One-pot deuteration and reduction of ketones in the synthesis of [16,16,17-2H3]-epitestosterone

Article abstract of DOI:10.1135/cccc19961037

5α-Androstan-17-one and 6β-methoxy-3α,5-cyclo-5α-androstan-17-one were reduced by sodium in deuterium oxide to [16,16,17-²H₃]-17β-alcohols. The 17β-tosyloxy group of [16,16,17-²H ₃]-6β-methoxy-3α,5-cyclo-5α-andr

Full text of DOI:10.1135/cccc19961037

One-Pot Deuteration and Reduction of Ketones
1037
ONE-POT DEUTERATION AND REDUCTION OF KETONES IN THE
SYNTHESIS OF [16,16,17-2H3]-EPITESTOSTERONE*,**
1
2
3
4
Hana CHODOUNSKA , Alexander KASAL , David SAMAN and Karel UBIK
Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic,
166 10 Prague 6, Czech Republic; e-mail: 1 hchod@uochb.cas.cz, 2 kasal@marilyn.uochb.cas.cz,
3 saman@uochb.cas.cz, 4 ubik@uochb.cas.cz
Received March 12, 1996
Accepted June 8, 1996
5α-Androstan-17-one and 6β-methoxy-3α,5-cyclo-5α-androstan-17-one were reduced by sodium in
deuterium oxide to [16,16,17-2H3]-17β-alcohols. The 17β-tosyloxy group of [16,16,17-2H3]-6β-methoxy-
3α,5-cyclo-5α-androstan-17β-ol tosylate was found to be stable under the conditions of the i-steroid
rearrangement. The SN2 reaction of the 17β-tosyloxy group with potassium nitrite yielded the corre-
sponding 17α-hydroxy derivative without any loss of deuterium.
Key words: Steroid synthesis; Labelled epitestosterone.
Over the last 15 years the use of GC-MS for the determination of steroids in various
biological samples has increased3 considerably, and polydeuterated analytes have been
used as internal standards. Since mono- and dideuterated steroids are less useful in this
respect because of interference in the analysis by natural distribution of isotopes in
tested compounds, trideuterated steroids seem to be the optimal choice4. E.g., the tri-
deuterated testosterone standard has the labels in positions 16, 17 (refs3–5) or 19 (ref.6).
Deuterated epitestosterone is required as the standard for the determination of uri-
nary epitestosterone in drug abuse control7, further, it could be used in tracing a role of
epitestosterone in living organisms which has not yet been fully understood8.
[2,2,4,6,6-2H5]-Epitestosterone, prepared recently9, contains the highest content of
deuterium, though mostly in enolizable10 positions. Attempted synthesis9 of [16,16,17-
2H3]-epitestosterone (14) from commercialy available [16,16,17-2H3]-testosterone by
Mitsunobu reaction11 failed. Here we present its synthesis of from 6β-methoxy-3α,5-
cyclo-5α-androstan-17-one (5).
Model experiments, carried out with 5α-androstan-17-one (1) (see Scheme 1), were
devised to answer two questions: can a system of sodium and deuterium oxide sub-
* Part CCCLXXXIV in the series On steroids; Part CCCLXXXIII: see ref.1.
**Preliminary communication: ref.2.
Collect. Czech. Chem. Commun. (Vol. 61) (1996)

1038
Chodounska, Kasal, Saman, Ubik:
stitute for deuteride reagents in the reduction of ketones on a preparative scale? And
secondly, will solvolysis of a [17α-2H]-17β-tosyloxy derivative (e.g. 4) proceed without
loss of deuterium?
We found that two steps (isotope exchange and reduction) could conveniently be
combined together in one pot: sodium deuteroxide, generated from sodium and deute-
rium oxide, first facilitated the deuteration of ketone 1 in 1,2-dimethoxyethane, and
then the deuterated ketone was reduced by an additional lot of sodium. The metal was
added in several portions, until the TLC demonstrated the absence of the starting
ketone. Two alcohols were isolated, i.e. 2 (74%) and 3 (2%), no signals of H-17 were
found in their 1H NMR spectra. The major product 2 was converted in the 17β-tosyloxy
derivative 4 which on heating with potassium nitrite in DMF (refs12,13) afforded a pro-
duct 3 with the reversed configuration at carbon 17. No deuterium was lost during the
1
operation: H NMR spectrum of 4 revealed no signal due to H-17 proton.
