S. Broch et al. / European Journal of Medicinal Chemistry 45 (2010) 1657–1662
1661
IR (KBr): 3502; 3390; 1620 cmꢀ1
.
J2 ¼ 7.0 Hz, J3 ¼ 1.0 Hz), 7.70 (1H, ddd, J1 ¼ 8.5 Hz, J2 ¼ 7.0 Hz,
J3 ¼ 1.5 Hz), 7.79 (1H, dd, J1 ¼ 8.5 Hz, J2 ¼ 2.0 Hz), 7.80 (1H, dd,
J1 ¼ 8.5 Hz, J2 ¼ 1.5 Hz), 7.81–7.84 (1H, m), 7.97 (1H, d, J ¼ 8.5 Hz),
7.99 (1H, d, J ¼ 8.5 Hz), 8.05 (1H, d, J ¼ 8.5 Hz), 8.05 (1H, d,
J ¼ 1.5 Hz), 8.08 (1H, d, J ¼ 1.5 Hz), 8.28 (1H, d, J ¼ 9.0 Hz), 8.48 (1H,
s), 8.50 (1H, s).
HRMS (ESþ) calcd for C20H18BN2O4 [M þ H]þ 361.1360, found
361.1355.
1H NMR (400 MHz, DMSO-d6): 4.02 (3H, s), 4.06 (3H, s), 7.48 (1H,
ddd, J1 ¼8.0 Hz, J2 ¼ 7.0 Hz, J3 ¼ 1.0 Hz), 7.68–7.74 (2H, m), 7.85 (1H, d,
J ¼ 8.5 Hz), 7.96–8.02(3H, m), 8.13 (2H, s, OH), 8.45(1H, s), 8.46(1H, s).
13C NMR (100 MHz, DMSO-d6): 53.3, 53.7 (CH3), 124.5, 125.5,
126.4, 126.8, 127.7, 128.1, 130.0, 139.0, 145.2 (CHarom), 124.0, 125.2,
125.3, 138.0, 145.3, 146.5, 159.1, 164.6 (Carom). The signal of the
carbon bearing the boron atom was not visible on the spectrum.
13C NMR (100 MHz, DMSO-d6): 14.4 (2C), 14.5 (CH3), 61.3, 61.8
(2C) (CH2), 113.4, 124.4, 125.5, 125.8, 126.3, 126.6, 127.0, 127.3, 127.6,
128.1, 129.9, 138.6, 138.9 (2C) (CHarom), 124.1, 124.5, 125.2 (2C),
125.4, 137.7, 137.9, 145.3, 145.4, 145.9, 158.7, 159.0, 161.9 (Carom).
4.1.15. 2,20-Diethoxy-3,70-biquinoline-30-boronic acid 13
Following the general procedure described for 3 and 4 and using
diisopropylamine (0.50 mL, 3.54 mmol) in THF (3.5 mL), n-Buli
(2.5 M in hexane, 1.48 mL, 3.70 mmol), 11 (405 mg, 1.18 mmol) and
triisopropylborate (0.82 mL, 3.55 mmol) in THF (8 mL). The solid
was washed with minimum ether to give 13 (377 mg, 0.97 mmol,
83%) as a white solid (mp 210 ꢁC).
4.1.18. 200-butoxy-2,20-dimethoxy-3,70:30,700-terquinoline 16
Following the typical procedure described for 10, 11 and using 9
(360 mg, 1.28 mmol), THF (7 mL), PdCl2(PPh3)2 (45.1 mg,
64.3 mmol), Na2CO3 2 M (3.2 mL, 6.4 mmol),12 (602 mg,1.67 mmol).
Microwave irradiation (65 ꢁC, 50 W, Patm, 30 min). Purification by
chromatography (cyclohexane-EtOAc, 94:6) provided 16 (507 mg,
0.98 mmol, 77%) as a white solid (mp 113 ꢁC).
IR (KBr): 3527; 3413; 1620; 1609 cmꢀ1
.
IR (KBr): 1615 cmꢀ1
.
HRMS (ESþ) calcd for C22H22BN2O4 [M þ H]þ 389.1673, found
HRMS (ESþ) calcd for C33H30N3O3 [M þ H]þ 516.2287, found
389.1685.
