J. Jin et al. / European Journal of Medicinal Chemistry 62 (2013) 280e288
287
(1H, m), 3.10 (1H, m), 2.98 (1H, d, J ¼ 18 Hz), 2.61 (1H, m), 2.47 (5H,
m), 2.36 (3H, s), 2.05 (3H, m); 13C NMR (75 MHz; CDCl3; TMS)
Isomer 9b, mp: 146e148 ꢀC; 1H NMR (300 MHz; CDCl3; TMS)
d:
d:
7.60 (2H, m), 7.37 (2H, m), 6.72 (1H, s), 6.19 (1H, d, J ¼ 6.3 Hz), 5.47
(1H, m), 4.35 (1H, d, J ¼ 15.6 Hz), 4.05 (1H, d, J ¼ 6.3 Hz), 3.79 (3H, s),
3.77 (3H, s), 3.55 (3H, s), 3.33 (1H, m), 3.10 (1H, m), 2.97 (1H, d,
J ¼ 17.7 Hz), 2.62 (1H, m), 2.48 (5H, m), 2.06 (3H, m); 13C NMR
193.7,170.2,153.7,152.6,145.6,144.9,138.8,131.5,128.9,128.1,127.5,
127.3,126.1,124.1,123.4,114.4,106.1, 84.0, 61.9, 56.5, 56.2, 55.0, 53.4,
48.8, 47.3, 45.1, 42.8, 40.6, 35.3, 29.8, 21.5. ESI-MS m/z: 547
[M þ H]þ, HRESIMS m/z: found: 547.2463 [M þ H]þ (calcd for
C31H35N2O7, 547.2444).
(75 MHz; CDCl3; TMS) d: 193.6, 169.5, 152.4, 145.5, 144.9, 134.8,
130.4, 130.0 1299, 126.8, 126.7, 126.0, 124.8, 123.5, 114.2, 105.9, 84.3,
61.9, 56.2, 56.0, 54.8, 53.3, 48.6, 47.2, 47.1, 44.8, 42.5, 40.4, 35.0, 29.6,
21.3. ESI-MS m/z: 567, 569 [M þ H]þ, HRESIMS m/z: found: 567.1932
[M þ H]þ (calcd for C30H32ClN2O7, 567.1898).
5.1.4.4. Compound 7a and 7b yield 96%; isomer ratio: 7a:7b ¼ 34:66.
Isomer 7a, mp 158e160 ꢀC; 1H NMR (300 MHz; CDCl3; TMS)
d:
7.66 (2H, m), 6.92 (2H, m), 6.74 (1H, s), 6.04 (1H, d, J ¼ 5.7 Hz), 5.45
(1H, d, J ¼ 2.1 Hz), 4.38 (1H, d, J ¼ 15.6 Hz), 4.20 (1H, d, J ¼ 5.7 Hz),
3.84 (3H, s), 3.79 (3H, s), 3.78 (3H, s), 3.50 (3H, s), 3.30 (1H, m),
3.05 (1H, m), 2.82 (1H, d, J ¼ 17.7 Hz), 2.62 (2H, m), 2.47 (3H, s),
2.45 (1H, d, J ¼ 15.6 Hz), 1.95 (3H, m); 13C NMR (75 MHz; CDCl3;
5.1.4.7. Compound 10a and 10b yield 87%; isomer ratio: 10a:10b
¼ 34:66. Isomer 10a, mp 158e160 ꢀC; 1H NMR (300 MHz; CDCl3;
TMS)
d
: 7.57 (4H, m), 6.70 (1H, s), 6.10 (1H, d, J ¼ 6.0 Hz), 5.45 (1H, d,
J ¼ 2.1 Hz), 4.37 (1H, d, J ¼ 15.6 Hz), 4.20 (1H, d, J ¼ 6.0 Hz), 3.79 (3H,
s), 3.77 (3H, s), 3.50 (3H, s), 3.30 (1H, m), 3.05 (1H, m), 2.81 (1H, d,
J ¼ 17.7 Hz), 2.60 (2H, m), 2.46 (3H, s), 2.45 (1H, d, J ¼ 15.6 Hz), 1.95
TMS) d: 193.5, 170.1, 161.3, 152.8, 152.4, 145.3, 144.6, 128.4, 127.1,
125.9, 123.0, 120.6, 114.2, 106.0, 84.0, 62.0, 56.2, 56.0, 55.3, 54.7,
53.2, 49.1, 47.2, 45.2, 42.6, 40.5, 35.6, 21.4. ESI-MS m/z: 563
[M þ H]þ, HRESIMS m/z: found: 563.2416 [M þ H]þ (calcd for
C31H35N2O8, 563.2393).
