Journal of Medicinal Chemistry
Article
7.04 (d, J = 7.4 Hz, 1H), 7.35 (m, 1H), 7.61 (d, J = 8.0 Hz, 1H), 8.17
(s, 1H). 13C NMR (CDCl3): δ 20.2, 52.2, 56.0 (2×), 61.1, 95.3 (2×),
113.9, 120.8, 127.2, 127.9, 133.1, 133.6, 136.7, 140.4, 142.8, 146.7,
153.9 (2×), 165.3. MS (ESI): [M + 1]+ = 388.0. Anal. (C20H21NO5S)
C, H, N.
Ethyl 4-Methyl-3-[(3,4,5-trimethoxyphenyl)amino]-1-benzo[b]-
thiophene-2-carboxylate (4d). Following general procedure C, the
crude residue was purified by flash chromatography, using ethyl
acetate/petroleum ether (2:8, v/v) as the eluting solution, to furnish
4d as a yellow solid (48% yield), mp 120−122 °C. 1H NMR (CDCl3):
δ 1.36 (t, J = 7.0 Hz, 3H), 2.36 (s, 3H), 3.61 (s, 6H), 3.77 (s, 3H), 4.35
(q, J = 7.2 Hz, 2H), 6.00 (s, 2H), 7.02 (d, J = 7.4 Hz, 1H), 7.36 (m,
1H), 7.62 (d, J = 7.6 Hz, 1H), 8.18 (s, 1H). 13C NMR (CDCl3): δ
14.4, 20.1, 56.0 (2×), 61.1, 61.3, 95.3 (2×), 114.4, 120.8, 126.1, 127.2,
127.8, 133.7, 136.7, 140.4, 142.9, 146.5, 153.9 (2×), 164.9. MS (ESI):
[M + 1]+ = 402.0. Anal. (C21H23NO5S) C, H, N.
Methyl 5-Methyl-3-[(3,4,5-trimethoxyphenyl)amino]-1-benzo[b]-
thiophene-2-carboxylate (4e). Following general procedure C, the
crude residue was purified by flash chromatography, using ethyl
acetate/petroleum ether (2:8, v/v) as the eluting solution, to furnish
4e as a yellow solid (73% yield), mp 190−192 °C. 1H NMR (CDCl3):
δ 2.29 (s, 3H), 3.73 (s, 6H), 3.84 (s, 3H), 3.91 (s, 3H), 6.31 (s, 2H),
7.23 (d, J = 7.8 Hz, 1H), 7.27 (s, 1H), 7.65 (d, J = 7.8 Hz, 1H), 8.77
(s, 1H). 13C NMR (CDCl3): δ 21.5, 51.9, 56.2 (2×), 61.2, 99.4 (2×),
106.3, 122.9, 125.5, 129.8, 132.1, 133.3, 134.4, 137.5, 138.2, 146.0,
153.5 (2×), 166.0. MS (ESI): [M + 1]+ = 388.0. Anal. (C20H21NO5S)
C, H, N.
Ethyl 6-Methoxy-3-[(3,4,5-trimethoxyphenyl)amino]-1-benzo[b]-
thiophene-2-carboxylate (4j). Following general procedure C, the
crude residue was purified by flash chromatography, using ethyl
acetate/petroleum ether (2:8, v/v) as the eluting solution, to furnish 4j
1
as a yellow solid (69% yield), mp 155−157 °C. H NMR (CDCl3): δ
1.40 (t, J = 7.2 Hz, 3H), 3.75 (s, 6H), 3.81 (s, 3H), 3.87 (s, 3H), 4.36
(q, J = 7.2 Hz, 2H), 6.33 (s, 2H), 6.84 (dd, J = 9.0 and 2.4 Hz, 1H),
7.14 (d, J = 2.4 Hz, 1H), 7.32 (d, J = 9.0 Hz, 1H), 8.80 (s, 1H). 13C
NMR (CDCl3): δ 14.6, 55.6, 56.2 (2×), 60.8, 61.2, 99.9 (2×), 104.7,
110.1, 114.0, 125.5, 126.8, 130.5, 134.6, 138.1, 142.4, 153.6 (2×),
159.9, 165.6. MS (ESI): [M + 1]+ = 418.1. Anal. (C21H23NO6S) C, H,
N.
