Y.-X. Wang et al. / Bioorg. Med. Chem. 20 (2012) 6552–6558
6557
5.4.5. 5,6-Dihydro-9-fluoro-10-methoxy-benzo[g]-1,3-
benzodioxolo[5,6-a] quinolizinium (7e)
(s, 3H), 3.27 (t, J = 6.4 Hz, 2H); 13C NMR d: 154.6, 150.3, 145.3,
144.9, 143.6, 137.6, 133.0, 128.9, 126.7, 123.6, 122.5, 121.5,
120.6, 120.1, 112.4, 61.9, 60.3, 57.0, 56.1, 54.8, 20.6; IR (KBr) c:
3358, 2944, 1601, 1508, 1455, 1358, 1338, 1284, 1228, 1125,
1036 cmÀ1; HRMS-ESI: Calcd for C21H22NO4Cl (MÀCl)+ 352.1549,
found 352.1525.
Yield: 73%; Orange solid; mp 251–253 °C (decomp); 1H NMR d:
10.08 (s, 1H), 9.03 (s, 1H), 8.29 (t, J = 8.8 Hz, 1H), 8.09 (d, J = 8.8 Hz,
1H), 7.80 (s, 1H), 7.09 (s, 1H), 6.18 (s, 2H), 4.93 (t, J = 6.4 Hz, 2H),
4.10 (s, 3H), 3.21 (t, J = 6.4 Hz, 2H); 13C NMR d: 150.8, 148.5,
146.3, 144.7, 138.9, 133.1, 131.6, 127.9, 125.3, 121.0, 120.9,
117.7, 109.2, 106.2, 102.9, 58.2, 56.1, 49.3, 26.9; IR (KBr)
c: 3187,
5.4.11. 2-Hydroxyl-3,9,10-trimethoxy-5,6-
3004, 1623, 1611, 1504, 1393, 1367, 1283, 1225, 1103,
1035 cmÀ1; HRMS-ESI: Calcd for C19H15NO3FCl (MÀCl)+ 324.1036,
found 324.1019.
dihydroisoquinolino[2,1-b]isoquinolin-7-ium (7k)
Yield: 43%; Brown solid; mp 219À221 °C (decomp); 1H NMR d:
9.35 (s, 1H), 8.57 (s, 1H), 7.97 (s, 1H), 7.56 (s, 1H), 7.50 (s, 1H), 7.44
(s, 1H), 6.54 (s, 1H), 4.66 (t, J = 6.0 Hz, 2H), 4.03 (s, 3H), 3.96 (s, 3H),
3.85 (s, 3H), 3.04 (t, J = 6.0 Hz, 2H); 13C NMR d: 150.6, 150.2, 146.4,
145.4, 143.5, 137.8, 133.1, 127.0, 126.7, 123.6, 121.3, 119.5, 119.1,
5.4.6. 5,6-Dihydro-10-methoxy-12-bromo-benzo[g]-1,3-
benzodioxolo[5,6-a] quinolizinium-9-ol (7f)
Yield: 51%; Brown solid; mp 135À137 °C (decomp); 1H NMR d:
10.00 (s, 1H), 8.41 (s, 1H), 8.38 (s, 1H), 7.89 (s, 1H), 7.07 (s, 1H),
6.17 (s, 2H), 4.89 (t, J = 6.0 Hz, 2H), 4.06 (s, 3H), 3.17 (t, J = 6.0 Hz,
2H); 13C NMR d: 156.3, 153.0, 151.0, 138.7, 136.4, 133.1, 130.3,
129.2, 127.3, 126.9, 123.3, 122.1, 117.0, 107.6, 104.5, 59.6, 55.3,
112.2, 111.4, 61.9, 57.0, 55.9, 55.5, 25.9; IR (KBr) c: 2939, 2840,
1604, 1363, 1274, 1246, 1105, 1026 cmÀ1; HRMS-ESI: Calcd for
C20H20NO4Cl (MÀCl)+ 338.1392, found 338.1361.
