM) dissolved in THF (5 ml), was added to the blue
3
(d, 1H, Ar-4H), 6.74 (dd, 1H, Ar-2H), 7.21 (d, 1H, Ar-1H).
0
2
.1.20. (7␤)-7-Hydroxy-3-methoxy-7-methylestra-1,3,5(10)-
solution over a 5 min interval. The dry ice bath was re-
moved, and the reaction mixture was refluxed at Ϫ35°C for
3 h. Then, isopropanol ( Ϯ 10 ml) was added over 15 min
until the color of the reaction mixture changed from blue to
white. The excess ammonia evaporated upon standing, wa-
ter was added, and the reaction mixture was extracted three
times with ethyl acetate. The organic layers were combined,
washed with saturated sodium chloride solution, dried
(Na SO ), and evaporated to dryness to give (7␤)-7-Hy-
trien-17-one cyclic 1,2-ethanediyl acetal (28)
Compound 24 (1 g, 3.2 mM), ethylene glycol (4 ml), and
p-toluenesulfonic acid (30 mg) were dissolved in triethylortho-
formate (7 ml) and stirred for 30 min at 60°C. Water was
added, and the mixture was extracted with ethyl acetate and
washed successively with water and saturated sodium chloride
solution. The organic portion was dried (Na SO ), evaporated
to dryness, and filtered over a short silica gel column using
heptane/ethyl acetate 9/1 (v/v) as eluent to give crude 3-me-
thoxy-7-methylestra-1,3,5(10),6-tetraen-17-one cyclic 1,2-
2
4
2
4
droxy-3-methoxy-7-methylestra-2,5(10) -dien-17-dione cy-
clic (1,2-ethanediyl acetal) 29 (258 mg, 0.72 mM, 92%), Rf
0.5 [heptane/ethyl acetate (1:1 v/v)]. 29 (256 mg, 0.71 mM)
ethanediyl acetal 25, R 0.85 [heptane/ethyl acetate (8:2 v/v)].
f
25 (1 g, crude) was dissolved in THF (10 ml). Water (1 ml)
was dissolved in methylene chloride (3 ml) and SiO (49
2
was added, and the solution was cooled to 0°C. N-Chlorosuc-
cinimide (400 mg, 3 mM) was added, the ice bath was re-
moved, and the reaction mixture was stirred for 18 h at room
temperature. Then, water was added, and the solution was
extracted with ethyl acetate, dried (Na SO ), and evaporated to
mg) was added. Then, a solution of oxalic acid (63 mg, 0.7
mM) and 1 drop of water in methanol (1 ml) was added, and
the reaction mixture was stirred for 15 min at room tem-
perature. The suspension was filtered and rinsed with meth-
ylene chloride, and the filtrate was washed successively
2
4
dryness to give crude (6␤,7␣)-7-chloro-6-hydroxy-3-methoxy-
-methylestra-1,3,5(10)-trien-17-one cyclic 1,2-ethanediyl ac-
with saturated NaHCO solution and saturated sodium chlo-
3
7
ride solution. The organic layer was dried (Na SO ) and
2
4
etal 26 (R 0.15 [heptane/ethyl acetate (8:2 v/v)], which was
evaporated to dryness to give 240 mg of a 1/1 mixture of 30,
f
used in the next step without further purification. Crude 26 (1.2
g) was dissolved in THF (20 ml), potassium tert-butoxide (337
mg, 3 mM) was added and the reaction mixture was stirred for
R 0.25 [heptane/ethyl acetate (1:1 v/v)], and (7␤)-7-hy-
f
droxy-7-methylestr-5(10)-ene-3,17-dione 31, R 0.15 [hep-
f
tane/ethyl acetate (1:1 v/v)]. This mixture was used in the
next step without further purification.
