Y. Yamazaki et al. / Bioorg. Med. Chem. 20 (2012) 4279–4289
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1H), 6.71 (d, J = 2.3 Hz, 1H), 6.66 (dd, J = 8.3, 2.3 Hz, 1H), 3.86 (s,
3H), 3.69 (s, 3H), 1.38 (s, 9H); 13C NMR (150 MHz, DMSO-d6) d
194.1, 163.0, 159.0, 157.4, 156.1, 140.2, 138.4, 134.2, 133.3,
133.2, 131.5, 129.5, 128.6, 127.3, 120.4, 112.9, 105.4, 98.8, 55.6,
55.4, 31.8, 30.4; HRMS (EI): m/z 500.2051 [M+] (Calcd for
115.7, 115.6, 112.8, 104.6, 31.8, 30.4; HRMS (EI): m/z 458.1762
[M+] (Calcd for C26H23FN4O3: 458.1754).
4.2.57. (3Z,6Z)-3-((5-tert-Butyl-1H-imidazol-4-yl)methylene)-6-
(3-(2-chlorobenzoyl)benzylidene)piperazine-2,5-dione (16k)
7% Yield from 15; 1H NMR (600 MHz, DMSO-d6) d 12.50 (br s,
1H), 12.13 (br s, 1H), 10.42 (s, 1H), 7.99 (br s, 1H), 7.85 (s, 1H),
7.83–7.81 (m, 1H), 7.63–7.60 (m, 2H), 7.59–7.58 (m, 2H), 7.53–
7.52 (m, 2H), 6.83 (s, 1H), 6.77 (s, 1H), 1.38 (s, 9H); 13C NMR
(150 MHz, DMSO-d6) d 194.1, 157.4, 156.1, 140.2, 137.7, 135.9,
134.6, 134.2, 134.0, 131.8, 129.9, 129.8, 129.8, 129.2, 129.2,
129.1, 127.7, 127.3, 112.5, 104.7, 31.8, 30.4; HRMS (EI): m/z
474.1453 [M+] (Calcd for C26H23ClN4O3: 474.1458).
C28H28N4O5: 500.2059).
4.2.52. (3Z,6Z)-3-((5-tert-Butyl-1H-imidazol-4-yl)methylene)-6-
(3-(3,4-dimethoxybenzoyl)benzylidene)piperazine-2,5-dione
(16f)
32% Yield from 15; 1H NMR (600 MHz, DMSO-d6) d 12.60 (br s,
1H), 12.05 (br s, 1H), 10.41 (s, 1H), 8.05 (br s, 1H), 7.79 (s, 1H), 7.73
(d, J = 7.7 Hz, 1H), 7.61 (d, J = 7.6 Hz, 1H), 7.57 (dd, J = 7.6, 7.6 Hz,
1H), 7.42 (d, J = 2.0 Hz, 1H), 7.39 (dd, J = 8.3, 2.1 Hz, 1H), 7.11 (d,
J = 8.4 Hz, 1H), 6.82 (s, 1H), 6.80 (s, 1H), 3.87 (s, 3H), 3.83 (s, 3H),
1.38 (s, 9H); 13C NMR (150 MHz, DMSO-d6) d 194.1, 157.4, 156.3,
152.9, 148.6, 140.1, 138.1, 134.2, 133.3, 132.7, 129.8, 129.1,
128.6, 128.5, 127.4, 125.2, 113.1, 111.6, 110.6, 104.4, 55.7, 55.5,
31.8, 30.3; HRMS (EI): m/z 500.2066 [M+] (Calcd for C28H28N4O5:
500.2059).
4.2.58. (3Z,6Z)-3-((5-tert-Butyl-1H-imidazol-4-yl)methylene)-6-
(3-(3-chlorobenzoyl)benzylidene)piperazine-2,5-dione (16l)
49% Yield from 15; 1H NMR (600 MHz, DMSO-d6) d 12.55 (br s,
1H), 12.10 (br s, 1H), 10.40 (s, 1H), 8.02 (br s, 1H), 7.84 (s, 1H),
7.79–7.76 (m, 4H), 7.66–7.63 (m, 1H), 7.62–7.59 (m, 2H), 6.82 (s,
2H), 1.38 (s, 9H); 13C NMR (150 MHz, DMSO-d6) d 194.2, 157.4,
156.2, 140.1, 138.8, 136.7, 134.2, 133.6, 133.5, 133.4, 132.5,
130.5, 130.1, 128.9, 128.8, 128.8, 128.4, 127.6, 112.8, 104.6, 31.8,
30.4; HRMS (EI): m/z 474.1455 [M+] (Calcd for C26H23ClN4O3:
474.1458).