The above results were utilized for the synthesis of [16,16,17-2H3]-epitestosterone
(14): readily available14 6β-methoxy-3α,5-cyclo-5α-androstan-17-one (5) (see Scheme 2)
was used as starting material. The above mentioned one-pot deuteration and reduction
afforded a mixture composed mainly of 17β-alcohol 6; the mixture was converted into
tosylate 7 without purification. The acid-sensitive15 i-steroid grouping in 7 was
isomerized by the action of perchloric acid in acetone16 under the formation of a 3β-hy-
droxy compound 8 which was acetylated to 9 prior to the next reaction step.
The solvolysis of the 17β-tosylate 9 was carried out using the above described con-
ditions. The major product 10 was isolated by means of column chromatography: its RF
OH
O
D
D
D
H
H
1
2
D
OTs
OH
D
D
D
D
D
H
H
3
4
SCHEME 1
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One-Pot Deuteration and Reduction of Ketones
1039
on a thin layer of silica gel was found identical with that of androst-5-ene-3β,17α-diol
3-acetate17, its NMR spectra were compatible with the proposed structure 10.
The following reaction sequence consisted of the protection of the 17α-hydroxy
group (by benzoylation to 11), deprotection of the 3β-hydroxy group (by selective hy-
drolysis to 12) and Oppenauer oxidation18 to ketone 13. Hydrolysis of benzoate 13
afforded [16,16,17-2H3]-epitestosterone (14) (yield 8% from ketone 5). Its NMR spec-
tra correlate well with spectra of unlabelled epitestosterone 15 (see Tables I to III) apart
from absent signals of H-17 (δ 3.76), H-16α and H-16β (δ 2.18 and 1.49). Further, the
OR
O
D
D
D
OMe
OMe
R = H
R = Ts
5
6,
7,
OTs
D
OR2
D
D
D
D
D
R1O
RO
R1 = Ac; R2 = H
10,
11,
12,
R = H
R = Ac
8,
9,
R1 = Ac; R2 = Bz
R1 = H; R2 = Bz
R1
OR2
R1
R1
O
R1 = D; R2 =Bz
R1 = D; R2 = H
R1 = H; R2 = H
13,
14,
15,
SCHEME 2
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1040
Chodounska, Kasal, Saman, Ubik:
multiplicity of signals of H-15α (δ 1.80) and H-15β (δ 1.22) collapsed due to the ab-
sence of vicinal coupling with H-16 protons. No residual signal of H-17 was observed.
For unambiguous assignment of 13C NMR spectra of compounds 14 and 15, 2D NMR
1
one bond (HMQC) H, 13C heteronuclear chemical shift correlated spectra was used19.
Interpretation of electron impact mass spectra of compound 14 reveals the pre-
valence of the desired [16,16,17-2H3]-epitestosterone. The mass spectrum is in full ac-
cord with fragmentation patterns of 4-ene-3-ketones20. The structure characteristic ions
2
and isotopic distribution in the molecular group are as follows: 1.6% H0 (m/z 288),
2
2
2
0.4% H1 (m/z 289), 8.1% 2H2 (m/z 290), 86.9% H3 (m/z 291), 1.2% H4 (m/z 292) and
2
1.8% H5 (m/z 293).