1
h at room temperature. Then, another portion potassium
tert-butoxide (300 mg, 2.7 mM) was added, and the reaction
mixture was stirred for another hour. The solution was poured
into water, extracted with ethyl acetate, and washed with sat-
urated sodium chloride solution. The organic layer was dried
2.1.22. (3␣,7␤)-3,7-Dihydroxy-7-methylestr-5(10)-en-17-
one (34)
Lithium tri-tert-butoxy aluminum hydride (406 mg, 1.6
mM) in THF (4 ml) was added dropwise to a solution of 30
and 31 (240 mg, 0.7 mM) dissolved in THF (4 ml). The
reaction mixture was stirred for 1.5 h at room temperature,
poured into saturated ammonium chloride solution, and
extracted two times with ethyl acetate. The combined or-
ganic layers were washed successively with water and sat-
urated sodium chloride solution, dried (Na SO ), and evap-
(
Na SO ) and evaporated to dryness to give crude (6␤,7␤)-
2
4
6,7-epoxy-3-methoxy-7-methylestra-1,3,5(10) -trien-17-one
cyclic 1,2-ethanediyl acetal 27 (770 mg, crude), R 0.3 [hep-
f
tane/ethyl acetate (8:2 v/v)]. 27 (770 mg, crude), dissolved in
dry THF (7 ml), was added to a suspension of lithium alumi-
num hydride (569 mg, 15 mM) in dry THF (15 ml). The
reaction mixture was heated to reflux for 0.5 h. Then, saturated
sodium sulfate solution was added dropwise until the excess of
2
4
orated to dryness to give 236 mg of a 1/1 mixture of
LiAlH was quenched. Water was added, and the reaction
(3␣,7␤)-3,7-Dihydroxy-7-methylestr-5(10)-en-17-one cy-
4
mixture was extracted with ethyl acetate and washed with
saturated sodium chloride solution. The organic portion was
dried (Na SO ) and evaporated to dryness to give the crude
clic 1,2-ethanediyl acetal 33, R 0.55 (ethyl acetate), and
f
(3␣,7␤,17␤)-7-methylestr-5(10)-ene-3,7,17-triol 32, R 0.4
f
(ethyl acetate). This mixture was dissolved in acetone (5
ml), and a 0.1 N HCl (0.5 ml) solution was added. The
solution was stirred for 5 h at room temperature, saturated
sodium bicarbonate solution was added, and the mixture
was extracted with ethyl acetate. The organic layer was
dried (Na SO ) and evaporated to dryness to give the crude
2
4
product, which was purified by column chromatography [silica
gel, heptane/ethyl acetate 8/2 (v/v)] to give 28 (280 mg, 0.78
1
mM, 26%), R 0.1 [heptane/ethyl acetate (8:2 v/v)]. H NMR:
f
(
CDCl , 400 MHz) ␦ 0.94 (s, 3H, 18CH ), 1.18 (s, 3H, 7-CH ),
3
3
3
2.77 (d, J ϭ 16 Hz, 1H, 6␣H), 2.97 (d, J ϭ 16 Hz, 1H, 6␤H),
2
4
3
.77 (s, 3H, Ar-OCH ), 3.92 (m, 4H, ketal) 6.57 (d, 1H,
product mixture, which was separated by column chroma-
3
Ar-4H), 6.73 (dd, 1H, Ar-2H), 7.21 (d, 1H, Ar-1H).
tography [silica gel, toluene/ethanol 95/5 (v/v)] to give 34
(
75 mg, 0.25 mM, 37%), R 0.35 [toluene/ethanol (8:2 v/v)].
H NMR: (CD OD, 400 MHz) ␦ 0.92 (s, 3H, 18-CH ), 1.11
3 3
f
1
2
.1.21. (7␤)-7-Hydroxy-7-methylestr-5(10)-ene-3,17-dione
cyclic 17-(1,2-ethanediyl acetal) (30)
Ammonia (ca. 20 ml) was condensed in a three-necked
flask equipped with a thermometer, dry ice cooler and ad-
(s, 3H, 7-CH ), 3.70 (m, 1H, 3␤H), exact mass calculated
for [MϩH] is 305.2117, Found: 305.2090, and 32 (25 mg,
3
ϩ
0.08 mM, 12%), R 0.2 [toluene/ethanol (8:2 v/v)], m.p.
