Clinical Candidate of PI3 Kinase
Journal of Medicinal Chemistry, 2008, Vol. 51, No. 18 5529
1
to give 14 as an off-white crystalline solid (51 mg, 45%). H NMR
400 MHz, CDCl ) δ 2.21 (s, 3H), 2.40-2.55 (m, br, 8H), 3.72 (s,
H), 3.75-3.79 (m, 4H), 3.93-3.96 (m, 4H), 7.11 (s, 1H).
-Chloro-6-(4-methanesulfonyl-piperazin-1-ylmethyl)-4-morpho-
2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine (20).
To a solution of 19 (80 mg, 0.25 mmol) in chloroform (8 mL) and
acetic acid (4 mL) was added isoamyl nitrite (36 µL, 1.1 equiv).
The reaction mixture was stirred for 1 day at room temperature.
The mixture was then quenched with sodium bicarbonate solution
and extracted in to chloroform and reduced in vacuo. The residue
was purified using flash chromatography (EtOAC/hexanes to
(
2
3
2
lin-4-yl-thieno[3,2-d]pyrimidine (15). A mixture of 13 (5.56 g, 19.6
mmol), 1-methanesulfonyl-piperazine hydrochloride (4.72 g, 1.2
equiv), and trimethylorthoformate (6.42 mL, 3 equival) was stirred
in 1,2-dichloroethane (100 mL) for 6 h at room temperature. To
this was added sodium triacetoxyborohydride (10.39 g, 2.5 equiv),
and the reaction mixture was stirred for 24 h at room temperature.
The mixture was then quenched with brine, extracted with dichlo-
1
EtOAc) to yield 20 (17 mg, 20%). H NMR (400 MHz, DMSO-
d
6
) δ 3.86-3.91 (m, 4H), 4.05-4.09 (m, 4H), 7.48-7.52 (m, 1H),
7
.63-7.70 (m, 2H), 8.26 (d, 1H), 8.33 (d, 1H), 8.95 (s, 1H), 13.20
+
(
s, br, 1H). MS (ESI): m/z (M + H) 338. Analytical LC-MS using
Waters XBridge Phenyl analytical column and H20/MeCN modified
with 0.1% formic acid running a linear gradient from 10% MeCN
to 100% MeCN monitored by UV wavelength 210 nm and ESI+
TIC MS showed 98.2% purity.
2-(1H-Indazol-6-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine (22).
A mixture of 3 (806 mg, 3.15 mmol), 3-amino-4-methylbenzenebo-
ronic acid (524 mg, 1.1 equiv), DME (10 mL), water (5 mL),
romethane, dried (MgSO
4
), and the solvent removed in vacuo. The
residue was triturated with hot ethyl acetate to yield 15 as a white
1
solid (1.01 g). NMR H NMR (400 MHz, CDCl
m, 4H), 2.79 (s, 3H), 3.30-3.33 (m, 4H), 3.82-3.86 (m, 6H),
.98-4.01 (m, 4H), 7.18 (s, 1H).
-(1H-Indazol-4-yl)-6-(4-methyl-piperazin-1-ylmethyl)-4-mor-
3
) δ 2.69-2.71
(
3
2
pholin-4-yl-thieno[3,2-d]pyrimidine (16). Compound 16 was pre-
pared from 14 according to the same procedure described for 17.
2 3 2
sodium carbonate (640 mg, 2 equiv), and PdCl (PPh ) (100 mg,
1
5%) was heated to reflux for 16 h. The reaction mixture was then
cooled, diluted with ethyl acetate, and reduced in vacuo. The residue
was purified using flash chromatography to yield 2-methyl-5-(4-
morpholin-4-yl-thieno[3,2-d]pyrimidin-2-yl)-phenylamine (21) (840
mg, 82%). To a solution of 21 (99 mg, 0.30 mmol) in chloroform
H NMR (400 MHz, DMSO-d ) δ 2.18 (s, 3H), 2.30-2.45 (m,
6
4
3
8
H), 2.48-2.55 (m, 4H), 3.82-3.84 (m, 4H), 3.86 (s, 2H),
.98-4.00 (m, 4H), 7.44-7.47 (m, 2H), 7.65 (d, J ) 8.2 Hz, 1H),
.21 (d, J ) 7.2 Hz, 1H), 8.87 (s, 1H), 13.16 (s, br,1H). MS (ESI):
+
m/z (M + H) 450. Analytical LC-MS using Waters XBridge
Phenyl analytical column and H20/MeCN modified with 0.1%
formic acid running a linear gradient from 10% MeCN to 100%
MeCN monitored by UV wavelength 210 nm and ESI+ TIC MS
showed 98.4% purity.
