Structural modifications and biological evaluation of 3-isoxazolylvinylcephalosporins

Article abstract of DOI:10.1016/S0040-4020(00)00415-4

Structural modifications and biological evaluations of 3-isoxazolylvinylcephalosporins (I) were performed. The replacement of a hydrogen atom at the 7-aminothiazole group by a chlorine resulted in an improvement of the activity against resistant Gram-posi

Full text of DOI:10.1016/S0040-4020(00)00415-4

TETRAHEDRON
Pergamon
Tetrahedron 56 (2000) 5657±5665
Structural Modi®cations and Biological Evaluation of
3-Isoxazolylvinylcephalosporins
Ae Nim Pae,a Jie Eun Lee,a Bo Hyung Kim,a Joo Hwan Cha,a Hea Yeon Kim,a Yong Seo Cho,a
b
c
Kyung Il Choi,a Hun Yeong Koh,a, Eun Lee and Je Hak Kim
*
aBiochemicals Research Center, Korea Institute of Science and Technology, P.O. Box 131, Cheongryang, Seoul 130-650, South Korea
bDepartment of Chemistry, College of Natural Sciences, Seoul National University, Seoul 151-742, South Korea
cInstitute of Science and Technology, Cheil Jedang Corporation, Ichon, Kyonggi-Do, 467-810, South Korea
Received 4 October 1999; accepted 20 December 1999
AbstractÐStructural modi®cations and biological evaluations of 3-isoxazolylvinylcephalosporins (I) were performed. The replacement of a
hydrogen atom at the 7-aminothiazole group by a chlorine resulted in an improvement of the activity against resistant Gram-positive bacterial
strains including the methicillin-resistant Staphylococcus aureus (MRSA) and the cipro¯oxacin-resistant Staphylococcus aureus (CRSA).
The introduction of other heterocycles such as an isothiazole or a thiadiazole in place of the isoxazole moiety gave slightly decreased in vitro
activities. q 2000 Elsevier Science Ltd. All rights reserved.
Introduction
In recent years, the worldwide incidence of antibiotic resis-
tance among Gram-positive bacteria, e.g. Staphylococcus,
Streptococcus and Enterococcus species, has increased.
Consequently, great concern has been focused on the
development of novel cephalosporin antibiotics having
improved antibacterial activities against resistant bacterial
strains. During the last several years we have synthesized
various 3-isoxazolylvinylcephalosporins I1 and investigated
their antibacterial activities. There we found that the introduc-
tion of an isoxazole moiety into the C-3 position of a cephem
nucleus gave a remarkable enhancement in the activities
against Gram-positive bacteria including Streptococcus
pyogenes and Staphylococcus aureus. As the C-7 substituent,
2-(2-aminothiazol-4-yl)-2-hydroxyiminoacetamido group was
a good match.
In 1997, Microcide Pharmaceutical Inc. (MPI) reported that
the chlorine-substituted aminothiazole moiety of the
cephalosporin derivative MC-02479 was responsible for
its excellent activity against resistant Staphylococcus
aureus.2 While working on optimizing in vitro activities
and pharmacokinetic properties of 3-isoxazolylvinyl-
cephalosporins I against Gram-positive bacteria, we became
interested in the chlorine-substituted aminothiazole, i.e.
2-amino-5-chlorothiazole, and selected it as the major struc-
tural modi®cation at the C-7 position. The replacement of
isoxazole moiety by other heterocycles such as the isothia-
zole or the thiadiazole was also carried out. Herein we report
the synthesis and biological evaluation of modi®ed 3-isoxa-
zolylvinylcephalosporins 1 and 3±6 focusing on Gram-posi-
tive bacteria (Fig. 1).
Synthesis
The cephalosporins 1 and 2 were synthesized following the
reaction pathway depicted in Scheme 1. The cephalosporins
2 had been reported previously,1d and here they were freshly
Keywords: antibacterials; cephalosporins; isoxazoles; thiazoles.
* Corresponding author. E-mail: hykoh@kist.re.kr
0040±4020/00/$ - see front matter q 2000 Elsevier Science Ltd. All rights reserved.
PII: S0040-4020(00)00415-4

5658
A. N. Pae et al. / Tetrahedron 56 (2000) 5657±5665
Figure 1.
resynthesized for comparison. The amine function of 2-(2-
amino-5-chlorothiazol-4-yl)-2-trityloxyiminoacetic acid (7),
which was prepared by a known method,2 and 2-(2-
aminothiazol-4-yl)-2-trityloxyiminoacetic acid (8) each
was protected (Ph3CCl/Et3N). The completely protected
carboxylic acids 9 and 10 each was reacted with p-methoxy-
benzyl 7-amino-3-chloromethylcephalosporanate 11 using
phosphorus oxychloride (POCl3) and pyridine at 2408C to
afford the key intermediates 12 and 13, respectively. Then
they were coupled with the isoxazole aldehydes3 14 via
phosphonium salts to produce the compounds 15 and 16,
respectively in ca. 50% yield as mixtures of geometrical
isomers (Z/E,9:1). The major (Z)-isomer was separated
from the mixture by column chromatography using a mixed
solvent of hexane and ethyl acetate (3:2) as the eluent.
Through the sequential deprotection by tri¯uoroacetic acid
and formic acid, crude free acids of the ®nal products 1 and
2 were obtained. Sodium salt formation by the treatment
with aqueous sodium bicarbonate solution followed by
puri®cation by reverse phase column chromatography
(Lichrosorbw RP-18, 20%, aq. MeOH) and lyophilization
afforded the pure ®nal products 1 and 2 in ca. 40% yield.
The synthesis of cephalosporin compounds 3±6 is outlined
in Scheme 2. It utilized basically the same protocol adopted
in the preparation of 1 and 2. The intermediates 12 and 13
were each coupled with the isothiazole or the thiadiazole
aldehyde4 (17 and 18) via phosphonium salts to produce the
compounds 19 and 20, or 21 and 22, respectively, in ca. 50%
yield as mixtures of geometrical isomers (Z/E,2:1). The
Scheme 1. Synthesis of 3-isoxazolylvinylcephalosporins.

A. N. Pae et al. / Tetrahedron 56 (2000) 5657±5665
5659
Scheme 2. Synthesis of 3-isothiazolylvinyl- and 3-thiadiazolylvinylcephalosporins.
(Z)- and (E)-isomer were separated, if possible, by column
chromatography using hexane and ethyl acetate (3:2) as the
eluent. Following the same procedure described above for
the preparation of compounds 1 and 2, the compounds 3±6
could be obtained in ca. 40% yield.
