Ergosteroids III. Syntheses and biological activity of seco-steroids related to dehydroepiandrosterone

Article abstract of DOI:10.1016/S0039-128X(98)00066-X

The unusual activity of some D-ring-seco estrogens led us to prepare several seco steroids related to dehydroepiandrosterone (DHEA) and to test for their ability to mimic thyroid hormone and 7-oxo-DHEA (1) as inducers of thermogenic enzymes in rats' livers. Only one, 3β-acetoxy-17a-oxa-androst- 5-ene-7,17-dione (17), was capable of inducing both mitochondrial glycerophosphate dehydrogenase and malic enzyme. The closely related 3β- hydroxy-17a-oxa-androsta-5,15-diene- 7,17-diones (both 14α and 14β, 14 and 15) induce the formation of malic enzyme but not of glycerophosphate dehydrogenase. The 3β-propionyl ester of the above 14α steroid was not active, presumably because it was not deacylated in vivo. The 16,17 dicarboxylic acid (9) produced by opening the D-ring also induced the formation of malic enzyme but not of glycerophosphate dehydrogenase. 3β- Acetoxyandrost-5-ene-7,16,17-trione, an intermediate in the synthesis of D- ring seco compounds enhanced the formation of both enzymes. Twelve other D- ring seco compounds were not active. Seco androstanes oxygenated at position 7 and with expanded A or B rings were not active.

Full text of DOI:10.1016/S0039-128X(98)00066-X

Ergosteroids III. Syntheses and
biological activity of seco-steroids
related to dehydroepiandrosterone
Ieva L. Reich,*† Henry Lardy,* Yong Wei,* Padma Marwah,* Nancy Kneer,*
Douglas R. Powell,† and Hans J. Reich†
*Institute for Enzyme Research and †Department of Chemistry, University of Wisconsin-Madison,
Madison, Wisconsin 53705
The unusual activity of some D-ring-seco estrogens led us to prepare several seco steroids related to dehydro-
epiandrosterone (DHEA) and to test for their ability to mimic thyroid hormone and 7-oxo-DHEA (1) as inducers
of thermogenic enzymes in rats’ livers. Only one, 3␤-acetoxy-17a-oxa-androst-5-ene-7,17-dione (17), was
capable of inducing both mitochondrial glycerophosphate dehydrogenase and malic enzyme. The closely related
3␤-hydroxy-17a-oxa-androsta-5,15-diene-7,17-diones (both 14␣ and 14␤, 14 and 15) induce the formation of
malic enzyme but not of glycerophosphate dehydrogenase. The 3␤-propionyl ester of the above 14␣ steroid was
not active, presumably because it was not deacylated in vivo. The 16,17 dicarboxylic acid (9) produced by
opening the D-ring also induced the formation of malic enzyme but not of glycerophosphate dehydrogenase.
3␤-Acetoxyandrost-5-ene-7,16,17-trione, an intermediate in the synthesis of D-ring seco compounds enhanced
the formation of both enzymes. Twelve other D-ring seco compounds were not active. Seco androstanes
oxygenated at position 7 and with expanded A or B rings were not active. (Steroids 63:542–553, 1998) © 1998
by Elsevier Science Inc.
Keywords: ergosteroids; 7-oxo-DHEA; DHEA; seco-steroids; thermogenic enzymes
manuscript in preparation). This was an inducement to
examine the seco compounds as well. We report here the
synthesis of several known and new A-, B- and D-ring seco
derivatives of DHEA. The ability of these steroids, and of
intermediates in their chemical synthesis, to induce thermo-
genic enzymes in rats has been tested. Some expanded
D-ring steroids retain the ability to induce one or both of the
thermogenic enzymes whereas androstanes with expanded
A or B rings were not effective.
Introduction
In 1932 Marrian and Haselwood,1 and independently, Doisy
and coworkers2 reported that alkali fusion of theelol (estriol)
yielded a dicarboxylic acid. Soon thereafter the Doisy
group3 demonstrated that similar treatment of theelin (es-
trone) yielded a monocarboxylic acid and that both of these
products possessed several times the estrogen potency of
estrone. The structures of these fusion products were estab-
lished by Miescher4,5 who named them marrianolic acid and
doisynolic acid, respectively. The latter is 10 to 20 times as
active as estrone and a levorotatory bisdehydrodoisynolic
acid is more than 100 times as active when given orally.5
Heer and Miescher6 prepared a series of D-ring seco
compounds beginning with dehydroepiandrosterone (here-
after DHEA) but found them devoid of androgen or estro-
gen activity.5 In our search for possible metabolites of
DHEA with greater ability than the parent steroid to induce
the formation of thermogenic enzymes when administered
to rats, we have found that activity is retained in compounds
with wide variations of substituents on the D-ring7 (and
Experimental
Nuclear magnetic resonance (NMR) spectra were obtained on
Bruker WP-200, WP-270, AM-300 or AM-500 spectrometers.
Unless otherwise indicated, the spectra were measured in CDCl3
using tetramethylsilane (␦ 0) as reference for the 1H NMR spectra,
and the CDCl3 triplet (␦ 77.0) for the 13C spectra. Infrared (IR)
spectra in CHCl3 were recorded on a Mattson Polaris spectropho-
tometer. A Kratos MS-80RFA spectrometer was employed to
obtain mass spectra (MS). Elemental analyses were done by Gal-
braith Laboratories. Other conditions were as described.8
Diethyl ether and tetrahydrofuran (THF) were freshly distilled
from sodium benzophenone ketyl. Dry CH2Cl2 was distilled from
CaH2 as were diisopropylamine and triethylamine. Pyridine was
distilled from NaOH. n-Butyllithium in hexane was obtained from
Foote Mineral Co. and titrated using n-propanol with 1,10-
phenanthroline as indicator.9 Column chromatography was carried
Address reprint requests to Ieva L Reich, Department of Chemistry, 1101
University Avenue, Madison, WI 53706 or Henry Lardy, Institute for
Enzyme Research, 1710 University Avenue, Madison, WI 3705, USA.
Received May 12, 1998; accepted July 6, 1998.
Steroids 63:542–553, 1998
© 1998 by Elsevier Science Inc. All rights reserved.
655 Avenue of the Americas, New York, NY 10010
0039-128X/98/$19.00
PII S0039-128X(98)00066-X

Seco-steroids related to dehydroepiandrosterone: Reich et al.
out with Baker silica gel (60–200 mesh). Preparative thin-layer
chromatography was done on Brinkmann silica gel MN PUV254.
Assays of compounds for their ability to induce mitochondrial
glycerophosphate dehydrogenase and cytosolic malic enzyme in
rat livers have been described.8,10 Again, we emphasize that the
assays are not quantitative, but serve as a guide to further work.
Compounds are considered active only if they enhance the assayed
activity of these thermogenic enzymes to greater than 150% of that
in livers of control rats.
Scheme 2
Syntheses
Synthesis of 16,17-dione and D-ring seco
androsterones
acetoxyselenide 6 as a mixture of diastereomers. Oxidative
elimination formed the enol acetate 7 in 60% yield from 5.
A small amount (7%) of the enol m-chlorobenzoate was also
formed and separated in the chromatography. The enol
acetate and enol benzoate were hydrolyzed in methanol-
triethylamine to form 58% of the yellow solid 16, 17-dione
8. Oxidative cleavage with sodium metaperiodate gave the
crystalline diacid 9 in 49% yield.
The related secoandroectenenoic acid 12a was prepared
from the doisynolic acid analog 11a6 as shown in Scheme 2.
The half-esters of 3␤-hydroxy-16,17-secoandrost-5-ene-
16,17-dioic acid 11b and 11c, prepared according to litera-
ture procedures,13 were converted to the corresponding
7-oxo compounds 12b and 12c by chromic acid oxidation as
described for 11a.
16,17-Diones in the 7-oxo-dehydroepiandrosterone or 3␤-
hydroxyandrost-5-ene-7,17-dione (7-oxo-DHEA) series
have not been reported, although structures without the
7-oxo group have been prepared in poor yield.11 Scheme 1
presents a new approach in which a seleno–Pummerer re-
action12 is used to oxidize the 16-position. This route is
more efficient than the previously published procedure.
The starting material for the synthesis of 8 was 1, 3␤-
acetoxyandrost-5-ene-7,17-dione (7-oxo-DHEA acetate),
which was prepared from dehydroepiandrosterone acetate
as described.8 Treatment of 1 with 2.4 equivalents of lithium
diisopropylamide in the presence of trimethylchlorosilane
(TMSCl) gave the 17-silyl enol ether (70% of the material
was also C-silylated on the 3␤-acetoxy group). This mixture
was selenenylated with benzeneselenenyl chloride and de-
silylated with tetra-n-butyl ammonium fluoride (nBu4NF) to
give the 16␣-selenide 5 in ϳ90% yield from 1. Oxidation of
the selenide 5 to the selenoxide and treatement with acetic
anhydride (the seleno-Pummerer reaction) gave the 16-
3␤-(trimethylsilyl)acetoxy-17-hydroxyandrosta-5,16-
dien-7-one trimethylsilyl ether (2)
To a solution of 1.00 g (2.90 mmol) of 3␤-acetoxyandrost-
5-ene-7,17-dione (1) in 20 mL of dry THF under N2, was
added 1.07 mL (8.43 mmol) of TMSCl. The reaction mix-
ture was cooled to Ϫ78°C. Lithium diisopropylamide
(LDA) was prepared at Ϫ78°C under N2 from 1.07 mL
(7.66 mmol) of diisopropylamine and 3.60 mL of a 1.94 M
solution of n-butyllithium in hexane (6.96 mmol) in 4 mL of
THF. The LDA solution was added under N2 via cannula to
the cold (Ϫ78°C) THF solution of 1. The reaction mixture
was allowed to warm to room temperature, poured into a
separatory funnel with 40 mL of hexane and 40 mL of
saturated NaHCO3 solution, and extracted. The organic
extract was washed with saturated NaCl solution, dried
(Na2SO4) and solvent was removed under reduced pressure
to yield 1.63 g of a solid which NMR showed to be a 70:30
mixture of the 17-enol silyl ether with C- (2) and
O-silylation (ketene acetal) on the 3-acetate. This mixture
was stirred rapidly in 60 mL of ether and 60 mL of H2O for
15 min to hydrolyze the ketene acetal, yielding a mixture of
2 and 3. 1H NMR of 2 in mixture (300 MHz) ␦ 5.76 (d, J ϭ
2 Hz, 6-H), 4.73 (tt, J ϭ 11, 5 Hz, 3␣-H), 4.53 (dd, J ϭ 3,
1.5 Hz, 16-H), 2.73 (ddd, J ϭ 14.5, 6, 3 Hz, 15␣-H), 2.15
(ddd, J ϭ 14.5, 10.5, 1.5 Hz, 15␤-H), 1.90 (s, CH2-Si), 1.22
(s, 19-CH3), 0.86 (s, 18-CH3), 0.20 (s, OSiMe3), 0.14 (s,
C-SiMe3). NMR of 3 in the mixture is essentially the same
as 2 except that the peak at ␦ 1.90 is replaced by a peak at
␦ 2.06 (s, CH3COO) and the peak at ␦ 0.14 is absent.