f

R. Plate et al. / Steroids 65 (2000) 497–504
503
1
2
02°C. H NMR: (CD OD, 400 MHz) ␦ 0.78 (s, 3H, 18-
3.1. Synthesis of (3␣,7␣,17␣ )-7-methyl-19-norpregn-
5(10)-en-20-yne-3,7,17-triol (11)
3
CH ), 1.03 (s, 3H, 7-CH ), 1.10 (m, 1H, 12␣H), 1.20 (m,
3
3
1
H, 11␤H), 1.29 (m, 1H, 14␣H), 1.36 (m, 1H, 2␣H), 1.42
(m, 1H, 16␤H), 1.53 (t, 1H, 8␤H), 1.60 (m, 1H, 9␣H), 1.67
(m, 1H, 15␤H), 1.77 (m, 1H, 4␣H), 1.82 (m, 1H, 6␣H), 1.83
(m, 1H, 12␤H), 1.85 (m, 1H, 15␣H), 1.90 (m, 1H, 1␣H),
In our first approach toward 7-hydroxy Org OD 14 de-
rivatives, a synthetic route via the key intermediate 5 was
6
elaborated commencing with the ⌬ -estrone derivative 1
1
1
4
.92 (m, 1H, 2␤H), 1.93 (m, 1H, 11␣H), 1.95 (m, 1H,
(Scheme 2). Ketalization of the readily available starting
6␣H), 2.11 (m, 1␤H), 2.12 (m, 1H, 6␤H), 2.14 (m, 1H,
material 1 gave the corresponding 17-acetal 2. Epoxidation
1
3
6
␤H), 3.58 (t, 1H, 17␣H), 3.70 (m,1H, 3␤H). C-NMR
of the ⌬ -double bond, using 3-chloro-peroxybenzoic acid
(
CD OD, 100 MHz) 12.2 (q, 18-CH ), 20.7 (q, 7-CH ), 26.7
as a reagent, gave the (6␣,7␣)-6,7-epoxy-3-methoxyestra-
1,3,5(10)-trien-17-one cyclic 1,2-ethanediyl acetal (3). Ring
opening of the epoxy function, using lithium aluminum
hydride as a reagent, gave the (7␣)-7-hydroxy-3-me-
thoxyestra-1,3,5 (10)-trien-17-one cyclic 1,2-ethanediyl ac-
etal (4). Oxidation of 4, using the Jones reagent, gave the
3
3
3
(t, C15), 27.2 (t, C11), 31.3 (t, C16), 33.6 (t, C2), 38.4 (t,
C1), 38.8 (t, C12), 40.9 (t, C4), 45.8 (s, 13C), 47.9 (d, C14),
8.4 (d, C9), 48.6 (d, C8), 49.7 (t, C6), 68.9 (d, C3), 72.9 (s,
C7), 82.6 (d, C17), 126.4 (s, C5), 130.1 (s, C10). Exact mass
4
ϩ
calculated for [MϩHϩGlyc.]
is 399.2747, Found:
3
99.2740.
desired
17,17-ethylenedioxy-3-methoxyestra-1,3,5(10)-
trien-7-one (5) in moderate yield. Attempts to introduce a
methyl group at the 7-keto position of derivative 5, either
using methyllithium or methylmagnesium chloride as re-
agents, failed in that no reaction occurred. This failure may
be attributed to complete enolization of the keto function
under these conditions. Application of a modified method
[6] gave the desired addition product 17,17-ethylenedioxy-
3-methoxy-7␤-methylestra-1,3,5(10)-trien-7␣-ol (6) in 55%
yield. The stereochemistry observed (7␣-OH, 7␤-Me) was
surprising and could not be rationalized with the Felkin–
Anh principle [7,8]. Birch reduction of 6 gave 7, which
upon mild, selective hydrolysis, gave 17,17-ethylenedioxy-
7␣-hydroxy-7␤-methylestr-5(10)-en-3-one (8). Reduction
of the 3-ketofunction of 8, using lithium tri-tert-butoxy
aluminum hydride as a reagent, gave the 3␣-hydroxy deriv-
ative 9. Finally, deprotection of the 17-ketal function, gave
10, which upon ethynylation, gave the desired (3␣,7␣,17␣)-
7-methyl-19-norpregn-5(10)-en-20-yne-3,7,17-triol (11).