(3H, m); 13C NMR (75 MHz; CDCl3; TMS)
d: 193.6, 169.7, 152.5, 152.4,
145.4, 144.9, 132.0, 128.2, 127.2, 126.5, 126.0, 124.8, 123.1, 114.3, 105.9,
84.6, 61.4, 56.1, 56.0, 54.7, 53.3, 49.1, 47.2, 45.2, 42.6, 40.5, 35.5, 21.5.
ESI-MS m/z: 611, 613 [M þ H]þ, HRESIMS m/z: found: 611.1433
[M þ H]þ (calcd for C30H32BrN2O7, 611.1393).
Isomer 7b, mp 150e152 ꢀC; 1H NMR (300 MHz; CDCl3; TMS)
d:
7.60 (2H, d, J ¼ 9 Hz), 6.91 (2H, d, J ¼ 9 Hz), 6.76 (1H, s), 6.14 (1H, d,
J ¼ 6.3 Hz), 5.48 (1H, d, J ¼ 1.8 Hz), 4.35 (1H, d, J ¼ 15.6 Hz), 4.04 (1H,
d, J ¼ 6.3 Hz), 3.83 (3H, s), 3.79 (3H, s), 3.77 (3H, s), 3.56 (3H, s), 3.33
(1H, m), 3.09 (1H, m), 2.97 (1H, d, J ¼ 18.3 Hz), 2.61 (1H, m), 2.47
Isomer 10b, mp 153e155 ꢀC; 1H NMR (300 MHz; CDCl3; TMS)
d:
7.53 (4H, m), 6.72 (1H, s), 6.19 (1H, d, J ¼ 6.3 Hz), 5.47 (1H, d,
J ¼ 2.1 Hz), 4.35 (1H, d, J ¼ 15.6 Hz), 4.05 (1H, d, J ¼ 6.3 Hz), 3.78 (3H,
s), 3.77 (3H, s), 3.55 (3H, s), 3.33 (1H, m), 3.10 (1H, m), 2.97 (1H, d,
J ¼ 18.3 Hz), 2.61 (1H, m), 2.47 (5H, m), 20.2 (3H, m); 13C NMR
(5H, s), 2.06 (3H, m); 13C NMR (75 MHz; CDCl3; TMS)
d: 193.6, 170.1,
161.2, 152.9, 152.3, 145.4, 144.8, 128.3, 127.2, 125.9, 123.3, 120.5,
114.3, 114.2, 105.9, 83.6, 61.9, 56.2, 55.9, 55.3, 54.8, 53.2, 48.7, 47.1,
44.9, 42.6, 40.4, 35.1, 29.6, 21.3. ESI-MS m/z: 563 [M þ H]þ, HRESIMS
m/z: found: 563.2428 [M þ H]þ (calcd for C31H35N2O8, 563.2393).
(75 MHz; CDCl3; TMS) d: 193.7, 169.6, 152.6, 152.3, 145.6, 145.1,
132.0, 128.2, 127.1, 126.5, 126.2, 124.7, 123.7, 114.5, 105.8, 84.2, 61.3,
56.0, 55.9, 54.8, 53.3, 48.7, 47.0, 44.8, 42.5, 40.5, 35.0, 29.6, 21.3. ESI-
MS m/z: 611, 613 [M þ H]þ, HRESIMS m/z: found: 611.1361 [M þ H]þ
(calcd for C30H32BrN2O7, 611.1393).