Scheme 1 Preparation of 3␤-acetoxy-7-oxo-16,17-secoandrost-
5-ene-16,17-dioic acid (9).
Steroids, 1998, vol. 63, October 543

Papers
and evaporated under reduced pressure to yield 0.713 g of
selenoacetate (85:15 mixture of diastereomers). NMR
showed some incorporation of m-chlorobenzoate, ϳ7%. 1H
NMR of chromatographed material, major isomer in mix-
ture; (300 MHz) ␦ 7.65 (dm, J ϭ 7.5 Hz, o-SePh), 7.35 (m,
SePh, 3H), 5.72 (d, J ϭ 1.5 Hz, 6-H), 4.71 (tt, J ϭ 11, 5 Hz,
3␣-H), 3.15 (dd, J ϭ 14.5, 6.5 Hz, 15␣-H), 2.06 (s,
3-CH3COO), 2.03 (s, 16-CH3COO), 1.35 (s, 19-CH3), 1.23
(s, 18-CH3); minor isomer in mixture ␦ 7.61 (dm, J ϭ 7.5
Hz, o-SePh), 7.35 (m, SePh), 5.76 (d, J ϭ 1.5 Hz, 6-H), 4.71
(tt, J ϭ 11, 5 Hz, 3␣-H), 3.29 (dd, J ϭ 13.5, 4.5 Hz, 15␣-H),
2.10 (s, 16-CH3COO), 2.07 (s, 3-CH3COO), 1.35 (s, 19-
CH3), 1.24 (s, 18-CH3).
3␤-acetoxy-16␣-phenyl selenoandrost-5-ene-7,17-
dione (5)
To a solution of 2.90 mmol of enol silyl ether (2 ϩ 3) in 20
mL of dry THF and 0.320 mL (4.00 mmol) of dry pyridine
at Ϫ78°C under N2 was added via cannula a THF solution
of 0.710 g (3.71 mmol) of PhSeCl in 4 mL of THF. The
reaction mixture was immediately poured into 60 mL of
CH2Cl2 and 60 mL of 0.5 N HCl solution and extracted. The
organic extract was washed with water and saturated
NaHCO3 solution. After drying (Na2SO4), solvent was re-
moved under reduced pressure yielding crude selenide (4 ϩ
5), 1.72 g. Chromatography on 100 g of silica eluting with
hexane-ethyl acetate (90:10 to 50:50, 50 mL fractions)
separated 4 and 5 from diphenyl diselenide and small
amounts of unreacted 1. Fractions 14–17 were combined to
give 1.44 g of 4 ϩ 5. 1H NMR (300 MHz) of 4: ␦ 7.68 (m,
o-SePh), 7.32 (m, SePh), 5.73 (d, J ϭ 1.5 Hz, 6-H), 4.72 (tt,
J ϭ 10.5, 5 Hz, 3␣-H), 4.10 (d, J ϭ 7.5 Hz, JSeH ϭ 5 Hz,
16␤-H), 2.98 (ddd, J ϭ 14, 5.5, 0.5 Hz, 15␣-H), 1.90 (s,
CH2Si), 1.20 (s, 19-CH3), 0.91 (s, 18-CH3), 0.14 (s,
C-SiMe3).
3␤,16-diacetoxyandrosta-5,15-diene-7,17-dione (7)
To a solution of 1.1 mmol selenoacetate 6 in 25 mL of CCl4
was added 0.12 mL of pyridine and 10 mg of benzenesele-
ninic acid. The mixture was stirred vigorously and 3.4 mL
of 15% aqueous H2O2 was added. After stirring for 45 min
(mild exotherm), the yellow color faded. The CCl4 solution
was washed twice with saturated NaHCO3, dried and evap-
orated under reduced pressure to yield 0.390 g of a crude
solid containing 7% of the enol m-chlorobenzoate. Chroma-
tography on 50 g of silica gel with 50% ethyl acetate/hexane
(25 mL fractions), gave 0.236 g (54% yield) of pure enol
acetate (fractions 9 and 10), m.p. 208–210°C. 1H NMR (300
MHz) ␦ 7.81 (broad s, 15-H), 5.81 (d, J ϭ 1.5 Hz, 6-H),
4.74 (tt, J ϭ 11.5, 4.5 Hz, 3␣-H), 2.25 (s, 16-CH3COO),
2.07 (s, 3-CH3COO), 1.29 (s, 19-CH3), 1.23 (s, 18-CH3).
HRMS Calcd. for C23H28O6: 400.1886; Found: 400.1889.
To a solution of 1.19 mmol of the C-silylated acetate 4
(containing some 5) in 10 mL of CH2Cl2 and 15 mL of H2O
was added 4 mL of a 1 M solution of nBu4NF in THF. The
mixture was stirred vigorously for 45 min and then poured
into 20 mL of ether-20 mL of hexane-40 mL of H2O and
extracted. The organic extract was washed with H2O two
times, saturated NaCl, dried (Na2SO4) and the solvent was
removed under reduced pressure to yield a white solid, 95%
1
pure by H NMR, 0.594 g, ϳ95% yield; m.p. 123–124°C.
1
H NMR (300 MHz) of 5: ␦ 7.68 (m, o-SePh), 7.31 (m,
SePh), 5.73 (d, J ϭ 1.5 Hz, 6-H), 4.72 (tt, J ϭ 11, 5 Hz,
3␣-H), 4.10 (d, J ϭ 7.5 Hz, JSeH ϭ 5 Hz, 16␤-H), 2.98 (ddd,
J ϭ 14, 5.5, 0.5 Hz, 15␣-H), 2.59 (ddd, J ϭ 14.5, 5, 2 Hz,
4␣-H), 2.47 (ddd, J ϭ 14.5, 11.5, 2 Hz, 4␤-H), 2.06 (s,
CH3COO), 1.21 (s, 19-CH3), 0.91 (s, 18-CH3). 13C NMR
(62.9 MHz) ␦ 215.8 (17-CO), 199.8 (7-CO), 170.1
(CH3COO), 164.7 (5-C), 135.6 (o-CH), 129.0 (m-CH),
128.8 (i-C), 128.3 (p-CH), 126.3 (6-CH), 71.8 (3-CH), 49.8
(CH), 48.0 (13-C), 44.5 (CH), 43.9 (CH), 43.7 (CH), 38.2
(10-C), 37.7 (CH2), 35.8 (CH2), 33.1 (CH2), 31.1 (CH2),
27.1 (CH2), 21.1 (CH3COO), 20.4 (11-CH2), 17.3 (19-
CH3), 14.2 (18-CH3). HRMS: Calcd. for C27H32O4Se:
500.1470; Found: 500.1476.
3␤-acetoxyandrost-5-ene-7,16,17-trione (8)
Crude enol acetate 7 (2.90 mmol) was dissolved in 17 mL
of THF and 43 mL of methanol. Triethylamine (1.8 mL,
12.9 mmol) was added and the solution was kept under N2
for 16 h. The mixture was then extracted between CHCl3
and 0.5 N HCl solution. The organic layer was washed with
50% saturated NaCl solution, dried (Na2SO4) and the sol-
vent was evaporated to yield a yellow solid. The solid was
crystallized from CHCl3-CCl4-ether mixture to yield yellow
crystals, 0.534 g, m.p. 221–222°C. The mother liquor was
chromatographed on preparative TLC plates to yield 0.076
1
g. The total yield was 0.61 g, 58% from 1 (6 steps). H
NMR (300 MHz) ␦ 5.78 (d, J ϭ 1.5 Hz, 6-H), 4.73 (tt, J ϭ
11.5, 4.5 Hz, 3␣-H), 3.42 (dd, J ϭ 19.5, 6.5 Hz, 15␣-H),
2.43 (dd, J ϭ 19.5, 13.5 Hz, 15␤-H), 2.07 (s, 3-CH3COO),
1.29 (s, 19-CH3), 1.06 (s, 18-CH3). 13C NMR (75.4 MHz) ␦
204.5 (CO), 203.1 (CO), 199.4 (7-CO), 170.1 (CH3COO),
165.4 (5-C), 126.0 (6-CH), 71.7 (3-CH), 49.4 (CH), 47.8
(13-C), 43.5 (CH), 38.9 (CH2), 38.6 (CH), 38.5 (10-C), 37.8
(CH2), 35.6 (CH2), 29.9 (CH2), 27.1 (CH2), 21.1
(CH3COO), 20.1 (11-CH2), 17.3 (19-CH3), 13.6 (18-CH3).
IR (cmϪ1) 3029, 2953, 1750, 1731, 1671, 1250, 1036.
HRMS Calcd. for C21H26O5: 358.1780; Found: 358.1778.
3␤,16-diacetoxy-16-phenylselenoandrost-5-ene-7,17-
dione (6)
A solution of 1.1 mmol of selenide 5 in 25 mL of CH2Cl2
was cooled to 0°C. To the well-stirred solution was added
0.296 g (1.2 mmol) of m-chloroperbenzoic acid, 70%
(MCPBA). After 5 min, 0.40 mL of dimethyl sulfide was
added to quench the oxidant. The CH2Cl2 solution was
stirred with 50 mL of cold saturated NaHCO3 solution and
filtered through Na2SO4 at 0°C. Acetic anhydride (1.2 mL)
and pyridine (1.2 mL) were added and the solution was
stirred at room temperature for 1 h. A solution of saturated
NaHCO3 was added and the mixture was stirred until bub-
bling ceased. Layers were separated and the organic layer
was washed again with saturated NaHCO3, dried (Na2SO4)
3␤-acetoxy-7-oxo-16,17-secoandrost-5-ene-16,17-
dioic acid (9)
To a solution of the pure triketone 8 (0.400 g, 1.12 mmol)
in 16 mL of MeOH was added 0.263 g (1.23 mmol) of
544 Steroids, 1998, vol. 63, October

Seco-steroids related to dehydroepiandrosterone: Reich et al.
sodium metaperiodate, 4 mL of H2O and 2 mL of 1 N HCl
(7 mL), a solution of CrO3 (690 mg) in acetic anhydride (3.8
mL) was added dropwise during a period of 15 min at room
temperature. The reaction mixture was stirred at 45°C under
N2 for 2.5 h and poured into ice-water (250 mL). The
product was extracted from the ice-water with ethyl acetate
(60 mL ϫ 6). The extracts were combined, washed with
brine (60 mL), dried (MgSO4), and concentrated to dryness.
