R. P. Tanpure et al. / Bioorg. Med. Chem. 17 (2009) 6993–7001
6999
Melting point: 23 (150–151 °C), 24 (126–127 °C).
23 E-isomer: 1H NMR (DMSO-d6, 360 MHz): d 8.75 (s, 1H, OH),
6.99–7.10 (m, 3H, PhH), 6.91–6.94 (m, 2H, PhH), 6.72 (d, 1H,
J = 8.6 Hz, ArH), 6.56 (d, 1H, J = 2.2 Hz, ArH), 6.50 (dd, 1H,
J = 8.3 Hz, J = 2.2 Hz, ArH), 6.45 (s, 2H, ArH), 3.74 (s, 6H, OCH3),
3.70 (s, 3H, OCH3), 3.68 (s, 3H, OCH3), 2.33 (q, 2H, J = 7.2 Hz
CH2CH3), 0.88 (t, 3H, J = 7.2 Hz, CH2CH3).
2H, ArH), 3.77 (s, 3H, OCH3), 3.57 (s, 3H, OCH3), 3.51 (s, 6H,
OCH3), 2.11 (s, 3H, CH3).
13C NMR (CDCl3, 125 MHz): d 152.5, 145.3, 145.2, 143.5, 139.2,
139.0, 136.9, 136.4, 134.9, 130.4, 127.5, 125.8, 121.8, 116.3, 110.2,
106.9, 60.9, 55.92, 55.9, 22.8.
Anal. Calcd for C25H26O5 27 E: C, 73.87, H, 6.45. Found: C, 73.78,
H, 6.36.
13C NMR (DMSO-d6, 90 MHz): d 152.7, 146.0, 145.7, 142.5,
141.2, 138.9, 137.4, 136.1, 133.9, 129.9, 127.5, 125.7, 120.4,
116.6, 111.4, 106.2, 60.0, 55.8, 55.3, 28.9, 13.4.
27 E-isomer: HRMS (EI+) m/z: 406.1769 (Calcd for C25H26O5—
406.1780).
28 Z-isomer: 1H NMR (DMSO-d6, 500 MHz): d 8.68 (s, 1H, OH),
7.36 (t, 2H, J = 7.4 Hz, PhH), 7.25 (t, 1H, J = 7.4 Hz, PhH), 7.19 (m,
2H, PhH), 6.65 (d, 1H, J = 8.4 Hz, ArH), 6.43 (s, 2H, ArH), 6.34 (d,
1H, J = 2.1 Hz, ArH), 6.29 (dd, 2H, J = 8.3 Hz, J = 2.2 Hz, ArH), 3.65
(s, 3H, OCH3), 3.60 (s, 3H, OCH3), 3.56 (s, 6H, OCH3), 2.03 (s, 3H,
CH3).
23 E-isomer: HRMS (EI+) m/z: 420.1940 (Calcd for C26H28O5—
420.1937).
24 Z-isomer: 1H NMR (DMSO-d6, 360 MHz): d 8.79 (s, 1H, OH),
7.37–7.42 (m, 2H, PhH), 7.28–7.32 (m, 1H, PhH), 7.23 (d, 2H,
J = 6.8 Hz, PhH), 6.79 (d, 1H, J = 7.9 Hz, ArH), 6.57 (d, 1H,
J = 1.8 Hz, ArH), 6.56 (dd, 1H, J = 7.9 Hz, J = 1.8 Hz, ArH), 6.11 (s,
2H, ArH), 3.72 (s, 3H, OCH3), 3.56 (s, 3H, OCH3), 3.44 (s, 6H,
OCH3), 2.30 (q, 2H, J = 7.2 Hz, CH2CH3), 0.87 (t, 3H, J = 7.2 Hz,
CH2CH3).
13C NMR (CDCl3, 125 MHz): d 152.5, 144.8, 144.7, 143.5, 139.4,
139.0, 136.8, 136.4, 134.7, 129.8, 128.1, 126.5, 122.5, 116.7, 109.8,
106.7, 60.9, 56.0, 55.8, 22.9.
Anal. Calcd for C25H26O5 28 Z: C, 73.87, H, 6.45. Found: C, 73.44,
H, 6.44.
