R.N. Hanson and N. Gajadeera
Steroids 145 (2019) 39–46
2
.88 (m, 2H), 2.56 – 2.39 (m, 2H), 2.38 – 2.35 (m, 1H), 2.24 (s, 3H),
.20 – 2.00 (m, 3H), 1.96 (dd, 1H), 1.72 – 1.43 (m, 3H), 0.45 (s, 3H).
C NMR (100 MHz, CDCl3) δ 220.20, 169.74, 155.51, 148.19, 137.53,
35.66, 130.58, 127.57,121.73, 119.24, 113.89, 66.74, 52.22, 50.28,
8.19, 47.69, 40.03, 38.12, 35.39, 34.94, 30.04, 27.18, 21.33, 15.25.
128.61, 128.03, 123.37, 123.07, 118.48, 115.28, 114.62, 66.04, 50.52,
49.64, 47.59, 45.39, 39.48, 38.79, 37.88, 37.82, 36.74, 35.36, 30.82,
26.70, 25.84, 21.80, 14.43. LC -MS – m/z - observed M + 1 – 720.27.
2
1
3
−1
−1
1
4
Retention time 2.66 min IR 1735.46 cm
(3C = O)
(17C = O), 1658.35 cm
LC -MS – m/z - observed M + 1 – 474.30 ; M + 18 – 491.32. Retention
time – 3.44 min.
2.10. Synthesis of 11β-(4-(2–4(-(((7-nitrobenzo[c][1,2,5] oxadiazol-4-yl)
amino)methyl-1H-1,2,3-triazol-1-yl)ethoxy)phenyl)-estra-4,9-diene-3,17-
dione (10b)
2.7. Synthesis of N-Dansyl propargylamine 8.
To dansyl chloride (55 mg, 0.21 mmol) dissolved in di-
chloromethane (4 mL) was added sequentially triethyl amine (20 µL,
.21 mmol) and propargyl amine (40 µL, 0.63 mmol). The reaction was
stirred at ambient temperature for 16 h and then evaporated to dryness.
The resultant crude material was purified by flash chromatography
11β-[4-(2-azidoethyoxy) phenyl]-estra-4,9-diene-3,17-dione, 6,
(16 mg 0.04 mmol) of was dissolved in a solution of tert butanol-water
(1:1, 5 mL v/v). 7-Nitro-N-(prop-2-yn-1-yl)benzo[c][1,2,5] oxadiazol-4-
amine 9 (15 mg, 0.06 mmol) sodium ascorbate (350 µL 0.035 mmol in a
solution of 0.1 mmol/mL), copper (II) sulfate, (70 µL, 0.007 mmol in a
solution of 0.1 mmol/mL) were added and the reaction vial was sealed.
The reaction was heated at 80 °C with stirring for 18 h, cooled to am-
bient temperature and partitioned between ethyl acetate and water.
The organic phase was washed with water (2 × 20 mL), brine, dried
over magnesium sulfate (anhydrous), filtered and evaporated to dry-
ness. The crude solid was purified by flash chromatography (ethyl
0
(
hexane:ethyl acetate 3:2) to yield 40 mg (0.14 mmol, 65%) of the final
product 8.
Rf = 0.5 (1:1 hexane: ethyl acetate) 1H NMR (400 MHz, CDCl
.56 (d, J = 8.5 Hz, 1H), 8.26 (t, J = 8.3 Hz, 2H), 7.55 (dd, J = 14.8,
.2 Hz, 2H), 7.19 (d, J = 7.5 Hz, 1H), 4.87 (t, J = 5.9 Hz, 1H), 3.77 (dd,
3
) δ
8
7
1
3
J = 6.0, 2.5 Hz, 2H), 2.89 (s, 6H), 1.91 (s, 1H) C NMR (100 MHz,
CDCl ) δ 152.19, 134.24, 130.97, 130.10, 129.98, 129.89, 128.73,
23.31, 118.61, 115.34, 77.86 72.84, 45.55, 33.16 LC -MS – m/z -
observed M + 1 – 289.17. Retention time 2.13 min
3
acetate : hexane, 9:1) to give the pure product 10b (12 mg,
1
0.018 mmol) in 46% isolated yield.