EXPERIMENTAL
The melting points were determined on a Kofler block and are uncorrected. Thin layer chromato-
graphy (ICN Silica G, TLC-60 A) was used for checking the purity of individual intermediates, un-
labelled standards were used for comparison. Column chromatography was carried out on silica gel
for thin layer chromatography (“Silpearl”, Kavalier, Czech Republic) using a slight overpressure. In-
frared spectra were recorded on a Brucker IFS 88 spectrometer in chloroform, wavenumbers are
given in cm–1. 1H and 13C NMR spectra were measured on a Varian UNITY-500 FT NMR spec-
trometer (499.8 MHz for 1H and 125.8 MHz for 13C) in deuteriochloroform. Chemical shifts were
referenced to internal tetramethylsilan (δ 0.0) and CDCl3 (δ 77.00) for 1H and 13C NMR, respec-
tively. Coupling constants (J) are given in Hz. For the HMQC heteronuclear correlation experiment
TABLE I
1H NMR parameters of unlabelled 15 and labelled 14 epitestosterone
Proton
14
15
Proton
14
15
1α
1β
2α
2β
4
1.72 m
1.72 m
11α
1.66 ddt
1.48 dq
1.58 dtq
1.53 ddd
1.42 ddd
1.79 bdd
1.20 bt
–
1.66 ddt
1.48 dq
2.05 ddd
2.35 dddd
2.43 ddd
5.73 dt
2.04 ddd
2.34 dddd
2.42 ddd
5.73 dt
11β
12α
1.59 dtq
1.52 ddd
1.42 ddd
1.80 dddd
1.22 dddd
2.18 dddd
1.49 dddd
3.76 dd
12β
14α
6α
6β
7α
7β
8β
9α
2.28 bddd
2.40 dddd
1.11 dddd
1.89 dddd
1.55 ddt
0.98 ddd
2.28 bddd
2.40 dddd
1.11 dddd
1.89 dddd
1.55 ddt
0.98 ddd
15α
15β
16α
16β
–
17β
–
18 (3 H)
19 (3 H)
0.71 d
1.20 d
0.71 d
1.20 d
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One-Pot Deuteration and Reduction of Ketones
1041
the standard pulse sequence with following parameters was applied: spectral width in f1 and f2 25 000
and 4 200 Hz, respectively, 2 048 × 2 048 Hz data matrix, 256 increments of 16 each and relaxation
delay 1 s. The Fourier transformations were performed with shifted and unshifted sine-bell apodiza-
tion functions in the f1 and f2 dimensions, respectively. Mass spectra and isotopic distribution in 14
was determined by a ZAB-EQ mass spectrometer (VG Analytical, Manchester) using a direct inlet
probe. The electron energy was 70 eV and the ion source temperature was 140 °C.
[16,16,17-2H3]-5α-Androstan-17β-ol (2)
Sodium (0.2 g, 8.7 mmol) was dissolved in a mixture of 1,2-dimethoxyethane (10 ml) and deuterium
oxide (3 ml, 97.4%) under nitrogen. 5α-Androstan-17-one (1; 150 mg, 0.55 mmol) was added and
the mixture was stirred at 90 °C for 7 h. Additional sodium (2.6 g, 113 mmol) was added to the
solution in seven portions during 16 h, until TLC certified the disappearance of the starting ketone.
After addition of ethanol (4 ml), the mixture was diluted with a saturated solution of ammonium
TABLE II
1
Proton–proton coupling constants (J, Hz) of protons in H NMR spectra of labelled 14 and unlabelled
15 epitestosterone
J(H,H)
14
15
J(H,H)
14
15
1α,1β
1α,2α
1α,2β
1α,19
1β,2α
1β,2β
2α,2β
2α,4
13.4
4.8
13.4
4.9
9α,11α
4.2
12.1
12.7
4.1
2.5
12.8
4.3
13.4
0.8
7.3
12.3
12.5
–
4.2
12.2
12.8
4.1
9α,11β
14.3
0.8
14.4
0.8
11α,11β
11α,12α
11α,12β
11β,12α
11β,12β
12α,12β
12α,18
3.1
3.2
2.6
5.1
5.1
12.4
4.2
16.8
0.8
16.9
0.9
13.4
0.8
4,6α
0.7
0.6
4,6β
1.9
1.8
14α,15α
14α,15β
15α,15β
15α,16α
15α,16β
15β,16α
15β,16β
16α,16β
16α,17β
16β,17β
7.2
6α,6β
6α,7α
6α,7β
6β,7α
6β,7β
7α,7β
7α,8β
7β,8β
8β,9α
8β,14α
14.6
4.3
14.7
4.2
12.2
12.2
2.7
2.5
2.5
13.7
5.4
13.9
5.4
–
9.5
–
10.9
6.7
12.8
11.7
3.5
12.8
11.6
3.6
–
–
15.1
5.9
–
10.6
10.9
10.6
10.8
–
0.8
Collect. Czech. Chem. Commun. (Vol. 61) (1996)

1042
Chodounska, Kasal, Saman, Ubik:
chloride (30 ml), and extracted with toluene (3 × 15 ml). Combined organic extracts were washed
with hydrochloric acid (5%, 2 × 5 ml), saturated solution of sodium hydrogencarbonate (5 ml), water
(5 ml) and dried over anhydrous magnesium sulfate. Evaporation and chromatography on silica gel
column (50 g, toluene–ether, 9 : 1) afforded 115 mg (74%) of (2), m.p. 165–167 °C (ref.21 gives m.p.
