R. He, B. Xu, L. Ping et al.
European Journal of Medicinal Chemistry 214 (2021) 113249
12.3 Hz), 159.81 (dd, JC-F ¼ 255.7, 12.9 Hz), 158.01/157.99*, 150.94,
150.39/150.16*, 147.89, 143.12/143.05*, 142.63/142.31*, 135.14/
135.11*, 134.35/134.21*, 132.33 (d, JC-F ¼ 10.9 Hz), 131.01, 129.66/
129.63*, 128.90, 125.98/125.88*, 125.83*/125.78, 125.55 (dd, JC-
135.70, 132.17 (d, JC-F ¼ 10.3 Hz), 131.56, 130.93, 129.48, 128.84,
125.81, 125.52 (dd, JC-F ¼ 14.0, 3.7 Hz), 125.21, 124.93, 122.43,
120.43, 111.99 (dd, JC-F ¼ 22.0, 3.4 Hz), 106.06 (t, JC-F ¼ 25.9 Hz),
55.36, 54.76, 53.73, 46.06, 46.03, 41.25, 20.79; HRMS: m/z calcd for
¼ 14.2, 3.8 Hz), 122.75/122.64*, 120.46, 120.32, 112.38 (dd, JC-
C
30H30F2N5O4S [M þ H]þ 594.1986, found 594.2008. HPLC (0.1%
F
¼ 24.7, 2.5 Hz), 106.31 (t, JC-F ¼ 26.6 Hz), 59.04*/58.80, 53.95/
TFA): tR ¼ 8.31 min (96.7% purity).
F
52.45*, 49.77, 37.08*/33.08, 20.81/20.71*; HRMS: m/z calcd for
C
28H27F2N4O5S [M
tR ¼ 7.35 min (97.2% purity).
þ
H]þ 569.1670, found 569.1665. HPLC:
4.1.9.5. (E)-2,4-difluoro-N-(5-(4-(4-(4-hydroxypiperidin-1-yl)-4-
oxobut-2-en-2-yl)quinolin-6-yl)-2-methoxypyridin-3-yl)benzene-
sulfonamide (14e). The compound was prepared from 12e (60 mg,
0.16 mmol) and 13 (68 mg, 0.16 mmol) according to the general
synthetic procedure B to afford the title compound as an off-white
solid (18 mg, 0.030 mmol, 19% yield). 1H NMR (400 MHz, DMSO‑d6)
4.1.9.2. (E)-2,4-difluoro-N-(2-methoxy-5-(4-(4-oxo-4-(pyrrolidin-1-
yl)but-2-en-2-yl)quinolin-6-yl)pyridin-3-yl)benzenesulfonamide
(14b). The compound was prepared from 12b (55 mg, 0.16 mmol)
and 13 (68 mg, 0.16 mmol) according to the general synthetic
procedure B to afford the title compound as an off-white solid
(28 mg, 0.050 mmol, 31% yield). 1H NMR (400 MHz, DMSO‑d6)
d
10.38 (s, 1H, NH), 8.91 (d, J ¼ 4.4 Hz, 1H, AreH), 8.41 (d, J ¼ 2.1 Hz,
1H, AreH), 8.17 (d, J ¼ 9.4 Hz, 1H, AreH), 8.05 (m, 2H, AreH), 7.95
(d, J ¼ 2.1 Hz, 1H, AreH), 7.80 (m, 1H, AreH), 7.59 (m, 1H, AreH),
7.48 (d, J ¼ 4.4 Hz, 1H, AreH), 7.23 (m, 1H, AreH), 6.37 (s, 1H, alkene
hydrogen), 4.77 (d, J ¼ 4.0 Hz,1H, OH), 4.01 (m,1H, CH), 3.77 (m, 2H,
CH2), 3.69 (s, 3H, OCH3), 3.41 (s, 1H, CH2), 3.17 (m, 1H, CH2), 2.29 (s,
3H, CH3), 1.76 (m, 2H, CH2), 1.42e1.31 (m, 2H, CH2); 13C NMR
d
10.40 (s, 1H, NH), 8.91 (d, J ¼ 3.8 Hz, 1H, AreH), 8.45 (s, 1H, AreH),
8.17 (d, J ¼ 8.7 Hz, 1H, AreH), 8.09 (m, 2H, AreH), 7.97 (s, 1H, AreH),
7.79 (m, 1H, AreH), 7.59 (m, 1H, AreH), 7.46 (d, J ¼ 3.8 Hz, 1H,
AreH), 7.24 (m, 1H, AreH), 6.29 (s, 1H, alkene hydrogen), 3.69 (s,