5.4.12. 3,10,11-Trimethoxy-2-ethoxy-5,6-
36.1, 25.6; IR (KBr)
c
: 3150, 2922, 1596, 1503, 1493, 1315, 1277,
dihydroisoquinolino[2,1-b]isoquinolin-7-ium (7l)
1261, 1097, 1031 cmÀ1
;
HRMS-ESI: calcd for C19H15NO4BrCl
Yield: 71%; Yellow solid; mp 220–222 °C; 1H NMR d: 9.56 (s,
1H), 8.86 (s, 1H), 7.71 (s, 1H), 7.66 (s, 1H), 7.60 (s, 1H), 7.07 (s,
1H), 4.78 (t, J = 6.4 Hz, 2H), 4.12 (q, J = 6.8 Hz, 2H), 4.07 (s, 3H),
3.99 (s, 3H), 3.93 (s, 3H), 3.20 (t, J = 6.4 Hz, 2H), 1.37 (t, J = 6.8 Hz,
3H); 13C NMR d: 157.4, 152.1, 150.7, 148.7, 145.5, 138.3, 136.6,
128.5, 122.0, 118.8, 117.9, 112.0, 108.5, 106.5, 105.3, 64.0, 56.6,
(MÀCl)+ 400.0184, found 400.0148.
5.4.7. 3,9,10-Trimethoxy-2-ethoxy-5,6-dihydroisoquinolino[2,1-
b]isoquinolin-7-ium (7g)
Yield: 69%; Orange solid; mp 224–226 °C; 1H NMR d: 9.89 (s,
1H), 9.08 (s, 1H), 8.20 (d, J = 9.2 Hz, 1H), 8.03 (d, J = 9.2 Hz, 1H),
7.71 (s, 1H), 7.06 (s, 1H), 4.95 (t, J = 6.0 Hz, 2H), 4.12 (q,
J = 6.8 Hz, 2H), 4.09 (s, 3H), 4.06 (s, 3H), 3.93 (s, 3H), 3.21 (t,
J = 6.0 Hz, 2H), 1.37 (t, J = 6.8 Hz, 3H); 13C NMR d: 150.7, 150.2,
148.8, 145.4, 143.6, 137.7, 133.1, 128.6, 126.8, 123.4, 121.3,
119.8, 118.8, 112.0, 108.8, 64.0, 61.9, 57.0, 56.0, 55.3, 25.9, 14.6;
56.3, 56.0, 54.6, 26.4, 14.6; IR (KBr) c: 3423, 2977, 1607, 1515,
1498, 1431, 1360, 1278, 1262, 1218, 1132 cmÀ1; HRMS-ESI: Calcd
for C22H24NO4Cl (MÀCl)+ 366.1705, found 366.1672.
5.4.13. 2,10,11-Trimethoxy-3-ethoxy-5,6-
dihydroisoquinolino[2,1-b]isoquinolin-7-ium (7m)
IR (KBr)
c
: 3338, 2978, 1605, 1525, 1511, 1456, 1364, 1274,
Yield: 86%; Yellow solid; mp 238À241 °C; 1H NMR d: 9.55 (s,
1H), 8.84 (s, 1H), 7.70 (s, 1H), 7.65 (s, 1H), 7.60 (s, 1H), 7.09 (s,
1H), 4.78 (t, J = 6.0 Hz, 2H), 4.19 (q, J = 6.8 Hz, 2H), 4.07 (s, 3H),
3.99 (s, 3H), 3.87 (s, 3H), 3.21 (t, J = 6.0 Hz, 2H), 1.40 (t, J = 6.8 Hz,
3H); 13C NMR d: 157.4, 152.2, 151.6, 147.8, 145.5, 138.4, 136.6,
128.5, 122.0, 118.9, 117.9, 111.4, 109.5, 106.5, 105.3, 64.2, 56.6,
1244, 1215, 1109 cmÀ1; HRMS-ESI: alcd for C22H24NO4Cl (MÀCl)+
366.1705, found 366.1675.
5.4.8. 2,9,10-Trimethoxy-3-ethoxy-5,6-dihydroisoquinolino[2,1-
b]isoquinolin-7-ium (7h)
Yield: 76%; Orange solid; mp 208À211 °C (decomp); 1H NMR d:
9.88 (s, 1H), 9.03 (s, 1H), 8.20 (d, J = 9.2 Hz, 1H), 8.02 (d, J = 9.2 Hz,
1H), 7.70 (s, 1H), 7.08 (s, 1H), 4.93 (t, J = 5.6 Hz, 2H), 4.20 (q,
J = 6.8 Hz, 2H), 4.09 (s, 3H), 4.07 (s, 3H), 3.86 (s, 3H), 3.23 (t,
J = 5.6 Hz, 2H), 1.40 (t, J = 6.8 Hz, 3H); 13C NMR d: 151.6, 150.2,
147.9, 145.4, 143.6, 137.7, 133.1, 128.6, 126.7, 123.4, 121.3,
119.8, 118.9, 111.3, 109.8, 64.4, 61.9, 57.0, 55.8, 55.3, 25.9, 14.6;
56.3, 55.8, 54.6, 26.1, 14.7; IR (KBr) c: 3389, 2975, 1607, 1515,
1497, 1465, 1430, 1360, 1277, 1216, 1128 cmÀ1; HRMS-ESI: Calcd
for C22H24NO4Cl (MÀCl)+ 366.1705, found 366.1678.
