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126.44 (C–14), 124.08 (C–11), 123.49 (C–12), 20.76 (Me–18), 23.62 and 23.66 (Me–17 and
Me–16), 24.77 (Me–19), 33.15 (C–15), 46.93 (C–5), 29.25 (C–25, C-26, C-27), 18.59 (C–2), 19.46
(C–6), 30.11 (C–7), 36.18 (C–28, C-29, C-30), 42.13 (C–22, C-23, C-24), 55.77 (C–4), 37.89 (C–1),
37.50 and 37.46 (C–10 and C-3), 50.04 (C-21). Found, m/z: 448.3445 [M]+. C30H44ON2.
Calculated, m/z: 448.3448.
N-2-adamantyl-N’-[(1R,4aS,10aR)-7-isopropyl-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-
octahydrophenanthren-1-yl]urea
ν
(
10). Yield 80%, white powder. M.p. 233 oC. IR (KBr)
1
max 3357, 2912, 1623, 1556 cm−1. H NMR (400MHz, CDCl3,
δ, ppm, J/Hz): 6.78 (1H, s,
H–14), 6.89 (1H, d, J11, 12 = 8.2, H–12), 7.07 (1H, d, J11, 12 = 8.2, H–11), 1.20 (3H, s, Me–18),
1.11 (6H, d, J16, 15 = 6.9, Me–16 and Me–17), 1.10 (3H, s, Me–19), 2.73 (1H, sept, J15, 16 = 6.9,
H–15), 2.74–2.87 (2H, m, 2H–7), 2.15 (1H, d, 2J = 12.3, H–1e), 3.64 (1H, s, H–22), 1.29–1.41
(1H, m, H–1a), 1.42–1.54 (2H, m, H–24, H–29), 1.94–2.08 (2H, m, H–23, H–28), 1.78–1.93 (2H,
m, H–24, H–29), 1.55–1.65 (4H, m, H–26, H–27, 2H–30), 1.65–1.78 (11H, m, 2H–2, 2H–3,
2H–6, H–5, H–21, H–25, H–23, H–28). 13C NMR (100MHz, DMSO-d6,
δ, ppm, J/Hz): 157.25
(C–20), 147.74 (C–9), 145.54 (C–13), 134.99 (C-8), 127.00 (C–14), 124.69 (C–11), 124.16 (C–12),
21.91 (Me–18), 24.14 and 24.48 (Me–17 and Me–16), 25.25 (Me–19), 46.53 (C–5), 30.43 (C–7),
18.89 (C–2), 20.06 (C–6), 55.76 (C–4), 53.25 (C-22), 27.46 and 27.52 (C-21, C-25), 31.80 and
31.92 (C-24, C-29), 32.86 and 32.94 (C-26, C-27), 33.43 (C–15), 37.57 (C-3), 37.73 (C–10), 38.32
and 38.36 (C–1, C-30), 37.97 and 37.92 (C-23, C-28). Found, m/z: 448.3449 [M]+. C30H44ON2.
Calculated, m/z: 448.3448.
