Journal of Medicinal Chemistry
Article
NMR (400 MHz, CDCl3) δ 8.98 (s, 1H), 8.14 (s, 1H), 8.06 (d, J =
8.4 Hz, 1H), 7.90 (s, 1H), 7.69 (d, J = 8.4 Hz, 1H), 7.19 (d, J = 2.2
Hz, 1H), 7.03−6.95 (m, 2H), 6.89 (dd, J = 8.4, 2.3 Hz, 1H), 6.70 (d, J
= 8.4 Hz, 1H), 6.60 (d, J = 8.5 Hz, 1H), 4.48 (t, J = 6.2 Hz, 2H), 3.79
(d, J = 12.5 Hz, 6H), 2.73 (t, J = 6.2 Hz, 2H), 2.13 (d, J = 22.9 Hz,
6H). 13C NMR (100 MHz, CDCl3) δ 168.1, 157.7, 157.0, 154.8,
152.1, 149.1, 143.9, 141.3, 132.2, 132.0, 131.6, 131.4, 131.3, 128.6,
128.0, 126.6, 126.2, 112.0, 109.7, 109.2, 55.3 × 2, 44.8, 29.7, 16.2 × 2.
ESI-MS: m/z = 458 [M + H]+. HRMS: calculated (M +H): 458.2192;
found: 458.2200.
4.1.74. 5,6-Bis(4-methoxy-3-methylphenyl)-N-(3-
morpholinopropyl)pyridin-2-amine (7f). To a mixture of LiAlH4
(0.63 mmol) in 5 mL anhydrous THF was added compound 7d (0.11
mmol) at 0 °C under a N2 atmosphere. Then, the mixture was moved
to room temperature and stirred for 10 h. The reaction was quenched
by adding 24 μL of H2O, 24 μL of 15% NaOH, and 72 μL of H2O in
sequence. Then, the mixture was filtered and the filtrate was
concentrated under vacuum. The residue was purified through
chromatography to afford the compound, yield: 63.2%; retention
time: 7.380 min; purity: 98.60%; 1H NMR (400 MHz, CDCl3) δ 7.50
(d, J = 8.4 Hz, 1H), 7.34−7.28 (m, 1H), 7.11 (dd, J = 8.4, 2.3 Hz,
1H), 7.00 (dd, J = 2.2, 0.9 Hz, 1H), 6.90 (dd, J = 8.4, 2.3 Hz, 1H),
6.69 (dd, J = 18.6, 8.4 Hz, 2H), 6.44 (d, J = 8.4 Hz, 1H), 3.84 (d, J =
6.4 Hz, 6H), 3.79 (t, J = 4.7 Hz, 4H), 3.46 (t, J = 6.6 Hz, 2H), 2.60−
2.50 (m, 6H), 2.20 (d, J = 5.0 Hz, 6H), 1.90 (p, J = 6.8 Hz, 2H). 13C
NMR (100 MHz, CDCl3) δ 157.3, 156.3, 154.5, 140.6, 132.9, 132.6,
132.1, 131.7, 128.6, 128.0, 126.2, 125.7, 124.8, 109.7, 109.1, 104.2,
67.0 × 2, 56.9, 55.3 × 2, 53.8 × 2, 41.1, 26.2, 16.2 × 2. ESI-MS: m/z =
462.2 [M + H]+. HRMS: calculated (M + H): 462.2757; found:
462.2765.
3H). 13C NMR (100 MHz, CDCl3) δ 164.6, 161.2, 146.6, 145.0,
134.0, 129.4, 127.4, 126.3, 125.7, 120.3, 119.0, 115.0, 109.8, 67.4 × 2,
55.6, 52.0 × 2, 16.5. ESI-MS: m/z = 372.0 [M + H]+. HRMS:
calculated (M + H): 372.1559; found: 372.1564.