The compounds 1m and 1n having a heterocyclic substitu-
tent on the isoxazole also showed good activities against
Gram-positive bacteria. The compound 1m was superior
to vancomycin in activity against all the strains tested.
Regarding the effect of the substitutents of the isoxazole,
halogen substituents conferred higher activity, and hydro-
philic substituents such as carbamoyl or hydroxyl groups
gave lower activity against Gram-positive bacteria.
Biological Activity
In vitro activities of the compounds 3±6, having an iso-
thiazole or a thiadiazole moiety instead of the isoxazole,
against selected strains are summarized in Table 2. Com-
paring the activities of 4a with 2a and 4e with 2e, the
replacement of the isoxazole by an isothiazole resulted in
reduced activities against Gram-positive bacteria (Tables 1
and 2). As was the case with cephalosporins 2, the introduc-
tion of a chlorine atom into the aminothiazole of the C-7
substituent of 3-isothiazolylvinylcephalosporins 4 enhanced
the activity against Gram-positive bacteria. Unlike other
series of compounds, the cephalosporins 4 showed con-
siderable difference in activity according to the geometry
of the C-3 vinyl group. Comparing 4k(E) with 4k(Z), and
4e(E) with 4e(Z), respectively, (E)-vinyl isomer of
3-isothiazolylvinylcephalosporins exhibited better anti-
bacterial activity than (Z)-isomer by factors of 2±8
against Gram-positive bacteria. Interestingly, in the case
In vitro antibacterial activities (MIC) of all the compounds
prepared and references were determined by the Mueller±
Hinton agar dilution method.5 The activities of the
compounds synthesized were compared with cefdnir,
cefpodoxime and vancomycin as references. In vitro
activities of the compounds 1 and references against
selected strains are summarized in Table 1. Most of the
compounds 1 showed good antibacterial activity against
Gram-positive bacterial strains including MRSA and
CRSA. Comparing the activities of 1a and 2a, 1b with 2b
and 1e with 2e, the introduction of a chlorine atom into the
aminothiazole of the C-7 substituent enhanced the activity
slightly against MRSA and considerably against CRSA. The
activities of 1a, 1b and 1e each was comparable or superior
to the corresponding compound 2 by factors of 2 against
MRSA, and 2±4 against CRSA. In our previous work, the
compound 2b showed the best activity of the series.1d

5660
A. N. Pae et al. / Tetrahedron 56 (2000) 5657±5665
Table 1. In vitro antibacterial activities of 3-isoxazolylvinylcephaloporins (MIC (agar dilution method, Mueller±Hinton agar, 104 CFU/Spot), mg/mL)
Compounds
Microorganismsa
S.a
MRSA1
MRSA2
MRSA3
CRSA
MSSA
S.e
E.f
S.p
E.c1
E.c2
1a(M)b
1b(Z)
1c(Z)
1d(Z)
1e(Z)
1f(Z)
1g(Z)
1h(Z)
1i(Z)
0.5
0.25
0.5
0.5
0.5
0.5
0.5
1
0.5
0.5
1
0.5
1
0.5
1
1
1
1
2
1
0.5
0.5
0.5
0.25
1
0.5
0.5
0.5
4
1
1
2
2
2
1
2
4
2
2
16
2
1
1
2
2
4
2
0.5
0.25
0.5
0.5
0.5
0.5
0.5
1
2
2
4
2
4
2
2
4
4
4
16
4
1
2
8
4
16
4
8
0.5
0.5
1
0.5
1
0.5
1
1
1
0.5
4
1
1
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
0.5
0.25
0.25
1
0.25
#0.12
0.25
#0.12
#0.12
0.25
#0.12
#0.12
#0.12
1
0.5
0.5
1
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
0.25
#0.12
0.5
0.5
#0.12
0.25
0.25
#0.12
0.5
#0.12
1
0.25
0.5
1
0.25
0.5
0.5
0.25
0.5
0.25
2
0.25
0.5
1
0.5
0.5
0.5
0.5
0.5
0.5
0.5
4
0.5
0.5
1
1
1
2
1
2
4
.32
2
1
0.5
2
0.5
1
2
1j(Z)
1k(Z)
1l(Z)
#0.12
0.25
0.5
1
1
1m(Z)
1n(Z)
2a(Z)
2b(Z)
2e(Z)
2m(Z)
2n(Z)
Cefdnir
Cefpd.c
VCMd
0.25
0.5
0.5
0.25
1
0.25
0.5
0.5
4
0.25
0.5
0.25
0.25
0.5
0.25
0.25
0.25
2
1
1
1
0.5
0.25
1
0.25
0.5
0.5
4
#0.12
#0.12
#0.12
0.25
#0.12
#0.12
0.25
0.5
0.25
#0.12
0.25
.32
4
8
.32
1
0.25
.32
.32
1
#0.12
#0.12
.32
1
1
1
1
0.5
a S.a: Staphylococcus aureus ATCC 29213, MRSA1: Methicillin-resistant Staphylococcus aureus C2207, MRSA2: Methicillin-resistant Staphylococcus
aureus C5100, MRSA3: Methicillin-resistant Staphylococcus aureus C6068, CRSA: cipro¯oxacin-resistant Staphylococcus aureus C1062, MSSA:
Methicillin-susceptible Staphylococcus aureus C7142, S.e: Staphylococcus epidermis ATCC 12228, E.f.: Enterococcus faecalis ATCC 29212, S.p:
Streptococcus pyogenes ATCC 8668, E.c1: Enterobacter cloacae C4008, E.c2: Escherichia coli ATCC 10536.
b Mixture of geometrical isomers: the (Z)-isomer was partly isomerized during lyophilization.
c Cefpodoxime.
d Vancomycin.