The crude product was purified over silica gel (CHCl3:
acetone ϭ 20:1) to afford compound 12a (400 mg, 48%);
solution. The solution was stirred for 2 h at which point a
white solid had formed and the yellow color of 8 was
absent. The mixture was partitioned between 200 mL of
H2O and 100 mL of CHCl3. The aqueous layer was ex-
tracted two more times with CHCl3. The organic layers
were combined and washed with 50% saturated NaCl solu-
tion, dried (Na2SO4) and evaporated under reduced pressure
to yield a white solid (0.390 g) which was crystallized from
acetone producing 0.215 g of pure crystals (49%), mp
1
m.p. 175–179°C; H NMR (300 MHz, CDCl3) ␦ 5.76 (s,
1
228–238°C d. H NMR (300 MHz) ␦ 5.77 (d, J ϭ 1.5 Hz,
1H, 6-H), 4.71 (m, 1H, 3-H), 2.07 (s, -CO2CH3), 1.21 (s,
CH3), 1.08 (s, CH3), 0.89 (t, J ϭ 7 Hz, CH3). HRMS
calculated for C21H30O5: 362.2093; Found: 362.2089.
6-H), 4.72 (tt, J ϭ 11.5, 5 Hz, 3␣-H), 2.91 (m, 2H), 2.57
(ddd, J ϭ 13.5, 5, 1.5 Hz, 4␣-H), 2.45 (ddd, J ϭ 13, 11.5,
1.5 Hz, 4␤-H), 2.06 (s, CH3COO), 1.24 (s, CH3), 1.21 (s,
CH3). 13C NMR (75.4 MHz) ␦ 199.8 (7-CO), 184.0 (17-
COOH), 179.5 (16-COOH), 170.2 (CH3COO), 163.1 (5-C),
126.2 (6-CH), 72.1 (3-CH), 48.7 (CH), 44.9 (13-C), 44.5
(CH), 39.0 (10-C), 37.5 (CH2), 37.2 (CH), 36.4 (CH2), 35.8
(CH2), 34.7 (CH2), 27.1 (CH2), 21.2 (CH3COO), 19.2 (11-
CH2), 16.6 (19-CH3), 15.4 (18-CH3). IR (cmϪ1) 3600,
3300–3700 (br), 3013, 2954, 1715, 1675, 1249, 1205.
3␤-acetoxy- 7-oxo-16,17-secoandrost-5-ene-16,17-
dioic acid, 16 and 17-methyl half-esters (12b and
12c)
The half-esters 11b and 11c (1.0 g samples of each, pre-
pared by the procedure of von Seeman and Grant13) were
oxidized as decribed for 11a. Chromatography yielded
0.460 g (44%) of 12b and 0.200 g (19%) of 12c. The
compounds were very hygroscopic and resisted crystalliza-
tion. 12b: 1H NMR (300 MHz, CDCl3) ␦ 5.72 (s, 1H), 4.72
(m, 1H), 3.68 (s, 3H), 2.05 (s, 3H), 1.20 (s, 3H), 1.12 (s,
3H). 12c: 1H NMR (300 MHz, CDCl3) ␦ 5.78 (s, 1H), 4.72
(m, 1H), 3.69 (s, 1H), 2.05 (s, 3H), 1.23 (s, 3H), 1.10 (s,
3H).
Dimethyl 3␤-acetoxy-7-oxo-16,17-secoandrost-5-ene-
16,17-dioate (10)
The mother liquor from the crystallization of diacid 9 was
evaporated under reduced pressure and the solid (0.158 g,
50% diacid) was dissolved in 3 mL of THF and treated with
1 mmol of diazomethane. Preparative TLC (6% MeOH/
CHCl3) yielded 75 mg of a white solid; Rf ϭ 0.36, 16%
yield from the triketone 8. Crystallization from ether-
hexane gave colorless crystals, 64 mg, m.p. 155–156°C. 1H
NMR (300 MHz) ␦ 5.73 (d, J ϭ 1.5 Hz, 6-H), 4.71 (tt, J ϭ
11, 5 Hz, 3␣-H), 3.693 (s, OCH3), 3.685 (s, OCH3), 2.73
(dt, J ϭ 11, 4.5 Hz, 14␣-H), 2.56 (dd, J ϭ 13.5, 5.5, 2 Hz,
4␣-H), 2.46 (ddd, J ϭ 13.5, 11.5, 2 Hz, 4␤-H), 2.41 (t, J ϭ
11 Hz, 8-H), 2.36 (d, J ϭ 5 Hz, 15␣,␤-H), 2.05 (s,
CH3COO), 1.22 (s, 19-CH3), 1.14 (s, 18-CH3). 13C NMR
(75.4 MHz) ␦ 199.9 (7-CO), 177.3 (17-COOMe), 173.1
(16-COOMe), 170.1 (CH3COO), 163.4 (5-C), 126.2 (6-
CH), 72.0 (3-CH), 52.0 (OCH3), 51.4 (OCH3), 48.8 (CH),
46.5 (13-C), 45.4 (CH), 38.6 (10-C), 37.53 (CH), 37.48
(CH2), 37.3 (CH2), 35.7 (CH2), 34.7 (CH2), 27.1 (CH2),
21.1 (CH3COO), 19.6 (11-CH2), 16.9 (19-CH3), 15.6 (18-
CH3). IR (cmϪ1) 3025, 2953, 1731, 1672, 1437, 1367, 1248,
1175, 1035. HRMS calculated for C23H32O7: 420.2148;
Found: 420.2141. Analysis calculated for C23H32O7: C,
65.69; H, 7.67. Found: C, 65.55; H, 7.89.
Preparation of D-ring lactones
When the 16-␣ selenide 5a (prepared from 3␤-propionoxy-
7-oxo-DHEA,8 by the reactions in Scheme 1) was oxidized
under the normal conditions for selenoxide elimination,14,15
the lactone 13 was formed instead of the expected 15,16-
enone (Scheme 3). Similar observations have been reported
by Williams & Leber16 in the case of the selenide derived
from DHEA acetate, and by Fetizon et al.17 in the 17-oxo-
androstane series. This occurs because the selenoxide elim-
ination, which would place two more sp2 centers in the
already strained trans-fused five-membered ring, is slower
than the Baeyer-Villiger reaction to form the six-membered
ring lactone. After the ring expansion, the selenoxide elim-
inates rapidly to form the double bond in the six-membered
3␤-acetoxy-7-oxo-16,17-secoandrost-5-en-17-oic
acid (12a)
The doisynolic acid analog 11a (prepared by the method of
Heer and Miescher, m.p. 203–205°C; lit.6 204–206°C) (700
mg, 2.28 mmol) was stirred with acetic anhydride (4.5 mL),
triethylamine (3 mL), and 4-dimethylaminopyridine (42
mg) in CH2Cl2 for 2.5 h at room temperature. After removal
of the solvent the residue was dissolved in ethyl acetate and
the solution was washed with 0.1 N HCl and then with
brine, dried (MgSO4) and concentrated to dryness. To the
crude product (850 mg) and NaOAc (550 mg) in acetic acid
Scheme 3 Preparation of 3␤-hydroxy-17a-oxa-androsta-5,15-
diene-7,17-dione (15).
Steroids, 1998, vol. 63, October 545

Papers
ring. We have been able to form the five-membered ring
enone by doing the oxidation with MCPBA at 0°C, remov-
ing the acid and pyrolyzing rapidly in refluxing carbon
tetrachloride.15
al.19 and involved the use of N-hydroxyphthalimide as a
catalyst.
The 3-propionoxy group can be hydrolyzed to the alco-
hol by careful treatment with sodium carbonate in methanol.
Under more strongly basic conditions (sodium hydroxide in
methanol), the lactone isomerizes at the 14 position to give
the cis ring juncture. Ring opening does not occur. Minor
products include isomers at the 8 position and methoxide
addition to the enone.
3␤-propionoxy-17a-oxa-14␣-androsta-5,15-diene-
7,17-dione (13)
The crude selenide 5a (3.29 mmol), dissolved in 40 mL of
CH2Cl2, was stirred rapidly and treated with 10 mL of 15%
H2O2. After about 15 min, reaction began and the mixture
warmed slightly. After 45 min the yellow color of diphenyl
diselenide disappeared and the reaction mixture was poured
into ethyl acetate (200 mL) and H2O (60 mL) and extracted.
The organic layer was washed with 100 mL of saturated
NaHCO3 solution and 30 mL of saturated NaCl solution.
The crude solid (ϳ80% pure) was crystallized from MeOH
It is clear that 14 (14␣-H) is the kinetic product which
isomerizes under basic conditions to 15 (14␤-H). This is
1
confirmed by both the H NMR and 13C NMR data. In 14
the 14␣-H is axial and upfield, ␦ 2.80 (shielding by 7
carbonyl) with axial-axial coupling (J ϭ 11.5 Hz). The
15-H is downfield, ␦ 7.27 (deshielding by 7 carbonyl). In 15
the 14␤-H is equatorial with a small coupling and shifted
downfield since it is in the deshielding cone of the 7
carbonyl (␦ 3.51). The 15-H is upfield (␦ 6.38) as expected.
In the 13C NMR the major chemical shift changes occur in
the C- and D-rings. Most notably the C-18 methyl is sig-
nificantly upfield (8 ppm) for 14 due to the greater number
of ␥-gauche interactions present in the trans-fused rings.
The C-15 vinyl carbon is upfield in 15 (2 ppm) for the same
reason since it occupies an axial position. The C-14 tertiary
carbon having an axial substituent is shifted upfield (4
ppm).
A precedent for the formation of the lactone 17 (Scheme
4) was the demonstration by Westerfeld18 that treating es-
trone with H2O2 converted it to the corresponding 17 3 13
lactone. In the commercial synthesis of 7-oxo-DHEA-
acetate by the procedure described earlier,8 prolonged reac-
tion time results in the formation of by-products. Compound
17 was separated by chromatography from the 7-oxo-
DHEA-acetate in such a reaction mixture.