13C NMR (DMSO-d6, 90 MHz): d 151.7, 146.1, 145.9, 142.7,
141.3, 138.0, 137.3, 135.4, 134.4, 128.9, 128.2, 126.6, 119.8,
116.4, 111.7, 107.9, 59.8, 55.45, 55.36, 28.6, 13.3.
Anal. Calcd for C26H28O5 24 Z: C, 74.26, H, 6.71. Found: C, 74.09,
H, 6.79.
28 Z-isomer: HRMS (EI+) m/z: 406.1763 (Calcd for C25H26O5—
406.1780).
4.1.9.5. 1-{30-Hydroxy-40-methoxyphenyl}-1-phenyl-2-(300,400,500-
trimethoxyphenyl)-but-1-ene (E = 29, Z = 30). Purification by
flash chromatography (silica gel, 5:95, EtOAc/hexanes) afforded
29 (0.130 g, 0.309 mmol, 19%, E-isomer) and 30 (0.262 g,
0.623 mmol, 38%, Z-isomer) as white solids.
24 Z-isomer: HRMS (EI+) m/z: 420.1939 (Calcd for C26H28O5—
420.1937).
4.1.9.3. 2-{30-Hydroxy-40-methoxyphenyl}-1,2-bis-phenyl-1-
(300,400,500-trimethoxyphenyl)-ethylene (E = 25, Z = 26). Purifica-
tion by flash chromatography (silica gel, 5:95, EtOAc/hexanes)
afforded 25 (0.14 g, 0.30 mmol, 65%, E-isomer) and 26 (0.04 g,
0.09 mmol, 20%, Z-isomer) as white solids.
Melting point: 29 (140–142 °C), 30 (165–166 °C).
29 E-isomer: 1H NMR (DMSO-d6, 500 MHz): d 8.95 (s, 1H, OH),
7.07 (t, 2H, J = 7.3 Hz, PhH), 7.01 (t, 1H, J = 7.3 Hz, PhH), 6.90 (d,
1H, J = 8.3 Hz, ArH), 6.86 (d, 2H, J = 7.1 Hz, ArH), 6.63 (dd, 1H,
J = 8.2 Hz, J = 2.0 Hz, ArH), 6.59 (d, 1H, J = 2.0 Hz, ArH), 6.34 (s,
2H, ArH), 3.77 (s, 3H, OCH3), 3.58 (s, 3H, OCH3), 3.52 (s, 6H,
OCH3), 2.48 (q, 2H, J = 7.4 Hz, CH2CH3), 0.92 (t, 3H, J = 7.4 Hz,
CH2CH3).
Melting point: 25 (198–199 °C), 26 (184–186 °C).
25 E-isomer: 1H NMR (CDCl3, 500 MHz): d 7.02–7.16 (m, 10H,
PhH), 6.59 (d, 1H, J = 2.0 Hz, ArH), 6.57 (d, 1H, J = 8.4 Hz, ArH),
6.50 (dd, 1H, J = 8.4 Hz, J = 2.0 Hz, ArH), 6.18 (s, 2H, ArH), 5.37 (s,
1H, OH), 3.81 (s, 3H, OCH3), 3.79 (s, 3H, OCH3), 3.49 (s, 6H, OCH3).
13C NMR (DMSO-d6, 90 MHz): d 151.9, 146.2, 145.6, 144.0,
143.0, 140.2, 139.2, 138.6, 135.9, 135.6, 130.6, 130.2, 127.7,
126.4, 126.2, 121.9, 117.9, 111.2, 108.5, 59.9, 55.4, 55.2.
Anal. Calcd for C30H28O5 25 E: C, 76.90, H, 6.02. Found: C, 76.55,
H, 6.05.
13C NMR (CDCl3, 125 MHz): d 152.5, 145.3, 145.2, 143.5, 141.6,
138.4, 137.4, 136.9, 136.4, 130.3, 127.4, 125.7, 121.1, 115.8, 110.2,
107.2, 60.9, 56.0, 55.9, 28.5, 13.8.
Anal. Calcd for C26H28O5 29 E: C, 74.26, H, 6.71, O, 19.02. Found:
C, 74.14, H, 6.53, O, 18.85.
29 E-isomer: HRMS (EI+) m/z 420.1933 (Calcd for C26H28O5—
420.1937).
