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415
(KBr, cmꢀ1): 3417, 3234 (NH2), 3155 (NH), 2952(CH2), 1541 (C]N),
1519, 1325 (NO2), 1267 (CeOeC), 1033 (C]S). 1H NMR (400 MHz,
4.1.1.9. 3,4-Methylenedioxyde-6-cyano-benzaldehyde-thio-
semicarbazone (23). White crystals; yield: 93%; m.p. 210 ꢁC. IR (KBr,
cmꢀ1): 3477, 3332 (NH 2), 3143 (NH); 2976 (CH2), 2230 (C^N),1539
(C]N), 1240 (CeOeC), 1089 (C]S). 1H NMR (400 MHz, DMSO-d6)
DMSO-d6)
d
: 11.65 (s, 1H, H2), 8.46 (s, 1H, H70), 8.32 and 8.24 (broad
s, 2H, H4 and H5), 7.99 (s, 1H, H50), 7.54 (s, 1H, H20), 6.25 (s, 2H, H80).
13C NMR (DMSO-d6, 100 MHz)
d
: 178.8 (C3), 152.2 (C30), 149.0 (C60),
d
: 11.45 (s, 1H, H2), 8.35 (s, 1H, H70), 8.21 and 8.17 (broad s, 2H, H4
143.6 (C40), 137.9 (C70), 126.1 (C10), 106.3 (C20), 105.1 (C50), 104.1
(C80). Anal. Calcd. for C9H8N4O4S: C, 40.30, H, 3.01, N, 20.89. Found:
C, 40.15, H, 2.97, N, 20.97.
and H5), 7.52 (s, 1H, H50), 7.19 (s, 1H, H20), 6.12 (s, 2H, H80). 13C NMR
(DMSO-d6, 100 MHz) d
: 178.9 (C3), 154.6 (C30), 151.7 (C40), 144.5
(C70), 131.9 (C10), 116.1 (C90), 114.6 (C50), 111.5 (C20), 1085 (C60), 101.6
(C80). Anal. Calcd. for C10H8N4O2S: C, 48.38, H, 3.25, N, 22.57. Found:
C, 48.42, H, 3.31, N, 22.64.
4.1.1.4. 3,4-Methylenedioxyde-6-amino-benzaldehyde-thio-
semicarbazone (18). Greenish-yellow crystals; yield: 93%; m.p.
201 ꢁC. IR (KBr, cmꢀ1): 3402, 3275 (NH2), 3167 (NH), 2966 (CH2),
1541 (C]N),1259 (CeOeC),1076 (C]S). 1H NMR (400 MHz, DMSO-
4.1.1.10. 3,4-Methylenedioxyde-6-hydroxy-benzaldehyde-thio-
semicarbazone (24). White crystals; yield: 93%; m.p. 212 ꢁC. IR (KBr,
cmꢀ1): 3446, 3330 (NH2), 3134 (NH), 2947 (CH2), 2966 (OeH), 1541
(C]N), 1269 (CeOeC), 1064 (C]S). 1H NMR (400 MHz, DMSO-d6)
d6)
d
: 10.94 (s, 1H, H2), 8.06 (s, 1H, H70), 7.96 and 7.72 (broad s, 2H,
H4 and H5), 6.68 (s,1H, H20), 6.38 (s,1H, H50), 6.25 (broad s, 2H, H60)
d
: 11.25 (s,1H, H2), 9,84 (broad s,1H, H60), 8.27 (s, 1H, H70), 7.98 and
5.89 (s, 2H, H80). 13C NMR (DMSO-d6, 100 MHz)
d: 176.7 (C3), 150.2
7.95 (broad s, 2H, H4 and H5), 7.55 (s,1H, H20), 6.45 (s,1H, H50), 5.96
(C30), 147.5 (C70), 145.4 (C40), 138.5 (C60), 110.0 (C20), 107.0 (C10),
101.1 (C80), 96.5 (C50). Anal. Calcd. for C9H10N4O2S: C, 45.37, H, 4.23,
N, 23.51. Found: C, 45.42, H, 4.19, N, 23.47.