5.4.14. 3,4,10,11-Tetramethoxy-5,6-dihydroisoquinolino[2,1-
b]isoquinolin-7-ium (7n)
Yield: 78%; Bright green solid; mp 241À243 °C; 1H NMR d: 9.60
(s, 1H), 8.80 (s, 1H), 7.93 (d, J = 8.8 Hz, 1H), 7.27 (d, J = 8.8 Hz, 1H),
7.71 (s, 1H), 7.66 (s, 1H), 4.78 (t, J = 6.4 Hz, 2H), 4.06 (s, 3H), 3.99 (s,
3H), 3.93 (s, 3H), 3.79 (s, 3H), 3.25 (t, J = 6.4 Hz, 2H); 13C NMR d:
157.4, 154.6, 152.2, 145.4, 145.0, 138.3, 136.6, 128.9, 122.3,
122.2, 120.1, 118.1, 112.4, 106.5, 105.4, 60.3, 56.7, 56.3, 56.1,
IR (KBr)
c: 3358, 2975, 2942, 1605, 1511, 1455, 1363, 1274,
1242, 1108 cmÀ1
;
HRMS-ESI: Calcd for C22H24NO4Cl (MÀCl)+
366.1705, found 366.1676.
5.4.9. 1,2,3,9,10-Pentamethoxy-5,6-dihydroisoquinolino[2,1-
b]isoquinolin-7-ium (7i)
54.2, 20.7; IR (KBr) c: 3386, 3016, 2946, 1600, 1501, 1477, 1460,
Yield: 39%; Brown solid; mp 191À194 °C (decomp); 1H NMR d:
9.93 (s, 1H), 9.00 (s, 1H), 8.18 (s, 2H), 6.99 (s, 1H), 4.90 (t, J = 6.4 Hz,
2H), 4.09 (s, 3H), 4.07 (s, 3H), 3.90 (s, 3H), 3.87 (s, 3H), 3.83 (s, 3H),
3.19 (t, J = 6.4 Hz, 2H); 13C NMR d: 155.4, 152.1, 150.5, 145.4, 143.5,
141.2, 135.0, 133.2, 132.7, 126.5, 124.1, 123.0, 121.1, 113.0, 107.7,
1432, 1348, 1292, 1279, 1216, 1149, 1065, 1001 cmÀ1; HRMS-
ESI: Calcd for C21H22NO4Cl (MÀCl)+ 352.1549, found 352.1521.
5.5. Biology
61.9, 61.2, 60.7, 57.1, 56.2, 55.4, 27.3; IR (KBr)
c
: 3374, 2941, 2839,
5.5.1. Cell culture
1596, 1504, 1461, 1358, 1273, 1113, 1041 cmÀ1; HRMS-ESI: Calcd
HepG2 cells were maintained in minimum essential medium
(Gibco-Invitrogen, Grand Island, NY) containing 10% fetal bovine
serum (FBS) and antibiotics. Cells were cultured in an atmosphere
of 5% CO2 at 37 °C. One day before experiment, cells were trypsin-
ized and allowed to grow to about 70% confluence.
for C22H24NO5Cl (MÀCl)+ 382.1654, found 382.1658.
5.4.10. 3,4,9,10-Tetramethoxy-5,6-dihydroisoquinolino[2,1-
b]isoquinolin-7-ium (7j)
Yield: 72%; Bright yellow solid; mp 216–218 °C (decomp); 1H
NMR d: 9.94 (s, 1H), 9.01 (s, 1H), 8.21 (d, J = 9.2 Hz, 1H), 8.07 (d,
J = 9.2 Hz, 1H), 8.00 (d, J = 8.8 Hz, 1H), 7.28 (d, J = 8.8 Hz, 1H),
4.96 (t, J = 6.4 Hz, 2H), 4.10 (s, 3H), 4.08 (s, 3H), 3.94 (s, 3H), 3.81
5.5.2. Western blot
After treatment, cells were harvested; total protein content of
the cells was extracted as previously reported.14 The whole cell