56
steroids 7 1 ( 2 0 0 6 ) 54–60
1, 1H, s,), 5.4 (H-9ꢀ&H-10ꢀ, H-12ꢀ&H-13ꢀ and H-15ꢀ&H-16ꢀ, 6H,
m), 2.9 (H-6, 2H, m), 2.81 (H-11ꢀ&H-14ꢀ, 4H, m), 2.5 (H-2ꢀ, 2H, t,
J = 7.4 Hz), 1.4–2.4 (m), 2.0 (H-8ꢀ&H-17ꢀ, 4H, m), 0.97 (H-18ꢀ, 3H,
t, J = 7.6 Hz), 0.91 (H-18, 3H, s). 13C NMR ı: 126.2 (C-1), 118 (C-2,
t), 148.6 (C-3), 122 (C-4, t), 137.2 (C-5), 29 (C-6), 26.4 (C-7), 38.0
(C-8), 44.2 (C-9), 137.9 (C-10), 25.8 (C-11), 31.6 (C-12), 48.0 (C-13),
50.4 (C-14), 21.4 (C-15), 35.8 (C-16, m), 220.8 (C-17), 13.8 (C-18),
172.6 (C-1ꢀ), 34.4 (C-2ꢀ), 25.0 (C-3ꢀ), 29.1–29.8 (C-4ꢀ–C-7ꢀ), 27.2 (C-
8ꢀ), 130.3 (C-9ꢀ), 127.8 (C-10ꢀ), 25.6 (C-11ꢀ), 128.3 (C-12ꢀ), 128.3
(C-13ꢀ) 25.5 (C-14ꢀ), 127.1 (C-15ꢀ), 132.0 (C-16ꢀ), 21.4 (C-17ꢀ), 14.3
(C-18ꢀ).
Purification by preparative TLC gave 9 as white solid 48 mg,
83% yield. Recrystallization from methanol–CHCl3 gave white
crystals of 2,4,16,16,17␣-D4-estradiol-3,17-distearate 9, m.p.
73 ◦C (lit. 71–72 ◦C, unlabeled [21]). IR 1732 and 1760 cm−1
(C O). 1H NMR ı: 7.27 (H-1, 1H, s,), 2.85 (H-6, 2H, m), 2.53 (H-2ꢀ,
2H, t, J = 7.3 Hz), 2.31 (H-2ꢀ, 2H, t, J = 7.5 Hz), 2.1–2.3 (m), 1.1–1.9
(m), 0.88 (H-16ꢀ, 3H, t, J = 6.6 Hz), 0.82 (H-18, 3H, s).
13C NMR ı: 126.2 (C-1), 118.3 (C-2, t), 148.4 (C-3), 121.5 (C-4, t),
137.2 (C-5), 29 (C-6), 27.0 (C-7), 38.2 (C-8), 44.0 (C-9), 138.0 (C-10),
26.0 (C-11), 36.8 (C-12), 42.8 (C-13), 49.8 (C-14), 23.0 (C-15), 27.5
(C-16, m), 81.8 (C-17, t), 12.0 (C-18), 172.6 (C-1ꢀ) 173.9 (C-1ꢀ), 34.4
(C-2ꢀ), 34.6 (C-2ꢀ), 25.0 (C-3ꢀ), 25.1 (C-3ꢀ), 29.1–29.7 (C-4ꢀ–C-15ꢀ),
31.9 (C-16ꢀ), 22.7 (C-17ꢀ), 14.1 (C-18ꢀ).
2.4.
2,4,16,16,17␣-D5-Estradiol 7
2,4,16,16-D2-Estrone 1 180 mg (0.66 mmol) was reduced with
LiAlD4 138 mg (3.3 mmol) in 20 ml dry THF at room temper-
ature for 2.5 h. Saturated aq NH4Cl and water were added
and the mixture was extracted three times with 20 ml diethyl
ether. Combined organic phase was washed three times with
10 ml water, dried with anhydrous Na2SO4, and evaporated.
2,4,16,16,17␣-D5-Estradiol 7 was obtained as white crystals in
95% yield, recrystallized from hexane-acetone.