516.2300.
1H NMR (400 MHz, DMSO-d6): 1.37 (3H, t, J ¼ 7.0 Hz), 1.42 (3H, t,
J ¼ 7.0 Hz), 4.51 (2H, q, J ¼ 7.0 Hz), 4.56 (2H, q, J ¼ 7.0 Hz), 7.47 (1H,
ddd, J1 ¼ 8.0 Hz, J2 ¼ 7.0 Hz, J3 ¼ 1.0 Hz), 7.69 (1H, ddd, J1 ¼ 8.5 Hz,
J2 ¼ 7.0 Hz, J3 ¼ 1.5 Hz), 7.74 (1H, dd, J1 ¼ 8.5 Hz, J2 ¼ 1.5 Hz), 7.81
(1H, d, J ¼ 8.5 Hz), 7.94–8.00 (3H, m), 8.07 (2H, s, OH), 8.45 (2H, s).
13C NMR (100 MHz, DMSO-d6): 14.6 (2CH3), 61.7, 62.0 (CH2),
124.6, 125.6, 126.5, 126.9, 127.8, 128.3, 130.2, 139.1, 145.4 (CHarom),
124.1, 125.4 (2C), 138.4, 145.6, 146.8, 158.9, 164.4 (Carom). The signal of
the carbon bearing the boron atom was not visible on the spectrum.
1H NMR (400 MHz, DMSO-d6): 0.96 (3H, t, J ¼ 7.5 Hz), 1.47 (2H, sx,
J ¼ 7.5 Hz), 1.73–1.82 (2H, m), 4.07 (3H, s), 4.08 (3H, s), 4.44 (2H, t,
J ¼ 6.5 Hz), 7.04 (1H, d, J ¼ 9.0 Hz), 7.49 (1H, ddd, J1 ¼ 8.0 Hz,
J2 ¼ 7.0 Hz, J3 ¼ 1.0 Hz), 7.71 (1H, ddd, J1 ¼ 8.5 Hz, J2 ¼ 7.0 Hz,
J3 ¼ 1.5 Hz), 7.74 (1H, dd, J1 ¼ 8.5 Hz, J2 ¼ 1.5 Hz), 7.77 (1H, dd,
J1 ¼8.5 Hz, J2 ¼ 1.5 Hz), 7.85 (1H, d, J ¼ 8.5 Hz), 7.96 (1H, d, J ¼ 8.5 Hz),
7.99 (1H, dd, J1 ¼ 8.0 Hz, J2 ¼ 1.0 Hz), 8.03–8.06 (2H, m), 8.09 (1H, d,
J ¼ 1.5 Hz), 8.27 (1H, d, J ¼ 9.0 Hz), 8.47 (1H. s), 8.48 (1H, s).
13C NMR (100 MHz, DMSO-d6): 13.7, 53.7 (2C) (CH3), 18.8, 30.5,
65.3 (CH2), 113.4, 124.5, 125.4, 125.9, 126.4, 126.6, 127.0, 127.4, 127.7,
128.1, 129.9, 138.5, 138.9 (2C) (CHarom), 124.1, 124.6, 125.2, 125.3,
125.4, 137.5, 137.8, 145.1, 145.3, 145.8, 159.1, 159.4, 162.1 (Carom).
4.1.16. 2,20,200-trimethoxy-3,70:30,700-terquinoline 14
Following the general procedure described for 10 and 11 and
using 7 (71 mg, 0.30 mmol), THF (5 mL), PdCl2(PPh3)2 (10.5 mg,
15.0
mmol), Na2CO3 2 M (0.75 mL, 1.5 mmol), 12 (160 mg,
4.2. Antiproliferative activities
0.44 mmol). Microwave irradiation (65 ꢁC, 50 W, Patm, 15 min).
Compound 14 was obtained by filtration of the reaction mixture.
Purification was performed by washing with H2O and Et2O to give
14 (73 mg, 0.15 mmol, 52%) as a white solid (mp 261 ꢁC).
