N.J. Gumede, et al.
BioorganicChemistryxxx(xxxx)xxxx
57.1, 60.8, 79.6, 104.9, 105.0, 108.9, 109.4, 127.7, 127.9, 128.7,
129.2, 139.6, 140.2, 147.9, 171.3, 173.1 MS (APCI): m/z = 328
[M++H].
1H, aromatic), 7.47–7.51 (m, 1H, aromatic), 7.56–758 (1H, d,
3J = 7.78 Hz, aromatic), 7.83–7.85 (d, 1H, 3J = 7.63 Hz, aromatic),
7.94 (s, 1H, aromatic), 8.17–8.19 (m, 1H, pyridine-H), 8.30–8.32 (m,
1H, aromatic), 8.92 (d, 1H, 4J = 2.31 Hz, pyridine-H), 10.31 (s, 1H,
amide N-H), 10.47 (s, 1H, amide N-H); 13C NMR (100 MHz, DMSO-D6),
δ 67.2, 79.6, 100.7, 106.8, 116.7, 123.7, 126.9, 127.0, 127.7, 128.8,
130.9, 134.8, 136.2, 141.3, 142.4, 144.6, 144.9, 145.5, 164.3, 166.3;
MS (APCI): m/z = 375 [M++H].
4.1.7. 1-(4-butanamidophenyl)-1-oxopropan-2-yl-1H-indazole-3-
carboxylate (NCE6)
The reaction was carried out in 8 ml glass vial. The reactants were
loaded in view that 1.0 equivalent is equal to 1.4 mmol of the com-
pound. A vial was charged with 1H-Indazole-3-carboxylic acid (1.2
equiv.), DMF (2 ml), and DIPEA (1.2 equiv.). To the stirred mixture N-
[4-(2-Chloro-propionyl)-phenyl]-butyramide (1.0 equiv.) was added.
The vial was capped and heated under stirring for an 1hr. After 30 min,
the reaction mixture became clear and heating was continued for the
next 6hrs at 100 °C. Then the vial was cooled, diluted with water, and
extracted with chloroform. The combined organic layers were dried
over concentrated sodium sulfate. The crude product was purified with
CombiFlash chromatography on silica gel. The average yield was 50%;
mp, 205–206 °C; 1H NMR (400 MHz, DMSO-D6) δ 0.91–0.94 (t, 3H,
3J = 7.36 Hz, -CH3), 1.63–1.67 (m, 5H, -CH2 and -CH3), 2.32–2.36 (t,
2H, 3J = 7.46 Hz, -CH2), 6.31–6.41 (q, 1H, 3J = 6.86 Hz, -CH),
7.32–7.36 (m, 1H, aromatic), 7.46–7.50 (m, 1H, aromatic), 7.69–7.71
(d, 1H, 3J = 8.23 Hz, aromatic), 7.79–7.81 (d, 2H, 3J = 8.85 Hz,
aromatic), 8.05–8.09 (m, 3H, aromatic), 10.31 (s, 1H, -NH), 14.04 (s,
1H, -COOH): 13C NMR (100 MHz, DMSO-D6), δ 14.1, 17.8, 18.9, 72.1,
111.7, 118.9, 121.4, 122.7, 123.5, 127.2, 128.6, 130.3, 135.0, 141.4,
144.8, 161.9, 172.4, 195.5; MS (APCI): m/z = 380 [M++H].
4.1.12. N-(2-hydroxyphenyl)-3-{[(3-oxo-3,4-dihydro-2H-1,4-benzoxazin-
7-yl)amino]methyl}benzamide (NCE 8c).