M.p. 178–179 ◦C (lit. m.p. 174–176 ◦C, 16,16,17␣-D3-Estradiol
[19]). 1H NMR ı: 7.15 (H-1, 1H, s), 4.60 (ArOH, 1H, bs), 2.81 (H-
6, 2H, m), 2.30 (H-11, 1H, m), 2.16 (H-9, 1H, m), 1.94 (H-12, 1H,
m) 1.86 (H-7, 1H, m), 1.68 (H-15, 1H, m), 1.25–1.56 (5H, m), 1.18
(H-14, 1H, m), 0.78 (H-18, 3H, s). 13C NMR ı: 11.1 (C-18), 23.0 (C-
15), 26.3 (C-11), 27.2 (C-7), 29.6 (C-6), 30.6 (C-16, m), 36.6 (C-12),
38.8 (C-8), 43.1 (C-13), 43.3 (C-9), 50.0 (C-14), 81.9 (C-17, m), 113
(C-2, t), 115 (C-4, t), 126.4 (C-1), 132.5 (C-10), 138.2 (C-5), 153.4
(C-3).
2.5.3. 2,4,16,16,17␣-D5-Estradiol-3,17-dioleate 10
Purification by preparative TLC gave 10 as colorless oil 48 mg,
82% yield. IR 1734 and 1761 cm−1 (C O). 1H NMR ı: 7.27 (H-1,
1H, s,), 5.35 (H-9ꢀ&H-10ꢀ, 4H, m), 2.85 (H-6, 2H, m), 2.53 (H-2ꢀ,
2H, t, J = 7.3 Hz), 2.31 (H-2ꢀ, 2H, t, J = 7.5 Hz), 2.1–2.3 (m), 1.1–2.0
(m), 0.88 (H-16ꢀ, 3H, t, J = 6.4 Hz), 0.82 (H-18, 3H, s). 13C NMR ı:
126.2 (C-1), 118.3 (C-2, t), 148.4 (C-3), 121.5 (C-4, t), 137.2 (C-5),
29 (C-6), 27.0 (C-7), 38.2 (C-8), 44.0 (C-9), 138.0 (C-10), 26.0 (C-
11), 36.8 (C-12), 42.8 (C-13), 49.8 (C-14), 23.0 (C-15), 27.5 (C-16,
m), 81.8 (C-17, t), 12.0 (C-18), 172.6 (C-1ꢀ) 173.9 (C-1ꢀ), 34.4 (C-
2ꢀ), 34.6 (C-2ꢀ), 25.0 (C-3ꢀ), 25.1 (C-3ꢀ), 29.1–29.8 (C-4ꢀ–C-7ꢀ), 27.2
(C-8ꢀ), 129.7 (C-9ꢀ), 130.0 (C-10ꢀ), 27.2 (C-11ꢀ), 29.1–29.8 (C-12ꢀ–C-
15ꢀ), 31.9 (C-16ꢀ), 22.7 (C-17ꢀ), 14.1 (C-18ꢀ).
2.5.4. 2,4,16,16,17␣-D5-Estradiol-3,17-dilinoleate 11
Purification by preparative TLC gave 11 as colorless oil 43 mg,
79% yield. IR 1733 and 1760 cm−1 (C O). 1H NMR ı: 7.26 (H-1,
1H, s,), 5.36 (H-9ꢀ&H-10ꢀ and H-12ꢀ&H-13ꢀ, 8H, m), 2.85 (H-6, 2H,
m), 2.77 (H-11ꢀ, 4H, t, J = 6.0 Hz), 2.53 (H-2ꢀ, 2H, t, J = 7.5 Hz), 2.30
(H-2ꢀ, 2H, t, J = 7.5 Hz), 2.03 (H-8ꢀ&H-14ꢀ, 8H, m), 1.3–1.9 (m), 0.89
(H-18ꢀ, 3H, t, J = 6.9 Hz), 0.82 (H-18, 3H, s). 13C NMR ı: 126.2 (C-1),
118.3 (C-2, t), 148.4 (C-3), 121.5 (C-4, t), 137.2 (C-5), 29 (C-6), 27.0
(C-7), 38.2 (C-8), 44.0 (C-9), 138.0 (C-10), 26.0 (C-11), 36.8 (C-12),
42.8 (C-13), 49.8 (C-14), 23.0 (C-15), 27.5 (C-16, m), 81.8 (C-17, t),
12.0 (C-18), 172.5 (C-1ꢀ) 173.9 (C-1ꢀ), 34.4 (C-2ꢀ), 34.6 (C-2ꢀ), 25.0
(C-3ꢀ), 25.1 (C-3ꢀ), 29.1–29.6 (C-4ꢀ–C-7ꢀ), 27.2 (C-8ꢀ), 130.0 (C-9ꢀ),
128.1 (C-10ꢀ), 25.6 (C-11ꢀ), 127.9 (C-12ꢀ), 130.2 (C-13ꢀ) 27.2 (C-14ꢀ),
29 (C-15ꢀ), 31.5 (C-16ꢀ), 22.6 (C-17ꢀ), 14.1 (C-18ꢀ).