4.2.53. (3Z,6Z)-3-((5-tert-Butyl-1H-imidazol-4-yl)methylene)-6-
(3-(3,4,5-trimethoxybenzoyl)benzylidene)piperazine-2,5-dione
(16g)
32% Yield from 15; 1H NMR (600 MHz, DMSO-d6) d 12.50 (br s,
1H), 12.13 (br s, 1H), 10.37 (s, 1H), 7.99 (br s, 1H), 7.83 (s, 1H), 7.75
(d, J = 7.7 Hz, 1H), 7.67 (d, J = 7.6 Hz, 1H), 7.58 (dd, J = 7.7, 7.7 Hz,
1H), 7.08 (s, 2H), 6.82 (s, 2H), 3.82 (s, 6H), 3.78 (s, 3H), 1.38 (s,
9H); 13C NMR (150 MHz, DMSO-d6) d 194.5, 157.5, 156.2, 152.6,
141.5, 140.2, 137.5, 134.2, 133.5, 133.1, 131.9, 129.9, 128.8,
128.6, 127.5, 112.9, 107.3, 104.6, 60.1, 56.0, 31.8, 30.4; HRMS
(EI): m/z 530.2158 [M+] (Calcd for C29H30N4O6: 530.2165).
4.2.59. (3Z,6Z)-3-((5-tert-Butyl-1H-imidazol-4-yl)methylene)-6-
(3-(4-chlorobenzoyl)benzylidene)piperazine-2,5-dione (16m)
13% Yield from 15; 1H NMR (600 MHz, DMSO-d6) d 12.54 (br s,
1H), 12.11 (br s, 1H), 10.40 (s, 1H), 8.01 (br s, 1H), 7.84 (ddd, J = 8.5,
2.0, 2.0 Hz, 2H), 7.83 (s, 1H), 7.76 (d, J = 7.6 Hz, 1H), 7.65 (d,
J = 8.5 Hz, 3H), 7.59 (dd, J = 7.7, 7.7 Hz, 1H), 6.82 (s, 1H), 6.82 (s,
1H), 1.38 (s, 9H); 13C NMR (150 MHz, DMSO-d6) d 194.4, 157.5,
156.2, 140.2, 137.7, 136.9, 135.4, 134.2, 133.5, 133.4, 131.6,
130.1, 128.8, 128.7, 128.7, 127.6, 112.8, 104.7, 31.8, 30.4; HRMS
(EI): m/z 474.1450 [M+] (Calcd for C26H23ClN4O3: 474.1458).
4.2.54. (3Z,6Z)-3-((5-tert-Butyl-1H-imidazol-4-yl)methylene)-6-
(3-(2-fluorobenzoyl)benzylidene)piperazine-2,5-dione (16h)
20% Yield from 15; 1H NMR (600 MHz, DMSO-d6) d 12.55 (br s,
1H), 12.10 (br s, 1H), 10.43 (s, 1H), 8.02 (br s, 1H), 7.87 (s, 1H), 7.80
(d, J = 7.6 Hz, 1H), 7.71–7.67 (m, 1H), 7.65 (d, J = 7.6 Hz, 1H), 7.63
(ddd, J = 7.6, 7.6, 1.7 Hz, 1H), 7.59 (dd, J = 7.7, 7.7 Hz, 1H), 7.42–
7.39 (m, 2H), 6.82 (s, 1H), 6.79 (s, 1H), 1.38 (s, 9H); 13C NMR
(150 MHz, DMSO-d6) d 192.3, 160.1, 158.5, 157.4, 156.2, 140.2,
136.9, 134.3, 134.2, 133.7, 133.7, 130.6, 130.6, 129.8, 129.0,
128.9, 127.6, 126.2, 126.1, 124.7, 124.7, 116.4, 116.3, 112.6,
104.6, 31.8, 30.4; HRMS (EI): m/z 458.1759 [M+] (Calcd for
4.2.60. (3Z,6Z)-3-(3-(4-Bromobenzoyl)benzylidene)-6-((5-tert-
butyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (16n)
44% Yield from 15; 1H NMR (600 MHz, DMSO-d6) d 12.51 (br s,
1H), 12.13 (br s, 1H), 10.39 (s, 1H), 7.99 (br s, 1H), 7.83 (s, 1H), 7.80
(ddd, J = 8.6, 2.0, 2.0 Hz, 2H), 7.76 (ddd, J = 8.6, 1.9, 1.9 Hz, 3H), 7.65
(ddd, J = 7.7, 1.4, 1.4 Hz, 1H), 7.59 (dd, J = 7.6, 7.6 Hz, 1H), 6.83 (s,
1H), 6.81 (s, 1H), 1.38 (s, 9H); 13C NMR (150 MHz, DMSO-d6) d
194.5, 157.5, 156.2, 140.2, 136.9, 135.8, 134.2, 133.5, 133.4,
131.7, 131.6, 130.1, 128.8, 128.7, 127.6, 126.8, 112.8, 104.6, 31.8,
30.4; HRMS (EI): m/z 518.0945 [M+] (Calcd for C26H23Br N4O3:
518.0953).
