2386 J ournal of Medicinal Chemistry, 2002, Vol. 45, No. 12
Lesuisse et al.
1 M NaOH (4 mmol). The mixture was heated to 50 °C for 7.5
h and then poured into water and acidified with 13.25 mL
(13.25 mmol) of 1 N HCl. The mixture was extracted with
EtOAc, and the organic extracts were washed with brine,
dried, concentrated (1.47 g of crude), and purified by flash
chromatography (185 g of silica, CH2Cl2/MeOH, 96.5:3.5) to
give the first fraction of 285 mg (19%) of the recovered starting
material as a white foam. Then the product was eluted from
the column with CH2Cl2/MeOH/AcOH, 96.5:3.5:1.5 to give
897.8 mg (61%) of the expected material as a white foam. Rf
) 0.29 (RP-18F254Merck, MeOH/H2O, 85/15); HPLC (CH3CN/
H2O, 75:25, 0.01% TFA, 1.5 mL/min, 96 bar, 230 nm, 4.57 min,
92.2%; NMR 6:4 mixture of diastereoisomers δ 1.16 and 1.66
(2m, 2H), 1.39, 1.62-1.87 (m, 4H), 1.41 (s, 18H), 1.77 (s, 3H),
2.74 and 2.81 (2ld, 1H), 3.03 and 3.14 (2dd, 1H), 3.27 and 3.58
(2m, 2H), 4.31-4.76 (m, 3H), 5.43 (ls, 1H), 7.23 (m), 7.36, 7.47,
with EtOAc. The organic extracts were washed with brine,
dried, and concentrated. The residues were crystallized in Et2O
to give 59.7 mg (80.6%) and 29.6 mg (63.2%) of pure 28 (iso A)
and 29 (iso B), respectively. 28: HPLC, CH3CN/H2O, 75:25,
0.01% TFA, 1 mL/min, 60 bar, 230 nm, 3.14 min, 97.64%; NMR
δ 1.20-1.96 (m, 6H), 1.94 (s, 3H), 3.00 and 3.24 (dd, 2H), 3.32
and 3.58 (dd, 2H), 3.84 (s, 3H), 4.58 and 4.72 (m, 2H), 4.68
(m, 1H), 4.73 (m, 1H), 5.42 (s, 1H), 7.20-7.63 (m, 12H), 7.93
(m, 1H), 8.39 (m, 1H). 29: HPLC, CH3CN/H2O, 75:25, 0.01%
TFA, 1 mL/min, 59 bar, 230 nm, 2.88 min, 94.7%; NMR δ
1.23-1.96 (m, 6H), 3.04 and 3.17 (dd, 2H), 3.3 and 3.57 (m,
2H), 3.85 (s, 3H), 4.54 and 4.73 (m, 2H), 4.64 (m,1H), 4.72 (m,
1H), 5.37 (s, 1H), 7.34(m, 3H), 7.42 (m, 3H), 7.50 (dd, 1H), 7.59
(m, 4H), 7.91 (d, 1H).
Ack n ow led gm en t. This work was done during the
course of collaboration with ARIAD Pharmaceuticals.
We are grateful to Tom Sawyer, M. Weigele, R. Bohacek,
and the entire group of scientists there for stimulating
discussions and interactions. We thank M. R. Van
Schravendyck for providing the clones and procedures
for Src SH2 production and purification.
7.64, 7.88 (12H), 8.13, 8.19, 8.22 (3d, 2H), 13.04 (sl, 1H); MS
-
m/z 740 (M - H-)-, 666 (740, O - t-Bu)
.
[4-[2-(Ace t yla m in o)-3-[[(3S )-1-[[1,1′-b ip h e n yl)-4-yl]-
m et h yl]-2-oxoh exa h yd r o-1H -a zep in -3-yl]a m in o]-3-oxo-
1-p r op en yl]-2-ca r boxyp h en yl]p r op a n ed ioic Acid 20
a n d 4-[2-(Acetyla m in o)-3-[[(3S)-1-[[1,1′-bip h en yl)-4-yl]-
m et h yl]-2-oxoh exa h yd r o-1H -a zep in -3-yl]a m in o]-3-oxo-
1-p r op en yl]-2-ca r boxyben zen a cetic Acid 27. A total of 8.6
mg of [4-[2-(acetylamino)-3-[[(3S)-1-[[1,1′-biphenyl)-4-yl]methyl]-
2-oxo-hexahydro-1H-azepin-3-yl]amino]-3-oxo-1-propenyl]-2-
carboxyphenyl]propanedioic acid, 1,3-bis(1,1-dimethylethyl)
ester 26 (0.119 mmol) was dissolved in 2.2 mL of 98% HCOOH
(58.31 mmol). The solution was stirred for 2 h at room
temperature and then partitioned between water and EtOAc.