163–167 °C for unlabelled compound). 1H NMR spectrum: 0.79 s, 3 H (3 × H-19); 0.72 s, 3 H (3 × H-18).
[16,16,17-2H3]-5α-Androstan-17α-ol (3)
a) Lipophilic fractions of the above chromatography (3 mg, 2%) consisted of compound 3, with
RF identical with unlabelled compound22. 1H NMR spectrum: 0.79 s, 3 H (3 × H-19); 0.65 s, 3 H (3 × H-18).
b) The mixture of 4 (116 mg, 0.27 mmol) and sodium nitrite (1.00 g, 14.5 mmol) in N,N-di-
methylformamide (20 ml) and deuterium oxide (0.5 ml, 97.4%) was stirred at 140 °C for 8 h. The
mixture was cooled at room temperature, diluted with water (40 ml) and extracted with ether (3 × 15 ml).
The combined organic layers were washed with water (5 × 5 ml), dried over anhydrous magnesium
sulfate, and the solvent was evaporated in a vacuum. Chromatography of the residue (90 mg) on the
thin layer of silica gel (benzene–ether, 3 : 1) afforded 46 mg (59%) of 3, m.p. 144–147 °C (ref.22
records m.p. 142–146 °C for unlabelled compound). The product was identical with the sample pre-
pared sub a).
[16,16,17-2H3]-5α-Androstan-17β-ol Tosylate (4)
4-Toluenesulfonyl chloride (130 mg, 0.7 mmol) was added to a solution of 2 (100 mg, 0.36 mmol)
in pyridine (5 ml) cooled with an ice bath and the mixture was allowed to stand overnight at 40 °C.
Then it was poured onto ice, extracted with ethyl acetate (3 × 10 ml), the combined organic layers
were washed with hydrochloric acid (5%, 4 × 3 ml), water, saturated solution of sodium hydrogen-
carbonate (5 ml) and dried over anhydrous magnesium sulfate. Evaporation of the solvent in a va-
cuum afforded 116 mg (75%) of crude product 4. TLC (benzene–ether, 4 : 1) showed identity with
TABLE III
13C NMR chemical shifts (δ, ppm) of unlabelled 15 and labelled 14 epitestosterone
Carbon
14
15
Carbon
14
15
1
2
35.7
33.9
35.7
33.9
11
12
13
14
15
16
17
18
19
20.5
31.1
45.1
48.2
20.5
31.1
45.1
48.2
24.5
32.3
79.6
16.9
17.4
3
199.6
123.9
171.4
32.9
199.6
123.8
171.4
32.9
4
5
24.3
a
6
a
7
32.3
32.3
8
35.8
35.8
16.8
17.4
9
53.6
53.6
10
38.6
38.6
a
Value not determined.
