T. Ishikawa et al. / Bioorg. Med. Chem. 11 (2003) 2427–2437
2435
with MeOH (6 L), and dried under vacuum to give 16a
1
suspension were successively added 0.6 M NaHCO3 (26
mL) and 2-(5-amino-1,2,4-thiadiazol-3-yl)-2(Z)-ethoxy-
iminoacetyl chloride hydrochloride15 (2.0 g, 7.38 mmol),
and the reaction mixture was stirred at 5 ꢁC for 30 min.
After concentration under reduced pressure, the con-
centrate was purified by MCI gel SP-207 (300 mL) col-
umn chromatography. The fractions eluted by aqueous
20% EtOH were concentrated under reduced pressure
and the concentrate was lyophilized to give 2a (1.83 g,
(943 g, 81%): H NMR (CDCl3) d 3.41–3.73 (4H, m),
5.02 (1H, d, J=5 Hz), 5.84 (1H, dd, J=8 Hz, 5 Hz),
6.23 (1H, d, J=8 Hz), 6.97 (1H, s), 7.27–7.72 (17H, m),
7.73 (1H, s), 8.67 (2H, d, J=6 Hz). Anal. calcd for
.
C36H28N4O4S3 0.5H2O: C, 63.05; H, 4.11; N, 8.17; S,
14.02. Found: C, 63.16; H, 4.15; N, 8.27; S, 13.98.
Benzhydryl 7ꢀ-phenylacetamido-3-[4-(1-methyl-4-pyridi-
nio)-1,3-thiazol-2-yl]thio-3-cephem-4-carboxylate iodide
(18a). Iodomethane (324 g, 2.17 mol) was added drop-
wise to a solution of 16a (300 g, 0.43 mol) in DMF (0.6
L), and the reaction mixture was stirred at room tem-
perature for 8 h. The mixture was added dropwise to
EtOAc (6 L) with stirring, and the resulting precipitate
was collected by filtration, washed with EtOAc (500
mL) and diethyl ether (1 L) successively, and dried
under vacuum to give 18a (351 g, 97%): 1H NMR
(DMSO-d6) d 3.55 (2H, d, J=4 Hz), 3.70, 4.01 (2H,
ABq, J=18 Hz), 4.35 (3H, s), 5.33 (1H, d, J=5 Hz),
5.89 (1H, dd, J=8 Hz, 5 Hz), 6.99 (1H, s), 7.18–7.42
(15H, m), 8.54 (2H, d, J=7 Hz), 9.02 (1H, s), 9.03 (2H,
d, J=7 Hz), 9.32 (1H, d, J=8 Hz).
1
62%): H NMR (DMSO-d6) d 1.25 (3H, t, J=7 Hz),
3.34, 3.89 (2H, ABq, J=17 Hz), 4.18 (2H, q, J=7 Hz),
4.30 (3H, s), 5.18 (1H, d, J=5 Hz), 5.73 (1H, dd, J=8
Hz, 5 Hz), 8.13 (2H, br s), 8.49 (2H, d, J=7 Hz), 8.88
(1H, s), 8.95 (2H, d, J=7 Hz), 9.62 (1H, d, J=8 Hz);
IR (KBr) cmꢀ1 1775, 1640, 1615. Anal. calcd for
.
C22H20N8O5S4 4.5H2O: C, 38.53; H, 4.26; N, 16.34.
Found: C, 38.39; H, 4.06; N, 16.04.
Compounds 3a–d, 4a–6a were obtained from 21a–d by a
similar procedure to that used for preparation of 2a.
1
3a (23% yield): H NMR (DMSO-d6) d 3.34, 3.89 (2H,
ABq, J=17 Hz), 4.30 (3H, s), 5.19 (1H, d, J=5 Hz),
5.74 (1H, dd, J=8 Hz, 5 Hz), 5.79 (2H, d, J=55 Hz),
8.22 (2H, br s), 8.49 (2H, d, J=7 Hz), 8.88 (1H, s), 8.95
(2H, d, J=7 Hz), 9.84 (1H, d, J=8 Hz); IR (KBr) cmꢀ1
Benzhydryl
7ꢀ-amino-3-[4-(1-methyl-4-pyridinio)-1,3-
thiazol-2-yl]thio-3-cephem-4-carboxylate chloride (20a).
Under ice-cooling, pyridine (115 g, 1.44 mol) was
added dropwise to a suspension of phosphorus penta-
chloride (312 g, 1.44 mol) in CH2Cl2 (2.8 L), and the
mixture was stirred at 5 ꢁC for 30 min. To the mixture
18a (400 g, 0.48 mol) was added portionwise, and the
reaction mixture was stirred at 5 ꢁC for 1 h. Under
cooling at ꢀ10 ꢁC, the mixture was added dropwise to
iso-butanol (5.6 L), and stirred at room temperature for
3 h. The resulting precipitate was collected by filtration,
washed with EtOAc (500 mL) and diethyl ether (1L)
successively, and dried under vacuum to give 20a (270 g,
93%): 1H NMR (DMSO-d6) d 3.94 (2H, s), 4.35 (3H, s),
5.32 (1H, d, J=5 Hz), 5.45 (1H, d, J=5 Hz), 6.99 (1H,
s), 7.25–7.40 (10H, m), 8.59 (2H, d, J=7 Hz), 9.07 (2H,
d, J=7 Hz), 9.19 (1H, s).