NCE 8c was synthesized by using the general procedure of method
A; 192–193 °C; 1H NMR (400 MHz, DMSO-D6) δ 4.28–4.29 (d, 2H,
3J = 5.67 Hz, -CH2-NH), 4.40 (s, 2H, -CH2-O), 6.18–6.27 (m, 2H,
aromatic), 6.59–6.61 (d, 1H, 3J = 8.36 Hz, aromatic), 6.73–6.82 (m,
1H, aromatic), 6.89–6.92 (dd, 1H, 3J = 7.99 Hz and 4J = 1.23 Hz,
aromatic), 7.04–7.09 (m, 1H, aromatic), 7.44–7.48 (m, 1H, aromatic),
7.53–7.55 (d, 1H, 3J
= 7.60 Hz, aromatic), 7.67–7.69 (d, 1H,
3J = 7.78 Hz, aromatic), 7.81–7.82 (d, 1H, 3J = 7.60 Hz, aromatic),
9.49 (s, 1H, amide N-H), 10.29 (s, 1H, amide N-H); 13C NMR (100 MHz,
DMSO-D6), δ 67.3, 79.8, 100.8, 106.9, 116.4, 116.8, 117.2, 119.4,
126.1, 126.3, 126.8, 128.9, 130.9, 134.9, 141.4, 144.7. 145.6, 164.3,
165.7; MS (APCI): m/z = 390 [M++H].
4.1.13. N-{4-chloro-3-[(3-oxo-3,4-dihydro-2H-1,4-benzoxazin-7-yl)
carbamoyl]phenyl}-1H-pyrazole-4-carboxamide (NCE8d).
NCE 8d was synthesized by using the general procedure for method
B; mp, 308–309 °C; 1H NMR (400 MHz, DMSO‑d6) δ 4.56 (s, 2H, -CH2-
O), 6.85–6.87 (d, 1H, 3J = 8.48 Hz, aromatic), 7.26–7.29 (dd, 1H,
3J = 8.01 Hz and 4J = 2.18 Hz, aromatic), 7.39–7.40 (d, 1H,
4J = 2.18 Hz, aromatic), 7.50–7.52 (d, 1H, 3J = 8.75 Hz, aromatic),
7.86–7.89 (m, 2H, aromatic), 8.07 (s, 1H, pyrazoe-H), 8.38 (s, 1H,
pyrazole-H), 10.06 (s, 1H, amide N-H), 10.49 (s, 1H, amide N-H), 10.69
(s, 1H, amide N-H), 10.33 (s, 1H, pyrazole N-H); 13C NMR (100 MHz,
DMSO-D6), δ 62.3, 108.3, 114.0, 116.2, 118.0, 120.0, 122.3, 123.7,
123.8, 130.4, 134.7, 137.3, 138.8, 143.6, 161.1, 164.9, 165.0; MS
(APCI): m/z = 412 [M++H].
4.1.8. N-(2-hydroxyphenyl)-4-{3-[(2-hydroxyphenyl)carbamoyl]
phenoxy}benzamide (NCE7).
NCE 7 was synthesized by using the general procedure in method C;
mp, 266–267 °C; 1H NMR (400 MHz, DMSO‑d6) δ 6.81–6.85 (m, 2H,
aromatic), 6.91–6.93 (dd, 2H, 3J = 8.06 Hz and 4J = 1.27 Hz aro-
matic), 7.01–7.06 (m, 2H, aromatic), 7.20–7.22 (m, 4H, aromatic),
7.64–7.66 (dd, 2H, 3J = 8.06 Hz and 4J = 1.27 Hz, aromatic),
8.04–8.07 (m, 4H, aromatic) Hz δ 9.56 (s, 2H, -NH-amide); 13C NMR
(100 MHz, DMSO-D6), δ 116.5, 119.1, 119.4, 124.8, 126.2, 126.3,
130.3, 130.4, 150.0, 159.2, 164.9; MS (APCI): m/z = 340 [M++H].
4.1.9. 2-chloro-N-(3-{[(6-chloro-3-oxo-3,4-dihydro-2H-1,4-benzoxazin-7-
yl)amino]methyl}phenyl)benzamide (NCE8).
4.1.14. 3-{[(6-chloro-3-oxo-3,4-dihydro-2H-1,4-benzoxazin-7-yl)amino]
methyl}-N-(pyridin-3-yl)benzamide (NCE8e).