3H, OCH3), 3.54e3.42 (m, 4H, CH2 ꢀ 2), 2.46 (s, 3H, CH3), 1.86 (m,
(100 MHz, DMSO‑d6)
d
166.79 (dd, JC-F ¼ 252.6, 11.4 Hz), 165.34,
4H, CH2 ꢀ 2); 13C NMR (100 MHz, DMSO‑d6)
d 165.56 (dd, JC-
¼ 253.2, 12.1 Hz), 164.68, 159.82 (dd, JC-F ¼ 257.5, 13.3 Hz), 157.95,
159.82 (dd, JC-F ¼ 257.2, 13.7 Hz), 158.01, 150.92, 149.97, 147.90,
142.91, 141.77, 135.13, 134.15, 134.11, 132.32 (d, JC-F ¼ 10.4 Hz),
131.02,129.67,128.86,125.91, 125.85, 125.63 (dd, JC-F ¼ 14.8, 2.6 Hz),
122.67, 120.52, 112.36 (dd, JC-F ¼ 22.0, 3.4 Hz), 106.30 (t, JC-
F
150.91, 150.47, 147.91, 144.85, 143.07, 135.13, 134.14, 132.29 (d, JC-
¼ 10.8 Hz), 131.00, 129.56, 128.84, 125.69, 125.56 (dd, JC-F ¼ 15.8,
F
3.6 Hz), 125.07, 122.67, 120.44, 120.41, 112.38 (dd, JC-F ¼ 22.3,
3.6 Hz), 106.32 (t, JC-F ¼ 26.3 Hz), 53.95, 46.84, 45.81, 26.12, 24.34,
20.70; HRMS: m/z calcd for C29H27F2N4O4S [M þ H]þ 565.1721,
found 565.1709. HPLC: tR ¼ 9.19 min (95.4% purity).
¼ 25.9 Hz), 65.99, 53.95, 43.83, 38.88, 35.15, 34.38, 20.72; HRMS
F
(ESI) m/z calcd for C30H29F2N4O5S [M þ H]þ 595.1826, found
595.1823. HPLC: tR ¼ 7.31 min (97.8% purity).
4.1.9.3. (E)-2,4-difluoro-N-(2-methoxy-5-(4-(4-morpholino-4-
oxobut-2-en-2-yl)quinolin-6-yl)pyridin-3-yl)benzenesulfonamide
(14c). The compound was prepared from 12c (58 mg, 0.16 mmol)
and 13 (68 mg, 0.16 mmol) according to the general synthetic
procedure B to afford the title compound as an off-white solid
(22 mg, 0.038 mmol, 24% yield). 1H NMR (400 MHz, DMSO‑d6)
4.1.9.6. (Z)-2,4-difluoro-N-(2-methoxy-5-(4-(4-morpholino-4-
oxobut-2-en-2-yl)quinolin-6-yl)pyridin-3-yl)benzenesulfonamide
(14f). The compound was prepared from 12f (58 mg, 0.16 mmol)
and 13 (68 mg, 0.16 mmol) according to the general synthetic
procedure B to afford the title compound as an off-white solid
(20 mg, 0.034 mmol, 21% yield). 1H NMR (400 MHz, DMSO‑d6)
d
10.41 (s, 1H, NH), 8.91 (d, J ¼ 4.4 Hz, 1H, AreH), 8.43 (d, J ¼ 1.6 Hz,
d
10.38 (s, 1H, NH), 8.87 (d, J ¼ 4.4 Hz, 1H, AreH), 8.45 (s, 1H, AreH),
1H, AreH), 8.17 (d, J ¼ 8.8 Hz, 1H, AreH), 8.07 (d, J ¼ 8.8 Hz, 1H,
AreH), 8.06 (s, 1H, AreH), 7.96 (d, J ¼ 1.6 Hz, 1H, AreH), 7.79 (m, 1H,
AreH), 7.59 (m, 1H, AreH), 7.48 (d, J ¼ 4.4 Hz, 1H, AreH), 7.23 (m,
1H, AreH), 6.39 (s, 1H, alkene hydrogen), 3.68 (s, 3H, OCH3),
3.52e3.66 (m, 8H, CH2 ꢀ 4), 2.34 (s, 3H, CH3); 13C NMR (100 MHz,
8.12 (d, J ¼ 8.8 Hz, 1H, AreH), 8.03 (dd, J ¼ 8.8, 1.6 Hz, 1H, AreH),
7.95 (d, J ¼ 1.6 Hz, 1H, AreH), 7.92 (d, J ¼ 1.4 Hz, 1H, AreH), 7.78 (m,
1H, AreH), 7.64e7.54 (m, 1H, AreH), 7.35 (d, J ¼ 4.4 Hz, 1H, AreH),