1
to yield 0.518 g, m.p. 233–235°C, 42%. H NMR (300
MHz) ␦ 7.26 (dd, J ϭ 9.5, 2.5 Hz, 15-H), 6.02 (dd, J ϭ 9.5,
3 Hz, 16-H), 5.81 (d, J ϭ 1.5 Hz, 6-H), 4.74 (tt, J ϭ 11.5,
5 Hz, 3␣-H), 2.81 (dt, J ϭ 11.5, 2 Hz, 14␣-H), 2.62 (ddd,
J ϭ 13.5, 5, 2 Hz, 4␣-H), 2.49 (ddd, J ϭ 13.5, 11.5, 2 Hz,
4␤-H), 2.39 (t, J ϭ 12 Hz, 8␤-H), 2.33 (q, J ϭ 7.5 Hz,
CH3CH2), 1.35 (s, 18-CH3), 1.25 (s, 19-CH3), 1.15 (t, J ϭ
7.5 Hz, CH3CH2). 13C NMR (67.9 MHz) ␦ 198.9 (7-CO),
173.5 (CH3CH2CO), 164.9 (5-C), 163.6 (17-CO), 148.8
(15-CH), 125.7 (6-CH), 120.2 (16-CH), 82.4 (13-C), 71.5
(3-CH), 48.6 (CH), 43.0 (CH), 42.3 (CH), 38.6 (10-C), 37.6
(CH2), 37.0 (CH2), 35.6 (CH2), 27.6 (CH2), 27.0 (CH2),
21.5 (11-CH2), 18.3 (18-CH3), 16.9 (19-CH3), 8.9
(CH3CH2). IR (cmϪ1) 2950, 1674, 1722. HRMS calculated
for C22H26O5: 372.1937. Found: 372.1934.
3␤-hydroxy-17a-oxa-14␣-androsta-5,15-diene-7,17-
dione (14)
To 0.240 g (0.64 mmol) of propionate 13 dissolved in 24
mL of MeOH was added with stirring 12 mL of 5% Na2CO3
solution. Stirring was continued for 1 h and a sample was
removed to check the extent of hydrolysis. This was nec-
essary because in the heterogeneous mixture the reaction
rate depends on the effectiveness of stirring and can’t al-
ways be reproduced. After workup (vide infra), NMR
showed 10% propionate 13, 80% hydrolyzed 3␤-OH, 14␣
lactone 14, and 10% 14␤ lactone 15. The reaction mixture
was neutralized with acetic acid, diluted with 100 mL of
H2O and extracted with CHCl3 two times. The organic
layers were combined, washed with 50% saturated NaCl,
dried (Na2SO4) and solvent evaporated. The residue could
be used directly or purified by preparative TLC (6% MeOH/
The air oxidation of 18 and 19 to their respective 7-oxo-
derivatives (Scheme 4) was based on a patent by Foricher et
1
CHCl3, Rf ϭ 0.20). H NMR (270 MHz) ␦ 7.27 (d, J ϭ 9,
2.5 Hz, 15-H), 6.03 (d, J ϭ 9, 3 Hz, 16-H), 5.80 (d, J ϭ 1.5
Hz, 6-H), 3.72 (tt, J ϭ 11, 5 Hz, 3␣-H), 2.80 (dt, J ϭ 11.5,
2.5 Hz, 14␣-H), 2.60 (ddd, J ϭ 13.5, 5, 2 Hz, 4␣-H), 2.45
(ddd, J ϭ 13.5, 11, 2 Hz, 4␤-H), 2.41 (t, J ϭ 12 Hz, 8␤-H),
1.36 (s, 18-CH3), 1.23 (s, 19-CH3). 13C NMR (90.6 MHz) ␦
199.4 (7-CO), 166.6 (5-C), 163.9 (17-CO), 149.1 (15-CH),
125.1 (6-CH), 120.2 (16-CH), 82.7 (13-C), 70.0 (3-CH),
48.8 (CH), 43.0 (CH), 42.3 (CH), 41.6 (CH2), 38.6 (10-C),
37.0 (CH2), 36.0 (CH2), 30.7 (CH2), 21.5 (11-CH2), 18.2
(18-CH3), 16.9 (19-CH3).
Scheme 4 Preparation of additional D-ring lactones related to
3␤-Acetoxyandrost-5-ene-7,17-dione.
546 Steroids, 1998, vol. 63, October

Seco-steroids related to dehydroepiandrosterone: Reich et al.
NMR, IR and MS; these data were not previously available.
18 1H NMR (500 MHz, CDCl3): ␦ 5.38 (d, J ϭ 4 Hz, 6-H),
4.62 (m, 1H, 3␣-H), 4.49 (m, 1H, 16-H), 4.28 (m, 1H,
16-H), 2.04 (s, 3H, CH3COO), 1.23 (s, 3H, 19-CH3), 1.04
(s, 3H, 18-CH3). IR (cmϪ1): 2941, 1750, 1735, 1430, 1376,
1255, 1115, 903, 737. MS m/z: 347 (M ϩ1), 331 (M -CH3),
304 (M -CH2CO) 302 (M -CO2), 286 (M -AcOH), 271 (286
-CH3). 19 1H NMR (500 MHz, CDCl3): ␦ 5.38 (d, J ϭ 4 Hz,
6-H), 4.62 (m, 3␣-H), 3.99 (d, J ϭ 11 Hz, 17-H), 3.91 (d,
J ϭ 11 Hz, 17-H), 2.72 (dd, J ϭ 19, 5 Hz, 15-H), 2.13 (dd,
J ϭ 19, 13 Hz, 15-H), 2.05 (s, CH3COO), 1.04 (s, CH3),
1.03 (s, CH3). IR (cmϪ1): 2949, 1730, 1467, 1376, 1249,
1199, 1034, 912, 733. MS m/e: 346 (Mϩ), 330 (M -CH3,
-H), 304 (M -CH2CO), 302 (M -CO2), 286 (M -AcOH), 271
(286 -CH3), 242 (286 -CO2), 227 (271 -CO2).
3␤-hydroxy-17a-oxa-14␤-androsta-5,15-diene-7,17-
dione (15)
The crude 14␣ lactone 14 (0.896 g, 2.50 mmol) was dis-
solved in 30 mL of MeOH and treated under N2 with 5 mL
of 2 M NaOH solution. After 1 h 1.4 mL of AcOH was
added and most of the MeOH was removed under vacuum.
Water (60 mL) and CHCl3 (80 mL) were added and the
mixture was extracted. The aqueous layer was washed two
more times with CHCl3. The organic layers were combined,
washed with 50% saturated NaCl, dried (Na2SO4) and sol-
vent was evaporated to yield 0.610 g of crude 14␤ lactone
15. The lactone can be purified by preparative TLC (8%
MeOH/CHCl3, Rf ϭ 0.45). To obtain Ͼ95% purity, three
consecutive chromatographies were necessary. The enzyme
induction assays were performed on ϳ80% pure material
after one chromatography. 1H NMR (300 MHz) ␦ 6.38 (dd,
J ϭ 10, 2 Hz, 15-H), 6.02 (dd, J ϭ 10, 3 Hz, 16-H), 5.91 (d,
J ϭ 2 Hz, 6-H), 3.74 (tt, J ϭ 11.5, 4.5 Hz, 3␣-H), 3.51
(broad s, 14␤-H), 2.62 (ddd, J ϭ 14.5, 5, 2 Hz, 4␣-H), 2.54
(dd, J ϭ 13, 4.5 Hz, 8␤-H), 2.49 (ddd, J ϭ 14.5, 11.5, 2 Hz,
4␤-H), 1.56 (s, 18-CH3), 1.16 (s, 19-CH3). 13C NMR (90.6
MHz) ␦ 197.4 (7-CO), 168.9 (5-C), 163.2 (17-CO), 147.3
(15-CH), 126.0 (6-CH), 121.7 (16-CH), 82.9 (13-C), 69.7
(3-CH), 45.2 (CH), 44.4 (CH), 41.8 (CH2), 39.9 (CH), 37.6
(10-C), 35.7 (CH2), 31.4 (CH2), 30.6 (CH2), 25.0 (18-CH3),
23.0 (11-CH2), 18.0 (19-CH3). HRMS calculated for
C19H24O4: 316.1674; Found: 316.1615.
3␤-acetoxy-17-oxa-androst-5-ene-7,17a-dione (20)
and 3␤-acetoxy-17a-homo-17-oxa-androst-5-ene-
7,16-dione (21)
The oxidation of 18 and 19 to their respective products
(Scheme 4) was based on a patent of Foricher et al.19 (cf ref.
8). To a refluxing solution of lactone 18 or 19 (1.0 g, 2.9
mmol) and N-hydroxyphthalimide (0.5 g, 2.9 mmol) in
acetone, dibenzoyl peroxide (30 mg) was added and a slow
stream of compressed air was passed into the solution for
8–9 h. Acetone was removed under vacuum and the result-
ant mass was taken up in CCl4. After stirring for some time
the precipitated N-hydroxyphthalimide was filtered and
washed with two 5 mL portions of CCl4. The combined
solutions were washed thoroughly with 1% NaHCO3, water
and brine and dried over MgSO4. After evaporating the
solvent under vacuum, the product was crystallized from
acetone-hexane or may be further purified by chromatogra-
phy on silica gel using 25% ethyl acetate in hexane. The
white crystalline compounds were obtained in 58–60%
yields.
3␤-acetoxy-17a-oxa-14␤-androsta-5,15-diene-7,17-
dione (16)
To 5 mg of alcohol 15 was added 0.1 mL of acetic anhy-
dride, 0.1 mL of dry pyridine and 1 mg of dimethylamin-
opyridine (DMAP). The solution was allowed to stand for
16 h and was then extracted between 0.5 N HCl solution and
CH2Cl2. The organic layer was washed with 50% saturated
NaCl solution, dried (Na2SO4) and solvent was removed to
yield 5 mg of the acetate 16. TLC (6% MeOH/CHCl3, Rf ϭ
0.50) gave 4 mg of pure acetate. 1H NMR (300 MHz) ␦ 6.37
(dd, J ϭ 10, 2 Hz, 15-H), 6.03 (dd, J ϭ 10, 3 Hz, 16-H),
5.92 (d, J ϭ 2 Hz, 6-H), 4.76 (tt, J ϭ 11.5, 4.5 Hz, 3␣-H),
3.51 (broad s, 14␤-H), 2.67 (ddd, J ϭ 14.5, 5, 2 Hz, 4␣-H),
2.54 (ddd, J ϭ 14.5, 9, 2 Hz, 4␤-H), 2.48, 2.51(d,d, J ϭ 13,
4.5 Hz, 8␤-H), 2.07 (s, CH3COO), 1.56 (s, 18-CH3), 1.17 (s,
19-CH3).
