E.F. da Silva et al. / European Journal of Medicinal Chemistry 37 (2002) 979ꢂ
/
984
983
(C11), 216.21 (C15), 123.35 (C9), 118.49 (C7), 114.83
(C12), 108.83 (Cb), 55.66 (OCH3).
2j (Xꢁ
cmꢃ1) 3050, 1600 and 1500, 1570, 1620; 1H-NMR
(DMSO-d6) d 12.35 (s, NH), 7.86 (d, Ha, Jꢁ15.5),
7.80ꢂ7.57 (m, Harom), 7.40 (m, H11, 11?), 7.12 (m, H12,
12?), 6.80 (d, Hb, Jꢁ
15.5); 13C-NMR (CDCl3/CD3OD)
/
OH): Yield 65%; m.p. 205 8C; IR (KBr,
2d (Xꢁ
/
NO2): Yield 89%; m.p. 232ꢂ
/
233 8C; IR (KBr,
/
cmꢃ1) 3410, 3044, 2720, 1615, 1565, 1522, 1450, 1341,
1
956; H-NMR (DMSO-d6) d 12.35 (s, NH), 8.396 (d,
/
/
H12, 12?, Jꢁ
/
8.2), 8.26 (d, Ha, Jꢁ
/
16.6), 8.151 (d, H11,
d 162.58 (C5), 159.00 (C13), 157.94 (C2), 149.93
(Ca),139.49 (C6), 138.07 (C14), 131.67 (C17), 131.45
(C11), 130.18 (C8), 129.00 (C16), 125.42 (C15), 124.24
(C9), 124.00 (C14), 118.85 (C7), 116.42 (C12), 105.86
(Cb).
11?, Jꢁ8.3), 8.00ꢂ
/
/
7.87 (m, Harom), 7.41 (d, Hb, Jꢁ
/
16.2); 13C-NMR (DMSO-d6) d 162.05 (C5), 159.68
(C2), 149.59 (C13), 144.67 (Ca), 139.99 (C10), 138.54
(C14), 137.05 (C6), 131.90 (C17), 130.25 (C16), 130.15
(C8), 129.56 (C11), 126.23 (C15), 124.32 (C9), 124.20
(C12), 118.71 (C7), 115.52 (Cb).
2k (Yꢁ
NMR (DMSO-d6) d 12.38 (s, NH), 7.92 (d, Ha, Jꢁ
16.0), 3.76
/
OCH3):Yield 68%; m.p. 284ꢂ
285 8C; 1H-
/
/
2e (Xꢁ
/
F): Yield 85%; m.p. 251ꢂ
/
253 8C; IR (KBr,
16.1), 7.86ꢂ
/
7.31 (m, Harom), 7.06 (d, Hb, Jꢁ
/
cmꢃ1) 3442, 3007, 2669, 1618, 1570, 1512, 1448, 1315,
(s, CH3O); 13C-NMR (DMSO-d6) d 162.82 (C12?),
159.66 (C5), 159.15 (C2), 147.96 (Ca), 137.02 (C6),
138.55 (C14), 135.22 (C10), 121.12 (C11), 131.74 (C17),
130.19 (C16), 117.62 (C11?), 126.16 (C15), 129.54 (C8),
124.22 (C9), 114.49 (C13), 118.64 (C7), 118.89 (Cb),
55.38 (CH3O).
1
1230; H-NMR (DMSO-d6) d 12.33 (s, NH), 8.13 (d,
Ha, Jꢁ
H11, 11?), 7.54 (d, H11, 11?, Jꢁ
16.2); 13C-NMR (DMSO-d6) d 164.83 (C5), 159.04 (C2),
/
16.2), 7.69 (d, H12, 12?, Jꢁ
/
8.4, 1Hꢄ1
H-COSY
/
/
8.4), 7.18 (d, Hb, Jꢁ
/
144.58 (Ca), 138.69 (C14), 136.96 (C6), 131.80 (1JCÃF
ꢁ
/
251.8, C10), 131.67 (C17), 130.57 (3JCÃF 3.8, C11, C11?),
130.16 (C8), 129.51 (C16), 126.15 (C12), 124.12 (C9),
118.62 (C7), 116.47 (2JCÃF 21.9, C12), 111.39 (Cb).
2l (YꢁNO2): Yield 73%; m.p. 185 8C; IR (KBr,
/
cmꢃ1) 3050, 1620, 1570, 1335; d 13C-NMR (CDCl3/
CD3OD) d 145.17 (Ca), 137.4 (C6), 136.3 (C14), 134.5
(C10), 133.42 (C11), 131.58 (C17), 129.75 (C12), 128.69
(C16), 125.21 (C11?), 124.96 (C15), 124.63 (C8), 124.36
2f (Xꢁ
/
Cl): Yield 83%; m.p. 264ꢂ2658C; IR (KBr,
/
cmꢃ1) 3423, 3046, 2653, 1615, 1566, 1538, 1497, 1450,
1
1326; H-NMR (DMSO-d6) d 12.61 (s, NH), 8.11 (d,
(C9), 122.7 (C13), 118.44 (C7), 112.79 (Cb).