4.1.80. N-(2-(4-Hydroxypiperidin-1-yl)-5-nitrophenyl)benzamide
(8d). General procedure L, yield: 93.2%; melting point: 188−190 °C;
1H NMR (400 MHz, DMSO-d6) δ 9.68 (s, 1H), 8.68 (d, J = 2.7 Hz,
1H), 8.03 (dd, J = 9.1, 2.9 Hz, 2H), 7.99 (d, J = 1.5 Hz, 1H), 7.68−
7.61 (m, 1H), 7.57 (dd, J = 11.4, 4.4 Hz, 2H), 7.30 (d, J = 9.0 Hz,
1H), 4.79 (d, J = 3.9 Hz, 1H), 3.67 (dt, J = 11.4, 3.7 Hz, 1H), 3.31−
3.18 (m, 2H), 2.96−2.77 (m, 2H), 1.93−1.78 (m, 2H), 1.65−1.49
(m, 2H). 13C NMR (100 MHz, DMSO-d6) δ 165.4, 152.1, 141.8,
134.4, 132.6, 131.2, 129.2 × 2, 127.8 × 2, 121.5, 120.2, 119.8, 65.8,
48.8 × 2, 34.8 × 2. ESI-MS: m/z = 342.0 [M + H]+. HRMS:
calculated (M + H): 342.1454; found: 342.1460.
4.1.81. N-(2-(4-Methylpiperazin-1-yl)-5-nitrophenyl)benzamide
(8e). General procedure L, yield: 73.6%; melting point: 166−169
°C; 1H NMR (400 MHz, CDCl3) δ 9.40 (d, J = 2.6 Hz, 1H), 9.12 (s,
1H), 7.97 (dd, J = 8.8, 2.7 Hz, 1H), 7.95−7.88 (m, 2H), 7.63−7.58
(m, 1H), 7.55 (t, J = 7.3 Hz, 2H), 7.30−7.26 (m, 1H), 3.03 (t, J = 4.7
Hz, 4H), 2.65 (m, 4H), 2.40 (s, 3H). 13C NMR (100 MHz, CDCl3) δ
165.0, 147.3, 145.0, 134.3, 133.8, 132.5, 129.2 × 2, 127.1 × 2, 120.5,
119.5, 115.1, 55.8 × 2, 51.9 × 2, 46.2. ESI-MS: m/z = 341.0 [M +
H]+. HRMS: calculated (M + H): 341.1614; found: 341.1622.
4.1.82. N-(2-Morpholino-5-nitrophenyl)benzamide (8f). General
1
procedure L, yield: 37.3%; melting point: 218−220 °C; H NMR
(400 MHz, DMSO-d6) δ 9.81 (s, 1H), 8.71 (d, J = 2.7 Hz, 1H), 8.06
(dt, J = 8.2, 4.1 Hz, 1H), 8.01−7.97 (m, 2H), 7.64 (dd, J = 8.4, 6.1
Hz, 1H), 7.57 (dd, J = 15.2, 8.2 Hz, 2H), 7.35 (d, J = 9.0 Hz, 1H),
3.84−3.72 (m, 4H), 3.13−3.01 (m, 4H). 13C NMR (100 MHz,
DMSO-d6) δ 165.0, 150.8, 141.9, 133.8, 132.1, 131.1, 128.8 × 2,
127.4 × 2, 121.1, 120.0, 119.5, 66.1 × 2, 50.6 × 2. ESI-MS: m/z =
328.0 [M + H]+.
4.1.75. N-(2-Fluoro-5-nitrophenyl)benzamide (31a). General
procedure K, yield: 56.8%; 1H NMR (400 MHz, DMSO-d6) δ
10.49 (s, 1H), 8.67 (dd, J = 6.5, 2.9 Hz, 1H), 8.17 (ddd, J = 9.0, 4.0,
3.0 Hz, 1H), 8.03−7.95 (m, 2H), 7.65 (td, J = 5.2, 2.2 Hz, 1H), 7.63−
7.51 (m, 3H). ESI-MS: m/z = 260.8 [M + H]+.