Table 2. In vitro antibacterial activities of 3-isothiazolyl- and 3-thiadiazolylvinylcephaloporins (MIC (agar dilution method, Mueller±Hinton agar, 104 CFU/
Spot) mg/mL)
Compounds
Microorganismsa
S.a
MRSA1
MRSA2
MRSA3
CRSA
MSSA
S.e
E.f
S.p
E.c1
E.c2
3a(M)b
3a(E)
3e(Z)
0.5
0.5
0.25
0.5
0.5
0.5
1
0.5
1
0.5
2
0.25
0.5
0.5
0.25
0.5
4
0.5
1
0.5
1
0.5
1
1
1
1
1
2
0.5
0.5
0.5
0.5
0.5
4
1
1
1
1
2
4
8
2
8
2
8
2
4
4
4
8
.32
1
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
2
0.5
0.25
0.5
0.25
0.25
2
2
2
2
2
4
8
32
4
32
4
16
4
8
8
16
.32
.32
1
0.5
0.5
1
0.5
0.5
0.5
1
0.5
1
0.5
2
0.5
0.5
0.5
0.25
0.5
4
0.25
0.25
#0.12
0.25
0.25
0.25
0.25
0.5
0.25
0.5
0.5
#0.12
#0.12
#0.12
#0.12
#0.12
1
1
1
0.5
0.25
2
4
4
0.25
4
0.5
4
1
2
1
1
4
.32
2
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
0.25
0.5
0.5
0.5
1
#0.12
#0.12
0.5
#0.12
#0.12
#0.12
1
#0.12
#0.12
#0.12
#0.12
#0.12
#0.12
.32
0.5
0.5
0.5
0.5
#0.12
#0.12
1
#0.12
#0.12
#0.12
1
#0.12
#0.12
#0.12
0.25
#0.12
0.25
.32
3e(E)
3k(Z)
4a(M)b
4e(Z)
4e(E)
4k(Z)
4k(E)
5a(Z)
5d(E,Z)c
6a(Z)
6a(E)
6d(E,Z)c
Cefdnir
Cefpd.d
VCMe
1
1
1
1
0.5
a S.a: Staphylococcus aureus ATCC 29213, MRSA1: Methicillin-resistant Staphylococcus aureus C2207, MRSA2: Methicillin-resistant Staphylococcus
aureus C5100, MRSA3: Methicillin-resistant Staphylococcus aureus C6068, CRSA: cipro¯oxacin-resistant Staphylococcus aureus C1062, MSSA:
Methicillin-susceptible Staphylococcus aureus C7142, S.e: Staphylococcus epidermis ATCC 12228, E.f.: Enterococcus faecalis ATCC 29212, S.p:
Streptococcus pyogenes ATCC 8668, E.c1: Enterobacter cloacae C4008, E.c2: Escherichia coli ATCC 10536.
b Mixture of geometrical isomers: the (Z)-isomer was partly isomerized during lyophilization.
c The isomers could not be separated by column chromatography.
d Cefpodoxime.
e Vancomycin.

A. N. Pae et al. / Tetrahedron 56 (2000) 5657±5665
5661
of 3-thiadiazolyvinyl cephalosporins, the effect of intro-
ducing a chlorine atom into the aminothiazole was reversed.
Comparing 5a(Z) with 6a(Z), chlorine substitution at the
aminothiazole lowered the activity of the cephalosporin
compound against almost all the strains tested.
(d, 2H, phenyl-H), 6.06 (m, 1H, C7-H), 5.26 (s, 2H,
±CO2CH2Ph), 5.04 (d, 1H, Jˆ5.0 Hz, C6-H), 4.48 (dd,
2H, C3±CH2Cl), 3.79 (s, 3H, ±OCH3), 3.41 (dd, 2H, C2-H).
p-Methoxybenzyl
(6R,7R)-7-(Z)-[2-(5-chloro-2-trityl-
aminothiazol-4-yl)-2-trityloxyiminoacetamido]-3-[(3-
chloroisoxazol-5-yl)vinyl]-3-cephem-4-carboxylate
(15b(Z)). To the solution of p-methoxybenzyl (6R,7R)-7-
(Z)-[2-(5-chloro-2-tritylaminothiazol-4-yl)-2-trityloxyimino-
acetamido]-3-chloromethyl-3-cephem-4-carboxylate (12, 1.5 g,
1.42 mmol) in acetone (4 mL) were added sodium iodide
(223 mg, 1.49 mmol) and triphenylphosphine (391 mg,
1.49 mmol) in acetone (4 mL) were added sodium iodide
(223 mg, 1.49 mmol) and triphenylphosphine (391 mg,
1.49 mmol). The mixture was stirred at room temperature
for 1 h and concentrated under reduced pressure. The
residue was dissolved in CH2Cl2 (9 mL), and to the solution
were added 3-chloro-5-formylisoxazole (255 mg, 1.94
mmol) and 5% aq NaHCO3 (6 mL). After stirring at room
temperature for 1 h, the reaction mixture was extracted with
ethyl acetate (5 mL£2). The organic layer was washed with
brine, dried over MgSO4 and evaporated. The residue was
puri®ed on silica gel column chromatography (hexane/ethyl
acetateˆ3:2) to afford the product 15b(Z) (823 mg, 51%).
IR (KBr) 3396, 3052, 1796, 1720, 1683, 1500, 1248, 979,
Conclusion
For the 3-isoxazolylvinyl- and the 3-isothiazolylvinyl-
cephalosporins, the introduction of a chlorine atom into
the aminothiazole of the C-7 substitutent enhanced the
activity slightly against MRSA and considerably against
CRSA. But the tendency was reversed for the 3-thidiazolyl-
vinylcephalosporins. Replacing the isoxazole of the
3-isoxazolylvinylcephalosporins by an isothiazole or a thia-
diazole did not give better results. Up to this point, the series
of cephalosporins 1 looks like the best combination. Of
them, the compounds, 1a, 1b, 1f, 1m and 1n showed
promising in vitro activities. Their in vivo ef®cacy and
pharmacokinetic properties are now under evaluation. The
oral bioavailability of their C-4 esters is also being studied.
Experimental
1
700 cm21; H NMR (300 MHz, CDCl3) d 7.29 (m, 32H,
General
phenyl-H), 7.12 (d, 1H, ±NH), 6.91 (d, 2H, phenyl-H),
6.78 (d, 1H, Jˆ12.3 Hz, vinyl-H), 6.39 (d, 1H,
Jˆ12.3 Hz, vinyl-H), 6.18 (m, 1H, C7-H), 6.09 (s, 1H,
isoxazole), 5.19 (s,2H,±OCH2Ph), 5.17 (d, 1H, C6-H),
3.82 (s, 3H, ±OCH3), 3.51 (d, 1H, Jˆ18.4 Hz, C2-H),
3.08 (d, 1H, Jˆ18.4 Hz, C2-H).