5.1.4.5. Compound 8a and 8b yield 81%; isomer ratio: 8a:8b ¼ 28:72.
Isomer 8a, mp 157e159 ꢀC; 1H NMR (300 MHz; CDCl3; TMS)
d: 7.66
(2H, m), 7.39 (2H, m), 6.73 (1H, s), 6.09 (1H, d, J ¼ 5.7 Hz), 5.44 (1H,
d, J ¼ 2.1 Hz), 4.38 (1H, d, J ¼ 15.6 Hz), 4.25 (1H, d, J ¼ 5.7 Hz), 3.79
(6H, s), 3.51 (4H, m), 3.28 (1H, m), 2.95 (1H, m), 2.78 (2H, m), 2.59
(3H, s), 2.58 (1H, d, J ¼ 15.6 Hz), 2.04 (3H, m); 13C NMR (75 MHz;
5.1.4.8. Compound 11a and 11b yield 86%; isomer ratio 11a:11b
¼ 31:69. Isomer 11a, mp 157e159 ꢀC; 1H NMR (300 MHz; CDCl3;
TMS) d: 7.91 (1H, m), 7.59 (2H, m), 7.29 (1H, m), 6.69 (1H, s), 6.10
(1H, d, J ¼ 6.0 Hz), 5.44 (1H, d, J ¼ 1.8 Hz), 4.37 (1H, d, J ¼ 15.6 Hz),
4.20 (1H, d, J ¼ 6.0 Hz), 3.80 (3H, s), 3.78 (3H, s), 3.51 (3H, s), 3.32
(1H, m), 3.08 (1H, m), 2.83 (1H, d, J ¼ 17.7 Hz), 2.63 (2H, m), 2.48
(3H, s), 2.46 (1H, d, J ¼ 15.6 Hz), 1.96 (3H, m); 13C NMR (75 MHz;
CDCl3; TMS) d: 192.9, 169.7, 152.6, 152.5, 145.6, 144.8, 136.6, 129.1,
128.1, 126.9, 126.7, 124.8, 122.4, 112.9, 106.4, 84.4, 61.5, 56.8, 56.1,
54.9, 53.4, 48.6, 47.5, 44.0, 42.2, 40.0, 34.5, 21.8. ESI-MS m/z: 567,
569 [M þ H]þ, HRESIMS m/z: found: 567.1904 [M þ H]þ (calcd for
C30H32ClN2O7, 567.1898).
CDCl3; TMS) d: 193.5, 169.6, 152.4, 152.2, 145.4, 144.8, 133.3, 130.3,
130.2, 129.7, 126.5, 125.9, 125.3, 123.0, 122.9, 114.1, 105.9, 84.7, 61.3,
56.2, 56.0, 54.8, 53.3, 49.0, 47.2, 45.0, 42.5, 40.4, 35.4, 21.5. ESI-MS
m/z: 611, 613 [M þ H]þ, HRESIMS m/z: found: 611.1367 [M þ H]þ
(calcd for C30H32BrN2O7, 611.1393).