2
.1.23. (3␣,7␤,17␣ )-7-Methyl-19-norpregn-5(10)-en-20-
yne-3,7,17-triol (35)
Potassium tert-butoxide (258 mg, 2.3 mM) was dis-
solved in dry THF (3 ml). The solution was cooled to 0°C
and stirred for 30 min whereas nitrogen was passed through.
Then, although the temperature was kept at 0°C, acetylene
was passed through the solution for 1 h. 34 (70 mg, 0.23
mM), dissolved in THF (3 ml), was added to the white
suspension, and the reaction mixture was stirred for 1 h at
0
°C. Then, the yellow suspension was poured into saturated
ammonium chloride solution and extracted with ethyl ace-
tate. The organic layer was dried (Na SO ) and evaporated
to dryness to give the crude product, which was purified by
column chromatography [silica gel, toluene/ethanol 95/5
2
4
(v/v)] to give 35 (40 mg, 0.12 mM, 53%), R 0.3 [toluene/
f
1
ethanol (8:2 v/v)], m.p. 175°C. H NMR: (CDCl , 600
MHz) ␦ 0.86 (s, 3H, 18-CH ), 1.03 (s, 3H, 7-CH ), 1.19 (m,
3
3
3
1
1
H, 11␤H), 1.38 (m, 1H, 2␤H), 1.52 (t, 1H, 8␤H), 1.58 (m,
H, 9␣H),1.61 (m, 1H, 12␤H), 1.73 (m, 1H, 15␤H), 1.78
3.2. Synthesis of (3␣,7␤,17␣ )-7-methyl-19-norpregn-
5(10)-en-20-yne-3,7,17-triol (35)
(m, 1H, 4␤H), 1.82 (m, 1H, 12␣H), 1.83 (dd, 1H, 6␣H),
1
1
1
4
3
2
3
.86 (m, 1H, 14␣H), 1.90 (m, 1H, 15␣H), 1.90 (m, 1H,
6␤H), 1.92 (m, 1H, 1␤H), 1.92 (m, 1H, 2␣H), 1.99 (m, 1H,
1␣H), 2.13 (m, 1␣H), 2.13 (dd, 1H, 6␤H), 2.15 (m, 1H,
␣H), 2.20 (m, 1H, 16␣H), 2.87 (s,1H, CϵCH), 3.7 (m, 1H,
In our second approach toward 7-hydroxy Org OD 14
derivatives, a route commencing with 17␤-hydroxyestr-4-
en-3-one (12, 19-nortestosterone) was devised (Scheme 3)
[9]. The oxidation of 12 with oxygen in dimethylform-
amide, followed by in situ methylation with methyliodide,
1
3
␤H). C-NMR (CD OD, 150 MHz) 13.9 (q, 18-CH ),
3
3
0.8 (q, 7-CH ), 26.3 (t, C15), 27.4 (t, C11), 28.6 (t, C1),
3
3.7 (t, C2), 34.8 (t, C12), 40.6 (t, C16), 41.1 (t, C4), 47.0
gave
17␤-hydroxy-3-methoxyestra-1,3,5(10)-trien-6-one
(
d, C9), 47.6 (d, C14), 49.3 (s, C13), 49.5 (d, C8), 49.6 (t,
(13). Protection of the 17-hydroxy function of 13 with the
tert-butyldimethylsilyl group gave 17␤-(tert-butyldimethyl-
silyl)oxy-3-methoxyestra-1,3,5(10)-trien-6-one (14). Selec-
tive methylation at the 7-position, using methyliodide in
dimethoxyethane as a reagent, gave 17␤-(tert-butyldimeth-
ylsilyl)oxy-3-methoxy-7␣-methyl-estra-1,3,5(10)-trien-6-
one (15). Reduction of the keto function of 15 with lithium
aluminum hydride gave 17␤-(tert-butyldimethylsilyl)oxy-3-
methoxy-7␣-methyl-estra-1,3,5(10)-trien-6␣-ol (16). Dehy-
dration of 16 into 17␤-(tert-butyldimethylsilyl)oxy-3-me-
thoxy-7-methylestra-1,3,5(10),6-tetraene (17) was carried
out using Amberlyst 15 in refluxing toluene at high dilution.