Methyl 7-Methoxy-3-[(3,4,5-trimethoxyphenyl)amino]-1-benzo-
[b]thiophene-2-carboxylate (4k). Following general procedure C,
the crude residue was purified by flash chromatography, using ethyl
acetate/petroleum ether (2:8, v/v) as the eluting solution, to furnish
4k as a yellow solid (74% yield), mp 176−178 °C. 1H NMR (CDCl3):
δ 3.72 (s, 6H), 3.84 (s, 3H), 3.92 (s, 3H), 3.99 (s, 3H), 6.33 (s, 2H),
6.80 (dd, J = 7.2 and 1.4 Hz, 1H), 7.06 (m, 2H), 8.76 (s, 1H). 13C
NMR (CDCl3): δ 52.0, 55.8, 56.1 (2×), 61.2, 99.8 (2×), 104.2, 106.9,
118.4, 121.6, 124.8, 133.4, 134.5, 138.1, 146.8, 153.5 (2×), 154.5,
166.1. MS (ESI): [M + 1]+ = 404.0. Anal. (C20H21NO6S) C, H, N.
Methyl 3-[(3,4,5-Trimethoxyphenyl)amino]thieno[2,3-b]pyridine-
2-carboxylate (4l). Following general procedure C, the crude residue
was purified by flash chromatography, using ethyl acetate/petroleum
ether (3:7, v/v) as the eluting solution, to furnish 4l as a yellow solid
(52% yield), mp 176−178 °C. 1H NMR (CDCl3): δ 3.74 (s, 6H), 3.86
(s, 3H), 3.94 (s, 3H), 6.35 (s, 2H), 7.09 (m, 1H), 7.61 (dd, J = 8.4 and
1.6 Hz, 1H), 8.61 (dd, J = 4.4 and 1.6 Hz, 1H), 8.88 (s, 1H). 13C NMR
(CDCl3): δ 52.1, 56.2 (2×), 61.2, 100.6 (2×), 110.3, 118.4, 125.6,
133.5, 135.3, 137.4, 144.6, 145.7, 150.3, 153.8 (2×), 166.0. MS (ESI):
[M + 1]+ = 375.1. Anal. (C18H18N2O5S) C, H, N.
Methyl 6-Methyl-3-[(3,4,5-trimethoxyphenyl)amino]-1-benzo[b]-
thiophene-2-carboxylate (4f). Following general procedure C, the
crude residue was purified by flash chromatography, using ethyl
acetate/petroleum ether (2:8, v/v) as the eluting solution, to furnish 4f
1
as a yellow solid (64% yield), mp 168−170 °C. H NMR (CDCl3): δ
Ethyl 3-[(3,4,5-Trimethoxyphenyl)amino]thieno[2,3-b]pyridine-2-
carboxylate (4m). Following general procedure C, the crude residue
was purified by flash chromatography, using ethyl acetate/petroleum
ether (3:7, v/v) as the eluting solution, to furnish 4m as a yellow solid
2.43 (s, 3H), 3.73 (s, 6H), 3.84 (s, 3H), 3.91 (s, 3H), 6.33 (s, 2H),
7.02 (d, J = 8.6 Hz, 1H), 7.31 (d, J = 8.6 Hz, 1H), 7.53 (s, 1H), 8.80
(s, 1H). 13C NMR (CDCl3): δ 21.7, 51.9, 56.2 (2×), 61.2, 99.9 (2×),
104.3, 123.0, 125.3, 125.6, 129.5, 134.6, 138.1, 138.4, 140.7, 146.6,
153.5 (2×), 166.1. MS (ESI): [M + 1]+ = 388.0. Anal. (C20H21NO5S)
C, H, N.
1
(76% yield), mp 167−168 °C. H NMR (CDCl3): δ 1.42 (t, J = 7.2
Hz, 3H), 3.74 (s, 6H), 3.86 (s, 3H), 4.42 (q, J = 7.2 Hz, 2H), 6.34 (s,
2H), 7.09 (m, 1H), 7.62 (dd, J = 8.2 and 1.6 Hz, 1H), 8.62 (dd, J = 4.6
and 1.6 Hz, 1H), 8.90 (s, 1H). 13C NMR (CDCl3): δ14.5, 56.2 (2×),
61.2, 61.3, 100.4 (2×), 104.4, 118.4, 125.7, 133.4, 135.2, 137.5, 144.4,
150.2, 153.7 (2×), 161.1, 165.7. MS (ESI): [M + 1]+ = 389.1. Anal.
(C19H20N2O5S) C, H, N.
Ethyl 6-Methyl-3-[(3,4,5-trimethoxyphenyl)amino]-1-benzo[b]-
thiophene-2-carboxylate (4g). Following general procedure C, the
crude residue was purified by flash chromatography, using ethyl
acetate/petroleum ether (2:8, v/v) as the eluting solution, to furnish
4g as a yellow solid (79% yield), mp 125−127 °C. 1H NMR (CDCl3):
δ 1.37 (t, J = 7.0 Hz, 3H), 2.43 (s, 3H), 3.73 (s, 6H), 3.84 (s, 3H), 4.39
(q, J = 7.0 Hz, 2H), 6.32 (s, 2H), 6.96 (d, J = 8.4 Hz, 1H), 7.31 (d, J =
8.6 Hz, 1H), 7.53 (s, 1H), 8.82 (s, 1H). 13C NMR (CDCl3): δ 14.5,
21.8, 56.1 (2×), 60.9, 61.2, 99.7 (2×), 104.9, 121.7, 122.9, 125.3,
125.5, 129.6, 138.2, 138.3, 140.6, 146.4, 153.5 (2×), 165.8.. MS (ESI):
[M + 1]+ = 402.2. Anal. (C21H23NO5S) C, H, N.