C26H23FN4O3: 458.1754).
4.2.55. (3Z,6Z)-3-((5-tert-Butyl-1H-imidazol-4-yl)methylene)-6-
(3-(3-fluorobenzoyl)benzylidene)piperazine-2,5-dione (16i)
20% Yield from 15; 1H NMR (600 MHz, DMSO-d6) d 12.52 (br s,
1H), 12.12 (br s, 1H), 10.39 (s, 1H), 8.00 (br s, 1H), 7.84 (s, 1H), 7.77
(d, J = 7.7 Hz, 1H), 7.67–7.63 (m, 3H), 7.60 (dd, J = 7.8, 7.8 Hz, 1H),
7.58–7.55 (m, 2H), 6.82 (s, 2H), 1.38 (s, 9H); 13C NMR (150 MHz,
DMSO-d6) d 194.2, 162.6, 161.0, 157.5, 156.1, 140.3, 139.1, 139.0,
136.8, 134.2, 133.6, 133.5, 130.8, 130.7, 130.1, 128.8, 127.6,
126.1, 126.0, 119.7, 119.6, 116.0, 115.8, 112.6, 104.9, 31.8, 30.5;
HRMS (EI): m/z 458.1747 [M+] (Calcd for C26H23FN4O3: 458.1754).
4.2.61. (3Z,6Z)-3-((5-tert-Butyl-1H-imidazol-4-yl)methylene)-6-
(3-(4-(trifluoromethyl)benzoyl)benzylidene)piperazine-2,5-
dione (16o)
25% Yield from 15; 1H NMR (600 MHz, DMSO-d6) d 12.51 (br s,
1H), 12.12 (br s, 1H), 10.40 (s, 1H), 8.00 (d, J = 8.0 Hz, 2H), 7.95 (s,
1H), 7.95 (d, J = 8.2 Hz, 2H), 7.87 (s, 1H), 7.79 (d, J = 7.9 Hz, 1H),
7.68 (d, J = 7.7 Hz, 1H), 7.61 (dd, J = 7.7, 7.7 Hz, 1H), 6.83 (s, 1H),
6.82 (s, 1H), 1.38 (s, 9H); 13C NMR (150 MHz, DMSO-d6) d 194.6,
157.5, 156.1, 140.4, 140.2, 136.4, 134.2, 133.8, 133.6, 132.1,
131.9, 130.4, 130.3, 128.9, 127.7, 125.5, 125.5, 124.6, 122.8,
122.7, 104.7, 31.8, 30.4; HRMS (EI): m/z 508.1727 [M+] (Calcd for
4.2.56. (3Z,6Z)-3-((5-tert-Butyl-1H-imidazol-4-yl)methylene)-6-
(3-(4-fluorobenzoyl)benzylidene)piperazine-2,5-dione (16j)
14% Yield from 15; 1H NMR (600 MHz, DMSO-d6) d 12.52 (br s,
1H), 12.12 (br s, 1H), 10.39 (s, 1H), 8.00 (br s, 1H), 7.93–7.90 (m,
2H), 7.82 (s, 1H), 7.76 (d, J = 7.7 Hz, 1H), 7.64 (ddd, J = 7.7, 1.3,
1.3 Hz, 1H), 7.59 (dd, J = 7.6, 7.6 Hz, 1H), 7.42–7.39 (m, 2H), 6.82
(s, 1H), 6.81 (s, 1H), 1.38 (s, 9H); 13C NMR (150 MHz, DMSO-d6) d
194.1, 165.5, 163.8, 157.5, 156.2, 140.2, 137.2, 134.2, 133.4,
133.4, 133.3, 133.3, 132.7, 132.7, 130.0, 128.8, 128.7, 127.5,
C27H23F 3N4O3: 508.1722).
4.3. Biological evaluation
4.3.1. Tubulin binding assay
Fluorescence spectra were measured at 37 °C as described pre-
viously.21,24 Porcine tubulin (0.5
lM) in MES buffer (0.1 M MES,