M.M. Gamal El-Din et al. / European Journal of Medicinal Chemistry 90 (2015) 45e52
51
(
2
1
1
m, 2H), 6.87 (d, 2H, J ¼ 9.2 Hz), 6.78 (d, 1H, J ¼ 8.2 Hz), 5.32 (s, 2H),
.34 (s, 3H); 13C NMR (100 MHz, CDCl
166.0, 163.2, 161.9, 154.2,
51.7, 143.1 (d, JCF ¼ 11 Hz), 132.2 (d, JCF ¼ 8 Hz), 130.2, 129.4, 129.1,
1.07 (t, 2H, J ¼ 8.2 Hz); 13C NMR (100 MHz, CDCl
3
)
d
166.0, 162.7,
3
)
d
161.4, 154.5, 151.9, 142.8 (d, JCF ¼ 11 Hz), 132.7 (d, JCF ¼ 9 Hz), 128.9,
117.9, 116.8 (d, JCF ¼ 3 Hz), 114.6, 110.4, 110.2, 61.9, 55.5, 53.4, 17.2,
12.8.
27.2, 123.3, 118.1 (d, JCF ¼ 3 Hz), 117.5, 114.3, 111.4, 111.2, 61.9, 55.6;
MS [m/z þ Na] 478.0.
Compounds 1gek were prepared with the same method from
b and the appropriate sulfonyl chlorides.
4.5.17. N-(3-fluoro-4-((5-(4-methoxyphenyl)-1,3,4eoxadiazol-2-
yl)methoxy)phenyl)benzenesulfonamide (1n)
6
ꢁ
1
3
Yield 30%; mp 138e140 C; H NMR (400 MHz, CDCl ) d 8.01 (d,
4
3
.5.10. N-(4-((5-(4-Chlorophenyl)-1,3,4eoxadiazol-2-yl)methoxy)-
-fluorophenyl)methanesulfonamide (1g)
Yield 30%; mp 130e132 C; H NMR (400 MHz, CD OD) d 8.05 (d,
3
2H, J ¼ 8.2 Hz), 7.75 (d, 2H, J ¼ 8.2 Hz), 7.50 (s, 1H), 7.48e7.35 (m,
2H), 6.92e7.09 (m, 4H), 5.38 (s, 2H), 7.81e7.71 (m, 1H), 5.32 (s, 2H),
3.89 (s, 3H); MS [m/z þ Na] 478.2.
ꢁ
1
2
1
H, J ¼ 6.8 Hz), 7.66e7.58 (m, 3H), 7.29 (t, 1H, J ¼ 8.4 Hz), 7.13 (dd,
H, J ¼ 2.4, J ¼ 12.4 Hz), 7.0e7.03 (m, 1H), 5.47 (s, 2H), 2.97 (s, 3H);
4.5.18. 4-Chloro-N-(3-fluoro-4-((5-(4-methoxyphenyl)-
MS [m/z þ Na] 419.9.
1,3,4eoxadiazol-2-yl)methoxy)phenyl)benzenesulfonamide (1o)
ꢁ
1
3
Yield 32%; mp 164e166 C; H NMR (400 MHz, CDCl ) d 8.01 (d,
4
3
.5.11. N-(4-((5-(4-Chlorophenyl)-1,3,4eoxadiazol-2-yl)methoxy)-
-fluorophenyl)benzenesulfonamide (1h)
2H, J ¼ 7.6 Hz), 7.69 (d, 2H, J ¼ 8.4 Hz), 7.39 (d, 2H, J ¼ 8.4 Hz),
7.06e7.02 (m, 4H), 6.81 (d, 1H, J ¼ 7.6 Hz), 5.35 (s, 2H), 3.89 (s, 3H);
ꢁ
1
13
Yield 27%; mp 170e172 C; H NMR (400 MHz, CD
3
OD)
d
8.05 (d,
3
C NMR (100 MHz, CDCl ) d 166.1, 162.8, 161.2, 154.3, 150.6, 144.0,
2
H, J ¼ 8 Hz), 7.74 (d, 2H, J ¼ 6.8 Hz), 7.63e7.58 (m, 3H), 7.50 (d, 2H,
139.8 (d, JCF ¼ 11 Hz), 137.2, 131.3 (d, JCF ¼ 9 Hz), 129.4, 129.0, 128.6,
127.2, 118.5 (d, JCF ¼ 4 Hz), 117.4, 115.6, 114.6, 112.0, 111.7, 61.7, 55.5;
MS [m/z þ Na] 512.0.
