4
46
D. Bastien et al. / European Journal of Medicinal Chemistry 64 (2013) 442e447
ꢀ1
þ
þ
(
(
M þ H) calculated for C25
H36ClO
3
¼ 419.2348; found ¼ 419.2346
IR(NaCl, nmax, cm ): 3449 (OeH), 1656 (C]O), 1615 (C]C), 1077
1
M ꢀ H) .
(CeO); H NMR (CDCl
CH), 5.02e4.94 (2H, dd, J ¼ 3.1 Hz and J ¼ 1.2 Hz, 22-CH
J ¼ 11.3 Hz and J ¼ 2.0 Hz, 22-CH ), 3.64 (1H, t, J ¼ 16.8 Hz, 17-CH),
1.19 (3H, s, 19-CH ), 0.78 (3H, s, 18-CH
3
,
d
ppm): 5.69 (1H, s, 4-CH), 5.60 (1H, m, 21-
a
and dt,
4
.3.2. Synthesis of 7
a
-(4-chloro-but-2-enyl)-4-androsten-17
b-ol-3-
b
13
one (5)
3
3 3
); C NMR (CDCl , d ppm):
The steroid 4 (0.65 g, 1.55 mmol) was initially dissolved in
methanol (12 mL). To this solution, 5 N aqueous HCl (4 mL) was
added and the mixture was gently heated for 2 h. Of note, the re-
action can occur at room temperature but must be stirred for a
longer period of time. The reaction mixture was cooled down and
transferred into a separatory funnel with ether (50 mL) and water
199.4 (C-3), 169.8 (C-5), 137.1 (C-21), 126.4 (C-4), 117.0 (C-22), 81.8
(C-17), 47.4, 46.4, 43.1, 38.9, 38.7, 36.5, 36.3, 36.2, 36.1, 34.3, 30.5,
þ
þ
30.4, 22.9, 21.1, 18.2, 11.2; MS (m/e): 328 (M ), 287 (M ꢀ C
3 5
H ).
exact mass: calculated for C22
32 2
H O
¼ 328.2402; found ¼ 328.2395.
4.4.2. Synthesis of 4-{4-[bis-(2-chloro-ethyl)-amino]-phenyl}-butyric
(
25 mL). The ethereal phase was washed once with a saturated
sodium bicarbonate (NaHCO ) solution (10 mL) and with water
4 ꢂ 10 mL). The organic solution was dried, filtered and evaporated
to a solid. The final product 5 (0.54 g) was obtained with 94% yield
acid allyl ester (7)
Chlorambucil (350 mg, 1.15 mmol) was added into a 10 mL pear
shaped flask. Oxalyl chloride ((COCl) , 3.0 g, 2 mL, 23.6 mmol) was
then added to the solid, and the mixture stirred for a period of 15 min
at room temperature (20 C), under an inert atmosphere of nitrogen.
3
(
2
ꢁ
(
90% trans) and excellent purity. As a result, it was used without
ꢁ
ꢀ1
purification at the next step. MP: 142e145 C; IR(NaCl,
3
n
max, cm ):
Afterwards, the excess (COCl)
nitrogen. The resulting oil was dissolved in CH
2
was evaporated using a flow of dry
Cl (2 mL) and
1
423 (OeH),1663 (C]O),1611 (C]C); H NMR (CDCl
3
,
d
ppm): 5.69
2
2
(
3
CH
2
4
1
1H, s, 4-CH), 5.61 (2H, m, 21-CH and 22-CH), 3.99 (2H, m, 23-CH
2
),
), 0.78 (3H, s, 18-
ppm): 199.3 (C-3), 169.4 (C-5), 134.2 (C-
evaporated once more with nitrogen. The acid chloride intermediate
was treated with a solution of allyl alcohol (667 mg, 0.78 mL,
11.5 mmol) dissolved in DCM (2 mL) and pyridine (1.82 g, 1.86 mL,
23.0 mmol). The resulting solution was stirred at room temperature
for 30 min. Then, the reaction mixture was transferred into a sepa-
ratory funnel with diethyl ether (40 mL) and washed several times
with water (5 ꢂ 10 mL). The organic phase was dried, filtered and
evaporated to a crude solid. Flash chromatography with hexanes/
acetone (4:1) as the eluent gave the final pure ester 7 (373 mg) with
.63 (1H, t, J ¼ 8.2 Hz, 17-CH), 1.19 (3H, s, 19-CH
3
13
3
); C NMR (CDCl
3
,
d
1), 128.4 (C-22), 126.4 (C-4), 81.8 (C-17), 47.4, 46.4, 45.2 (C-23),
3.2, 38.9, 38.8, 36.9, 36.5, 36.4, 36.2, 34.2, 30.5, 28.8, 23.0, 21.1,18.2,
þ
þ
1.2; ESI þ HRMS: (M þ H) calculated for C23
H34ClO
2
¼ 377.2242;
found ¼ 377.2240 (M þ H) .