4.1.76. N-(2-Fluoro-5-nitrophenyl)-4-methoxy-3-methylbenza-
mide (31b). General procedure K, yield: 63.8%; 1H NMR (400
MHz, CDCl3) δ 9.46 (dd, J = 6.8, 2.7 Hz, 1H), 8.08 (s, 1H), 7.99
(ddd, J = 7.5, 4.0, 3.1 Hz, 1H), 7.76 (dd, J = 8.5, 2.2 Hz, 1H), 7.69 (s,
1H), 7.27 (t, J = 9.5 Hz, 1H), 6.91 (d, J = 8.5 Hz, 1H), 3.91 (s, 3H),
2.28 (s, 3H). ESI-MS: m/z = 305.0 [M + H]+.
4.1.83. N-(5-Amino-2-(4-hydroxypiperidin-1-yl)phenyl)-4-me-
thoxy-3-methylbenzamide (8g). General procedure M, yield:
1
43.6%; melting point: 181−183 °C; H NMR (400 MHz, DMSO-
d6) δ 9.60 (s, 1H), 7.72 (dd, J = 8.5, 2.0 Hz, 1H), 7.68 (d, J = 8.0 Hz,
1H), 7.66 (d, J = 2.5 Hz, 1H), 7.11 (d, J = 8.6 Hz, 1H), 6.96 (d, J =
8.4 Hz, 1H), 6.26 (dd, J = 8.4, 2.5 Hz, 1H), 5.00 (s, 2H), 4.75 (d, J =
3.9 Hz, 1H), 3.87 (s, 3H), 3.61 (dt, J = 26.9, 13.4 Hz, 1H), 2.82 (d, J
= 11.8 Hz, 2H), 2.63 (t, J = 9.7 Hz, 2H), 2.23 (s, 3H), 1.87 (d, J = 9.9
Hz, 2H), 1.57 (td, J = 12.5, 3.4 Hz, 2H). 13C NMR (100 MHz,
DMSO-d6) δ 163.6, 160.6, 146.5, 134.4, 131.9, 129.6, 127.1, 126.5,
126.5, 121.8, 110.7, 109.1, 104.9, 66.2, 56.1 × 2, 51.4, 36.2, 16.7 × 2.
ESI-MS: m/z = 356.0 [M + H]+. HRMS: calculated (M + H):
356.1974; found: 356.1986.
4.1.77. N-(2-(4-Hydroxypiperidin-1-yl)-5-nitrophenyl)-4-me-
thoxy-3-methylbenzamide (8a). General procedure L, yield:
1
94.2%; melting point: 188−190 °C; H NMR (400 MHz, DMSO-
d6) δ 9.45 (s, 1H), 8.72 (d, J = 2.1 Hz, 1H), 7.99 (dd, J = 8.7, 1.9 Hz,
1H), 7.86 (d, J = 8.0 Hz, 1H), 7.79 (s, 1H), 7.30 (d, J = 8.9 Hz, 1H),
7.10 (d, J = 8.5 Hz, 1H), 4.57 (s, 1H), 3.88 (s, 3H), 3.77−3.57 (m,
1H), 3.22 (d, J = 11.6 Hz, 2H), 2.84 (t, J = 9.8 Hz, 2H), 2.22 (s, 3H),
1.87 (d, J = 9.5 Hz, 2H), 1.72−1.48 (m, 2H). 13C NMR (100 MHz,
DMSO-d6) δ 164.9, 160.9, 151.6, 142.1, 131.8, 130.2, 127.3, 126.3,
125.9, 121.0, 120.3, 118.9, 110.6, 65.8, 56.1, 48.9 × 2, 34.9 × 2, 16.6.
ESI-MS: m/z = 386.0 [M + H]+. HRMS: calculated (M + H):
386.1716; found: 386.1719.
4.1.84. N-(5-Amino-2-(4-methylpiperazin-1-yl)phenyl)-4-me-
thoxy-3-methylbenzamide (8h). General procedure M, yield:
1
44.8%; H NMR (400 MHz, CDCl3) δ 9.66 (s, 1H), 8.01 (d, J =
2.6 Hz, 1H), 7.78 (dt, J = 10.6, 5.3 Hz, 1H), 7.73 (d, J = 1.8 Hz, 1H),
7.03 (d, J = 8.4 Hz, 1H), 6.92 (d, J = 8.5 Hz, 1H), 6.38 (dd, J = 8.4,
2.7 Hz, 1H), 3.90 (s, 3H), 3.10−2.87 (m, 4H), 2.86−2.40 (m, 4H),
2.38 (s, 3H), 2.30 (s, 3H). 13C NMR (100 MHz, CDCl3) δ 164.5,
160.6, 144.6, 135.0, 132.4, 129.4, 127.0, 126.9, 126.5, 121.9, 109.6,
109.5, 105.7, 56.5 × 2, 55.5, 52.7 × 2, 46.3, 16.4. ESI-MS: m/z =
355.1 [M + H]+. HRMS: calculated (M + H): 355.2134; found:
355.2136.