1H NMR and 13C NMR spectra were obtained on a Varian
Gemini 300 spectrometer and a Bruker Avance 300 spectro-
meter at 300 and 75 MHz, respectively. Chemical shifts of
protons were recorded in ppm relative to tetramethylsilane
as an internal standard unless stated otherwise. Infrared
spectra were recorded on a Perkin Elmer 16F PC FT-IR
using the potassium bromide pellet. HRMS spectra were
obtained on a VG70-VSEQ (VG ANALYTICAL, UK)
mass spectrometer. All the commerically available reagents
were obtained from Aldrich, Fluka, and Tokyo Kasei chemi-
cal company and generally used without further puri®cation.
All the organic solvents were obtained from Orienta and
Samchun. Methylene chloride was distilled from calcium
hydride. Flash column chromatography was conducted
with silica gel grad 230±400 mesh (Merck Kiesegel 60
Art 9385). Reverse phase column chromatography was
conducted with LiChroprepw RP-18 (40±63 mm).
Sodium (6R,7R)-7-(Z)-[2-(2-amino-5-chlorothiazol-4-yl)-
2-hydroxyiminoacetamido]-3-[(3-chloroisoxazol-5-yl)-
vinyl]-3-cephem-4-carboxylate (1b(Z)). The compound
15b(Z) (720 mg, 0.63 mmol) was dissolved in anisole
(2.1 mL, 19 mmol) and added tri¯uoroacetic acid (TFA
7.3 mL, 95 mmol) under ice cooling. The reaction mixture
was stirred for 30 min at the same temperature followed by
stirring for 1 h at room temperature, and triturated with
isopropyl ether (15 mL). The resulting precipitate was
dissolved in formic acid (500 mL). After stirring for 1 h at
room temperature, the reaction mixture was triturated with
isopropyl ether (15 mL) again. The crude free acid obtained
by ®ltration was dissolved with aq. sodium bicarbonate
solution. The sodium salt was puri®ed by a reverse phase
column chromatography (LiChrosorbw RP-18, 20% aq.
MeOH) followed by the freeze drying to yield the pure
product 1b(Z) (68 mg, 19%). IR (KBr) 3363, 1769, 1651,
Representative procedures
p-Methoxybenzyl
(6R,7R)-7-(Z)-[2-(5-chloro-2-trityl-
aminothiazol-4-yl)-2-trityloxyiminoacetamido]-3-chloro-
methyl-3-cephem-4-carboxylate (12). To a mixture of
(Z)-2-(5-chloro-2-tritylaminothiazol-4-yl)-2-trityloxyimino-
acetic acid (9, 2.00 g, 2.83 mmol) and p-methoxybenzyl
(6R,7R)-7-amino-3-chloromethyl-3-cephem-4-carboxylate
hydrochloride (11, 1.15 g, 2.83 mmol) in CH2Cl2 (40 mL)
were added pyridine (920 mL), 1.13 mmol) and POCl3
(53 mL, 0.56 mmol) at 2408C. After stirring for 1 h, the
reaction mixture was diluted with ethyl acetate, washed
with brine, dried over MgSO4 and evaporated to give the
crude protected product. The crude product was puri®ed on
silica gel column chromatography (hexane/ethyl acetate/
1
1618, 1543, 1366 cm21; H NMR (300 MHz, D2O) d 6.52
(d, 1H, Jˆ12.1 Hz, vinyl-H), 6.33 (s, 1H, isoxazole), 6.29
(d, 1H, Jˆ12.1 Hz, vinyl-H), 5.77 (d, 1H, C7-H), 5.20 (d,
1H, C6-H), 3.52 (d, 1H, Jˆ18.0 Hz, C2-H), 3.23 (d, 1H,
Jˆ18.0 Hz, C2-H); 13C NMR (300 MHz, D2O) d 169.6,
168.7, 167.2, 165.2, 164.7, 154.2, 147.2, 136.0, 135.2,
130.7, 119.6, 116.0, 115.3, 104.9, 59.0, 57.9, 26.8; HRMS
(M1H), FAB): calcd for C17H13Cl2N6O6S2 530.9715, found
530.9716.
Physicochemical properties of the ®nal products
1
chloroformˆ2:1:1) to afford 12 (2.24 g, 75%). H NMR
(300 MHz CDCl3) d: 7.48±7.16 (m, 32H, phenyl-H), 6.92
Sodium (6R,7R)-7-[2-(2-amino-5-chlorothiazol-4-yl)-2-

5662
A. N. Pae et al. / Tetrahedron 56 (2000) 5657±5665
hydroxyiminoacetamido]-3-[(isoxazol-5-yl)vinyl]-3-
cephem-4-carboxylate (1a(E,Z). 1H NMR (300 MHz,
D2O) d (E): 8.39 (s, 1H, isoxazole), 7.43 (d, 1H,
Jˆ16.5 Hz, vinyl-H), 6.78 (d, 1H, Jˆ16.5 Hz, vinyl-H),
6.51 (s, 1H, isoxazole), 5.93 (d, 1H, C7-H), 5.32 (d, 1H,
C6-H), 3.84 (s, 1H, Jˆ15.8 Hz, C2-H), 3.72 (d, 1H,
Jˆ15.8 Hz, C2-H), (Z): 8.39 (s, 1H, isoxazole), 6.28 (d,
1H, Jˆ11.2 Hz, vinyl-H), 6.54 (d, 1H, Jˆ11.2 Hz, vinyl-
H), 6.41 (s, 1H, isoxazole), 5.93 (d, 1H, C7-H), 5.38 (d,
1H, C6-H), 3.68 (s, 1H, Jˆ18.8 Hz, C2-H), 3.40 (d, 1H,
Jˆ18.8 Hz, C2-H); 13C NMR (300 MHz, D2O) d 168.9,
167.1, 166.8, 165.1, 164.7, 151.8, 147.1, 136.1, 133.5,
130.5, 120.4, 117.2, 116.3, 115.2, 104.2, 101.8; HRMS
(M1H, FAB): calcd for C17H14ClN6O6S2 497.0105, found
497.0106.
130.9, 119.2, 115.1, 110.7, 106.8, 59.0, 26.9; HRMS
(M1H, FAB): calcd for C18H13ClN7O6S2 522.0057, found
522.0055.