4
.3.3. Synthesis of 7 -(4-(4-(bis-(2-chloro-ethyl)-amino)-phenyl-
a
propylcarbonyloxy)-but-2-enyl)-4-androsten-17 -ol-3-one (1)
b
max
95% yield (see numbering system on Scheme 2). IR(NaCl, n ,
ꢀ1
1
Steroid 5 (50 mg, 0.133 mmol) was initially dissolved in DMF
1 mL) in a pear shaped flask. In another flask, chlorambucil (50 mg,
cm ): 1734 (C]O, ester); 1617 (C]C); H NMR (CDCl
(2H, d, J ¼ 8.5 Hz, 9-CH), 6.63 (2H, d, J ¼ 8.5 Hz,10-CH), 5.90 (1H, m, 2-
CH), 5.25 (2H, m, 1-CH ),
3.66 (8H, m, 12-CH and 13-CH ), 2.36
(t, 2H, J ¼ 7.4 Hz, 5-CH
(CDCl
3
, d ppm): 7.08
(
0
.164 mmol) and NaHCO
3
(10 mg, 0.16 mmol) were dissolved in
2
), 4.58 (2H, apparent d, J ¼ 2.3 Hz, 3-CH
2
DMF (4 mL) and water (0.15 mL). The resulting mixture was
warmed up to help dissolution. Then, the steroid solution was
added and, the resulting mixture, stirred at reflux for 9 h. After-
wards, the solution was cooled down, transferred into a separatory
funnel with ether (30 mL) and water (10 mL). The ethereal phase
was washed with water (6 ꢂ 20 mL), dried, filtered and evaporated
to a crude solid. Flash chromatography with hexanes/acetone
2
2
), 2.57 (2H, t, J ¼ 7.4 Hz, 7-CH
2
1
3
2
), 1.94 (2H, q, J ¼ 7.4 Hz, 6-CH
2
); C NMR
3
,
d
ppm): 173.3 (C-4), 144.5 (C-11), 132.5 (C-2), 130.7 (C-9),
129.9 (C-8), 118.3 (C-1), 112.4 (C-10), 65.1 (C-3), 53.8 (C-12), 40.7 (C-
þ
13), 34.1 (C-7), 33.7 (C-5), 26.9 (C-6); ESI þ HRMS: (M þ H) calcu-
þ
lated for C17H24Cl NO
2 2
¼ 344.1179; found ¼ 344.1182 (M þ H) .