1
20. Mp 154–155°. H NMR (500 MHz, CDCl3): ␦ 5.74
(s, 6-H), 4.72 (m, 3␣-H), 4.45 (m, 2H, 16-H), 2.82 (br d, J ϭ
15 Hz 15␣-H), 2.59 (dd, J ϭ 15, 4 Hz, 4␣-H), 2.47 (t, J ϭ
13 Hz, 4␤-H), 2.33 (t, J ϭ 11 Hz, 8-H), 2.22 (br d, J ϭ 13
Hz, 15␤-H), 2.08 (s, CH3COO), 1.25 (s, CH3), 1.24 (s,
CH3). IR (cmϪ1): 2953, 1734, 1673, 1630, 1456, 1377,
1246, 1053, 736. MS m/z: 360 (Mϩ), 344 (M -CH3, -H),
316 (M -CH2CH2O or CO2), 300 (M-AcOH), 299 (M -H,
-AcOH), 284 (300 -CH3, -H), 255 (299 -CH2CH2O or CO2).
21 m.p. 213–214°C. 1H NMR (500 MHz, CDCl3): ␦ 5.76 (s,
6-H), 4.72 (m, 3␣-H), 3.98 (s, 2H, 17-H), 3.75 (dd, J ϭ 20,
6 Hz, 15␣-H), 2.59 (dd, J ϭ 14, 5 Hz, 4␣-H), 2.49 (t, J ϭ
13 Hz, 4␤-H), 2.17 (dd, J ϭ 20, 13 Hz, 15␤-H), 2.16 (t, J ϭ
9 Hz, 8-H), 2.08 (s, CH3COO), 1.27 (s, 19-CH3), 1.04 (s,
18-CH3). IR (cmϪ1): 2960, 1725, 1670, 1650, 1372, 1237,
1031. MS m/z: 360 (Mϩ), 344 (M -CH3, -H), 318 (M
-CH2CO), 316 (M -CO2), 300 (M -AcOH), 285 (300 -CH3),
272 (300 -CO), 270 (300 -CH2O), 256 (300 -CO2).
3␤-acetoxy-17a-oxa-androst-5-ene-7,17-dione (17)
Compound 17 was isolated as a by-product in the commer-
cial synthesis of 7-oxo-DHEA-acetate; m.p. 262–263°C;
[␣]D24°C ϭ 206.3° (c ϭ 0.52, CHCl3). 1H NMR (200 MHz,
CDCl3): ␦ 5.75 (d, J ϭ 1.5 Hz, 6-H), 4.71 (m, 3␣-H), 2.06
(s, OCOCH3), 1.33 (s, 18-CH3), 1.22 (s, 19-CH3). Calcu-
lated for C21H28O5: C ϭ 69.98; H ϭ 7.83. Found: C ϭ
69.77; H ϭ 7.70.
3␤-acetoxy-17-oxa-androst-5-en-17a-one (18) and
3␤-acetoxy-17a-homo-17-oxa-androst-5-en-16-one
(19)
Degradation of the chenodeoxycholic acid
side chain
Compounds 18 and 19 were prepared according to von
Scheme 5 presents a new sequence for removal of the
chenodeoxycholic acid side chain, related to a procedure
Seeman and Grant.13 Their structures were confirmed by
Steroids, 1998, vol. 63, October 547

Papers
for 20 min. After cooling, the organic product was extracted
into CH2Cl2. The organic layer was washed with 5% satu-
rated NaHCO3 solution. NMR showed quantitative conver-
1
sion to the ester 23. H NMR (270 MHz) ␦ 3.86 (q, J ϭ 3
Hz, 7␤-H), 3.68 (s, OCH3), 3.47 (tt, J ϭ 10.5, 4.5 Hz,
3␤-H), 0.94 (d, J ϭ 6.5 Hz, 21-CH3), 0.91 (s, 19-CH3), 0.66
(s, 18-CH3).
Methyl 3␣,7␣-dihydroxy-5␤-chol-22-en-24-oate (25)
To 4.9 mL of diisopropylamine (35 mmol) in 18 mL of dry
THF at Ϫ78°C under N2 was added 14.6 mL of nBuLi (2.38
M in hexane, 34.5 mmol) followed by 5.1 mL of TMSCl (40
mmol). The ester 23 (10.2 mmol) in 35 mL of THF was
added slowly via cannula to the LDA-TMSCl solution at
Ϫ78°C. The mixture was warmed to 0°C for 15 min and
then cooled again to Ϫ78°C. To the cooled solution PhSeCl
(2.35 g, 12.2 mmol) was added in 25 mL of THF rapidly via
cannula. Triethylamine (5.0 mL) was added and the mixture
was extracted between CH2Cl2 and 5% NaHCO3 solution.
The organic layer was dried (Na2SO4) and evaporated to
yield the crude 24 as a mixture of epimers at C-23, but
1
otherwise pure. H NMR (200 MHz) ␦ 7.61 (m, o-SePh),
7.29 (m, SePh), 3.87 (m, 7␤-H), 3.80 (dd, J ϭ 12, 3.5 Hz,
23-H), 3.70 (dd, J ϭ 9.5, 5 Hz, 23-H), 3.62, 3.60 (s, OMe),
3.42 (m, 3␤-H), 0.92 (d, J ϭ 6.5 Hz, 21-CH3), 0.89, 0.87 (s,
19-CH3), 0.63, 0.59, (s, 18-CH3), 0.12 (s, 3-OSiMe3), 0.07,
0.06, (s, 7-OSiMe3).
To 10.2 mmol of the selenide mixture in 65 mL of
CH2Cl2 was added 1.6 mL of pyridine and 7 mL of 15%
H2O2 (40 mmol). The mixture was stirred vigorously until
the yellow color was gone (15 min). The mixture was
partitioned between CH2Cl2 and 5% NaHCO3 solution. The
solvent was evaporated and the residue dissolved in 70 mL
of THF and 70 mL of 1 N HCl solution. The mixture was
stirred vigorously for 1 h then extracted between H2O and
EtOAc. The aqueous layer was washed two more times with
EtOAc. The organic layers were combined, dried (Na2SO4)
and evaporated. A small sample of 25 was purified by
preparative TLC (6% MeOH/CHCl3, Rf ϭ 0.18). 1H NMR
(300 MHz) ␦ 6.84 (dd, J ϭ 15.5, 9 Hz, 22-H), 5.74 (dd, J ϭ
15.5, 0.5 Hz, 23-H), 3.84 (q, J ϭ 2.5 Hz, 7␤-H), 3.72 (s,
OMe), 3.46 (tt, J ϭ 11, 4.5 Hz, 3␤-H), 1.09 (d, J ϭ 6.5 Hz,
21-CH3), 0.91 (s, 19-CH3), 0.70 (s, 18-CH3). 13C NMR
(75.4 MHz) ␦ 167.4 (24-CO), 154.8 (22-CH), 118.6 (23-
CH), 71.6 (3-CH), 68.3 (7-CH), 54.9 (CH), 51.3 (OCH3),
50.2 (CH), 42.9 (13-C), 41.5 (CH), 39.8 (CH2), 39.7 (CH2),
39.5 (CH), 39.4 (CH), 35.3 (CH2), 35.0 (CH2), 35.0 (10-C),
34.7 (CH2), 32.8 (CH), 30.6 (CH2), 28.1 (CH2), 23.7 (CH2),
22.8 (21-CH3), 20.5 (11-CH2), 19.2 (19-CH3), 12.0 (18-
CH3). IR (cmϪ1) 3613, 3410, 2934, 2869, 1711, 1437, 1283,
1238, 1075, 908. HRMS calculated for C25H40O4:
404.2927; Found: 404.2934.
Scheme
5
Preparation of 4-oxa-4a-homo-7␣-hydroxy-17␤-
acetoxy-5␤-androstan-3-one and 3␣-hydroxy-7a-oxa-17␤-ace-
toxy-5␤-androstan-7-one.
reported by Barton et al. with 11-oxolithocholic acid.20 The
methyl ester 23 was selenenylated via the silyl enol ether
formed in situ. Oxidation with hydrogen peroxide resulted
in selenoxide elimination to form the ␣,␤-unsaturated side
chain compound 25 in nearly quantitative yield. Attempted
deconjugation of the 22, 23 double bond by base-catalyzed
enolization-reprotonation of 25 with amide bases such as
lithium diisopropylamide and hexamethylphosphoridri-
amide (HMPA) was unsuccessful. A successful procedure
involved conversion of 25 to the ␤,␥-unsaturated ketene
acetal with t-butyldimethylsilyl trifluoromethanesulfonate
(TBSOTf), followed by hydrolysis to 26. Ozonolysis and
reductive workup gave the 17-acetyl compound 27. Baeyer–
Villiger oxidation produced the 17-acetoxy compound 28 in
69% yield and the two lactones 29 and 30 in 7% and 9%
yield, respectively, after chromatographic separation.
The two minor products 29 and 30 are formed by Bae-
yer–Villiger oxidation of ketone precursors. One can envi-
sion formation of these ketones by a direct oxidation of the
silyl ether or cleavage of one of the silyl groups by the
peracid yielding the alcohol which is subsequently oxidized.
Methyl 3␣,7␣-bis(tert-butyldimethylsilyloxy)-5␤-chol-
20-en-24-oate (26)
Methyl 3␣,7␣-dihydroxy-5␤-cholan-24-oate (23)
To 6.38 g (15.8 mmol) of diol 25 in 50 mL of freshly
distilled CH2Cl2 at 0°C was added, under N2, 12 mL of
triethylamine (86 mmol) and 13 mL of tert-butyl-
dimethylsilyl triflate (56.8 mmol). The mixture was allowed
To 4.0 g (10.2 mmol) of 3␣,7␣-dihydroxy-5␤-cholan-24-
oic acid (22) in 15 mL of dry MeOH was added 0.80 mL of
TMSCl (6.31 mmol). The solution was heated under reflux
548 Steroids, 1998, vol. 63, October

Seco-steroids related to dehydroepiandrosterone: Reich et al.
to stand at room temperature for 48 h after which NMR
1H NMR (300 MHz) ␦ 4.57 (dd, J ϭ 9, 7.5 Hz, 17␣-H), 3.80
(q, J ϭ 2.5 Hz, 7␤-H), 3.40 (tt, J ϭ 11, 4.5 Hz, 3␤-H), 2.26
(td, J ϭ 13, 11 Hz, 4␣-H), 0.90 (s, t-BuSiO), 0.88 (s,
19-CH3), 0.87 (s, t-BuSiO), 0.75 (s, 18-CH3), 0.088 (s,
CH3SiO), 0.026 (s, CH3SiO), 0.022 (s, CH3SiO), 0.018 (s,
CH3SiO). 13C NMR (75.4 MHz) ␦ 171.2 (CH3COO), 82.9
(17-CH), 72.6 (3-CH), 69.0 (7-CH), 44.5 (CH), 42.5 (13-C),
41.9 (CH), 40.5 (CH2), 36.6 (CH2), 35.7 (CH2), 35.1 (10-C),
34.5 (CH2), 32.5 (CH), 30.9 (CH2), 27.6 (CH2), 25.9 (t-Bu-
CH3), 23.4 (CH2), 22.8 (19-CH3), 21.2 (CH3COO), 20.0
(11-CH2), 18.3 (t-Bu-C), 18.1 (t-Bu-C), 12.0 (18-CH3),
Ϫ2.38 (7-OSiCH3), -4.61 (3-OSiCH3), -5.51 (7-OSiCH3).