Collect. Czech. Chem. Commun. (Vol. 61) (1996)

One-Pot Deuteration and Reduction of Ketones
1043
1
unlabelled compound23. Compound 4 was used for the next conversion without purification. H NMR
spectrum: 7.75 d, 2 H, J = 8.5 (H-2 and H-6, tosylate); 7.32 d, 2 H, J = 8.5 (H-3 and H-5, tosylate);
2.44 s, 3 H (CH3, tosylate); 0.78 s, 3 H (3 × H-19); 0.69 s, 3 H (3 × H-18).
[16,16,17-2H3]-6β-Methoxy-3α,5-cyclo-5α-androstan-17β-ol (6)
Potassium tert-butoxide (2.1 g, 18.7 mmol) was dissolved in 1,2-dimethoxyethane (25 ml) and deuterium
oxide (3 ml, 94.7%) and then 6β-methoxy-3α,5-cyclo-5α-androstan-17-one4 (5; 1.0 g, 3.31 mmol)
was added. The mixture was stirred at 90 °C for 7 h under nitrogen. The mixture was further diluted
with deuterium oxide (3.5 ml, 94.7%) and sodium (2.7 g, 129 mmol) was introduced into this solu-
tion at room temperature in seven portions over a period of 16 h, until the TLC showed the disap-
pearance of the starting ketone. The reaction mixture was poured onto ice and left in a refrigerator
overnight. The precipitate was filtered off, dissolved in toluene, washed with water, the solution was
dried over anhydrous magnesium sulfate and the solvent evaporated. The residue (890 mg, 79%) con-
sisted of compound 6 (1H NMR and TLC of unlabelled compound24) was used for the next step without
1
purification. H NMR spectrum: 3.37 s, 3 H (CH3O); 2.83 t, 1 H, J = 2.8 (H-6α); 1.05 s, 3 H (3 × H-19);
0.92 s, 3 H (3 × H-18); 0.68 dd, 1 H, J = 4.5, J′ = 5.6 (H-4α); 0.47 dd, 1 H, J = 5.7, J′ = 8.9 (H-4β).
[16,16,17-2H3]-6β-Methoxy-3α,5-cyclo-5α-androstan-17β-ol Tosylate (7)
4-Toluenesulfonyl chloride (1.5 g, 7.9 mmol) was added to a solution of 6 (890 mg, 2.61 mmol) in
pyridine (10 ml) cooled with an ice bath. The mixture was allowed to stand for 40 h at 40 °C. Then
it was poured onto ice and left in a refrigerator overnight. The precipitate was filtered, washed with
water and dried at 20 °C. The yield of tosylate 7, m.p. 120–122 °C (ref.24 gives m.p. 124 °C for
unlabelled compound) was 1.0 g (79%). 1H NMR spectrum: 7.75 d, 2 H, J = 8.5 (H-2 and H-6,
tosylate); 7.33 d, 2 H, J = 8.5 (H-3 and H-5, tosylate); 3.36 s, 3 H (3 × H, OCH3); 2.80 t, 1 H, J = 3
(H-6α); 2.45 s, 3 H (CH3, tosylate); 1.02 s, 3 H (3 × H-19); 0.89 s, 3 H (3 × H-18); 0.66 dd, 1 H, J = 4.5,
J′ = 5.5 (H-4α).
[16,16,17-2H3]-Androst-5-ene-3β,17β-diol 17-Tosylate (8)
Tosylate 7 (1.0 g, 2.05 mmol) in acetone (25 ml) was treated with mixture of water (0.25 ml) and
perchloric acid (73%, 0.25 ml) for 5 min. Saturated hydrogencarbonate solution (0.6 ml) was added
and acetone was removed in a stream of argon. Water (5 ml) was added and the product was taken
up in ethyl acetate, the extract was dried with magnesium sulfate and evaporated to yield 890 mg
1
(98%) of 8 (TLC showed identity with unlabelled standard25). H NMR spectrum: 7.76 d, 2 H, J = 8.5
(H-2 and H-6, tosylate); 7.34 d, 2 H, J = 8.5 (H-3 and H-5, tosylate); 5.39 d, 1 H, J = 4.8 (H-6); 3.50 m, 1 H
(H-3α ); 2.44 s, 3 H (CH3, tosylate); 0.98 s, 3 H (3 × H-19); 0.79 s, 3 H (3 × H-18).