25 E-isomer: HRMS (EI+) m/z 468.1928 (Calcd for C30H28O5—
468.1937).
30 Z-isomer: 1H NMR (DMSO-d6, 500 MHz): d 8.65 (s, 1H, OH),
7.36 (t, 2H, J = 7.5 Hz, PhH), 7.26 (m, 1H, J = 7.5 Hz, PhH), 7.18 (d,
2H, J = 7.0 Hz, ArH), 6.62 (d, 1H, J = 8.4 Hz, ArH), 6.40 (s, 2H, ArH),
6.32 (d, 1H, J = 2.1 Hz, ArH), 6.27 (dd, 1H, J = 8.3 Hz, J = 2.1 Hz,
ArH), 3.63 (s, 3H, OCH3), 3.61 (s, 3H, OCH3), 3.57 (s, 6H, OCH3),
2.37 (q, 2H, J = 7.4 Hz, CH2CH3), 0.89 (t, 3H, J = 7.4 Hz, CH2CH3).
13C NMR (CDCl3, 125 MHz): d 152.6, 144.66, 144.65, 143.5,
141.3, 138.4, 137.6, 136.8, 136.4, 129.3, 128.1, 126.5, 122.4,
116.6, 109.7, 107.0, 60.9, 56.0, 55.8, 28.6, 13.7.
26 Z-isomer: 1H NMR (CDCl3, 500 MHz): d 7.00–7.12 (m, 10H,
PhH), 6.62 (d, 1H, J = 2.0 Hz, ArH), 6.61 (d, 1H, J = 8.2 Hz, ArH),
6.54 (dd, 1H, J = 8.3 Hz, 2.1 Hz, ArH), 6.25 (s, 2H, ArH), 5.40 (s,
1H, OH), 3.83 (s, 3H, OCH3), 3.81 (s, 3H, OCH3), 3.55 (s, 6H, OCH3).
13C NMR (DMSO-d6, 90 MHz): d 151.9, 146.3, 145.8, 143.4,
142.8, 140.2, 139.2, 138.6, 136.2, 136.1, 130.7, 130.5, 127.61,
127.56, 126.4, 126.2, 121.4, 117.6, 111.6, 108.4, 59.9, 55.5.
Anal. Calcd for C30H28O5 26 Z: C, 76.90, H, 6.02. Found: C, 76.38,
H, 6.07.
Anal. Calcd for C26H28O5 30 Z: C, 74.26, H, 6.71. Found: C, 74.34,
H, 6.67.
26 Z-isomer: HRMS (EI+) m/z 468.1934 (Calcd for C30H28O5—
468.1937).
30 Z-isomer: HRMS (EI+) m/z 420.1936 (Calcd for C26H28O5—
420.1937).
4.1.9.4. 1-{30-Hydroxy-40-methoxyphenyl}-1-phenyl-2-(300,400,500-
trimethoxyphenyl)-prop-1-ene (E = 27, Z = 28). Purification by
flash chromatography (silica gel, 5:95, EtOAc/hexanes) afforded
27 (0.116 g, 0.285 mmol 22%, E-isomer) and 28 (0.200 g,
0.492 mmol, 37%, Z-isomer) as white solids.
4.2. Biology
4.2.1. Effects on tubulin polymerization
Bovine brain tubulin was purified as described previously.33 To
evaluate the effect of the compounds on tubulin assembly in vitro,
Melting point: 27 (134–135 °C), 28 (167–168 °C).
27 E-isomer: 1H NMR (DMSO-d6, 500 MHz): d 8.93 (s, 1H, OH),
7.09 (t, 2H, J = 7.0 Hz, PhH), 7.03 (t, 1H, J = 7.2 Hz, PhH), 6.90 (d,
1H, J = 8.2 Hz, ArH), 6.88 (d, 1H, J = 7.3 Hz, PhH), 6.63 (dd, 1H,
J = 8.2 Hz, J = 2.0 Hz, ArH), 6.59 (d, 1H, J = 2.0 Hz, ArH), 6.37 (s,
varying concentrations were preincubated with 10
lM tubulin
(1.0 g/mL) in glutamate buffer at 30 °C and then cooled to 0 °C.
l
After the addition of GTP, the mixtures were transferred to 0 °C
cuvettes in a recording spectrophotometer and warmed to 30 °C.