Rf = 0.04 (9:1 ethyl acetate: hexane) 1H NMR (400 MHz, CDCl
) δ
3
8
.45 (d, J = 8.5 Hz, 1H), 7.85 (s, 1H), 7.26 (s, 1H), 7.08 (d, J = 8.3 Hz,
2
.8. Synthesis of 7-nitro-N-(prop-2-yn-1-yl)benzo[c][1,2,5] oxadiazol-4-
2H), 6.97 (s, 1H), 6.74 (d, J = 8.4 Hz, 2H), 6.35 (d, J = 8.5 Hz, 1H),
4.81 (d, J = 18.5 Hz, 2H), 4.44 – 4.25 (m, 2H), 3.34 (s, 1H), 2.77 – 2.68
amine (9)
(
m, 1H), 2.54 – 2.0 (m, 9H), 1.96 – 1.88 (dd, 1H), 1.92 (dd, J = 13.7,
1
3
To 4-chloro-7-nitrobenzo[c][1,2,5] oxadiazole, (75 mg 0.37 mmol)
dissolved in 3 mL THF, were added propargyl amine (26 μL,
.40 mmol), cesium carbonate (136 mg, 0.42 mmol). The reaction
mixture was stirred at ambient temperature for 18 h. The reaction
mixture was poured into ethyl acetate (50 mL) and washed with water
7.2 Hz, 1H), 1.61 – 1.50 (m, 3H), 0.96 – 0.78 (m, 3H), 0.51 (s, 3H).
C
NMR (100 MHz, acetone) δ 217.99, 198.12, 157.14, 156.54, 145.82,
138.39, 137.65, 130.66, 129.08, 124.69, 123.58, 115.42, 67.28, 60.53,
51.36, 50.42, 48.16, 40.36, 39.01, 38.76, 37.51, 35.68, 31.33, 27.73,
26.65, 22.41, 20.83, 14.85, 14.50. LC -MS – m/z - observed M + 1 –
0
−1
(
2 × 25 mL), dried over magnesium sulfate (anhydrous), filtered and
650.21. Retention time 2.34 min. IR = 1733.58 cm
(17C=O),
−
1
evaporated to dryness. The crude material was purified by flash chro-
1657.42 cm
(3C=O)
matography (hexane : ethyl acetate 3:2) to give 53 mg (0.24 mmol, 65%
yield) of 9.
2.11. Synthesis of 3-hydroxy-11β-(4-(2-(4((5-dimethylamino)
naphthalene)-1-sulfonamido)-methyl)-1H-1,2,3- triazol-1-yl)-ethoxy)
phenyl) estra-1,3,5(10)-triene-17-one-diene (11a)
1
Rf = 0.17 (hexane : ethyl acetate 7:3) H NMR (400 MHz, CDCl
3
) δ
8
3
.54 (d, J = 8.5 Hz, 1H), 6.37 (t, J = 12.5 Hz, 2H), 4.31 (d, J = 3.7 Hz,
H), 2.43 (s, 1H), 1.62 (s, 1H)
3
-Hydroxy-11β-[-4-(2-azidoethoxy)-phenyl]-estra-1,3,5(10)-triene-
2
.9. Synthesis of 11β-(4-(2-(4-(5-dimethylamino) naphthalene)-1-
17-one acetate 7 (36 mg, 0.08 mmol) was dissolved in tert-butanol –
water (6 mL, 1:1 solution). N-Dansyl propargylamine, 8a, (33 mg,
0.1 mmol), sodium ascorbate (350 µL, 0.035 mmol in a solution of
sulfonamido)-methyl)-1H-1,2,3- triazol-1-yl)-ethoxy)phenyl) estra 4,9-
diene-3,17-dione (10a)
0.1 M), copper (II) sulfate (70 µL, 0.007 mmol in a solution of 0.1 M)
1
1β-[4-(2-azidoethyoxy)phenyl]-estra-4,9-diene-3,17-dione,
13 mg, 0.03 mmol) was dissolved in tert-butanol : water (1:1, 4 mL). N-
Dansyl propargylamine, 8, (10 mg, 0.03 mmol), sodium ascorbate
350 µL, 0.035 mmol in a 0.1 mmol/mL solution) and copper (II) sul-
6,
were added to the reaction vial, sealed and the reaction was heated at
90 °C for 18 h. The reaction was cooled and partitioned between ethyl
acetate and water. The organic phase was washed sequentially with
water and brine, dried over magnesium sulfate (anhyd), filtered and
evaporated to dryness. The crude material was purified by flash chro-
matography (step gradient from 1:1 to 4:1 ethyl acetate: hexane) to give
the pure acetylated intermediate (40 mg, 0.052 mmol, 75%).