2.5.
General procedure for acyl chloride esterification of
2,4,16,16,17␣-D5-estradiol 7
2,4,16,16,17␣-D5-Estradiol 7 (20 mg/0.07 mmol) and DMAP
(17.8 mg/0.14 mmol) were dissolved in 1 ml of pyridine. Palmi-
toyl, steaoryl, oleoyl, or linoleoyl chloride (0.15 mmol, 2.1 eq)
was added. The mixture was microwave irradiated (100 W) for
30 min at 60 ◦C, and then poured into 20 ml water, neutral-
ized with 2 M HCl and extracted three times with 15 ml diethyl
ether. Combined organic phase was washed with 10 ml water,
10 ml 5% NaHCO3, 3 × 10 ml water and dried with Na2SO4 and
evaporated. 2,4,16,16,17-D5-Estradiol-3,17 fatty acid diester
was purified by preparative TLC (hexane-ethyl acetate 2:1).
2.6.
2,4,16,16,17␣-D5-Estradiol-3,17-dilinolenate 12
(20 mg/0.07 mmol) and DMAP
Purification by preparative TLC gave 8 as white solid 52 mg,
96% yield. Recrystallization from methanol–CHCl3 gave white
crystals of 2,4,16,16,17␣-D4-estradiol-3,17-dipalmitate 8, m.p.
66–67 ◦C (lit. 65–67 ◦C, unlabeled [20]). IR 1730 and 1762 cm−1
(C O). 1H NMR ı: 7.27 (H-1, 1H, s,), 2.85 (H-6, 2H, m), 2.53 (H-2ꢀ,
2H, t, J = 7.3 Hz), 2.31 (H-2ꢀ, 2H, t, J = 7.5 Hz), 2.1–2.3 (m), 1.1–1.9
(m), 0.88 (H-16ꢀ, 3H, t, J = 7.0 Hz), 0.82 (H-18, 3H, s). 13C NMR ı:
126.2 (C-1), 118.3 (C-2, t), 148.4 (C-3), 121.5 (C-4, t), 137.2 (C-5),
29 (C-6), 27.0 (C-7), 38.2 (C-8), 44.0 (C-9), 138.0 (C-10), 26.0 (C-11),
36.8 (C-12), 42.8 (C-13), 49.8 (C-14), 23.0 (C-15), 27.5 (C-16, m),
81.8 (C-17, t), 12.0 (C-18), 172.6 (C-1ꢀ) 173.9 (C-1ꢀ), 34.4 (C-2ꢀ), 34.6
(C-2ꢀ), 25.0 (C-3ꢀ), 25.1 (C-3ꢀ), 29.1–29.7 (C-4ꢀ–C-13ꢀ), 31.9 (C-14ꢀ),
22.7 (C-15ꢀ), 14.1 (C-16ꢀ).
2,4,16,16,17␣-D5-Estradiol
7
(17.8 mg/0.14 mmol) were dissolved in 1 ml of pyridine.
Linolenic anhydride (0.15 mmol, 2.1 eq) was added. The mix-
ture was microwave irradiated (100 W) for 60 min at 60 ◦C. After
the reaction, mixture was poured to 20 ml water, neutralized
with 2 M HCl and extracted three times with 15 ml diethyl
ether. Combined organic phase was washed with 10 ml water,
10 ml 5% NaHCO3, 3 × 10 ml water and dried with Na2SO4 and
evaporated. 2,4,16,16,17-D5-Estradiol-3,17-dilinolenate 12 was
obtained as colorless oil 45 mg, 84% yield after purification
by preparative TLC (hexane-ethyl acetate 2:1). IR 1733 and
1760 cm−1 (C O). 1H NMR ı: 7.26 (H-1, 1H, s), 5.36 (H-9ꢀ&H-
10ꢀ, H-12ꢀ&H-13ꢀ, H-15ꢀ&H-16ꢀ, 12H, m), 2.85 (H-6, 2H, m), 2.81
(H-11ꢀ&H-14ꢀ, 8H, t, J = 6.0 Hz), 2.53 (H-2ꢀ, 2H, t, J = 7.5 Hz), 2.30