Sodium (6R,7R)-7-(Z)-[2-(2-amino-5-chlorothiazol-4-yl)-
2-hydroxyiminoacetamido]-3-[(3-carbamoylisoxazol-5-
yl)vinyl]-3-cephem-4-carboxylate (1g(Z)). IR (KBr) 3299,
1774, 1677, 1565, 1538, 1355, 1259 cm21 1H NMR
;
(300 MHz, D2O) d 6.60 (d, 1H, Jˆ12./1 Hz, vinyl-H),
6.59 (s, 1H, isoxazole), 6.42 (d, 1H, Jˆ12.1 Hz, vinyl-H),
5.83 (d, 1H, C7-H), 5.28 (d, 1H, C6-H), 3.57 (d, 1H,
Jˆ18.0 Hz, C2-H), 3.28 (d, 1H, Jˆ18.0 Hz, C2-H); 13C
NMR (300 MHz, D2O) d 169.5, 168.7, 167.0, 64.9, 164.8,
163.1, 158.5, 147.0, 136.2, 134.7, 130.7, 120.0, 115.8,
115.0, 103.6, 59.0, 58.1, 27.2; HRMS (M1H, FAB): calcd
for C18H15ClN7O7S2 540.0163, found 540.0162.
Sodium (6R,7R)-7-(Z)-[2-(2-amino-5-chlorothiazol-4-yl)-
2-hydroxyiminoacetamido]-3-[(3-bromoisoxazol-5-yl)-
vinyl]-3-cephem-4-carboxylate (1c(Z)). 1H NMR (300
MHz, D2O) d 6.72 (d, 1H, Jˆ12.0 Hz, vinyl-H), 6.55 (s,
1H, isoxazole), 6.48 (d, 1H, Jˆ11.8 Hz, vinyl-H), 5.94 (d,
1H, Jˆ4.7 Hz, C7-H), 5.37 (d, 1H, Jˆ4.6 Hz, C6-H), 3.68
(d, 1H, Jˆ18.0 Hz, C2-H), 3.38 (d, 1H, Jˆ18.1 Hz, C2-H);
HRMS (M1H, FAB): calcd for C17H13BrClN6O6S2
574.9210, found 574.9207.
Sodium (6R,7R)-7-(Z)-[2-(2-amino-5-chlorothiazol-4-yl)-
2-hydroxyiminoacetamido]-3-[(3-dimethylcarbamoyl-5-
yl)vinyl]-3-cephem-4-carboxylate (1h(Z)). IR (KBr) 3342,
1774, 1629, 1543, 1377, 1253, 1001 cm21 1H NMR
;
(300 MHz, D2O) d 6.74 (d, 1H, Jˆ12.1 Hz, vinyl-H), 6.57
(s, 1H, isoxazole), 6.55 (d, 1H, Jˆ12.1 Hz, vinyl-H), 5.93
(d, 1H, C7-H), 5.37 (d, 1H, C6-H), 3.70 (d, 1H, Jˆ17.9 Hz,
C2-H), 3.39 (d, 1H, Jˆ17.9 Hz, C2-H), 3.12 (s, 3H,
±NCH3±), 3.10 (s, 3H, ±NCH3±); 13C NMR (300 MHz,
D2O) d: 163.7, 163.5, 162.0, 160.0, 159.8, 157.8, 153.6,
142.4, 131.3, 130.0, 126.1, 114.9, 110.6, 110.0, 99.3, 54.0,
53.2, 34.4, 31.0, 22.3; HRMS (M1H, FAB): calcd for
C20H19ClN7O7S2 568.0476, found 568.0482.
Sodium (6R,7R)-7-(Z)-[2-(2-amino-5-chlorothiazol-4-yl)-
2-hydroxyiminoacetamido]-3-[(3-methylisoxazol-5-yl)-
vinyl]-3-cephem-4-carboxylate (1d(Z)). IR (KBr) 3320,
1
1774, 1656, 1543, 1355 cm21; H NMR (300 MHz, D2O)
d
6.45 (d, 1H, Jˆ12.1 Hz, vinyl-H), 6.27 (d, 1H,
Jˆ12.1 Hz, vinyl-H), 6.10 (s, 1H, isoxazole), 5.77 (d, 1H,
C7-H), 5.19 (d, 1H, C6-H), 3.54 (d, 1H, Jˆ17.8 Hz, C2-H),
3.20 (d, 1H, Jˆ17.8 Hz, C2-H), 2.09 (s, 3H, isoxazole-
CH3); 13C NMR (300 MHz, D2O) d 168.8, 167.1, 167.0,
165.0, 164.8, 161.9, 147.1, 136.2, 133.2, 130.6, 120.6,
116.4, 115.0, 105.6, 58.9, 58.0, 27.2, 10.9; HRMS (M1H,
FAB): calcd for C18H16ClN6O6S2 511.0261, found
511.0265.
Sodium (6R,7R)-7-(Z)-[2-(2-amino-5-chlorothiazol-4-yl)-
2-hydroxyiminoacetamido]-3-[(3-carbamoyloxymethyl-
isoxazol-5-yl)vinyl]-3-cephem-4-carboxylate (1i(Z)). IR
(KBr) 3310, 2365, 1769, 1715, 1656, 1543, 1393, 1334,
1098 cm21 1H NMR (300 MHz, D2O) d 6.72 (d, 1H,
;
Jˆ12.0 Hz, vinyl-H), 6.50 (d, 1H, Jˆ12.0 Hz, vinyl-H),
6.43 (s, 1H, isoxazole), 5.95 (d, 1H, Jˆ4.5 Hz, C7-H),
5.37 (d, 1H, Jˆ4.5 Hz, C6-H), 5.15 (s, 2H, ±CH2OCO±),
3.66 (d, 1H, Jˆ17.3 Hz, C2-H), 3.37 (d, 1H, Jˆ17.3 Hz,
C2-H); 13C NMR (300 MHz, D2O) d 168.7, 168.2, 167.1,
165.0, 162.8, 161.3, 158.8, 147.1, 136.2, 134.1, 130.8,
120.4, 116.1, 115.1, 103.7, 58.9, 58.7, 58.0, 27.1; HRMS
(M1H, FAB): calcd for C19H17ClN7O8S2 570.0268, found
570.0268.
Sodium (6R,7R)-7-(Z)-[2-(2-amino-5-chlorothiazol-4-yl)-
2-hydroxyiminoacetamido]-3-[(3-methoxyisoxazol-5-
yl)vinyl]-3-cephem-4-carboxylate (1e(Z)). IR (KBr) 3342,
1
1774, 1656, 1559, 1511, 1409 cm21; H NMR (300 MHz,
D2O) d 6.70 (d, 1H, Jˆ9.0 Hz, vinyl-H), 6.49 (d, 1H,
Jˆ9.0 Hz, vinyl-H), 6.18 (s, 1H, isoxazole), 5.96 (d, 1H,
C7-H), 5.48 (d, 1H, C6-H), 3.97 (s, 3H, ±OCH3), 3.68 (d,
1H, Jˆ18.0 Hz, C2-H), 3.40 (d, 1H, Jˆ18.0 Hz, C2-H); 13C
NMR (300 MHz, D2O) d 173.0, 168.7, 168.5, 167.1, 165.0,
164.8, 147.1, 136.2, 134.2, 134.1, 130.9, 120.2, 116.5,
116.1, 95.2, 58.9, 58.1, 57.9, 27.2; HRMS (M1H, FAB):
calcd for C18H16ClN6O7S2 527.0210, found 527.0208.