Methyl 6-Methyl-3-[(3,4,5-trimethoxyphenyl)amino]thieno[2,3-
b]pyridine-2-carboxylate (4n). Following general procedure C, the
crude residue was purified by flash chromatography, using ethyl
acetate/petroleum ether (3:7, v/v) as the eluting solution, to furnish
4n as a yellow solid (63% yield), mp 186−188 °C. 1H NMR (CDCl3):
δ 2.64 (s, 3H), 3.74 (s, 6H), 3.85 (s, 3H), 3.92 (s, 3H), 6.34 (s, 2H),
6.93 (d, J = 8.6 Hz, 1H), 7.50 (d, J = 8.6 Hz, 1H), 8.88 (s, 1H). 13C
NMR (CDCl3): δ 24.8, 52.0, 56.2 (2×), 61.2, 100.5 (2×), 102.1, 118.8,
122.3, 133.5, 135.2, 137.4, 144.8, 145.3, 153.7 (2×), 160.1, 166.2. MS
(ESI): [M + 1]+ = 389.0. Anal. (C19H20N2O5S) C, H, N.
Antiproliferative Assays. Human T-cell leukemia (Jurkat),
human B-cell leukemia (SEM), and human promyelocytic leukemia
(HL-60) cells were grown in RPMI-1640 medium (Gibco, Milano,
Italy). Breast adenocarcinoma (MCF-7), human non-small-cell lung
carcinoma (A549), human cervix carcinoma (HeLa), and human colon
adenocarcinoma (HT-29) cells were grown in DMEM medium
(Gibco). Both media were supplemented with 115 units/mL penicillin
G (Gibco), 115 μg/mL streptomycin (Invitrogen, Milano, Italy), and
10% fetal bovine serum (Invitrogen). These cell lines were purchased
from ATCC. CEMVbl‑100 cells are a multi-drug-resistant line selected
against vinblastine.24 LoVoDoxo cells are a doxorubicin-resistant
subclone of LoVo cells25 and were grown in complete Ham’s F12
medium supplemented with doxorubicin (0.1 μg/mL). LoVoDoxo and
CEMVbl‑100 were a kind gift of Dr. G. Arancia (Istituto Superiore di
Methyl 7-Methyl-3-[(3,4,5-trimethoxyphenyl)amino]-1-benzo[b]-
thiophene-2-carboxylate (4h). Following general procedure C, the
crude residue was purified by flash chromatography, using ethyl
acetate/petroleum ether (2:8, v/v) as the eluting solution, to furnish
1
4h as a cream-colored solid (70% yield), mp 170−172 °C. H NMR
(CDCl3): δ 2.52 (s, 3H), 3.72 (s, 6H), 3.85 (s, 3H), 3.93 (s, 3H), 6.34
(s, 2H), 7.10 (m, 1H), 7.33 (m, 2H), 8.80 (s, 1H). 13C NMR
(CDCl3): δ 19.7, 52.0, 56.1 (2×), 61.2, 99.8 (2×), 102.4, 123.6, 123.9,
127.9, 131.6, 132.5, 124.6, 138.1, 140.4, 147.2, 153.5 (2×), 166.1. MS
(ESI): [M + 1]+ = 388.0. Anal. (C20H21NO5S) C, H, N.
Methyl 6-Methoxy-3-[(3,4,5-trimethoxyphenyl)amino]-1-benzo-
[b]thiophene-2-carboxylate (4i). Following general procedure C,
the crude residue was purified by flash chromatography, using ethyl
acetate/petroleum ether (2:8, v/v) as the eluting solution, to furnish 4i
1
as a yellow solid (78% yield), mp 155−156 °C. H NMR (CDCl3): δ
3.73 (s, 6H), 3.84 (s, 3H), 3.87 (s, 3H), 3.90 (s, 3H), 6.34 (s, 2H),
6.76 (dd, J = 9.2 and 2.4 Hz, 1H), 7.16 (d, J = 2.4 Hz, 1H), 7.30 (d, J =
9.2 Hz, 1H), 8.81 (s, 1H). 13C NMR (CDCl3): δ 51.8, 55.6, 56.1 (2×),
61.2, 100.0 (2×), 104.7, 113.3, 114.1, 125.4, 126.8, 137.9, 138.2, 142.5,
146.7, 153.5 (2×), 159.9, 166.0. MS (ESI): [M + 1]+ = 404.2. Anal.
(C20H21NO6S) C, H, N.
̀
Sanita, Rome, Italy). A549-T12 cells are a non-small-cell lung
carcinoma line exhibiting resistance to paclitaxel29 and were kindly
donated by Prof. I. Castagliuolo (University of Padova). They were
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dx.doi.org/10.1021/jm400043d | J. Med. Chem. 2013, 56, 2606−2618