The acetylated intermediate (20 mg, 0.03 mmol) was dissolved in
methanol (4 mL) and saponified by the addition of 4.2 mg (0.07 mmol)
of potassium hydroxide. Isolation of the product yielded 10a (18 mg,
99%)
(
(
fate, (70 µL, 0.007 mmol in a 0.1 M solution) were added. The reaction
vessel was sealed and heated at 80 °C for 16 hrs. The reaction was
cooled and partitioned between ethyl acetate and water. The organic
phase was washed sequentially with water and brine, dried over mag-
nesium sulfate (anhydrous), filtered and evaporated to dryness. The
crude material was purified by flash chromatography (ethyl acetate :
hexane 9:1) to obtain 14 mg (0.019 mmol, 66% yield) of 11β-(4-(2-(4-
(
3
aminomethyl)-1H-1,2,3- triazol-1-yl)-ethoxy)phenyl) estra-4,9-diene-
Rf = 0.16 (7:3 hexane: ethyl acetate) 1H NMR (400 MHz, CDCl
3
) δ
,17-dione 10a.
8.42 (d, J = 8.5 Hz, 1H), 8.17 (t, J = 7.8 Hz, 2H), 7.44 (t, J = 8.0 Hz,
1H), 7.37 (t, J = 7.9 Hz, 1H), 7.32 (s, 1H), 7.11 (d, J = 7.5 Hz, 1H),
6.98 (d, J = 8.1 Hz, 2H), 6.75 (d, J = 8.4 Hz, 1H), 6.62 (s, 1H), 6.54 (d,
J = 8.2 Hz, 2H), 6.38 (d, J = 7.9 Hz, 1H), 5.74 (s, 1H), 5.29 (s, 1H),
5.11 (s, 1H), 4.43 (s, 2H), 4.15 (d, J = 5.5 Hz, 2H), 4.05 (d, J = 4.3 Hz,
2H), 3.97 (s, 1H), 2.85 (s, 6H), 2.47 (d, J = 14.9 Hz, 2H), 2.30 (d,
Rf = 0.13 (9:1 ethyl acetate: hexane) 1H NMR (400 MHz, CDCl
) δ
3
8
1
4
2
2
.45 (d, J = 8.5 Hz, 1H), 7.85 (s, 1H), 7.08 (d, J = 8.3 Hz, 2H), 6.97 (s,
H), 6.74 (d, J = 8.4 Hz, 2H), 6.34 (d, J = 8.5 Hz, 1H), 5.79 (s, 1H),
.83–4.78 (d, 2H), 4.40 – 4.27 (m, 2H), 3.34 (s, 1H), 2.77–2.67 (m, 1H),
.66 – 2.57 (m, 3H), 2.53 – 2.32 (m, 6H), 2.31–2.20 (m, 2H), 2.19 –
.12 (m, 2H), 1.96 – 1.88 (dd, 1H), 0.95 – 0.80 (m, 3H), 0.51 (s, 3H).
J = 11.2 Hz, 2H), 2.18 – 2.03 (m, 3H), 2.00 – 1.90 (m, 1H), 1.67 – 1.40
1
3
13
C NMR (100 MHz, CDCl
43.72, 137.06, 134.30, 130.12, 130.12, 129.80, 129.59, 129.49,
3
) δ 218.88, 199.22, 155.85, 151.96, 144.56,
3
(m, 3H), 0.43 (s, 3H). C NMR (100 MHz, CDCl ) 154.84, 153.54,
1
151.80, 137.65, 136.76, 134.60, 130.84, 130.62, 129.75, 129.65,
42