N-1-adamantyl-N’-[(1R,4aR,10aR)-7-isopropyl-1,4a-dimethyl-1,2,3,4,4a,4b,5,6,10,10a-
decahydrophenanthren-1-yl]urea
ν
(
11). Yield 90%, white powder. M.p. 224 oC. IR (KBr)
1
max 3346, 2906, 1633, 1560 cm−1. H NMR (400MHz, CDCl3 + CD3OD,
δ
, ppm, J/Hz):
5.65 (1H, s, H–14), 5.30 (1H, s, H–7), 0.88 and 0.89 (3H both, d, J16, 15=6.9, Me–16 and Me–17),
0.67 (3H, s, Me–18), 1.12 (3H, s, Me–19), 2.10 (1H, sept, J15, 16 = 6.9, H–15), 0.95–1.10 (2H, m,
H–1a, H–11a), 1.32–1.48 (2H, m, H–2a, H–2e), 1.63–1.73 (2H, m, H–3a, H–11e), 1.48–1.57
(6H, m, 2H–29, 2H–30, 2H–28), 1.72–1.87 (9H, m, 2H–22, 2H–23, 2H–24, H–1e, H–3e, H–9),
1.87–2.05 (8H, m, H-25, H-26, H–27, H–5, H–6a, H–6e, 2H–12). 13C NMR (100MHz, CDCl3
+ CD3OD, δ, ppm): 156.99 (C–20), 145.02 (C–13), 135.33 (C-8), 122.19 (C–14), 120.64 (C–7),
29.32 (C–25, C-26, C-27), 36.22 (C–28, C-29, C-30), 42.21 (C–22, C–23, C–24), 21.10 and
20.53 (Me–17 and Me–16), 21.38 (Me–18), 13.61 (Me–19), 34.65 (C–15), 50.65 (C–9), 47.00
(C–5), 19.15 (C–2), 27.20 (C–12), 35.44 (C–10), 38.14 (C–1), 37.92 (C–3), 22.49 (C–11), 23.50
(C–6), 55.45 (C–4), 50.17 (C–21). Found, m/z: 450.3600 [M]+. C30H46ON2. Calculated,
m/z: 450.3605.
N-2-adamantyl-N’-[(1R,4aR,10aR)-7-isopropyl-1,4a-dimethyl-1,2,3,4,4a,4b,5,6,10,10a-
decahydrophenanthren-1-yl]urea
ν
(
12). Yield 80%, white powder. M.p. 213 oC. IR (KBr)
1
max 3395, 2908, 1629, 1556 cm−1. H NMR (400MHz, CDCl3 + CD3OD,
δ
, ppm, J/Hz):
5.62 (1H, s, H–14), 5.27 (1H, s, H–7), 0.88 and 0.87 (3H both, d, J16, 15 = 6.9, Me–16 and
Me–17), 0.67 (3H, s, Me–18), 1.13 (3H, s, Me–19), 2.08 (1H, sept, J15, 16 = 6.9, H–15), 0.93–1.10
(2H, m, H–1a, H–11a), 1.32–1.48 (4H, m, H–2a, H–2e, H–24, H–29), 1.54–1.61 (2H, m, 2H–30),
1.89–2.05 (5H, m, H–5, H–6a, H–6e, 2H–12), 1.73–1.89 (3H, m, H–1e, H–3e, H–9), 1.60–1.75
(10H, m, H–3a, H–11e, H–26, H–27, H–21, H–25, 2H–23, 2H–28, H–24, H–29), 3.59 (1H, s,
H–22). 13C NMR (100MHz, CDCl3 + CD3OD,
δ, ppm): 157.29 (C–20), 144.82 (C–13), 135.25
(C–8), 122.11 (C–14), 120.52 (C–7), 13.53 (Me–19), 21.03 and 20.47 (Me–17 and Me–16), 21.28
(Me–18), 22.42 (C–11), 23.40 (C–6), 27.13 (C–12), 27.05 and 26.90 (C–21, C-25), 31.37 (C–24,
C-29), 32.38 and 32.40 (C–26, C–27), 34.55 (C–15), 35.35 (C–10), 37.05 and 37.03 (C–23, C–28),
38.10 (C–1), 37.92 (C–3), 37.35 (C–30), 46.87 (C–5), 50.68 (C–9), 55.33 (C–4), 53.10 (C–22).
Found, m/z: 450.3604 [M]+. C30H46ON2. Calculated, m/z: 450.3605.
3.2. TDP1 Assay
The recombinant TDP1 was purified to homogeneity by chromatography on Ni-
chelating resin and phosphocellulose P11 as previously described [45], using plasmid pET
16B-TDP1, kindly provided by Dr. K.W. Caldecott (University of Sussex, United Kingdom).
The TDP1 activity measurements were carried out as described [13]. Briefly, TDP1-biosensor