.
1775, 1640, 1610. Anal. calcd for C21H17FN8O5S4
5.0H2O: C, 36.10; H, 3.89; N, 16.04. Found: C, 36.14;
H, 3.83; N, 15.69.
1
3b (30% yield): H NMR (D2O) d 3.55, 4.00 (2H, ABq,
J=18 Hz), 4.35 (3H, s), 5.38 (1H, d, J=5 Hz), 5.79 (2H,
d, J=55 Hz), 5.88 (1H, d, J=5 Hz), 8.57 (2H, d, J=7
Hz), 8.82 (2H, d, J=7 Hz); IR (KBr) cmꢀ1 1770, 1670,
.
1630, 1610. Anal. calcd for C20H16FN9O5S4 4.0H2O: C,
35.24; H, 3.55; N, 18.49. Found: C, 35.15; H, 3.37; N,
18.21.
1
3c (52% yield): H NMR (DMSO-d6) d 3.19, 3.54 (2H,
ABq, J=17 Hz), 4.29 (3H, s), 5.02 (1H, d, J=5 Hz),
5.62 (1H, dd, J=8 Hz, 5 Hz), 5.76 (2H, d, J=56 Hz),
8.04 (1H, d, J=1 Hz), 8.25 (2H, br s), 8.43 (2H, d, J=7
Hz), 8.78 (1H, d, J=1 Hz), 8.95 (2H, d, J=7 Hz), 9.74
(1H, d, J=8 Hz); IR (KBr) cmꢀ1 1760, 1670, 1635,
Dihydrochloride salt of 7ꢀ-amino-3-[4-(1-methyl-4-pyri-
dinio) - 1,3 - thiazol - 2 - yl]thio - 3 - cephem - 4 - carboxylate
(21a). To a suspension of 20a (430 g, 0.66 mol) in ace-
tonitrile (3.5 L) was added dropwise concd HCl (3.5 L),
and the mixture was stirred at room temperature for 30
min. EtOAc (7 L) was added, and the mixture was stir-
red at room temperature for 5 h. The resulting crystals
were collected by filtration, washed twice with aceto-
nitrile (1 L), and dried under vacuum to give 21a as the
dihydrochloride form (236 g, 76%): MP 202 ꢁC; 1H
NMR (DMSO-d6) d 3.78, 3.98 (2H, ABq, J=17 Hz),
4.35 (3H, s), 5.26 (1H, d, J=5 Hz), 5.42 (1H, d, J=5
Hz), 8.61 (2H, d, J=7 Hz), 9.05 (2H, d, J=7 Hz), 9.17
.
1605. Anal. calcd for C22H18FN7O5S4 3.0H2O: C, 39.93;
H, 3.66; N, 14.82. Found: C, 39.82; H, 3.52; N, 14.56.
1
3d (14% yield): H NMR (DMSO-d6) d 3.12, 3.27 (2H,
ABq, J=17 Hz), 4.26 (3H, s), 5.01 (1H, d, J=5 Hz),
5.61 (1H, dd, J=9 Hz, 5 Hz), 5.75 (2H, d, J=55 Hz),
7.60 (1H, s), 8.20 (2H, br s), 8.33 (2H, d, J=6 Hz), 8.93
(2H, d, J=6 Hz), 8.96 (1H, s), 9.67 (1H, d, J=9 Hz); IR
(KBr) cmꢀ1 1765, 1670, 1640, 1610. Anal. calcd for
.
C22H18FN7O6S3 4.5H2O: C, 39.25; H, 4.01; N, 14.57.
Found: C, 39.67; H, 3.79; N, 14.30.
.
(1H, s). Anal. calcd for C16H14N4O3S3 2HCl: C, 40.08;
H, 3.36; N, 11.69. Found: C, 39.83; H, 3.43; N, 11.78.
1
4a (38% yield): H NMR (DMSO-d6) d 3.53, 3.91 (2H,
ABq, J=18 Hz), 3.92 (3H, s), 4.31 (3H, s), 5.17 (1H, d,
J=5 Hz), 5.74 (1H, dd, J=8 Hz, 5 Hz), 8.15 (2H, br s),
8.47 (2H, d, J=7 Hz), 8.89 (1H, s), 8.97 (2H, d, J=7
Hz), 9.66 (1H, d, J=8 Hz); IR (KBr) cmꢀ1 1770, 1640,
1610. Anal. calcd for C21H18N8O5S4 4.0H2O: C, 38.06;
H, 3.95; N, 16.91. Found: C, 38.26; H, 4.01; N, 16.87.
7ꢀ-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2(Z)-ethoxyimi-
noacetamido]-3-[4-(1-methyl-4-pyridinio)-1,3-thiazol-2-
yl]thio-3-cephem-4-carboxylate (2a, T-91825). Under
ice-cooling, 21a (2.0 g, 4.92 mmol) was suspended in a
mixture of THF (80 mL) and water (80 mL). To the
.