J ¼ 8 Hz), 7.15 (t, 2H, J ¼ 8.8 Hz), 6.97 (dd,1H, J ¼ 2.4, 2.4 Hz), 6.84 (d,
1
H, J ¼ 7.6 Hz), 5.40 (s, 2H); MS [m/z þ Na] 482.0.
4
.5.12. 4-Chloro-N-(4-((5-(4-chlorophenyl)-1,3,4eoxadiazol-2-yl)
4.5.19. N-(3-fluoro-4-((5-(4-methoxyphenyl)-1,3,4eoxadiazol-2-
yl)methoxy)phenyl)-4-methylbenzenesulfonamide (1p)
methoxy)-3-fluorophenyl)benzenesulfonamide (1i)
ꢁ
1
ꢁ
1
Yield 28%; mp 173e175 C; H NMR (400 MHz, CD
3
OD)
d
7.93 (d,
Yield 42%; mp 136e138 C; H NMR (400 MHz, CDCl ) d 8.99 (d,
3
2
H, J ¼ 8.4 Hz), 7.58 (d, 2H, J ¼ 8.4 Hz), 7.50 (d, 2H, J ¼ 8.4 Hz), 7.50
2H, J ¼ 8.8 Hz), 7.63 (d, 2H, J ¼ 10.8 Hz), 7.20 (d, 2H, J ¼ 7.6 Hz),
7.03e6.98 (m, 4H), 6.77 (d, 1H, J ¼ 9.6 Hz), 5.32 (s, 2H), 3.87 (s, 3H),
(
2
d, 2H, J ¼ 8.4 Hz), 7.05 (t, 2H, J ¼ 8.8 Hz), 6.75 (dd, 1H, J ¼ 2.4,
13
.4 Hz), 6.69e6.71 (m, 1H), 5.29 (s, 2H); MS [m/z þ Na] 516.0.
3
2.35 (s, 3H); C NMR (100 MHz, CDCl ) d 165.9, 162.7, 161.4, 154.1,
151.6, 144.0, 143.1 (d, JCF ¼ 11 Hz), 135.8, 132.1 (d, JCF ¼ 9 Hz), 129.7,
4
3
.5.13. N-(4-((5-(4-Chlorophenyl)-1,3,4eoxadiazol-2-yl)methoxy)-
-fluorophenyl)-4-methylbenzenesulfonamide (1j)
128.9, 127.2, 118.1 (d, JCF ¼ 4 Hz), 117.4, 115.7, 114.6, 111.4, 111.2, 61.8,
55.5, 21.5; MS [m/z þ Na] 492.0.
ꢁ
1
Yield 43%; mp 152e154 C; H NMR (400 MHz, CDCl
3
) d 8.00 (d,
2
H, J ¼ 8 Hz), 7.64 (d, 2H, J ¼ 8 Hz), 7.49 (d, 2H, J ¼ 8 Hz), 7.21 (d, 2H,
4.5.20. N-(3-fluoro-4-((5-(4-methoxyphenyl)-1,3,4eoxadiazol-2-
yl)methoxy)phenyl)-4-methoxybenzenesulfonamide (1q)
J ¼ 8 Hz), 7.03e6.98 (m, 2H), 6.80e6.78 (m, 1H), 5.32 (s, 2H), 2.36 (s,
13
ꢁ
1
3
1
1
H); C NMR (100 MHz, CDCl
43.0 (d, JCF ¼ 11 Hz), 138.8, 132.2 (d, JCF ¼ 9 Hz), 129.8, 129.6, 128.4,
27.2, 121.7, 117.9 (d, JCF ¼ 3 Hz), 117.5, 111.3, 111.1, 61.8, 21.5; MS [m/
z þ Na] 495.9.
3
)
d
165.2, 162.0, 154.1, 151.7, 144.1,
Yield 43%; mp 124e126 C; H NMR (300 MHz, CDCl
3
) d 8.99 (d,
2H, J ¼ 8.8 Hz), 7.68 (d, 2H, J ¼ 12 Hz), 7.47 (s, 1H), 7.10e6.95 (m,
13
4H), 6.87 (d, 2H,
NMR (100 MHz, CDCl
J
J
d
¼ 12 Hz), 5.31 (s, 2H), 3.87 (s, 3H), 3.81 (s, 3H);
165.9, 163.2, 162.7, 154.1, 151.7, 143.1 (d,
CF ¼ 11 Hz), 132.2 (d, JCF ¼ 9 Hz), 130.3, 129.4, 128.9, 118.0 (d,
CF ¼ 3 Hz), 117.4, 115.7, 114.6, 114.3, 111.4, 111.3, 61.9, 55.5, 55.4.