1
1
Ha, Jꢁ
/
16.5, Hꢄ1
/
H-COSY Hb), 7.91 (H12, 12?, Jꢁ
8.7),
16.2); 13C-NMR (DMSO-d6) d 159.13
/
2m (Yꢁ
/
Cl): Yield 72%; m.p. 261ꢂ
/
263 8C; H-NMR
8.7, Hꢄ1
/
H-COSY H11, 11?), 7.65 (H11, 11?, Jꢁ
/
(DMSO-d6) d 12.79 (s, NH), 7.98 (d, Ha, Jꢁ
/
16.4),
15.5); 13C-NMR
1
7.22 (d, Hb, Jꢁ
/
7.92ꢂ7.36 (m, Harom), 7.19 (d, Hb, Jꢁ
/
/
(C2), 146.2 (Ca), 136.97 (C6), 136.28 (C13), 132.76
(C17), 131.62 (C10), 130.69 (C8), 130.1 (C16), 129.45
(C12), 129.21 (C11), 126.08 (C15), 124.03 (C9), 118.67
(C7), 112.24 (Cb).
(CDCl3/CD3OD) d 162.45 (C5), 159.12 (C2), 145.94
(Ca), 137.02 (C6), 138.55 (C14), 131.71 (C17), 128.52
(C11?), 126.16 (C13), 129.43 (C8), 130.15 (C16), 127.73
(C15), 124.09 (C9), 118.55 (C7), 133.91 (C12?), 130.94
2g (Xꢁ
/
CN): Yield 82%; m.p. 185 8C; IR (KBr,
(C12), 113.21 (Cb).
cmꢃ1) 3404, 2932, 2224, 1623, 1564, 1537, 1502 and
1420, 1310; 13C-NMR (DMSO-d6) d 161.0 (C5), 145.4
(Ca), 137.0 (C6), 131.5 (C17), 128.5 (C8), 129.6 (C16),
132.9 (C12), 128.7 (C11), 124.9 (C15), 124.4 (C9), 118.4
2n (Yꢁ
/
Br): Yield 80%; m.p. 263ꢂ
/
264 8C; H-NMR
1
(DMSO-d6) d 12.84 (s, NH), 7.98 (d, Ha, Jꢁ
/
16.1),
16.2); 13C-NMR
7.87ꢂ7.27 (m, Harom), 7.19 (d, Hb, Jꢁ
/
/
(CDCl3/CD3OD) d 162.46 (C5), 159.13 (C2), 145.90
(Ca), 137.05 (C6), 138.59 (C14), 131.73 (C17), 131.29
(C11?), 134.17 (C13), 128.00 (C11), 129.43 (C8), 130.19
(C16), 126.28 (C15), 122.46 (C9), 118.56 (C7), 124.09
(C12?), 130.19 (C12), 113.17 (Cb).
(C7), 113.2 (Cb).
1
2h (Xꢁ
/
Br): Yield 70%; m.p. 283ꢂ
/
285 8C; H-NMR
(DMSO-d6) d 12.58 (s, NH), 8.00 (d, Ha, Jꢁ
1Hꢄ1
H-COSY Hb), 7.86ꢂ7.17 (m, Harom), 7.10 (d, Hb,
Jꢁ
15.9); 13C-NMR (DMSO-d6) d 162.63 (C5), 159.18
/16.0,
/
/
/
(C2), 146.42 (Ca), 136.95 (C6), 124.16 (C13), 131.68
(C17), 133.04 (C10), 129.45 (C8), 130.16 (C16), 132.12
(C12), 130.90 (C11), 126.11 (C15), 125.46 (C9), 118.58
(C7), 112.13 (Cb).
5.2. Pharmacology
5.2.1. Anti-leishmanial assay
L.amazonensis (MHOM/BR/77/LTB0016 strain) pro-
mastigotes were grown at 25 8C in Schneider’s Droso-
phila medium [29] supplemented with 10% (v/v) heat-
inactivated foetal calf serum (FCS). The axenic amasti-
gotes were also cultivated in Schneider’s medium with
2i (Xꢁ
/
OCH2CH3): Yield 85%; m.p. 148ꢂ
150 8C; 1H-
/
NMR (DMSO-d6) d 12.22 (s, NH), 7.96 (d, Ha, Jꢁ
/
1
15.9, Hꢄ1
(d, Hb, Jꢁ
(t, H3CÃCH2Ã
/
H-COSY Hb), 7.85ꢂ
15.9), 4.07 (q, H3CÃCH2Ã
O, Jꢁ
6.9); 13C-NMR (DMSO-d6) d
/
6.97 (m, Harom), 6.90
/
/
/
O, Jꢁ7.0), 1.31
/
/
/
/
20% FCS, pH 5.5 and in a temperature of 32 8C [30ꢂ33].
/
161.93 (C5), 158.54 (C2), 148.08 (Ca), 138.65 (C6),
163.38 (C13), 131.54 (C17), 137.07 (C10), 128.94 (C8),
130.24 (C16), 113.10 (C12), 127.95 (C11), 125.19 (C15),
Parasites (promastigotes/amastigotes) were harvested
from the medium, resuspended in fresh medium,
counted in Neubauer chamber and adjusted to a
124.79 (C9), 119.84 (C7), 108.57 (Cb), 63.68 (CH2Ã
/
O),
concentration of 4ꢄ
/
106 parasites mLꢃ1, for the drug
14.53 (CH3ÃCH2ÃO).
/
/
assay.