[16,16,17-2H3]-Androst-5-ene-3β,17β-diol 3-Acetate 17-Tosylate (9)
Alcohol 8 (890 mg, 2.0 mmol) was acetylated with acetic anhydride (3 ml) in pyridine (4 ml). After
18 h the reaction mixture was poured onto ice, and the precipitate formed was extracted with ethyl
acetate (4 × 15 ml). The combined extracts were washed with hydrochloric acid (5%, 4 × 15 ml) and
then with a saturated solution of sodium hydrogencarbonate (2 × 5 ml), dried over anhydrous mag-
nesium sulfate and evaporated. Crystallization from a mixture of light petroleum, dichloromethane
and ether yielded product 9 (960 mg, 98%) m.p. 158–161 °C (ref.25 gives 162–164 °C for unlabelled
compound). 1H NMR spectrum: 7.79 d, 2 H, J = 8.5 (H-2 and H-6, tosylate); 7.35 d, 2 H, J = 8.5
(H-3 and H-5, tosylate); 5.34 d, 1 H, J = 4.3 (H-6); 4.58 m, W = 32, 1 H (H-3α); 2.45 s, 3 H (CH3,
Collect. Czech. Chem. Commun. (Vol. 61) (1996)

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Chodounska, Kasal, Saman, Ubik:
tosylate); 2.03 s, 3 H (OOCCH3); 1.00 s, 3 H (3 × H-19); 0.81 s, 3 H (3 × H-18). IR spectrum: 2 233,
2 144, 2 200 (C–D); 1 725 (C=O); 1 669 (C=C); 1 599, 1 496, 1 361, 1 178 (OTs); 1 255, 1 028,
959 (C–O).
[16,16,17-2H3]-Androst-5-ene-3β,17α-diol 3-Acetate (10)
Tosylate 9 (960 mg, 1.96 mmol) was added to a solution of sodium nitrite (10 g, 114 mmol) in N,N-
dimethylformamide (40 ml) and deuterium oxide (1 ml, 97.4%). The mixture was stirred at 140 °C
for 24 h. After cooling the precipitate was taken up, diluted with water (40 ml) and extracted with
ether (3 × 15 ml). Combined organic layers were washed with hydrochloric acid (5%, 15 ml) a satu-
rated solution of sodium hydrogencarbonate (5 ml), water (2 × 5 ml), dried over anhydrous magne-
sium sulfate, and evaporated in a vacuum. Chromatography of the residue on a column of silica gel
(benzene–ether, 9 : 1) afforded 350 mg (54%) of hydroxy derivative 10, m.p. 115–117 °C (ref.17
gives 110–115 °C for unlabelled compound). 1H NMR spectrum: 5.40 d, 1 H, J = 4.8 (H-6); 4.5–4.7 m,
1 H (H-3α); 2.03 s, 3 H (OOCCH3); 1.03 s, 3 H (3 × H-19); 0.68 s, 1 H (3 × H-18).
[16,16,17-2H3]-Androst-5-ene-3β,17α-diol 3-Acetate 17-Benzoate (11)
Benzoyl chloride (4.0 ml, 35 mmol) was added to a solution of hydroxy derivative 10 (350 mg, 1.05 mmol)
in pyridine (10 ml) at 0 °C. After 24 h at room temperature the reaction mixture was poured into hot
water, after cooling, the precipitate was taken up in ether. The solution was washed with hydro-
chloric acid (5%, 4 × 20 ml), and a saturated solution of sodium hydrogencarbonate (2 × 10 ml). The
extract was dried over anhydrous magnesium sulfate and evaporated. Benzoyl derivative 11 (420 mg,
1
90%) was obtained, m.p. 134–136 °C (ref.17 records 130–132 °C for unlabelled compound). H NMR
spectrum: 8.00–8.09 m, 2 H (H-2 and H-6, benzoate); 7.40–7.60 m, 3 H (H-3, H-4 and H-5, benzoate);
5.39 d, 1 H, J = 4.4 (H-6); 4.54 m, 1 H (H-3α); 2.04 s, 3 H (OOCCH3); 1.04 s, 3 H (3 × H-19); 0.84 s,