Sodium (6R,7R)-7-(Z)-[2-(2-amino-5-chlorothiazol-4-yl)-
2-hydroxyiminoacetamido]-3-[(3-(N,N-dimethylcarba-
moyloxymethyl)isoxazol-5-yl)vinyl]-3-cephem-4-car-
boxylate (1j(Z)). IR (KBr) 3046, 2934, 1774, 1688, 1629,
1
1543, 1404, 1194 cm21; H NMR (300 MHz, D2O) d 6.69
(d, 1H, Jˆ7.5 Hz, vinyl-H), 6.48 (d, 1H, Jˆ7.5 Hz, vinyl-
H), 6.40 (s, 1H, isoxazole), 5.93 (d, 1H, C7-H), 5.32 (d, 1H,
C6-H), 5.15 (s, 2H, ±CH2OCON±), 3.60 (d, 1H, Jˆ18.0 Hz,
C2-H), 3.33 (d, 1H, Jˆ18.0 Hz, C2-H), 2.87 (br d, 6H,
±OCON(CH3)2); 13C NMR (300 MHz, D2O) d 168.6,
168.1, 166.8, 164.8, 164.7, 164.4, 161.3, 157.2, 147.0,
136.4, 134.3, 131.2, 120.4, 115.9, 114.6, 103.8, 59.1, 58.9,
58.2 36.5, 36.1, 27.2; HRMS (M1H, FAB): calcd for
C21H21ClN7O8S2 598.0581, found 598.0585.
Sodium (6R,7R)-7-(Z)-[2-(2-amino-5-chlorothiazol-4-yl)-
2-hydroxyiminoacetamido]-3-[(3-cyanoisoxazol-5-yl)-
vinyl]-3-cephem-4-carboxylate (1f(Z)). IR (KBr) 3331,
3181, 1779, 1672, 1618, 1538, 1409, 1253, 1193, 1055,
1006 cm21; 1H NMR (300 MHz, D2O) d 6.81 (s, 1H, isoxa-
zole), 6.75 (d, 1H, Jˆ12.1 Hz, vinyl-H), 6.53 (d, 1H,
Jˆ12.1 Hz, vinyl-H), 5.92 (d, 1H, J-4.8 Hz, C7-H), 5.36
(d, 1H, Jˆ4.8 Hz, C6-H), 3.19 (d, 1H, Jˆ17.9 Hz, C2-H),
3.38 (d, 1H, Jˆ17.9 Hz, C2-H); 13C NMR (300 MHz, D2O)
d 170.2, 168.6, 167.1, 165.0, 164.7, 147.1, 140.0, 136.2,
Sodium (6R,7R)-7-(Z)-[2-(2-amino-5-chlorothiazol-4-yl)-
2-hydroxyiminoacetamido]-3-[(3-hydroxyisoxazol-5-

A. N. Pae et al. / Tetrahedron 56 (2000) 5657±5665
5663
yl)vinyl]-3-cephem-4-carboxylate (1k(Z)). IR (KBr) 3320,
Sodium (6R,7R)-7-(Z)-[2-(2-aminothiazol-4-yl)-2-hydroxy-
iminoacetamido]-3-[(3-chloroisoxazol-5-yl)vinyl]-3-cephem-
4-carboxylate (2b(Z)). IR (KBr) 3365, 2947, 1666, 1638,
1
2375, 1769, 1656, 1543, 1463, 1366, 1275 cm21; H NMR
(300 MHz, D2O) d 6.52 (d, 1H, Jˆ12.1 Hz, vinyl-H), 6.22
(d, 1H, Jˆ12.1 Hz, vinyl-H), 5.82 (d, 1H, Jˆ4.8 Hz, C7-H),
5.80 (s, 1H, isoxazole), 5.24 (d, 1H, Jˆ4.9 Hz, C6-H), 3.55
(d, 1H, Jˆ17.9 Hz, C2-H), 3.29 (d, 1H, Jˆ17.9 Hz, C2-H);
13C NMR (300 MHz, D2O) d 172.5, 168.9, 168.2, 167.3,
165.3, 164.7, 147.2, 136.0, 133.6, 130.5, 120.4, 117.1,
115.3, 96.6, 59.0, 57.9, 26.9; HRMS (M1H, FAB): calcd
for C17H14ClN6O7S2 513.0054, found 513.0050.
1
1446, 1381, 1267 cm21; H NMR (300 MHz, D2O) d 7.04
(s, 1H, thiazole), 6.72 (d, 1H, vinyl-H), 6.52 (s, 1H, isoxa-
zole), 6.51 (d, 1H, vinyl-H), 5.92 (d, 1H, C7-H), 5.40 (d, 1H,
C6-H), 3.40 (2H, ABq, C2-H0; HRMS (M1H, FAB): calcd
for C17H14ClN6O6S2 497.0105, found 497.0112.
Sodium (6R,7R)-7-(Z)-[2-(2-aminothiazol-4-yl)-2-hydroxy-
iminoacetamido]-3-[(3-methoxylisoxazol-5-yl)vinyl]-3-
cephem-4-carboxylate (2e(Z)). IR (KBr) 3045, 2957, 1666,
Sodium (6R,7R)-7-(Z)-[2-(2-amino-5-chlorothiazol-4-yl)-
2-hydroxyiminoacetamido]-3-[(3-(2-¯uoroethoxy)isoxa-
1
1636, 1576, 1526, 1466, 1411 cm21; H NMR (300 MHz,
1
zol-5-yl)vinyl]-3-cephem-4-carboxylate (1l(Z)). H NMR
D2O) d 7.02 (s, 1H, thiazole), 6.64 (d, 1H, vinyl-H), 6.36 (d,
1H, vinyl-H), 6.07 (s, 1H, isoxazole), 5.88 (d, 1H, C7-H),
5.36 (d, 1H, C6-H), 3.95 (s, 3H, isoxazole±OCH3), 3.38
(ABq, 2H, C2-H); HRMS (M1H, FAB): calcd for
C18H17N6O7S2 493.0600, found 493.0606.