IR (cmϪ1) 2958, 2935, 2857, 1724, 1256, 1093, 1028, 837.
HRMS calculated for C33H62O4Si2: 578.4200; Found:
578.4198.
showed the reaction to be 92% complete. The reaction
mixture was poured into 200 mL of 1 N HCl solution and
extracted with CH2Cl2. The CH2Cl2 layer was washed with
5% NaHCO3 solution. The solvent was dried (Na2SO4), and
evaporated to yield 14.3 g. A small sample of 26 was
purified by preparative TLC (5% EtOAc-hexane, Rf ϭ
0.30). 1H NMR (270 MHz) ␦ 5.37 (tm, J ϭ 6.5 Hz, 23-H),
3.82 (q, J ϭ 2.5 Hz, 7␤-H), 3.68 (s, OMe), 3.41 (tt, J ϭ
11.5, 4.5 Hz, 3␤-H), 3.10 (d, J ϭ 6.5 Hz, 24-H, 2H), 2.28
(td, J ϭ 13, 11 Hz, 4␣-H), 2.04 (t J ϭ 10 Hz, 17␤-H), 1.64
(s, 21-H), 0.89 (s, t-BuSiO), 0.87 (s, t-BuSiO), 0.87 (s,
19-CH3), 0.50 (s, 18-CH3), 0.10 (s, CH3SiO, 6H), 0.04 (s,
CH3SiO, 3H), 0.01 (s, CH3SiO, 3H).
Fraction-11 yielded solid lactone 29, 0.55 g (7%). A
portion was crystallized from hexane-CH2Cl2, m.p. 222–
223°C. H NMR (300 MHz) ␦ 4.73 (dd, J ϭ 13.5, 10 Hz,
3␣,7␣-bis(tert-butyldimethylsilyloxy)-5␤-pregnan-20-
one (27)
1
A solution of the silyl-protected diol 26 (15.8 mmol) in 60
ml of CH2Cl2 and 25 mL of MeOH was divided in half for
ozonolysis. Each half was cooled to Ϫ78°C and ozone
bubbled in for about 0.5 h until the solution was blue. The
solution was purged of ozone with N2 at Ϫ78°C, dimethyl
sulfide (3.3 mL, 45 mmol) was added and the solution was
warmed to room temperature for 1 h. The mixture was
poured into CH2Cl2 and extracted with H2O two times. The
organic layer was washed with 5% NaHCO3 solution. A
small sample was purified by preparative TLC (5% EtOAc-
hexane, Rf ϭ 0.16). 1H NMR (300 MHz) ␦ 3.83 (q, J ϭ 2.8
Hz, 7␤-H), 3.42 (tt, J ϭ 11.5, 4.5 Hz, 3␤-H), 2.54 (t, J ϭ 9
Hz, 17␣-H), 2.28 (td, J ϭ 13, 11Hz, 4␣-H), 2.13 (s, 21-
CH3), 0.92 (s, 7-t-Bu), 0.88 (s, 19-CH3 ϩ 3-t-Bu), 0.58 (s,
18-CH3), 0.106 (s, 7-OSiCH3), 0.050 (s, 7-OSiCH3), 0.038
(s, 3-OSiCH3), 0.036 (s, 3-OSiCH3). 13C NMR (75.4 MHz)
␦ 72.5 (3-CH), 69.5 (7-CH), 63.8 (17-CH), 50.5 (CH), 44.1
(13-C), 41.8 (CH), 40.6 (CH), 40.5 (CH2), 38.8 (CH2), 35.6
(CH2), 35.1 (10-C), 34.5 (CH2), 32.4 (CH), 31.6 (21-CH3),
31.0 (CH2), 26.1 (t-Bu-CH3), 25.9 (t-Bu-CH3), 24.1 (CH2),
22.8 (19-CH3), 22.7 (CH2), 20.5 (11-CH2), 18.4 (C-tBu),
18.1 (C-tBu), 13.3 (18-CH3), Ϫ2.37 (7-OSiCH3), Ϫ4.62
(3-OSiCH3), Ϫ5.56 (7-OSiCH3). IR (cmϪ1) 2952, 2931,
2857, 1698, 1472, 1462, 1254, 1092, 1027. HRMS calcu-
lated for C33H61O3Si2: 561.4159; Found: 561.4159.
4a-H), 4.59 (dd, J ϭ 9, 7.5 Hz, 17␣-H), 3.93 (dd, J ϭ 13.5,
1.5 Hz, 4a-H), 3.79 (q, J ϭ 2.5 Hz, 7␤-H), 2.76 (dd, J ϭ
14.5, 12.5 Hz, 2-H), 2.35 (dd, J ϭ 14.5, 8 Hz, 2-H), 2.04 (s,
17-CH3COO), 1.01 (s, 19-CH3), 0.90 (s, t-BuSiO), 0.76 (s,
18-CH3), 0.09 (s, CH3SiO), 0.06 (s, CH3SiO). 13C NMR
(75.4 MHz) ␦ 176.4 (3-COO), 171.1 (CH3COO), 82.4 (17-
CH), 71.7 (4a-CH2), 68.0 (7-CH), 45.3 (CH), 44.2 (CH),
42.4 (13-C), 40.5 (CH), 37.0 (10-C), 36.4 (CH2), 34.3
(CH2), 33.3 (CH2), 32.8 (CH), 28.0 (CH2), 27.4 (CH2), 26.0
(t-Bu-CH3), 23.3 (CH2), 23.2 (CH3COO), 21.1 (19-CH3),
20.2 (11-CH2), 18.2 (t-Bu-C), 11.9 (18-CH3), -2.19 (7-
OSiCH3), -5.48 (7-OSiCH3). IR (cmϪ1) 2956, 2935, 2881,
2859, 1728, 1256, 1056, 1048, 1024. HRMS calculated for
C27H46O5Si: 478.3115. Found: 478.3096.
Fractions 14–16 yielded semi-solid lactone 30, 0.70 g
1
(9%). H NMR (300 MHz) ␦ 4.64 (dd, J ϭ 9, 7.5 Hz,
17␣-H), 4.21 (dd, J ϭ 10, 9.5 Hz, 8␤-H), 3.59 (tt, J ϭ 10.5,
5 Hz, 3␤-H), 2.98 (dd, J ϭ 14.5, 0.5 Hz, 6-H), 2.35 (dd, J ϭ
14.5, 6 Hz, 6-H), 2.02 (CH3COO), 1.04 (19-CH3), 0.84
(t-BuSiO), 0.74 (18-CH3), 0.007 (CH3SiO), 0.004
(CH3SiO). 13C NMR (75.4 MHz) ␦ 174.0 (7-COO), 170.8
(CH3COO), 81.8 (17-CH), 79.2 (8-CH), 71.3 (3-CH), 49.1
(CH), 43.1 (13-C), 41.7 (CH), 40.0 (CH), 37.5 (10-C), 36.0
(CH2), 35.8 (CH2), 35.60 (CH2), 35.55 (CH2), 30.8 (CH2),
27.2 (CH2), 25.8 (t-Bu-CH3), 24.2 (CH2), 23.3 (19-CH3),
22.1 (11-CH2), 21.0 (CH3COO), 18.1 (t-BuC), 11.5 (18-
CH3), -4.70 (CH3SiO), -4.77 (CH3SiO). IR (cmϪ1) 2962,
2956, 2947, 2940, 1728, 1472, 1361, 1099, 1015, 909, 868,
837. HRMS calculated for C27H46O5Si: 478.3115. Found:
478.3118.
3␣,7␣-bis(tert-butyldimethylsilyloxy)-17␤-acetoxy-
5␤-androstane (28), 4-oxa-4a-homo-7␣-tert-
butyldimethylsilyloxy-17␤-acetoxy-5␤-androstan-3-
one (29) and 3␣-tert-butyldimethylsilyloxy-7a-oxa-
17␤-acetoxy-5␤-androstan-7-one (30)
4-oxa-4a-homo-7␣-hydroxy-17␤-acetoxy-5␤-
androstan-3-one (31)
A solution of 15.8 mmol of the methyl ketone 27 in 80 mL
of CH2Cl2 was treated with 10 g (ϳ85%, 49.2 mmol) of
m-chloroperoxybenzoic acid and stirred at room tempera-
ture for 5 days. The mixture was diluted with more CH2Cl2
and extracted two times with 5% NaHCO3 solution. The
organic layer was dried (Na2SO4) and evaporated to yield
11.7 g of crude product, which was chromatographed on
250 g of silica gel packed with hexane. The column was
eluted with 1.2 L of 20% EtOAc-hexane, 200 mL of 50%
EtOAc-hexane and then EtOAc (100 mL fractions).
To a solution of 450 mg (0.94 mmol) of the lactone 29 in 15
mL of dry THF was added 5 mL of a 1 M solution of
nBu4NF in THF. The solution was kept at room temperature
for 17 h. The mixture was partitioned between CHCl3 and
water. The organic layer was washed three times with water,
50% saturated NaCl solution, dried (Na2SO4) and evapo-
rated. Preparative TLC yielded 149 mg (44%) of the alcohol
1
31, Rf ϭ 0.47 (5% MeOH/CHCl3). H NMR (300 MHz) ␦
Fractions 7–9 yielded 6.21 g (69%) of the 17-acetate 28.
4.82 (dd, J ϭ 13.5, 10 Hz, 4a-H), 4.63 (dd, J ϭ 9, 7.5 Hz,
Steroids, 1998, vol. 63, October 549

Papers
17␣-H), 3.96 (dd, J ϭ 14, 1.5 Hz, 4a-H), 3.82 (q, J ϭ 2.5
Hz, 7␤-H), 2.79 (t, J ϭ 14 Hz, 2-H), 2.35 (dd, J ϭ 14.5, 8
Hz, 2-H), 2.04 (s, 17-CH3COO), 1.00 (s, 19-CH3), 0.78 (s,
18-CH3). 13C NMR (75.4 MHz) ␦ 176.7 (3-COO), 171.2
(CH3COO), 82.5 (17-CH), 71.5 (4a-CH2), 66.6 (7-CH),
45.2 (CH), 44.7 (CH), 42.5 (13-C), 39.3 (CH), 37.0 (10-C),
36.4 (CH2), 34.1 (CH2), 33.4 (CH), 33.3 (CH2), 28.2 (CH2),
27.5 (CH2), 23.3 (CH3COO), 22.9 (CH2), 21.1 (19-CH3),
20.2 (11-CH2), 11.8 (18-CH3). IR (cmϪ1) 3700 (broad),
2943, 2878, 1727, 1375, 1291, 1258, 1213, 1055, 1030, 908.