(
1
10 mL) and acetic acid (2 mL) was added isoamyl nitrite (44 µL,
.1 equiv). The reaction mixture was stirred for 2 days at room
temperature. The mixture was then quenched with sodium bicar-
bonate solution and extracted into chloroform and reduced in vacuo.
The residue was purified using flash chromatography (EtOAc) to
2
-(1H-Indazol-4-yl)-6-(4-methanesulfonyl-piperazin-1-ylmethyl)-
-morpholin-4-yl-thieno[3,2-d]pyrimidine (17). A mixture of 15
2.00 g, 4.63 mmol), 25 (2.26 g, 2 equiv), toluene (24 mL), ethanol
12 mL), water (6 mL), sodium carbonate (1.72 g, 3.5 equiv), and
1
yield the title compound (14 mg, 14%). H NMR (400 MHz,
4
DMSO-d ) δ 3.85-3.89 (m, 4H), 4.05-4.09 (m, 4H), 7.58 (d, J )
6
(
5
8
1
.4 Hz, 1H), 7.86 (d, J ) 8.4 Hz, 1H), 8.12 (s, 1H), 8.12 (d, J )
(
.5 Hz, 1H), 8.19 (d, J ) 5.4 Hz, 1H), 8.21 (s, 1H), 13.22 (s, br,
PdCl
microwave for 90 min. The reaction mixture was cooled, diluted
with chloroform, washed with brine, dried (MgSO ), and the solvent
2
(PPh
3
)
2
(325 mg, 0.1 equiv) was heated to 130 °C in the
+
H). MS (ESI): m/z (M + H) 338. Analytical LC-MS using Waters
XBridge Phenyl analytical column and H20/MeCN modified with
4
0
1
.1% formic acid running a linear gradient from 10% MeCN to
00% MeCN monitored by UV wavelength 210 nm and ESI+ TIC
removed in vacuo. The residue was purified using flash chroma-
tography (chloroform to 10% methanol/chloroform) and then
trituration with ether, yielding the desired title compound as a white
MS showed 95.5% purity.
4-Bromo-1H-indazole (24). To a solution of 3-bromo-2-methy-
1
solid (1.42 g, 60%). H NMR (CDCl
3
) δ 2.67-2.71 (m, 4H), 2.81
laniline (5.00 g, 26.9 mmol) in chloroform (50 mL) was added
potassium acetate (2.77 g, 28.2 mmol). Acetic anhydride (5.07 mL,
(
4
s, 3H), 3.29-3.33 (m, 4H), 3.89 (s, 2H), 3.89-3.93 (m, 4H),
.08-4.12 (m, 4H), 7.41 (s, 1H), 7.51 (t, J ) 7.2 Hz, 1H), 7.60 (d,
5
3.7 mmol) was added with concurrent cooling in ice-water. The
J ) 8.3 Hz, 1H), 8.28 (d, J ) 7.5 Hz, 1H), 9.02 (s, 1H), 10.10 (s,
mixture was then stirred at room temperature for 10 min, after which
time a white gelatinous solid formed. 18-crown-6 (1.42 g, 5.37
mmol) was then added, followed by isoamyl nitrite (7.94 mL, 59.1
mmol), and the mixture was heated under reflux for 18 h. The
reaction mixture was allowed to cool and was partitioned between
chloroform and saturated aqueous sodium hydrogen carbonate. The
combined organic extracts were washed with brine, separated, and
+
br,1H). MS (ESI): m/z (M + H) 514. Anal. (C23
27 7 3 2
H N O S ) C,
H, N. Analytical LC-MS using Waters XBridge Phenyl analytical
column and H20/MeCN modified with 0.1% formic acid running
a linear gradient from 10% MeCN to 100% MeCN monitored by
UV wavelength 210 nm and ESI+ TIC MS showed 98% purity.