(300 MHz, D2O) d 6.70 (d, 1H, Jˆ12.1 Hz, vinyl-H), 6.40
(d, 1H, Jˆ12.1 Hz, vinyl-H), 6.12 (s, 1H, isoxazole), 5.96 (d,
1H, C7-H), 5.37 (d, 1H, C6-H), 4.90 (m, 1H, ±OCH2CH2F),
4.75 (m 1H, ±OCH2CH2F), 4.56 (m 1H, ±OCH2CH2F), 4.46
(m 1H, ±OCH2CH2F), 3.70 (d, 1H, Jˆ18.0 Hz, C2-H), 3.40
(d, 1H, Jˆ18.0 Hz, C2-H); 13C NMR (300 MHz, D2O) d
167.1, 163.7, 162.1, 160.0, 142.1, 131.2, 129.3, 126.2, 115.1,
111.3, 110.0, 90.4, 78.8, 76.6, 65.0, 64.8, 54.0, 53.2, 22.4.
Sodium (6R,7R)-7-(Z)-[2-(2-aminothiazol-4-yl)-2-hydroxy-
iminoacetamido]-3-[(3-(thiazol-2-yl)isoxazol-5-yl)vinyl]-3-
cephem-4-carboxylate (2m(Z)). IR (KBr) 3315, 1774,
1
1656, 1549, 1387, 936 cm21; H NMR (300 MHz, D2O) d
Sodium (6R,7R)-7-(Z)-[2-(2-amino-5-chlorothiazol-4-yl)-
2-hydroxyiminoacetamido]-3-[(3-(thiophen-2-yl)isoxa-
zol-5-yl)vinyl]-3-cephem-4-carboxylate (1m(Z)). IR
(KBr) 3353, 3170, 1779, 1661, 1618, 1565, 1425, 1393,
1259, 996 cm21; 1H NMR (300 MHz, CD3OD) d 7.61 (dd,
1H, Jˆ3.6 Hz, 1 Hz, thiophen), 7.56 (dd, 1H, Jˆ5.0, 1 Hz,
thiophen), 7.17 (dd, 1H, Jˆ5.0 Hz, 3.6 Hz, thiophen), 6.91
(d, 1H, Jˆ12.3 Hz, vinyl-H), 6.74 (s, 1H, isoxazole), 6.40
(d, 1H, Jˆ12.3 Hz, vinyl-H), 5.91 (d, 1H, Jˆ4.9 Hz, C7-H),
5.26 (d, 1H, Jˆ4.9 Hz, C6-H), 3.68 (d, 1H, Jˆ17.4 Hz, C2-
H), 3.32 (d, 1H, Jˆ17.4 Hz, C2-H); 13C NMR (300 MHz,
CD3OD) d 169.4, 169.1, 167.0, 165.5, 165.0, 159.3, 148.1,
138.7, 135.7, 134.9, 131.5, 129.5, 128.9, 128.9, 117.8,
115.0, 113.1, 103.0, 60.3, 59.9, 27.9; HRMS (M1H, FAB):
calcd for C12H16ClN6O6S3 578.9982, found 578.9983.
7.67 (d, 1H, Jˆ3.2 Hz, thiazole), 7.53 (d, 1H, Jˆ3.2 Hz,
thiazole), 6.75 (s, 1H, isoxazole), 6.64 (d, 1H, Jˆ12.0 Hz,
vinyl-H), 6.58 (s, 1H, thiazole), 7.53 (d, 1H, Jˆ3.2 Hz,
thiazole), 6.75 (s, 1H, isoxazole), 6.64 (d, 1H, Jˆ12.0 Hz,
vinyl-H), 6.58 (s, 1H, thiazole), 6.35 (d, 1H, Jˆ12.1 Hz,
vinyl-H), 5.76 (d, 1H, Jˆ4.6 Hz, C7-H), 5.22 (d, 1H,
Jˆ4.7 Hz, C6-H), 3.49 (d, 1H, Jˆ17.0 Hz, C2-H), 3.12 (d,
1H, Jˆ17.6 Hz, C2-H); 13C NMR (300 MHz, D2O) d 165.9,
163.8, 163.5, 159.5, 152.8, 151.4, 143.2, 138.6, 136.0,
130.0, 126.2, 117.9, 115.6, 110.5, 107.4, 97.3, 54.2, 53.4,
22.6; HRMS (M1H, FAB): calcd for C20H16N7O6S3
546.0324, found 546.0329.
Sodium (6R,7R)-7-(Z)-[2-(2-aminothiazol-4-yl)-2-hydroxy-
iminoacetamido]-3-[(3(thiophen-2-yl)isoxazol-5-yl)vinyl]-
3-cephem-4-carboxylate (2n(Z)). IR (KBr) 3375, 2871,
1
Sodium (6R,7R)-7-(Z)-[2-(2-amino-5-chlorothiazol-4-yl)-
2-hydroxyiminoacetamido]-3-[(3-(thiazol-2-yl)isoxazol-
5-yl)vinyl]-3-cephem-4-carboxylate (1n(Z)). IR (KBr)
3331, 3202, 2966, 1774, 1656, 1618, 1543, 1361, 1259,
1740, 1651, 1636, 1571, 1406, 1272, 729 cm21; H NMR
(300 MHz, D2O) d 7.23 (dd, 1H, thiophen), 7.15 (dd, 1H,
thiophen), 6.76 (s, 1H, thiophen), 6.68 (s, 1H, isoxazole),
6.58 (d, 1H, Jˆ12.0 Hz, vinyl-H), 6.30 (s, 1H, thiazole),
6.23 (d, 1H, Jˆ12.2 Hz, vinyl-H), 5.70 (d, 1H, Jˆ4.6 Hz,
C6-H), 3.31 (d, 1H, Jˆ17.6 Hz, C2-H), 2.95 (d, 1H,
Jˆ17.5 Hz, C2-H); 13C NMR (300 MHz, D2O) d 170.8,
168.4, 167.7, 165.2, 164.3, 158.1, 148.4, 134.5, 131.7,
129.5, 129.1, 128.7, 128.4, 115.5, 112.5, 112.5, 102.6,
59.4, 58.6, 27.5; HRMS (M1H FAB): calcd for
C21H17N6O6S3 545.0371, found 545.0373.