(
95:5) as the eluent gave the final pure hybrid 1 (50 mg) with 58%
4.4.3. Synthesis of 7
propylcarbonyloxy)-but-2-enyl)-4-androsten-17b
4-{4-[bis-(2-chloro-ethyl)-amino]-phenyl}-butyric acid 4-(4-{4-[bis-
a
-(4-(4-(bis-(2-chloro-ethyl)-amino)-phenyl-
ꢁ
ꢀ1
yield (85% trans). MP: 47e50 C; IR(NaCl,
1
n
max, cm ): 3422 (OeH),
-ol-3-one (1) and of
1
730 (C]O, ester),1664 (C]O, enone),1242 (CeO); H NMR (CDCl
3
,
d
ppm): 7.07 (2H, d, J ¼ 8.6 Hz, 29-CH), 6.60 (2H, d, J ¼ 8.6 Hz, 30-
(2-chloro-ethyl)-amino]-phenyl}-butyryloxy)-but-2-enyl ester (8)
CH), 5.68 (1H, s, 4-CH), 5.54 (2H, m, 21-CH and 22-CH), 4.48 (2H, d,
J ¼ 4.7 Hz, 23-CH ), 3.65 (9H, m, 17-CH, 32-CH , 33-CH ), 1.19 (3H, s,
9-CH ), 0.78 (3H, s, 18-CH ppm): 199.3 (C-3),
73.6 (C-24), 169.52 (C-5), 144.5 (C-31), 134.3 (C-21), 129.9 (C-28, C-
Under a nitrogen atmosphere, the 7a-allyltestosterone 6 (20 mg,
ꢀ
5
2
2
2
6.1 ꢂ10 mol) was dissolved in dry CH
2
Cl
2
(0.5 mL). The Hoveyda-
13
ꢀ6
1
3
3
); C NMR (CDCl
3
,
d
Grubbs 2nd generation catalyst (4 mg, 6.3 ꢂ 10 mol) dissolved in
1
2
5
2
C
2 2
dry CH Cl (0.5 mL) was added to the steroid solution. Finally,
ꢀ
4
9), 126.5 (C-22), 126.4 (C-4), 112.5 (C-30), 81.8 (C-17), 64.9 (C-23),
3.9, 47.4, 46.4, 43.1, 40.7, 38.9, 38.7, 36.5, 36.4, 36.2, 34.2, 33.8, 30.5,
excess allyl ester 7 (40 mg, 1.16 ꢂ 10 mol) was added to the
mixture. The resulting solution was stirred at reflux for 24 h. Af-
terwards, the mixture was evaporated to dryness. The crude
brownish product was purified by flash chromatography with
hexanes/acetone (4:1) as the eluent to give the pure steroid 1
(30 mg) with 78% yield (85% trans). This yield is based on derivative
6 initially used. Along with the desired cross-coupling derivative 1,
the dimer 8 was also isolated (23 mg) as a side product with 61%
yield. The latter yield is based on derivative 7 used in the reaction
and the maximum quantity of dimer this could produce. The
spectral data for compound 1 are identical as those previously
described above in Section 4.3.3. The spectral data for compound 8
þ
8.9, 26.9, 22.9, 21.1, 18.2, 11.2; ESI þ HRMS: (M þ H) calculated for
þ
37
H52Cl
2
NO
4
¼ 644.3268; found ¼ 644.3260 (M þ H) .
4
.4. Synthesis of 7a-testosteroneechlorambucil hybrid (1) via an
olefin cross-metathesis reaction (Scheme 2)
4
.4.1. Synthesis 7
a-allyl-4-androsten-17b-ol-3-one (6)
The steroid 3 (0.71 g, 1.9 mmol) was initially dissolved in
methanol (15 mL). To this solution, 5 N aqueous HCl (5 mL) was
added and the mixture was gently heated for 2 h. The reaction
mixture was cooled down and transferred into a separatory funnel
with ether (100 mL) and water (25 mL). The ethereal phase was
ꢀ
1
(see numbering system on Scheme 2): IR(NaCl,
n
max, cm ): 1734
ppm): 7.08 (2H, d,
J ¼ 8.2 Hz, 8-CH), 6.64 (2H, d, J ¼ 8.2 Hz, 9-CH), 5.86 (1H, s, 1-CH),
4.60 (2H, s, 2-CH ), 3.67 (8H, m, 11-CH and 12-CH ), 2.57 (2H, t,
J ¼ 7.4 Hz, 6-CH ), 2.36 (t, 2H, J ¼ 7.4 Hz, 4-CH ), 1.94 (2H, apparent
p, J ¼ 7.4 Hz, 5-CH ppm): 173.2 (C-3), 144.4 (C-
10), 130.8 (C-7), 129.9 (C-8), 128.3 (C-1), 112.4 (C-9), 63.9 (C-2),
1
3
(C]O, ester), 1615 (C]C); H NMR (CDCl , d
3
washed once with a saturated sodium bicarbonate (NaHCO ) so-
lution (10 mL) and with water (4 ꢂ 10 mL). The organic solution was
dried, filtered and evaporated to a solid. The final product 6 (0.60 g)
was obtained with 95% yield and excellent purity. As a result, it was
2
2
2
2
2
13
2
3
); C NMR (CDCl , d
ꢁ
used without purification at the next step. MP: 195e200 C;