HRMS calculated for C21H32O5: 364.2250; Found:
364.2250.
3␣-hydroxy-7a-oxa-17␤-acetoxy-5␤-androstan-7-one
(32)
The t-butyldimethylsilyl group was cleaved from 30 (30
mg, 0.063 mmol) by treatment with nBu4NF for 2.5 h as
described above. The yield after chromatography (Rf ϭ
1
0.20, 5% MeOH/CHCl3) was 12 mg (52%). H NMR (300
MHz) ␦ 4.68 (dd, J ϭ 9.5, 7.5 Hz, 17␣-H), 4.26 (t, J ϭ 9.5
Hz, 8␤-H), 3.67 (tt, J ϭ 10.5, 5 Hz, 3␤-H), 3.04 (dd, J ϭ
14.5, 1.0 Hz, 6-H), 2.40 (dd, J ϭ 14.5, 6.5 Hz, 2-H), 2.05 (s,
17-CH3COO), 1.09 (s, 19-CH3), 0.78 (s, 18-CH3). 13C
NMR (75.4 MHz) ␦ 174.0 (7-COO), 170.9 (CH3COO), 81.8
(17-CH), 79.3 (8-CH), 70.7 (3-CH), 49.2 (CH), 43.1 (13-C),
41.7 (CH), 40.2 (CH), 37.6 (10-C), 35.9 (CH2), 35.7 (CH2),
35.59 (CH2), 35.55 (CH2), 30.2 (CH2), 27.3 (CH2), 24.2
(CH2), 23.4 (19-CH3), 22.2 (11-CH2), 21.1 (CH3COO),
11.5 (18-CH3), IR (cmϪ1) 3600 (broad), 2964, 2879, 1727,
1359, 1250, 1214, 1068, 1005, 909. HRMS calculated for
C21H32O5: 364.2250; Found: 364.2243.
Scheme 7
39 were determined unambiguously by single crystal X-ray
crystallography. The ␣ configuration of the epoxide trans-
lates into the ␤ configuration for the formyl group in the
rearrangement of 38 to 39 after the Baeyer–Villiger reac-
tion.
Formation of B-ring lactones by peroxidation of
7-keto steroids
3␤,17␤-diacetoxy-7a-oxa-5␣-androstan-7-one (34)
Several B-ring lactones were prepared by Baeyer–Villiger
oxidation. The 7-keto androstane 33 gave the normal prod-
uct of migration (34) of the secondary carbon in preference
to the primary one on treatment with m-chloroperbenzoic
acid (Scheme 6). The enone 3␤,17␤-diacetoxyandrost-5-en-
7-one (35) showed more complex behavior. Treatment with
excess p-nitroperoxybenzoic acid (PNPBA) in dichlo-
romethane for several days at room temperature gave mix-
tures of the ␣-5,6 epoxide (36) and the two Baeyer–Villiger
products 37 and 38 (Scheme 7). When the solvent was
changed to refluxing chloroform, 70% of the aldehyde 39
was formed. When epoxide lactone 38 was submitted to
these reaction conditions the aldehyde 39 was the only
product. The structures of the epoxide 36, and the aldehyde
To a solution of 3␤,17␤-diacetoxyandrostan-7-one (33,
0.45 g; 1.15 mmol) in dry chloroform (5 mL), was added
m-chloroperbenzoic acid (MCPBA) (0.5 g). After stirring at
room temperature for 20 h the reaction mixture was poured
into water and the organic layer separated. The CHCl3
extract was washed with 10% NaHCO3 and water, dried
over MgSO4 and the solvent was removed in vacuo. The
residue was purified by chromatography on a column of
silica gel and eluted with acetone-petroleum ether (1.5:8.5).
The starting steroid (0.1 g, 22% recovered) eluted first
followed by the product which was crystallized from ether-
hexane. Yield 0.30 g (82% based on 78% conversion); m.p.
163–165°C. 1H NMR (200 MHz): ␦ 4.65 (m, 2H, 3␣-,17␣-
H), 4.22 (m, 1H, 8-H), 2.05 (s, 3H, OCOCH3), 2.03 (s, 3H,
OCOCH3), 1.03 (s, 3H, 19-CH3), 0.79 (s, 3H, 18-CH3). 13
C
NMR (300 MHz): ␦ 174.6 (CAO, C-7), 170.8 & 170.3
(acetate CAO), 81.79, 79.1, 72.05 (CH, C-3,8,17), 52.3,
49.1, 40.9 (CH, C-5,9,14), 37.5, 36.7, 35.8, 35.5, 27.2, 26.7,
24.2, 22.5 (CH2, C-1,2,4,6,11,12,15,16), 21.3, 21.0 (acetate-
CH3), 12.9, 11.6 (CH3, C-18,19).
3␤,17␤-diacetoxy-5␣,6-epoxy-androstan-7-one (36)
3␤,17␤-Diacetoxyandrost-5-en-7-one (0.349 g, 0.90 mmol)
in 12 mL of CH2Cl2 was treated with 0.474 g (1.56 mmol,
Scheme 6
550 Steroids, 1998, vol. 63, October

Seco-steroids related to dehydroepiandrosterone: Reich et al.
65%) of PNPBA and 4 mg of 3-tert-butyl-4-hydroxy-5-
3␤,17␤-diacetoxy-5␣,6-epoxy-7a-oxa-androstan-7-
one (37) and 3␤,17␤-diacetoxy-5␣,6-epoxy-7-oxa-
androstan-7a-one (38)
methylphenyl sulfide (radical inhibitor) and stirred for 24 h.
At this point NMR analysis indicated that 36% starting
material remained. More PNPBA (0.256 g, 1.09 mmol,
65%) was added and stirring was continued for 48 h. NMR
analysis showed 42% epoxide 36, 37% aldehyde 39, and
21% lactone 37. The crude mixture was chromatographed
on silica gel eluting with 50% ethyl acetate-hexane. The
first fraction (Rf ϭ 0.70) contained 0.086 g of 36 and some
methyl p-nitrobenzoate. Pure 36 was crystallized from
3␤,17␤-Diacetoxyandrost-5-en-7-one (35) (0.212 g, 0.55
mmol) in 7 mL of CH2Cl2 was treated with 0.328 g (65%,
1.16 mmol) of PNPBA and the mixture was stirred at room
temperature for 70 h. At this point NMR analysis showed
the starting material to be consumed and a mixture of 37
(49%) and 38 (36%) was formed along with minor amounts
of other oxidation products. The CH2Cl2 solution was ex-
tracted with 10% aqueous NaOH and washed with saturated
NaCl. Solvent was removed under reduced pressure. This
mixture, which also contained some methyl p-nitroben-
zoate, was used in the biological assay. A small sample was
chromatographed on silica gel, eluting with 50% ethyl
1
CH2Cl2-hexane to yield 27 mg, m.p. 207–208°C. H NMR
(300 MHz) ␦ 4.92 (tt, J ϭ 11.5, 4.5 Hz, 3␣-H), 4.60 (dd, J ϭ
9, 7.5 Hz, 17␣-H), 3.04 (s, 6␤-H), 2.28 (dd, J ϭ 13, 12 Hz,
15␣-H), 2.038 (s, CH3COO), 2.036 (s, CH3COO), 1.98 (t,
J ϭ 11.5, 8␤-H), 1.07 (s, 19-CH3), 0.80 (s, 18-CH3). 13C
NMR (75.4 MHz) ␦ 206.5 (7-CO), 171.0 (CH3COO), 170.0
(CH3COO), 81.9 (17-CH), 70.7 (3-CH), 67.5 (5-C), 62.5
(6-CH), 46.7 (CH), 46.2 (CH), 44.0 (13-C), 43.1 (CH), 36.6
(CH2), 35.2 (10-C), 35.1 (CH2), 32.6 (CH2), 27.6 (CH2),
27.1 (CH2), 24.6 (CH2), 21.2 (CH3COO), 21.1 (CH3COO),
20.6 (11-C), 15.5 (19-CH3), 12.2 (18-CH3).
1
acetate-hexane two times, to yield pure 38. H NMR (300
MHz) ␦ 4.85 (m, 3␣-H), 4.82 (s, 6␤-H), 4.62 (dd, J ϭ 9.5,
7.0 Hz, 17␣-H), 2.41 (t, J ϭ 10.0 Hz, 8␤-H), 2.04, 2.01 (s,
3-CH3COO, 17-CH3COO), 1.13 (s, 19-CH3), 0.82 (s, 18-
CH3). 13C NMR (75.4 MHz) ␦ 174.1 (7-CO), 170.8
(CH3COO), 170.1 (CH3COO), 82.5 (6-C), 81.7 (17-CH),
69.5 (3-CH), 67.5 (5-C), 48.8 (CH), 46.9 (CH), 43.1 (13-C),
40.0 (CH), 38.6 (10-C), 36.3 (CH2), 34.6 (CH2), 32.5 (CH2),
27.1 (CH2), 26.6 (CH2), 23.5 (CH2), 21.2 (CH3COO), 21.1
(CH3COO), 20.3 (11-CH2), 15.9 (19-CH3), 11.9 (18-CH3).
1H NMR of 37 in mixture (300 MHz) ␦ 4.85 (m, 3␣-H),
4.66 (dd, J ϭ 10, 7 Hz, 17␣-H), 3.27 (s, 6␤-H), 2.05, 2.03
(s, 3-CH3COO, 17-CH3COO), 1.17 (s, 19-H), 0.77 (s, 18-
H).