4.1.85. N-(5-Amino-2-morpholinophenyl)-4-methoxy-3-methyl-
benzamide (8i). General procedure M, yield: 73.9%; 1H NMR
(400 MHz, CDCl3) δ 9.66 (s, 1H), 8.06 (d, J = 2.6 Hz, 1H), 7.80 (dt,
J = 7.2, 3.6 Hz, 1H), 7.75 (d, J = 8.8 Hz, 1H), 7.07 (d, J = 8.4 Hz,
1H), 6.96 (d, J = 8.5 Hz, 1H), 6.44 (dd, J = 8.4, 2.6 Hz, 1H), 3.94 (s,
3H), 3.90 (t, J = 13.0 Hz, 4H), 2.99−2.84 (m, 4H), 2.34 (s, 3H). 13C
NMR (100 MHz, CDCl3) δ 164.6, 160.7, 144.9, 135.1, 132.3, 129.5,
127.2, 127.0, 126.3, 122.0, 109.7, 109.7, 105.9, 68.2 × 2, 55.6, 53.3 ×
2, 16.6. ESI-MS: m/z = 342.1 [M + H]+.
4.1.78. 4-Methoxy-3-methyl-N-(2-(4-methylpiperazin-1-yl)-5-
nitrophenyl)benzamide (8b). General procedure L, yield: 56.8%;
melting point: 187−189 °C; 1H NMR (400 MHz, CDCl3) δ 9.40 (d,
J = 2.6 Hz, 1H), 9.07 (s, 1H), 7.96 (dd, J = 8.7, 2.7 Hz, 1H), 7.79 (dd,
J = 8.5, 2.2 Hz, 1H), 7.74 (s, 1H), 7.28 (d, J = 1.8 Hz, 1H), 6.95 (d, J
= 8.5 Hz, 1H), 3.94 (s, 3H), 3.05 (t, J = 4.7 Hz, 4H), 2.96−2.53 (m,
4H), 2.44 (s, 3H), 2.32 (s, 3H). 13C NMR (100 MHz, CDCl3) δ
164.6, 161.1, 146.9, 144.9, 134.0, 129.4, 127.3, 126.5, 125.8, 120.3,
118.9, 114.8, 109.8, 55.7 × 2, 55.6, 51.6 × 2, 46.1, 16.4. ESI-MS: m/z
= 385.0 [M + H]+. HRMS: calculated (M + H): 385.1876; found:
385.1883.
4.1.79. 4-Methoxy-3-methyl-N-(2-morpholino-5-nitrophenyl)-
benzamide (8c). General procedure L, yield: 70.8%; melting point:
1
210−213 °C; H NMR (400 MHz, CDCl3) δ 9.37 (d, J = 2.6 Hz,
4.2. Cell Culture and Plasmids. The NCI-H1299, H460, KHOS,
A2780, and 293T cell lines were purchased from the Cell Bank of
Type Culture Collection of the Chinese Academy of Sciences
(Shanghai, China). All cells were cultured in Dulbecco’s modified
1H), 9.05 (s, 1H), 7.95 (dd, J = 8.7, 2.6 Hz, 1H), 7.75 (dd, J = 8.5, 2.1
Hz, 1H), 7.70 (s, 1H), 7.25 (d, J = 8.8 Hz, 1H), 6.92 (t, J = 7.3 Hz,
1H), 3.97−3.93 (m, 4H), 3.92 (s, 3H), 3.06−2.95 (m, 4H), 2.30 (s,
8640
J. Med. Chem. 2021, 64, 8621−8643