(s, 2H, H80). 13C NMR (DMSO-d6, 100 MHz) : 177.7 (C3), 153.2 (C60),
d
150.1 (C40), 141.3 (C30), 140.0 (C70), 113.3 (C10), 105.0 (C20), 101.7
(C80), 98.0 (C50). Anal. Calcd. for C9H9N3O3S: C, 45.18, H, 3.79, N,
17.56. Found: C, 45.23, H, 3.82, N, 17.66.
4.1.1.5. 3,4-Methylenedioxyde-6-fluoro-benzaldehyde-thio-
semicarbazone (19). White crystals; yield: 95%; m.p. 205 ꢁC. IR (KBr,
cmꢀ1): 3417, 3228 (NH2), 3145 (NH), 2977 (CH2), 1535 (C]N), 1325
4.1.1.11. 3,4-Methylenedioxyde-6-methoxy-benzaldehyde-thio-
semicarbazone (25). White crystals; yield: 91%; m.p. 207 ꢁC. IR (KBr,
cmꢀ1): 3446, 3359 (NH2), 3174 (NH), 2976 (CH2), 2929 (CeH), 1539
(CeF), 1240 (CeOeC), 1097 (C]S). 1H NMR (400 MHz, DMSO-d6)
d:
11.50 (s, 1H, H2), 8.09 (broad s, 1H, H4), 8.02 (s, 1H, H70), 7.77 (d, 1H
(C]N), 1244 (CeO), 1082 (C]S). 1H NMR (400 MHz, DMSO-d6)
d:
4
4
JH5 eH2 ¼ 1.5 Hz, H50), 7.59 (d, 1H, JH2 eH5 ¼ 1.5 Hz, H20), 7.36
0
0
0
0
11.45 (s, 1H, H2), 8.22 (s, 1H, H70), 8.05 and 7.98 (broad s, 2H, H4 and
H5), 7.65 (s, 1H, H50), 6.45 (s, 1H, H20), 5.99 (s, 2H, H80), 3.86 (s, 3H,
(broad s,1H, H5), 6.12 (s, 2H, H80). 13C NMR (DMSO-d6, 100 MHz)
d:
177.7 (C3), 158.3 (C60), 150.2 (C40), 147.8 (C30), 142.3 (C70), 122.2
(C10), 116.3 (C20), 102.1 (C80), 97.1 (C50). Anal. Calcd. for C9H8FN3O2S:
C, 44.81, H, 3.34, N, 17.42. Found: C, 44.87, H, 3.29, N, 17.51.
H90). 13C NMR (DMSO-d6, 100 MHz) : 177.5 (C3), 151.8 (C60), 149.1
d
(C40), 144.8 (C30), 141.2 (C70), 119.1 (C10), 110.2 (C20), 102.9 (C80), 92.9
(C50), 55.9 (C90).
4.1.1.12. 3,4-Methylenedioxyde-6-aminochloroacetyl-benzaldehyde-
thiosemicarbazone (26). White-Greenish crystals; yield: 95%; m.p.
257 ꢁC. IR (KBr, cmꢀ1): 3473, 3352 (NH2), 3168 (NH), 2974 (CH2),
1652 (C]O), 1506 (C]N), 1238 (CeOeC), 1099 (C]S), 667 (CeCl).
4.1.1.6. 3,4-Methylenedioxyde-6-chloro-benzaldehyde-thio-
semicarbazone (20). Greenish crystals; yield: 98%; m.p. 208 ꢁC. IR
(KBr, cmꢀ1): 3477, 3332 (NH2), 3143 (NH), 2970 (CH2), 1539 (C]N),
1245 (CeOeC), 1089 (C]S), 667 (CeCl). 1H NMR (400 MHz, DMSO-
1H NMR (400 MHz, DMSO-d6) : 11.60 (broad s, 1H, H60); 10,66 (s,
d
d6)
d
: 11.51 (s, 1H, H2), 8.37 (s, 1H, H70), 8.23 and 8.19 (broad s, 2H,
1H, H2); 8,38 (s, 1H, H50); 8,12 (s, 1H, H70); 7,99 e 7,72 (broad s, 2H,
H4 and H5), 6.60 (s, 1H, H20), 5.92 (s, 2H, H80), 4.21 (s, 2H, H90). 13C
H4 and H5), 7.92 (s, 1H, H50), 7.10 (s, 1H, H20), 6.12 (s, 2H, H80). 13C
NMR (DMSO-d6, 100 MHz) d
: 178.4 (C3), 1450.0 (C40), 147.8 (C30),
NMR (DMSO-d6, 100 MHz) d
: 178.1 (C3), 165.9 (C90), 151.9 (C40),
138.7 (C70), 126.9 (C10), 125.7 (C60), 109.9 (C20), 106.3 (C50), 102.9
(C80). Anal. Calcd. for C9H8ClN3O2S: C, 41.95, H, 3.13, N, 16.31. Found:
C, 41.81, H, 3.07, N, 16.42.