4.2.1. Cell cultures
Stock cell cultures were maintained as monolayers in 75-cm2
culture flasks in Glutamax Eagle’s minimum essential medium
(MEM) with Earle’s salts supplemented with 10% fetal calf serum,
5 mL 100 mM sodium pyruvate, 5 mL of 100X non-essential amino
acids and 2 mg gentamicin base. Cells were grown at 37 ꢁC in
a humidified incubator under an atmosphere containing 5% CO2.
IR (KBr): 1621 cmꢀ1
.
HRMS (ESþ) calcd for C30H24N3O3 [M þ H]þ 474.1818, found
474.1819.
1H NMR (400 MHz, DMSO-d6): 4.03 (3H, s), 4.08 (3H, s), 4.09
(3H, s), 7.08 (1H, d, J ¼ 9.0 Hz), 7.50 (1H, ddd, J1 ¼ 8.0 Hz, J2 ¼ 7.0 Hz,
J3 ¼ 1.0 Hz), 7.72 (1H, ddd, J1 ¼ 8.5 Hz, J2 ¼ 7.0 Hz, J3 ¼ 1.5 Hz), 7.77
(1H, dd, J1 ¼8.5 Hz, J2 ¼ 2.0 Hz), 7.79 (1H, dd, J1 ¼8.5 Hz, J2 ¼ 1.5 Hz),
7.86 (1H, d, J ¼ 8.0 Hz), 7.98–8.03 (2H, m), 8.08 (1H, d, J ¼ 8.5 Hz),
8.08 (1H, d, J ¼ 1.5 Hz), 8.11 (1H, d, J ¼ 1.5 Hz), 8.31 (1H, d,
J ¼ 9.0 Hz), 8.49 (1H, s), 8.52 (1H, s).
4.2.2. Survival assays
Cells were plated at a density of 5 ꢂ 103 cells in 150
mL culture
medium in each well of 96-well microplates and were allowed to
adhere for 16 h before treatment with tested drug. A stock solution
20 mM of each tested drug was prepared in DMSO and kept at
13C RMN: due to low solubility, the 13C NMR spectrum of this
compound could not be recorded.
ꢀ20 ꢁC until use. Then 50
mL of each tested solution were added to
the cultures. A 48 h continuous drug exposure protocol was used.
The antiproliferative effect of the tested drug was assessed by the
resazurin reduction test.
4.1.17. 2,20,200-triethoxy-3,70–30,700-terquinoline 15
Following the typical procedure described for 10 and 11 and
using 8 (120 mg, 0.48 mmol), THF (5 mL), PdCl2(PPh3)2 (16.7 mg,
4.2.3. Resazurin reduction test
Plates were rinsed with 200
overturning on absorbent towelling. Then 150
23.8
m
mol), Na2CO3 2 M (1.2 mL, 2.4 mmol),13 (240 mg, 0.62 mmol).
m
L PBS at 37 ꢁC and emptied by
L of a 25 g/mL
Microwave irradiation (65 ꢁC, 50 W, Patm, 20 min). Purification by
chromatography (cyclohexane-EtOAc, 98:2) provided 15 (201 mg,
0.39 mmol, 82%) as a white solid (mp 127 ꢁC).
m
m
solution of resazurin in MEM without phenol red was added to each
well. Plates were incubated for 1 h at 37 ꢁC in a humidified
atmosphere containing 5% CO2. Fluorescence was then measured
on an automated 96-well plate reader (Fluoroscan Ascent FL,
Labsystem) using an excitation wavelength of 530 nm and an
emission wavelength of 590 nm. Under the conditions used,
fluorescence was proportional to the number of living cells in the
well. The IC50, defined as the drug concentration required to inhibit
IR (KBr): 1615 cmꢀ1
.
HRMS (ESþ) calcd for C33H30N3O3 [M þ H]þ 516.2287, found
516.2289.
1H NMR (400 MHz, DMSO-d6): 1.39 (6H, t, J ¼ 7.0 Hz), 1.41 (3H, t,
J ¼ 7.0 Hz), 4.50 (2H, q, J ¼ 7.0 Hz), 4.57 (2H, q, J ¼ 7.0 Hz), 4.58 (2H,
q, J ¼ 7.0 Hz), 7.04 (1H, d, J ¼ 9.0 Hz), 7.48 (1H, ddd, J1 ¼ 8.0 Hz,