7.23 (td, J ¼ 8.6, 2.1 Hz, 1H, AreH), 6.67 (s, 1H, alkene hydrogen),
3.69 (s, 3H, OCH3), 3.47 (s, 2H, CH2), 3.20 (s, 4H, CH2 ꢀ 2), 3.10 (s, 2H,
DMSO‑d6)
d
165.60, 165.54 (dd, JC-F ¼ 251.1, 11.9 Hz), 159.83 (dd, JC-
CH2), 2.28 (s, 3H, CH3); 13C NMR (100 MHz, DMSO‑d6)
d 165.62 (dd,
¼ 256.4, 13.3 Hz), 158.04, 150.89, 149.99, 147.90, 143.32, 143.06,
JC-F ¼ 258.5, 13.2 Hz), 164.91, 159.84 (dd, JC-F ¼ 257.4, 14.1 Hz),
158.05, 150.71, 149.03, 147.49, 143.18, 142.10, 134.62, 134.43, 133.80,
132.27 (d, JC-F ¼ 10.0 Hz), 130.76, 129.68, 128.60, 125.65, 125.46 (dd,
JC-F ¼ 12.9, 4.0 Hz), 123.32, 122.79, 120.06, 112.34 (dd, JC-F ¼ 19.3,
2.1 Hz), 106.31 (t, JC-F ¼ 25.3 Hz), 66.42, 66.36, 53.91, 46.50, 41.57,
26.21; HRMS: m/z calcd for C29H27F2N4O5S [M þ H]þ 581.1670,
found 581.1678. HPLC: tR ¼ 7.97 min (98.5% purity).
F
135.10, 134.29, 132.31 (d, JC-F ¼ 10.5 Hz), 131.00, 129.62, 128.84,
125.77, 125.59 (dd, JC-F ¼ 14.0, 3.1 Hz), 124.88, 122.69, 120.48,
120.43, 112.36 (dd, JC-F ¼ 22.2, 3.1 Hz), 106.30 (t, JC-F ¼ 26.0 Hz),
66.74, 66.55, 53.95, 46.64, 41.84, 20.84; HRMS: m/z calcd for
C
29H27F2N4O5S [M
þ
H]þ 581.1670, found 581.1681. HPLC:
tR ¼ 8.40 min (95.9% purity).
4.1.9.4. (E)-2,4-difluoro-N-(2-methoxy-5-(4-(4-(4-methylpiperazin-
1-yl)-4-oxobut-2-en-2-yl)quinolin-6-yl)pyridin-3-yl)benzenesulfo-
namide (14d). The compound was prepared from 12d (60 mg,
0.16 mmol) and 13 (68 mg, 0.16 mmol) according to the general
synthetic procedure B to afford the title compound as an off-white
solid (25 mg, 0.042 mmol, 26% yield). 1H NMR (400 MHz, DMSO‑d6)
4.2. Biology
4.2.1. In Vitro enzymatic assays, anti-proliferative assays and
docking study
The inhibitory activity against class I PI3Ks and mTOR, and the
anti-proliferative efficacy against tumor cell lines, as well as the
docking study were carried out according to the protocol disclosed
in our previous study [35].
d
8.89 (d, J ¼ 4.4 Hz, 1H, AreH), 8.26 (d, J ¼ 2.0 Hz, 1H, AreH), 8.15
(d, J ¼ 9.4 Hz, 1H, AreH), 8.02 (m, 2H, AreH), 7.87 (d, J ¼ 2.0 Hz, 1H,
AreH), 7.80 (m, 1H, AreH), 7.54e7.47 (m, 1H, AreH), 7.46 (d,
J ¼ 4.4 Hz, 1H, AreH), 7.19 (m, 1H, AreH), 6.37 (s, 1H, alkene
hydrogen), 3.71 (s, 3H, OCH3), 3.58 (s, 4H, CH2 ꢀ 2), 2.34 (s, 4H,
CH2 ꢀ 2), 2.31 (s, 3H, CH3), 2.21 (s, 3H, CH3); 13C NMR (100 MHz,
4.2.2. KINOMEscan
™ assay
Kinase selectivity was screened at the concentration of 10
the KinomeScan binding assay (DiscoveRx) according to a reported
m
M by
DMSO‑d6)
d
164.99 (dd, JC-F ¼ 251.0, 11.6 Hz), 165.39, 159.75 (d, JC-
¼ 255.2, 12.9 Hz), 157.93, 150.76, 149.93, 147.82, 142.79, 140.35,
F
9