The organic extracts were washed with water and brine, dried,
and concentrated to afford 68.1 mg of crude material as a white
foam. This material was triturated with cold diethyl ether and
filtered to afford 64 mg (85.6%) of the expected material [4-[2-
(acetylamino)-3-[[(3S)-1-[[1,1′-biphenyl)-4-yl]methyl]-2-oxo-
hexahydro-1H-azepin-3-yl]amino]-3-oxo-1-propenyl]-2-carbox-
yphenyl]propanedioic acid 20 as a white powder. Rf ) 0.12
(SiO2F254Merck60, CH2Cl2/MeOH/AcOH, 80/20/5); HPLC, CH3-
CN/H2O, 65/35, 0.01% TFA, 1 mL/min, 77 bar, 230 nm, 2.82
min, 90%; NMR 1:1 mixture of diastereoisomers δ 1.12-1.89
(m, 6H), 1.78 (s, 3H), 2.77 (dd, 1H), 3.26-3.58 (m, 2H), 3.68
(dd, 1H), 4.31-4.77 (m, 3H), 5.41 (sl, 1H), 7.23 (dd), 7.36, 7.46,
7.66, 7.85 (12H), 8.17, 8.23 (sl, 2H); MS m/z 650 (MNa+ - 2H)
-, 628 (M - H)-; HRMS calcd 630.2451, found 630.2454.
A total of 17 mg (0.027 mmol) of [4-[2-(acetylamino)-3-[[(3S)-
1-[[1,1′-biphenyl)-4-yl]methyl]-2-oxohexahydro-1H-azepin-3-yl]-
amino]-3-oxo-1-propenyl]-2-carboxyphenyl]propanedioic acid
20 was dissolved in 4 mL of methanol and warmed for 3 h at
50-60 °C. The solution was filtered, concentrated, and recrys-
tallized in 20 mL of diethyl ether to afford 15.7 mg (99%) of
the expected compound 4-[2-(acetylamino)-3-[[(3S)-1-[[1,1′-
biphenyl)-4-yl]methyl]-2-oxohexahydro-1H-azepin-3-yl]amino]-
3-oxo-1-propenyl]-2-carboxybenzenacetic acid 27 as a white
powder. Rf ) 0.43 (SiO2F254Merck60, CH2Cl2/MeOH/AcOH, 80/
20/5); HPLC CH3CN/H2O, 65/35, 0.01% TFA, 1 mL/min, 77
bar, 230 nm, 3.2 min, 86%; NMR 6.4 mixture of diastereoiso-
mers δ 1.12-1.90 (m, 6H), 1.77 (s, 3H), 2.76 (dd, 1H), 3.02,
3.12 (dd, 1H), 3.26-3.58 (m, 2H), 3.84 (s, 2H), 4.31-4.78 (m,
4H), 7.19, 7.35, 7.36, 7.46, 7.63, 7.66, 7.81 (12H), 8.08 (sl, 1H),
8.29 (sl, 1H), 12.56 (sl, 2H); MS m/z 584- (M - H), 540- (M -
H- - CO2); HRMS calcd 586.2553, found 586.2584.
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[4-[2-(Ace t yla m in o)-3-[[(3S )-1-[[1,1′-b ip h e n yl)-4-yl]-
m et h yl]-2-oxoh exa h yd r o-1H -a zep in -3-yl]a m in o]-3-oxo-
1-p r op en yl]-2-(m et h oxyca r b on yl)p h en yl]p r op a n ed ioic
Acid s 28 (Iso A) a n d 29 (Iso B). The 2 diastereoisomers of
26 were separated by chiral HPLC before ester hydrolysis. The
following were used for the AD chiralpack column: heptane/
ethanol, 60/40, 1.5 mL/min, 210 nm, 6.89 min (iso A), 10.92
min (iso B). From 145 mg of 26, 87 mg (0.115 mmol) of iso A
and 55 mg (0.072 mmol) of iso B were obtained. Each of them
was hydrolyzed separately using 3.6 mL (95.4 mmol) and 2.26
mL (59.90 mmol) of 98-100% formic acid respectively in 2 and
0.4 mL of CH2Cl2, respectively. The suspensions were for
stirred 2 h at 20 °C and then poured into water and extracted