Isomer 8b, mp 151e153 ꢀC; 1H NMR (300 MHz; CDCl3; TMS)
d:
7.60 (2H, m), 7.38 (2H, m), 6.72 (1H, s), 6.19 (1H, d, J ¼ 6.3 Hz), 5.47
(1H, d, J ¼ 2.1 Hz), 4.35 (1H, d, J ¼ 15.6 Hz), 4.05 (1H, d, J ¼ 6.3 Hz),
3.78 (3H, s), 3.77 (3H, s), 3.55 (3H, s), 3.32 (1H, m), 3.08 (1H, m), 2.97
(1H, d, J ¼ 18 Hz), 2.60 (1H, m), 2.46 (5H, m), 2.01 (3H, m); 13C NMR
Isomer 11b, mp 149e151 ꢀC; 1H NMR (300 MHz; CDCl3; TMS)
d:
7.85 (1H, m), 7.56 (2H, m), 7.28 (1H, m), 6.74 (1H, s), 6.19 (1H, d,
J ¼ 6.0 Hz), 5.48 (1H, d, J ¼ 1.8 Hz), 4.36 (1H, d, J ¼ 15.6 Hz), 4.07 (1H,
d, J ¼ 6.0 Hz), 3.80 (3H, s), 3.79 (3H, s), 3.56 (4H, m), 3.27 (1H, m),
3.02 (1H, d, J ¼ 18.3 Hz), 2.79 (1H, m), 2.63 (1H, m), 2.56 (3H, s), 2.52
(75 MHz; CDCl3; TMS) d: 193.6, 169.7, 152.5, 152.4, 145.4, 144.8,
136.5, 129.1, 128.0, 126.8, 126.6, 126.0, 123.3, 114.2, 105.8, 84.2, 61.4,
56.2, 26.0, 54.8, 53.3, 48.6, 47.1, 44.9, 42.6, 40.4, 35.1, 21.3. ESI-MS m/
z: 567, 569 [M þ H]þ, HRESIMS m/z: found: 567.1932 [M þ H]þ
(calcd for C30H32ClN2O7, 567.1898).
(1H, d, J ¼ 15.6 Hz), 2.08 (3H, m); 13C NMR (75 MHz; CDCl3; TMS)
d:
193.2, 169.5, 152.5, 152.3, 145.6, 145.0, 133.4, 130.3, 130.0, 129.7,
126.8, 125.3, 125.1, 123.0, 122.9, 113.3, 106.1, 84.1, 61.1, 56.4, 56.0,
54.9, 53.4, 48.2, 47.2, 43.9, 42.1, 40.1, 34.4, 29.6, 21.5. ESI-MS m/z:
611, 613 [M þ H]þ, HRESIMS m/z: found: 611.1412 [M þ H]þ (calcd
for C30H32BrN2O7, 611.1393).
5.1.4.6. Compound 9a and 9b yield 84%; isomer ratio: 9a:9b ¼ 43:57.
Isomer 9a, mp: 152e154 ꢀC; 1H NMR (300 MHz; CDCl3; TMS)
d:
7.66 (2H, m), 7.39 (2H, m), 6.71 (1H, s), 6.09 (1H, d, J ¼ 6.0 Hz), 5.43
(1H, s), 4.37 (1H, d, J ¼ 15.6 Hz), 4.22 (1H, d, J ¼ 6.0 Hz), 3.79 (3H, s),
3.78 (3H, s), 3.51 (3H, s), 3.38 (1H, m), 3.15 (1H, m), 2.92 (1H, d,
J ¼ 17.7 Hz), 2.69 (2H, m), 2.52 (3H, s), 2.47 (1H, d, J ¼ 15.6 Hz), 1.99
5.2. Biological activity
(3H, m); 13C NMR (75 MHz; CDCl3; TMS)
d
: 193.2, 169.6, 152.5,
5.2.1. Cultivation of fibroblasts
145.4, 144.8, 134.9, 130.5, 130.1, 130.0, 126.8, 126.7, 125.5, 124.9,
122.8, 113.6, 106.1, 84.6, 61.3, 56.4, 56.1, 54.8, 53.3, 48.8, 47.3, 44.6,
42.4, 40.3, 35.0, 29.6, 21.6. ESI-MS m/z: 567, 569 [M þ H]þ, HRE-
SIMS m/z: found: 567.1906 [M þ H]þ (calcd for C30H32ClN2O7,
567.1898).
The mouse embryonic fibroblast (NIH/3T3) cells were cultured in
Dulbecco’s modified Eagle’s medium (DMEM) (Invitrogen, Carlsbad,
CA) supplemented with 10% fetal bovine serum (FBS) (Invitrogen),
100 units/mL penicillin, and 100
mg/mL streptomycin and incubated
at 37 ꢀC in a humidified atmosphere containing 5% CO2.