C
3
) d
4
3
.5.14. N-(4-((5-(4-Chlorophenyl)-1,3,4eoxadiazol-2-yl)methoxy)-
-fluorophenyl)-4-methoxybenzenesulfonamide (1k)
ꢁ
1
Yield 40%; mp 146e148 C; H NMR (400 MHz, CDCl
3
)
d
8.01 (d,
4.6. Cancer cell line screening at the NCI
2
H, J ¼ 8 Hz), 7.68 (d, 2H, J ¼ 8 Hz), 7.51 (d, 2H, J ¼ 8.6 Hz), 7.04e6.97
(
(
1
1
m, 2H), 6.89 (d, 2H, J ¼ 8 Hz), 6.76 (d, 2H, J ¼ 8 Hz), 5.32 (s, 2H), 3.83
s, 3H); 13C NMR (100 MHz, CDCl
163.5, 161.6, 160.3, 152.5, 150.0,
41.3 (d, JCF ¼ 11 Hz), 136.9, 130.7 (d, JCF ¼ 9 Hz), 128.5, 127.8, 126.7,
Screening against the cancer cell lines was carried out at the
National Cancer Institute (NCI), Bethesda, Maryland, USA, applying
the standard protocol of the NCI [27]. The human cell lines are
grown in RPMI 1640 medium containing 5% fetal bovine serum and
3
) d
19.9, 116.3 (d, JCF ¼ 3 Hz), 115.8, 112.6, 109.6, 60.1, 53.9; MS [m/
z þ Na] 508.0.
2 mM
L-glutamine. For a typical screening experiment, cells are
Compounds 1leq were prepared with the same method from 6c
and the appropriate sulfonyl chlorides.
inoculated into 96-well microtiter plates in 100
mL at plating den-
sities ranging from 5000 to 40,000 cells/well depending on the
doubling time of individual cell lines. After cell inoculation, the
ꢁ
4.5.15. N-(3-fluoro-4-((5-(4-methoxyphenyl)-1,3,4eoxadiazol-2-
microtiter plates are incubated at 37 C, 5% CO
2
, 95% air and 100%
yl)methoxy)phenyl)methanesulfonamide (1l)
relative humidity for 24 h prior to addition of experimental drugs.
After 24 h, two plates of each cell line are fixed in situ with TCA, to
represent a measurement of the cell population for each cell line at
ꢁ
1
Yield 36%; mp 122e124 C; H NMR (400 MHz, CDCl
3
) d 8.01 (d,
2
5
d
H, J ¼ 8 Hz), 7.44 (s, 1H), 7.15 (t, 1H, J ¼ 8 Hz), 7.02e6.96 (m, 3H),
13
.36 (s, 2H), 3.88 (s, 3H), 2.99 (s, 3H); C NMR (100 MHz, CDCl
3
)
z
the time of drug addition (T ). Experimental drugs are solubilized in
166.0, 162.8, 161.3, 154.5, 151.9, 143.1 (d, JCF ¼ 10 Hz), 132.4 (d,
dimethyl sulfoxide at 400-fold the desired final maximum test
concentration and stored frozen prior to use. At the time of drug
addition, an aliquot of frozen concentrate is thawed and diluted to
twice the desired final maximum test concentration with complete
J
CF ¼ 9 Hz), 129.0, 117.9, 117.4 (d, JCF ¼ 4 Hz), 114.6, 110.9, 110.7, 61.9,
5
5.5, 39.3.
4
.5.16. N-(3-fluoro-4-((5-(4-methoxyphenyl)-1,3,4eoxadiazol-2-
medium containing 50
or 1/2 log serial dilutions are made to provide a total of five drug
concentrations plus control. Aliquots of 100 L of these different
drug dilutions are added to the appropriate microtiter wells already
containing 100 L of medium, resulting in the required final drug
mg/mL gentamicin. Additional four, 10-fold
yl)methoxy)phenyl)propane-1-sulfonamide (1m)
ꢁ
1
Yield 38%; mp 132e134 C; H NMR (400 MHz, CDCl
3
)
d
8.10 (d,
m
2
5
H, J ¼ 8.2 Hz), 7.65 (s, 1H), 7.08e7.22 (m, 2H), 7.07e6.98 (m, 3H),
.38 (s, 2H), 3.92 (s, 3H), 3.15 (t, 2H, J ¼ 8.2 Hz), 2.00e1.82 (m, 2H),
m