-Methyl-3-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phe-
nylamine (18). To a solution of 3-bromo-2-methylaniline (1.00 g,
.38 mmol) in dioxane (15 mL) was added triethylamine (3.0 mL,
equiv), Pd(OAc) (60 mg, 5%), 2-dicyclohexylphosphinobiphenyl
377 mg, 20%), and pinacol borane (2.34 mL, 3 equiv), and the
reaction mixture was heated to 80 °C for 1 h. The reaction mixture
was then cooled, diluted with ethyl acetate, washed with brine, dried
4
), and the solvent reduced in vacuo. The residue was
purified using flash chromatography (EtOAc/hexane 1:4) to yield
8 as a brown oil (1.07 g, 85%). H NMR (400 MHz, CDCl
.27 (s, 12H), 2.31 (s, 3H), 3.52 (s, 2H), 6.68 (d, J ) 7.8 Hz, 1H),
.92-6.96 (m, 1H), 7.14 (d, J ) 7.4 Hz, 1H)
-Methyl-3-(4-morpholin-4-yl-thieno[3,2-d]pyrimidin-2-yl)-phe-
nylamine (19). A mixture of 3 (169 mg, 0.66 mmol), 18 (155 mg,
.66 mmol), sodium carbonate (140 mg, 2 equiv), DME (10 mL),
water (3 mL), and PdCl (PPh (23 mg, 5%) was heated to reflux.
After 16 h, the reaction mixture was cooled, diluted with ethyl
acetate, washed with brine, dried (MgSO ), and reduced in vacuo.
2
dried (MgSO ). The crude product was evaporated onto silica and
4
5
4
purified by chromatography eluting with 20% to 40% EtOAc-petrol
2
to provide 1-(4-bromo-indazol-1-yl)-ethanone 23 (3.14 g, 49%) and
4-bromo-1H-indazole 24 (2.13 g, 40%). To a solution of 23 (3.09
g, 12.9 mmol) in methanol (50 mL) was added 6M aqueous HCl
(30 mL), and the mixture was stirred at room temperature for 7 h.
The methanol was evaporated and the mixture partitioned between
ethyl acetate and water. The combined organic layers were washed
(
(
MgSO
1
1
1
6
3
) δ
with brine, separated, and dried (MgSO ). The solvent was removed
4
by evaporation under reduced pressure to give 24 (2.36 g, 93%).
1
H NMR (400 MHz, CDCl ) δ 7.25 (t, J ) 7.3 Hz, 1H), 7.33 (d,
3
2
J ) 7.3 Hz, 1H), 7.46 (d, J ) 7.3 Hz, 1H), 8.11 (s, 1H), 10.20 (s,
br,1H).
0
4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-indazole (25).
To a solution of 24 (500 mg, 2.54 mmol) and bis(pinacolato)diboron
(967 mg, 3.81 mmol) in DMSO (20 mL) was added potassium
2
3 2
)
4
2 2
acetate (747 mg, 7.61 mmol) and PdCl (dppf) (62 mg, 0.076 mmol,
The residue was purified using flash chromatography (EtOAc/
hexanes 1:1 to EtOAc) to yield 19 (148 mg, 69%). H NMR (400
3 mol%). The mixture was degassed with argon and heated at 80
°C for 40 h. The reaction mixture was allowed to cool and
partitioned between water and ether. The combined organic layers
1
MHz, CDCl
.01-4.06 (m, 4H), 6.73-6.76 (m, 1H), 7.08-7.13 (m, 2H), 7.51
d, J ) 5.5 Hz, 1H), 7.77 (d, J ) 5.5 Hz, 1H).
3
) δ 2.29 (s, 3H), 3.70 (s, br, 2H), 3.85-3.88 (m, 4H),
4
were washed with brine, separated, and dried (MgSO
4
). The crude
(
material was purified by chromatography eluting with 30% to 40%