3 H (3 × H-18).
[16,16,17-2H3]-Androst-5-ene-3β,17α-diol 17-Benzoate (12)
Benzoyl derivative 11 (420 mg, 0.9 mmol) was dissolved in methanol (8 ml), and chloroform (1 ml)
and hydrochloric acid (37%, 0.6 ml) were added. The mixture was heated for 7 h at 40 °C. Then it
was diluted with water (100 ml) and extracted with ether (4 × 25 ml). The combined extracts were
washed with water and a saturated solution of sodium hydrogencarbonate. Drying over anhydrous
magnesium sulfate and evaporation of solvent yielded 370 mg (100%) of 3-hydroxy derivative 12,
1
m.p. 145–148 °C (ref.26 gives 148–149 °C for unlabelled compound). H NMR spectrum: 8.00–8.09 m,
2 H (H-2 and H-6, benzoate); 7.40–7.60 m, 3 H (H-3, H-4 and H-5, benzoate); 5.37 d, 1 H, J = 4.6
(H-6); 3.46 m, 1 H (H-3α); 1.02 s, 3 H (3 × H-19); 0.83 s, 3 H (3 × H-18).
[16,16,17-2H3]-17α-Benzoyloxyandrost-4-en-3-one (13)
1-Methyl-4-piperidone (1.0 ml, 8.1 mmol) was added under argon to a solution of 12 (370 mg, 0.9 mmol)
in toluene (50 ml). A part (3 ml) of the toluene was distilled off and 1 M solution of aluminium
isopropoxide in toluene (2.5 ml) was added. After refluxing under argon for 9 h the mixture was
cooled, poured into hydrochloric acid (5%, 20 ml), water (20 ml) and a saturated solution of sodium
hydrogencarbonate (20 ml). The organic layer was dried over anhydrous magnesium sulfate. Eva-
poration of solvents afforded ketone 13 (240 mg, 65%), m.p. 138–140 °C (ref.25 records 138–139 °C
1
for unlabelled compound). H NMR spectrum: 8.00–8.09 m, 2 H (H-2 and H-6, benzoate); 7.40–7.60
m, 3 H (H-3, H-4 and H-5, benzoate); 5.75 bs, 1 H (H-4); 1.20 s, (3 × H-19); 0.87 s, 3 H (3 × H-18).
Collect. Czech. Chem. Commun. (Vol. 61) (1996)

One-Pot Deuteration and Reduction of Ketones
1045
[16,16,17-2H3]-17α-Hydroxyandrost-4-en-3-one (14)
The solution of benzoate 13 (240 mg, 0.61 mmol) and potassium hydroxide (250 mg) in ethanol (10 ml)
and benzene (10 ml) was heated to 40 °C for 4 h. The cooled mixture at room temperature was
diluted with hydrochloric acid (2%, 50 ml), extracted with ether (3 × 50 ml). The combined extracts
were washed with saturated solution of sodium hydrogencarbonate and dried over anhydrous magnesium
sulfate. Chromatography on the column of silica gel (15 g) yielded 65 mg (36%) of 2H3-epitesto-
sterone (14), m.p. 221–223 °C (ref.27 gives 216–218 °C for unlabelled compound). NMR spectra: see
Tables I–III. IR spectrum: 3 617, 3 457 (O–H); 2 222, 2 189, 2 143 (C–D); 1 660 (C=O); 1 615
(C=C). Mass spectrum, m/z (%): 291 (M, 100), 124 (98), 149 (59), 231 (50), 249 (37), 206 (27), 168
(22), 216 (13), 273 (12).
The authors are indebted to Dr L. Bednarikova for taking and interpretation of IR spectra. The
project was supported by a grant of the Grant Agency of the Czech Republic (505/94/0009).
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