1184
G. Dꢀzimbeg et al. / European Journal of Medicinal Chemistry 43 (2008) 1180e1187
Table 1 (continued)
Compd.
R1
R2
H
1H NMR (DMSO-d6, d ppm)
3h
8.54e8.53 (d, 1H, 16), 8.45e8.44 (d, 2H, 60, 70), 8.09e8.07 (d, 1H, 14), 7.45e7.41 (m, 1H,
15), 7.21e7.20 (d, 2H, 50, 80), 6.48 (s, 1H, 9), 6.41 (t, 1H, 20), 6.27 (s, 1H, 11), 6.14e6.11 (d,
2H, 7, 1), 4.22e4.20 (d, 2H, 30), 3.82 (s, 3H, 17), 3.71e3.56 (m, 1H, 5), 3.05 (q, 2H, 2),
1.77e1.53 (m, 2H, 3), 1.63e1.30 (m, 2H, 4), 1.22e1.20 (d, 3H, 6)
6'
5'
4'
N
7'
H2C
8'
3
3i
H
8.54e8.52 (dd, 1H, 16), 8.09e8.06 (dd, 1H, 14), 7.44e7.40 (m, 1H, 15), 7.31e7.18 (m, 10H,
40e150), 6.81e6.78 (d, 1H, 20), 6.47 (s, 1H, 9), 6.26 (s, 1H, 11), 6.13e6.10 (d, 1H, 7), 5.95e
5.92 (t, 1H, 1), 5.89e5.86 (d, 1H, 30), 3.82 (s, 3H, 17), 3.70e3.56 (m, 1H, CH-5), 3.06e2.99
(q, 2H, CH2-2), 1.70e1.57 (m, 2H, CH2-3), 1.57e1.44 (m, 2H, CH2-4), 1.21e1.19 (d, 3H, 6)
6'
9'
7'
8'
5'
4'
3'
HC
10'
11'
12'
15'
14'
13'
cultures for which the cell number was counted using a Coulter
counter.
activity was expressed as the concentration required to reduce
virus-induced cytopathogenicity by 50% (EC50).
2.2.4. Antiviral activity assays
Antiviral activity against vesicular stomatitis virus, Cox-
sackie virus B4, respiratory syncytial virus, herpes simplex vi-
rus type 1 (KOS) and type 2 (G), thymidine kinase-deficient
herpes simplex virus type 1 (TKꢂ KOS ACVr), vaccinia virus,
parainfluenza-3 virus, reovirus-1, Sindbis virus and Punta Toro
virus was determined essentially as described previously
[10,11]. After a 2 h incubation period, residual virus was re-
moved and the infected cells were further incubated with the
medium containing different concentration of the test com-
pounds. After incubation for 3 days at 37 ꢀC, virus-induced
cytopathogenicity was monitored microscopically. Antiviral
2.2.5. Cytotoxicity assays
Cytotoxicity measurements were based on the inhibition of
HEL, Vero or HeLa cell growth. Cells were seeded at
5 ꢁ 103 cells/well into 96-well microtiter plates. Then, me-
dium containing different concentrations of the test com-
pounds was added. After 3 days of incubation at 37 ꢀC, the
alteration of morphology of the cell cultures was recorded mi-
croscopically. Cytotoxicity was expressed as minimum cyto-
toxic concentration (MCC) or the compound concentration
that causes a microscopically detectable alteration of normal
cell morphology.
Table 2
13C NMR spectral data of primaquine urea derivatives 3aeia
Compd. no.
13C NMR (DMSO-d6, d ppm)
3a
159.15 (10), 156.84 (10), 144.74 (8), 144.37 (16), 134.93 (14), 134.67 (13), 129.71 (12), 122.23 (15), 96.25 (9),
91.70 (11), 55.11 (17), 47.19 (5), 40.14 (20, 40), 39.53 (2), 33.52 (4), 26.95 (3), 20.31 (6), 13.98 (30, 50)
3b
3c
3d
3e
3f
159.53 (10), 158.24 (10), 145.16 (8), 144.77 (16), 135.33 (14), 135.05 (13), 130.12 (12), 122.63 (15), 96.64 (9),
92.14 (11), 55.51 (17), 51.46 (30), 47.57 (5), 39.75 (2), 34.05 (4), 33.46 (40, 70), 27.45 (3), 23.65 (50, 60), 20.75 (6)
159.14 (10), 157.48 (10), 144.76 (8), 144.38 (16), 134.93 (14), 134.65 (13), 129.71 (12), 122.23 (15), 96.24 (9), 91.75 (11),
55.11 (17), 47.81 (30), 47.18 (5), 39.36 (2), 33.66 (4), 33.45 (40, 80), 27.05 (3), 25.45 (60), 24.64 (50, 70), 20.36 (6)
159.14 (10), 158.27 (10), 144.77 (8), 144.37 (16), 134.93 (14), 134.65 (13), 129.71 (12), 122.24 (15), 96.23 (9), 91.74 (11),
55.12 (17), 47.17 (5), 45.69 (30), 39.37 (2), 38.29 (40), 33.67 (4), 30.49 (50, 90), 27.08 (3), 26.26 (70), 25.56 (60, 80), 20.36 (6)
159.41 (10), 156.92 (10), 144.97 (8), 144.61 (16), 135.18 (14), 134.65 (13), 129.97 (12), 122.48 (15), 96.53 (9), 91.94 (11), 55.36 (17),
54.56 (20, 80), 47.45 (5), 40.51 (2), 33.78 (4), 31.47 (30, 70, 90, 130), 27.13 (3), 26.27 (40, 60, 100, 120), 25.45 (50, 110), 20.55 (6)
159.15 (10), 158.21 (10), 144.78 (8), 144.39 (16), 141.14 (40), 134.94 (14), 134.67 (13), 129.72 (12), 128.29 (60, 80), 127.08 (50, 90),
126.60 (70), 122.24 (15), 96.25 (9), 91.76 (11), 55.12 (17), 47.19 (5), 43.02 (30), 39.49 (2), 33.65 (4), 27.06 (3), 20.37 (6)
159.15 (10), 158.09 (10), 144.77 (8), 144.38 (16), 139.92 (50), 134.94 (14), 134.66 (13), 129.72 (12), 128.77 (70, 90), 128.39 (60, 100),
126.05 (80), 122.24 (15), 96.24 (9), 91.75 (11), 55.12 (17), 47.19 (5), 41.05 (30), 39.37 (2), 36.32 (40), 33.66 (4), 27.04 (3), 20.37 (6)
159.15 (10), 158.19 (10), 150.38 (40), 149.48 (60, 70), 144.78 (8), 144.39 (16), 134.95 (14), 134.66 (13), 129.72 (12), 122.25 (15),
121.99 (50, 80), 96.25 (9), 91.76 (11), 55.19 (17), 47.18 (5), 42.08 (30), 39.53 (2), 33.63 (4), 26.99 (3), 20.37 (6)
159.15 (10), 157.36 (10), 144.77 (8), 144.39 (16), 143.90 (40, 100), 134.94 (14), 134.66 (13), 129.72 (12), 128.44 (60, 80, 120, 140),
127.05 (50, 90, 110, 150), 126.80 (70, 130), 122.24 (15), 96.25 (9), 91.77 (11), 56.98 (30), 55.12 (17), 47.17 (5), 39.37 (2), 33.65 (4),
27.02 (3), 20.36 (6)
3g
3h
3i
a
For assignation of atoms see Table 1.