90
T. Kline et al. / Bioorg. Med. Chem. 8 (2000) 73±93
by TLC. Rf=0.18 (90:10 CH2Cl2:MeOH). A solution of
129 (50 mg, 0.04 mmol) in 2 mL of CH2Cl2 was treated
with 10:1 TFA:anisole for 3.5 h, and the product pre-
cipitated as for 96 to give 20 mg (48%) of 98 as a light
C29H30N7O7S3 calculated, 684.1368, found, 684.1348
(M+).
5-Bromosalicylaldehyde (200 mg, 1.0 mmol) was added
to a solution of 132 (24 mg, 0.037 mmol) in DMF
(0.5 mL) and the resulting mixture was allowed to stir at
room temperature for 72 h, diluted with CH2Cl2, and
®ltered. The ®ltrate was concentrated in vacuo, sus-
pended in warm EtOAc, and allowed to stand for 1 h to
1
brown solid. H NMR (400 MHz, DMSO-d6) d 1.41±
1.44 (6H, m), 3.51±3.55 (2H, m), 3.70±3.78 (2H), 4.16±
4.19 (2H, m), 5.03 (2H, s), 5.21±5.24 (2H, m), 5.59±5.62
(1H, dd, J=3.9, 8.0 Hz), 5.83±5.88 (2H, m), 6.87 (1H,
s), 7.50±7.64 (2H, m), 7.85±7.88 (1H, m), 7.93±7.98 (1H,
m), 8.01±8.08 (2H, m), 8.20±8.23 (2H, m), 8.38±8.41
(1H, m), 8.94±8.99 (2H, m), 9.43±9.46 (1H, m), 9.79
1
give 16 mg (50%) of 100 as a yellow solid. H NMR
(400 MHz, DMSO-d6) d 1.43 (3H, d, J=5.0 Hz), 1.48
(3H, d, J=5.5 Hz), 3.72±3.79 (2H, m), 4.37 (2H, s),
5.19±5.25 (2H, m), 5.71 (2H, s), 6.97 (2H, d, J=7.4 Hz),
7.26±7.29 (1H, m), 7.45 (2H, d, J=7.6 Hz), 7.53±7.60
(2H, m), 7.86±7.89 (1H, m), 8.01±8.06 (2H, m), 8.90±
8.92 (3H, m), 12.68 (1H, bs). HRMS, m/z for C36H33
N7O8S3Br calculated, 866.0736, found, 866.0750 (M+).
10
(1H, s). HRMS, m/z for C38H33N6O S3 calculated,
829.1420, found, 829.1429 (M+).
(6R,7R)-7-[2-(2-Aminothiazol-4-yl)-2-[(Z)-(1-carboxy-1-
methylethoxy)imino]acetamido]-3-[[[1-[N-2-[(2-hydroxy)-
benzylidene] aminobenzyl] pyridinium-4-yl]thio]methyl]-8-
oxo-1-aza-5-thiabicyclo[4.2.0]oct-2-ene-2-carboxylate
(99). Bromomethyl intermediate 40 (76 mg, 0.27 mmol)
was reacted with 89 (200 mg, 0.27 mmol) as described
for 123 to give 217 mg (79%) of a brown glassy residue,
of which 101 mg, (0.097 mmol.) was treated with 5:1
TFA:anisole at 10 ꢂ for 4 h. Precipitation with diethyl
ether gave 86 mg (79%) of 130 as a beige solid. 1H
NMR (400 MHz, CD3OD) d 1.48 (3H, s), 1.49 (3H, s),
3.52±3.58 (2H, m), 4.32±4.36 (2H, m), 5.08 (1H, d,
J=4.8 Hz), 5.45 (2H, s), 5.76 (1H, d, J=4.8 Hz), 6.64
(1H, t, J=7.3 Hz), 6.70 (1H, d, J=7.8 Hz), 6.96 (1H, s),
7.10 (2H, d, J=7.7 Hz), 7.76 (2H, d, J=7.0 Hz), 8.35
(2H, d, J=6.8 Hz). HRMS, m/z for C29H30N7O7S3 cal-
culated, 684.1368, found, 684.1352 (M+). A suspension
of 130 (25 mg, 0.036 mmol) in EtOH (1.0 mL) was trea-
ted with salicylaldehyde (39 mL, 0.36 mmol). After 48 h
the reaction was diluted with CH2Cl2 and ®ltered. The
resulting ®ltrate was concentrated in vacuo, suspended
in CH2Cl2, and ®ltered to aord 15 mg (53%) of 99 as
(6R,7R)-7-[2-(2-Aminothiazol-4-yl)-2-[(Z)-[1-carbonyl-1-
(methylethyloxy)imino]acetamido]-3-[[[1-[N-(3-hydroxy-4-
(benzoylhydrazonomethyl)benzyl)]pyridinium-4-yl]thio]-
methyl]-8-oxo-1-aza-5-thiabicyclo[4.2.0]oct-2-ene-2-car-
boxylate (101). Cephalosporin 89 (7.5 mg, 0.01 mmol)
was reacted with bromomethyl intermediate 45 (11 mg,
0.01 mmol) to give, after 5:1 TFA:anisole deprotection,
5 mg (59%) of 101. 1H NMR (400 MHz, CD3OD) d
1.50 (3H, s), 1.51 (3H, s), 3.47±3.51 (1H, m), 3.64±3.68
(1H, m), 4.30 (1H, d, J=13.7 Hz), 4.49 (1H, d, J=
12.8 Hz), 5.10 (1H, d, J=4.7 Hz), 5.47±5.54 (2H, m),
5.78 (1H, d, J=4.7 Hz), 6.86±6.88 (2H, m), 6.98 (1H, s),
7.41±7.43 (3H, m), 7.50±7.52 (1H, m), 7.83±7.85 (4H,
m), 8.42 (1H, s), 8.56 (2H, d, J=6.6 Hz). HRMS, m/z
for C37H36N8O9S3 calculated, 832.4577 found, 831.1689
(M ).