C6), 68.5 (D, C3), 73.0 (t, C7), 75.1 (d, ϵCH), 80.2 (s,
C17), 89.4 (s, Cϵ), 126.4 (s, C5), 130.0 (s, C10). Exact
mass calculated for [MϩHϩGlyc.] is 423.2747, Found:
ϩ
4
23.2748.
3
. Results and discussion
Here, we describe our approaches toward the synthesis of
the 7-␤-hydroxy metabolite of Org OD 14, 35, as well as its
isomeric 7-␣-hydroxy derivative 11.

5
04
R. Plate et al. / Steroids 65 (2000) 497–504
This was necessary to prevent the formation of dimers e.g.
these dimers are formed in concentrated solution as a result
and 7␤-hydroxy-7␣-methyl-estra-5(10)-diene-3,17-dione
(31). This mixture was reduced with lithium tri-tert-butoxy
aluminum hydride to 17,17-ethylenedioxy-7␣-methyl-estr-
5(10)-ene-3␣,7␤-diol (33) and 7␣-methyl-estr-5(10)-ene-
3␣,7␤,17␤-triol (32). Hydrolysis of 33 gave 7␣-methyl-
of the attack of the intermediate formed carbocation at the
6
6
-position with the A-ring aromat. Reaction of the ⌬ -
double bond of 17 with N-chlorosuccinimide in aqueous
THF gave 17␤-(tert-butyldimethylsilyl)oxy-7␣-chloro-3-
methoxy-7␤-methyl-estra-1,3,5(10)-trien-6␤-ol (18). Ring
closure of 18 gave the 17␤-(tert-butyldimethylsilyl)oxy-3-
methoxy-7␣-methyl-6␤,7␤-epoxyestra-1,3,5(10)-triene
estr-5(10)-en-3␣,7␤-diol-17-one
(34),
and
finally,
ethynylation gave the desired target molecule (3␣,7␤,17␣)-
7-methyl-19-norpregn-5(10)-en-20-yne-3,7,17-triol (35).
The pharmacological properties of the 7-hydroxy deriv-
atives of ORG OD14, namely 11 and 35, will be described
elsewhere.
(19). In small-scale experiments, the epoxide 19 was iso-
lated after column chromatography in 30% yield. In larger
scale experiments, only 5% of 19 was isolated, whereas the
major portion of the epoxide underwent ringopening during
column chromatography to give 17␤-(tert-
butyldimethylsilyl)oxy-3-methoxy-7␣-methyl-estra-
Acknowledgments
1
,3,5(10)-trien-6␣,7␤-diol (20). Epoxide 19 was found to be
We wish to thank F. Martens, R. Aspers (NMR-spec.),
and G. Schmeits (Mass-spec.) for their help in interpretation
of the spectral data.
unstable at room temperature. Reduction of the epoxide
function of 19, using lithium aluminum hydride as a re-
agent, gave 17␤-(tert-butyldimethylsilyl)oxy-3-methoxy-
7
␣-methylestra-1,3,5(10)-trien-7␤-ol (22). Alternatively, 22
was prepared from 20 by mesylation and subsequent reduc-
tion of 21. Deprotection of 22 gave 3-methoxy-7␣-
methylestra-1,3,5(10)-trien-7␤,17␤-diol (23).
Oxidation of 23, using N-methylmorpholine N-oxide and
a catalytic amount of tetra-n-propylammonium perruthenate
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3