148.9 (C70), 145.8 (C30), 132.8 (C60), 125.8 (C10), 113.1 (C20), 102.5
(C80), 91.6 (C50), 43.1 (C100). Anal. Calcd. for C11H11ClN4O3S: C, 41.98,
H, 3.52, N, 17.80. Found: C, 41.90, H, 3.48, N, 17.88.
4.1.1.7. 3,4-Methylenedioxyde-6-bromo-benzaldehyde-thio-
semicarbazone (21). White crystals; yield: 97%; m.p. 216 ꢁC. IR (KBr,
cmꢀ1): 3477, 3323 (NH2), 3151 (NH), 2972 (CH2), 1535 (C]N), 1245
4.2. Biological assays
4.2.1. Parasites
(CeOeC), 1093 (C]S), 568 (CeBr). 1H NMR (400 MHz, DMSO-d6)
d:
L. amazonensis (strain: MHOM/BR/77/LTB0016) was maintained
as promastigotes at 26 ꢁC in Schneider's insect medium (Sigma-
eAldrich, St Louis, MO, USA) with 10% serum, 100 mg/mL strepto-
mycin and 100 U/mL penicillin. Parasites were maintained until the
10th passage; subsequently, new cultures were obtained from
infected animals. The Animal Ethics Committee of the Oswaldo
Cruz Foundation (license number LW07/2010) approved this study.
11.53 (s,1H, H20), 8.34 (s,1H, H70), 8.21 and 8.18 (broad s, 2H, H4 and
H5), 7.91 (s, 1H, H50), 7.22 (s, 1H, H20), 6.12 (s, 2H, H80). 13C NMR
(DMSO-d6, 100 MHz) d
: 178.5 (C3), 150.2 (C40), 148.3 (C30), 141.2
(C70), 127.2 (C10), 116.1 (C60), 112.8 (C20), 106.9 (C50), 102.9 (C80).
Anal. Calcd. for C9H8BrN3O2S: C, 35.78, H, 2.67, N, 13.91. Found: C,
35.82, H, 2.61, N, 14.02.
4.1.1.8. 3,4-Methylenedioxyde-6-iodo-benzaldehyde-thio-
semicarbazone (22). White crystals; yield: 99%; m.p. 213 ꢁC. IR (KBr,
cmꢀ1): 3417, 3228 (NH2), 3145 (NH), 2977 (CH2), 1535 (C]N), 1240
4.2.2. Promastigote inhibition growth assay
Promastigotes of L. amazonensis (LTB0016, 1 ꢂ 106/mL) were
incubated with thiosemicarbazones up to 400 m
M for 72 h at 26 ꢁC
(CeOeC), 1097 (C]S), 561 (CeI). 1H NMR (400 MHz, DMSO-d6)
d:
in Schneider's medium supplemented with 10% FBS, penicillin (100
UI/mL) and streptomycin (100 g/mL). The assays were performed
in triplicate in 96-well plates (Costar, New York, USA). The inhibi-
tion growth assay was evaluated by adding 22 L of MTT (5 mg/mL,
Sigma, Saint Louis, USA) per well. After 2 h, 80 L of DMSO (Vetec,
11.56 (s, 1H, H2), 8.25 (s, 1H, H70), 8.19 and 8.15 (broad s, 2H, H4 and
m
H5), 7.87 (s, 1H, H50), 7.39 (s, 1H, H20), 6.10 (s, 2H, H80). 13C NMR
(DMSO-d6, 100 MHz)
d
: 178.3 (C3), 150.2 (C40), 149.0 (C30), 145.8
m
m
(C70), 130.1 (C10), 118.6 (C60), 107.2 (C20), 102.6 (C80), 90.7 (C50). Anal.
Calcd. for C9H8IN3O2S: C, 30.96, H, 2.31, N, 12.04. Found: C, 30.90, H,
2.38, N, 12.11.
Rio de Janeiro, Brazil) was added to each well, and the optical
density was measured at 570 nm using a spectrometer (Bio-Tek