(6R,7R)-7-[2-(2-Aminothiazol-4-yl)-2-[(Z)-[1-carbonyl-1-
(methylethyloxy)[N-(3-hydroxy-4-[(2-hydroxy-3-methyl-
benzoylhydrazonomethyl)benzyl)]pyridinium - 4-yl]thio] -
methyl]-8-oxo-1-aza-5-thiabicyclo[4.2.0]oct-2-ene-2-car-
boxylate (102). Cephalosporin 89 (91 mg, 0.12 mmol)
was reacted with bromomethyl intermediate 46 (49 mg,
0.12 mmol) in dry DMF to give, after 5:1 TFA:anisole
1
an amber solid. H NMR (400 MHz, DMSO-d6) d 1.43
(3H, d, J=6.0 Hz), 1.49 (3H, d, J=6.5 Hz), 3.63±3.79
(2H, m), 4.30±4.39 (2H, m), 5.17±5.30 (1H, m), 5.46
(1H, s), 5.82±5.90 (2H, m), 6.99 (2H, d, J=6.2 Hz), 7.35
(2H, d, J=7.0 Hz), 7.40±7.54 (3H, m), 7.76 (1H, d,
J=6.5 Hz), 7.84 (1H, d, J=5.8 Hz), 7.94±8.01 (1H, m),
8.69±8.75 (1H, m), 8.80±8.87 (2H, m), 11.62 (1H, s),
11.90 (1H, s), 12.74 (1H, bs). HRMS, m/z for C36H34
N7O8S3 calculated, 788.1631, found, 788.1632 (M+).
1
deprotection, 27 mg (26%) of 102. H NMR (400 MHz,
CD3OD) d 1.25 (6H, s), 2.05 (3H, s), 3.38±3.42 (1H, m),
3.57±3.59 (1H, m), 4.09±4.12 (1H, m), 4.21±4.22 (1H,
m), 5.01±5.03 (1H, m), 5.46±5.47 (2H, m), 5.71 (1H, d,
J=4.8 Hz), 6.61 (2H, d, J=8.6 Hz), 6.80±6.82 (1H, m),
7.12 (2H, d, J=8.6 Hz), 7.50±7.52 (1H, m), 7.64±7.66
(2H, m), 8.00 (1H, d, J=8.1 Hz), 8.45±8.47 (2H, m),
8.63±8.65 (1H, m). MS, m/z 861.5 (M ).
(6R,7R)-7-[2-(2-Aminothiazol-4-yl)-2-[(Z)-(1-carboxy-1-
methylethoxy)imino]acetamido]-3-[[[1-[N-3-[(2-hydroxy-5-
bromo)benzylidene]aminobenzyl]pyridinium-4-yl]thio]-
methyl]-8-oxo-1-aza-5-thiabicyclo [4.2.0]oct-2-ene-2-car-
boxylate (100). Bromomethyl intermediate 41 (225 mg,
0.79 mmol) was reacted with 89 (230 mg, 0.31 mmol) as
described for 123 to give 217 mg (68%) of quaternary
intermediate 131. The crude product (217 mg, 0.21
mmol) was treated with 10:1 TFA:anisole to give, upon
precipitation, 190 mg of a peach solid. HPLC puri®ca-
tion of 50 mg (30±70% B in A over 30 min.) provided
(6R,7R)-7-[2-(2-Aminothiazol-4-yl)-2-[(Z)-[1-carbonyl-1-
(methylethyloxy)imino]acetamido]-3-[[[1[N-(3-hydroxy-4-
hydrazonomethyl)benzyl)]pyridinium-4-yl]thio]methyl]-8-
oxo-1-aza-5-thiabicyclo[4.2.0]oct-2-ene-2-carboxylate (103).
Cephaolosporin 89 (87 mg, 0.115 mmol) was reacted
with 49 (43 mg, 0.115 mmol) in dry DMF to give, after
5:1 TFA:anisole deprotection, 5 mg (6%) of 103. 1H
NMR (400 MHz, DMSO-d6) d 1.42 (3H, s), 1.43 (3H, s),
3.55 (5H, bs), 3.74 (1H, d, J=5.3 Hz), 4.37±4.38 (2H,
m), 5.21±5.22 (1H, m), 5.67±5.68 (2H, m), 5.85±5.87
(1H, m), 6.71 (1H, s), 7.01±7.04 (2H, m), 7.28±7.29
(2H, m), 7.74 (1H, d, J=5.1 Hz), 8.02±8.03 (2H, m),
1
24 mg (64%) of 132. H NMR (400 MHz, CD3OD) d
1.60 (3H, s), 1.61 (3H, s), 3.65±3.69 (2H, m), 4.46±4.50
(2H, m), 5.24 (1H, d, J=4.9 Hz), 5.54 (2H, s), 5.88 (1H,
d, J=4.9 Hz), 6.97±7.03 (2H, m), 7.09 (1H, s), 7.27±7.35
(1H, m), 7.33 (1H, t, J=8.0 Hz), 7.93 (2H, d,
J=7.2 Hz), 8.63 (2H, d, J=7.1 Hz). HRMS, m/z