The stereochemistry was determined by X-ray crystal-
lography. A colorless transparent plate-shaped crystal of
dimensions 0.58 ϫ 0.24 ϫ 0.04 mm was selected for
structural analysis. Intensity data for this compound were
collected using a Siemens SMART ccd area detector21,22
mounted on a Siemens P4 diffractometer equipped with
graphite-monochromated Mo K␣ radiation (␭ ϭ 0.71073
Å). The sample was cooled to 133 Ϯ 2°K. The intensity
data, which nominally covered one and a half hemispheres
of reciprocal space, were measured as a series of ␾ oscil-
lation frames each of 0.4° for 60 sec/frame. The detector
was operated in 512 ϫ 512 mode and was positioned 5.26
cm from the sample. Cell parameters were determined from
a non-linear least-squares fit of 5794 peaks in the range
3.0 Ͻ ␪ Ͻ 25.0°. The first 50 frames were repeated at the
end of data collection and yielded a total of 249 peaks
showing a variation of 0.05% during the data collection. A
total of 10361 data were measured in the range 2.50 Ͻ ␪ Ͻ
29.19°. The data were corrected for absorption by the em-
piric method23 giving minimum and maximum transmis-
sions of 0.9498 and 0.9964. The data were merged to form
a set of 5126 independent data with Rint ϭ 0.0234. Over
99.8% of the unique data were measured to 25° in ␪.
The orthorhombic space group P212121 was determined
by systematic absences and statistical tests and verified by
subsequent refinement. The structure was solved by direct
methods and refined by full-matrix least-squares methods
on F2.24,25 Hydrogen atom positions were initially deter-
mined by geometry and refined by a riding model. Non-
hydrogen atoms were refined with anisotropic displacement
parameters. A total of 263 parameters were refined against
5126 data to give wR(F2) ϭ 0.0884 and S ϭ 0.985 for
weights of w ϭ 1/[␴2(F2) ϩ (0.0500P)2], where P ϭ [Fo2 ϩ
3␤,17␤-diacetoxy-5␣-formyl-6-oxa-androstan-7-one
(39)
3␤,17␤-Diacetoxyandrost-5-en-7-one (0.346 g, 0.89 mmol)
was dissolved in 12 mL of CHCl3 and treated with 0.474 g
(65%, 1.68 mmol) of PNPBA and 4 mg of 3-tert-butyl-4-
hydroxy-5-methylphenyl sulfide (radical inhibitor) and re-
fluxed for 2 h. More PNPBA (0.450 g, 1.90 mmol, 65%)
was added and the mixture was refluxed for 2 more h. The
CHCl3 solution was extracted with 10% aqueous NaOH and
washed with saturated NaCl. Solvent was removed under
reduced pressure to yield a white solid. A small sample was
purified by preparative TLC on silica gel eluting with 5%
MeOH-CHCl3 to give 15 mg of pure aldehyde 39 which
was crystallized from ether; m.p. 143–145°C. 1H NMR (300
MHz) ␦ 9.66 (s, HCO), 5.16 (apparent quintet, J ϳ 5 Hz,
3␣-H), 4.65 (dd, J ϭ 9, 7 Hz, 6␣-H0, 2.05, 2.04 (s,
3-CH3COO, 17-CH3COO), 1.22 (s, 19-CH3), 0.79 (s, 18-
CH3). 13C NMR␦ 197.3 (HCO), 171.0 (CH3COO), 170.9
(CH3COO), 86.6 (5-C), 81.4 (17-CH), 68.3 (3-CH), 46.1
(CH), 43.4 (13-C), 43.3 (CH), 41.5 (CH), 37.6 (10-C), 35.9
(CH2), 33.9 (CH2), 32.7 (CH2), 27.3 (CH2), 25.0 (CH2),
24.7 (CH2), 21.22 (11-CH2), 21.14 (CH3COO), 21.09
(CH3COO), 17.6 (19-CH3), 12.1 (18-CH3). IR (cmϪ1) 3026,
2953, 1736, 1375, 1246, 1030, 908.
2
2Fc ]/3. The final R(F) was 0.0402 for the 4290 observed,
[F Ͼ 4␴(F)], data. The largest shift/s.u. was 0.002 in the
final refinement cycle. The final difference map had max-
ima and minima of 0.284 and Ϫ0.165, respectively. The
absolute structure was determined by refinement of the
Flack parameter.26
The crude mixture containing 65% aldehyde by NMR
was used in the biological assay. The stereochemistry was
determined by x-ray crystallography of a colorless transpar-
ent crystal of dimensions 0.60 ϫ 0.40 ϫ 0.02 mm. Intensity
data for this compound were collected as described for
Steroids, 1998, vol. 63, October 551

Papers
Table 1. Induction of thermogenic enzymes in rats’ livers by seco analogs of DHEA and related steroids
% of Control
GPDH
Number
of rats
Concentration
in diet %
Steroid
ME
3␤-Acetoxyandrost-5-en-17-one(DHEA-acetate)a
42
9
0.057
0.06
241 Ϯ 55
432 Ϯ 114
328
329 Ϯ75
416 Ϯ 47
485
3␤-Acetoxyandrost-5-en-7,17-dione (1)a
3␤-Acetoxy-17a-oxa-androst-5-ene-7,17-dione (17)
3␤-Hydroxy-17a-oxa-androsta-5,15-diene-7,17-dione (14␣) (14)
3␤-Propionoxy-17a-oxa-androsta-5,15-diene-7,17-dione(14␣) (13)
3␤-Hydroxy-17a-oxa-androsta-5,15-diene-7,17-dione (14␤) (15)
3␤-Hydroxy-7-oxo-16,17-secoandrost-5-ene-16,17-dioic acid (9)
3␤-Acetoxyandrost-5-ene-7,16,17-trione (8)b
4
0.064
0.052
0.065
0.068
0.063
0.066
4
128
396
5
100
120
2
112
207
2
80
205
3
253
460
Other inactive compounds tested at 0.05–0.1% of the diet: 3␤-Hydroxy-7-oxo-16,17-secoandrost-5-en-17-oic acid (12a); 3␤-hydroxy-
16,17 secoandrost-5-en-17-oic acid (11); methyl 3␤-hydroxy-7-oxo-17-oic-16,17-secoandrost-5-en-16-oate; methyl 3␤-hydroxy-17-oic-
16,17-secoandrost-5-en-16-oate; methyl 3␤-acetoxy-7-oxo-16-oic-16,17-secoandrost-5-en-17-oate; methyl 3␤-acetoxy-16-oic-16,17-
secoandrost-5-en-17-oate; dimethyl 3␤-hydroxy-7-oxo-16,17-secoandrost-5-ene-16,17-dioate (10); dimethyl 3␤-hydroxy-16,17-
secoandrost-5-ene-16,17-dioate; 3␤-acetoxy-16a-oxa-androst-5-ene-7,17-dione; 3␤-acetoxy-16a-oxa-androst-5-ene-7,16,-dione (21); 3␣-
hydroxy-7a-oxa-17␤-acetoxy-5␤-androstan-7-one (32); 4-oxa-4a-homo-7␣-hydroxy-17␤-acetoxy-5␤-androstan-3-one (31); 3␤,17␤-
diacetoxy-5␣,6-epoxy-6a-oxa-androstan-7-one and the corresponding 7a-oxa steroid (38, 37); 3␤,17␤-diacetoxy-5␣-formyl-6-oxa-
androstan-7-one; 3␤,17␤-diacetoxy-7a-oxa-androstan-7-one (34).
Two to 5 rats were used in each assay of the inactive compounds.
aThese data from ref. 8 are shown for comparative purposes.
bIn assays not shown, the 3-propionyl ester of this trione was as active as equimolar 7-oxo-DHEA.
compound 36 except that the temperature was 293 Ϯ 2°K.
Cell parameters were determined from a non-linear least-
squares fit of 6523 peaks in the range 3.0 Ͻ ␪ Ͻ 25.0°. The
first 50 frames were repeated at the end of data collection
and yielded a total of 270 peaks showing a variation of
0.04% during the data collection. A total of 13927 data were
measured in the range 5.36 Ͻ ␪ Ͻ 24.99°. The data were
corrected for absorption by the empiric method23 giving
minimum and maximum transmissions of 0.9466 and
0.9982. The data were merged to form a set of 7763 inde-
pendent data with Rint ϭ 0.0461. Over 97.0% of the unique
data were measured to 25° in ␪. A total of 571 anisotropic
displacement parameters were refined against 1 restraint
and 7763 data to give wR(F2) ϭ 0.3113 and S ϭ 1.244 for
in ␣- and ␤- configuration respectively, also induced the
formation of malic enzyme. The 3-␤ propionyl ester of the
former was apparently not hydrolyzed in vivo for it was
inactive. Compound 17, the 17 3 13 lactone of 7-oxo-
DHEA acetate, induced both enzymes as did 3␤-
acetoxyandrost-5-ene-7,16,17-trione, an intermediate in the
synthesis of the 16, 17 dicarboxylic acid. Some ability to
induce liver enzymes is retained despite extensive alter-
ations of the D-ring. Of special interest is the finding that
introduction of a double bond at 15, 16 abolishes the ability
to induce glycerophosphate dehydrogenase while activity
toward malic enzyme is retained (14 and 15 vs 17; other
examples will be presented: I. L. Reich et al., in prepara-
tion).
2
weights of w ϭ 1/[␴ (F2) ϩ (0.1182P)2 ϩ 11.8340P],
We presume that DHEA, its two 7-hydroxy derivatives,
and 7-oxo-DHEA are not hormones but are precursors of
one or more steroid hormones with higher specific activity
and for which specific receptors will be found. The enzyme-
2
where P ϭ [FO ϩ 2FC2]/3. The final R(F) was 0.1269 for
the 5784 observed, [F Ͼ 4␴(F)], data. The largest shift/s.u.
was 0.028 in the final refinement cycle. The final difference
map had maxima and minima of 0.724 and -0.448, respec-
tively. The absolute structure was determined by refinement
of the Flack parameter.26
inducing activities of compounds reported here, in Refer-
8,10
ence
and I. L. Reich et al. (in preparation) indicate that
the D ring of a hormone they form probably does not
participate in binding as specifically to a receptor as do
some other parts of the steroid nucleus.
Biological activity: Results and discussion
D-Ring seco estrogens have unusual chemical4,27,28 and
biologic properties.3,4,29 Despite low affinity for estrogen
receptors, some of the seco compounds have much greater
oral estrogenic activity than estrone or estradiol.5,29 D-ring
seco steroids related to DHEA have been made and were
found to have no androgen or estrogen activity.6 Because
DHEA has a wide variety of biologic effects we thought it
worthwhile to assay these derivatives for an activity other
than those of the sex hormones. Of the open ring structures
synthesized only the 16,17 dicarboxylic acid (9) induced the
formation of cytosolic malic enzyme but not of glycero-
phosphate dehydrogenase when fed to rats (Table 1). Two
5,15-diene-17 3 13 lactones, with their 14-hydrogen atom
Acknowledgments
This work was supported by Humanetics Corporation, St.
Louis Park, MN. The laboratory of D.R.P. is supported by
grants from the NSF and the University of Wisconsin.
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