NCE 8 was synthesized by using the general procedure for method
B; mp, 231–232 °C; 1H NMR (400 MHz, DMSO-D6) δ 4.36 (s, 2H, -CH2-N
and 2°), 4.42 (s, 2H, -CH2-O), 4.45 (s, 1H, 2° amine N-H) 6.14 (s, 1H,
aromatic), 6.82 (s, 1H, aromatic), 7.08–7.09 (m, 1H, aromatic),
7.27–7.31 (m, 1H, aromatic), 7.42–7.74 (m, 6H, aromatic), 10.41 (s,
1H, amide N-H), 10.50 (s, 1H, amide N-H); 13C NMR (100 MHz, DMSO-
D6), δ 67.3, 79.7, 100.5, 103.6, 110.6, 111.1, 116.5, 122.7, 127.9,
129.3, 129.4, 130.1, 130.6, 131.5, 137.6, 139.8, 140.7, 141.2, 143.8,
164.2, 165.4; MS (APCI): m/z = 443 [M++H].
NCE 8e was synthesized by using the general procedure for method
A; mp, 230–231 °C; 1H NMR (400 MHz, DMSO-D6), δ 4.42(s, 2H, -CH2-
N), 4.44 (s, 2H, –CH2-O), 6.13 (s, 1H, aromatic), 6.79 (s, 1H, aromatic),
7.38–7.41 (dd, 1H, 3J = 7.89 Hz and 3J = 4.68 Hz, aromatic),
7.48–7.56 (m, 2H, aromatic), 7.83–7.84 (d, 1H, 3J = 7.69 Hz, aro-
matic), 7.92 (s, 1H, aromatic), 8.16–8.19 (m, 1H, aromatic), 8.30–8.31
(dd, 1H, 3J = 4.68 Hz and 4J = 1.43 Hz, pyridine-H), 8.90–8.91 (d, 1H,
4J = 2.17 Hz, pyridine-H), 10.41 (s, 1H, amide N-H), 10.48 (s, 1H,
amide N-H); 13C NMR (100 MHz, DMSO-D6), δ 67.2, 79.6, 100.5, 111.0,
124.0, 126.5, 126.9, 127.8, 128.9, 135.0, 136.2, 140.4, 140.9, 142.4,
143.6, 145.0, 164.2, 166.5; MS (APCI): m/z = 409 [M++H].
4.1.10. 3-{[(6-hydroxy-3-oxo-3,4-dihydro-2H-1,4-benzoxazin-7-yl)
amino]methyl}-N-(pyridin-3-yl)benzamide (NCE8a).
NCE 8a was synthesized by using the general procedure for method
A; mp, 231–232 °C; 1H NMR (400 MHz, DMSO-D6) δ 4.39 (s, 2H, -CH2-
N), 4.45 (s, 2H, -CH2-O), 4.50 (s, 1H, 2° amine N-H), 6.54 (s, 2H, aro-
matic), 7.55–7.59 (m, 1H, aromatic), 7.65–7.67 (m, 1H, aromatic),
7.94–8.11 (m, 3H, aromatic), 8.56–8.64 (m, 2H, aromatic), 9.34–9.35
(d, 1H, 4J = 2.08 Hz, pyridine-H), 10.56 (s, 1H, amide N-H), 11.23 (s,
1H, amide N-H); MS (APCI): m/z = 391 [M++H].
4.2. Computational data
Table 1 shows the 78 studied inhibitors. They have ten different
enzyme (in vitro experimental IC50 between 13 and 20000 nM collected
were converted into pIC50 data for 3D-QSAR partial least squares (PLS)
modeling purposes. The inhibitors (in decreasing order of pIC50) are
named as in the original references. They were randomly divided into
63 training (subset 1) and 15 test (subset 2) compounds for model
validation.
4.1.11. 3-{[(3-oxo-3,4-dihydro-2H-1,4-benzoxazin-7-yl)amino]methyl}-
N-(pyridin-3-yl)benzamide (NCE 8b).
NCE 8b was synthesized by using the general procedure for method
A; mp, 118–119 °C; 1H NMR (400 MHz, DMSO-D6) δ 4.29–4.30 (d, 2H,
3J = 5.45 Hz, -CH2-NH), 4.41 (s, 2H, -CH2-O), 6.17–6.25 (m, 2H,
aromatic), 6.59–6.13 (d, 1H, 3J = 8.33 Hz, aromatic), 7.34–7.41 (m,
4.2.1. Computational software program
a graphical user interface (GUI) in
12