1
1055, 818 cm21; H NMR (300 MHz, D2O) d 7.86 (d, 1H,
Jˆ3.3 Hz, Jˆ3.3 Hz, thiazole), 7.67 (d, 1H, Jˆ12.0 Hz,
thiazole), 6.77 (s, 1H, isoxazole), 6.66 (d, 1H, Jˆ12.0 Hz,
vinyl-H), 6.45 (d, 1H, Jˆ12.0 Hz, vinyl-H), 5.85 (d, 1H,
Jˆ4.8 Hz, C7-H), 4.29 (d, 1H, Jˆ4.8 Hz, C6-H0, 3.59 (d,
1H, Jˆ18.0 Hz, C2-H), 3.25 (d, 1H, Jˆ18.0 Hz, C2-H); 13C
NMR (300 MHz, CD3OD) d 169.6, 168.1, 166.4, 164.6,
164.4, 158.4, 156.8, 147.1, 143.9, 137.2, 135.7, 132.9,
122.7, 117.9, 114.5, 113.3, 102.0, 59.1, 58.4, 27.0; HRMS
(M1H, FAB): calcd for C20H15ClN7O6S3 579.9934, found
479.9936.
Sodium (6R,7R)-7-[2-(2-amino-5-chlorothiazol-4-yl)-2-
hydroxyiminoacetamido]-3-[(isothiazol-5-yl)vinyl]-3-
cephem-4-carboxylate (3a(E,Z)). IR (KBr) 3320, 2365,
1
1769, 1667, 1624, 1543, 1371, 1307, 1253, 1001 cm21; H
Sodium (6R,7R)-7-(Z)-[2-(2-aminothiazol-4-yl)-2-hydroxy-
iminoacetamido]-3-[(isoxazol-5-yl)vinyl]-3-cephem-4-
carboxylate (2a(Z)). IR (KBr) (300 MHz, D2O) d 8.36 (s,
1H, isozazole), 7.00 (s, 1H, thiazole), 6.62 (d, 1H, vinyl-H),
6.50 (d, 1H, vinyl-H), 6.46 (s, 1H, isoxazole), 5.84 (d, 1H,
C7-H), 5.34 (d, 1H, C6-H), 4.03 (s, 3H, ±NOCH3), 3.35
(ABq, 2H, C2-H); HRMS (M1H, FAB): calcd for
C17H15N6O6S2 463.0494, found 463.0487.
NMR (300MHz, D2O) d E; 8.28 (s, 1H, isothiazole), 7.50 (s,
1H, isothiazole), 7.22 (d, 1H, Jˆ16.0 Hz, vinyl-H), 6.87 (d,
1H, Jˆ16.0 Hz, vinyl-H), 5.80 (d, 1H, Jˆ4.8 Hz, C7-H),
5.18 (d, 1H, Jˆ4.8 Hz, C6-H), 3.68 (d, 1H, Jˆ17.3 Hz,
C2-H), 3.59 (d, 1H, Jˆ17.3 Hz, C2-H) Z; 8.28 (s, 1H,
isothiazole), 7.15 (s, 1H, isothiazole), 6.68 (d, 1H,
Jˆ4.8 Hz, C6-H), 3.53 (d, 1H, Jˆ18.3 Hz, C2-H), 3.26 (d,
1H, Jˆ18.3 Hz, C2-H); 13C NMR (300 MHz, D2O) d 168.6,

5664
A. N. Pae et al. / Tetrahedron 56 (2000) 5657±5665
168.4, 166.8, 165.1, 164.6, 164.4, 159.2, 158.1, 146.9,
136.3, 132.9, 130.8, 122.1, 117.8, 114.9, 59.1, 58.0, 24.3;
HRMS (M1H, FAB): calcd for C17H14ClN6O5S3 512.9876,
found 512.9875.
(d, 1H, Jˆ4.1 Hz, vinyl-H), 5.81 (d, 1H, Jˆ4.7 Hz,
C7-H), 5.32 (d, 1H, Jˆ4.7 Hz, C6-H), 3.63 (s, 1H, C2-H),
Z: d 8.23 (s, 1H, isothiazole), 7.07 (s, 1H0, 6.67 (d, 1H,
Jˆ11.4 Hz, vinyl-H), 6.38 (d, 1H, Jˆ11.1 Hz, vinyl-H),
5.76 (d, 1H, Jˆ4.7 Hz, C7-H), 5.19 (d, 1H, Jˆ4.7 Hz,
C6-H), 3.51 (d, 1H, Jˆ16.7 Hz, C2-H), 3.24 (d, 1H,
Jˆ18.0 Hz, C2-H); 13C NMR (300 MHz, D2O) d 171.1,
171.0, 168.8, 168.5, 165.5, 165.2, 164.2, 163.9, 161.4,
159.2, 158.1, 148.6, 148.5, 141.4, 132.7, 141.4, 132.7,
132.0, 131.1, 130.6, 125.4, 121.9, 121.7, 121.0, 117.9,
112.7, 59.3, 59.1, 57.5, 26.5, 24.0; HRMS (M1H, FAB):
calcd for C17H15N6O5S3 479.0266, found 479.0269.
Sodium (6R,7R)-7-(E)-[2-(2-amino-5-chlorothiazol-4-yl)-
2-hydroxyiminoacetamido]-3-[(isothiazol-5-yl)vinyl]-3-
cephem-4-carboxylate (3a(E)). IR (KBr) 3320, 3181,
1
1774, 1672, 1543, 1371, 1307, 1253, 1001 cm21; H NMR
(300 MHz, D2O) d 8.28 (s, 1H, isothiazole), 7.50 (s, 1H,
isothiazole), 7.22 (d, 1H, Jˆ16.0 Hz, vinyl-H), 6.87 (d, 1H,
Jˆ16.0 Hz, vinyl-H), 5.80 (d, 1H, Jˆ4.8 Hz, C7-H), 5.18
(d, 1H, Jˆ4.8 Hz, C6-H), 3.68 (d, 1H, Jˆ18.3 Hz, C2-H),
3.59 (d, 1H, Jˆ18.3 Hz, C2-H); HRMS (M1H, FAB): calcd
for C17H14ClN6O5S3 512.9876, 512.9874.
Sodium (6R,7R)-7-(E)-[2-(2-aminothiazol-4-yl)-2-hydroxy-
iminoacetamido]-3-[(3-methoxyisothiazol-5-yl)vinyl]-3-
cephem-4-carboxylate (4e(E)). 1H NMR (300 MHz, D2O)
d 7.19 (d, 1H, Jˆ15.9 Hz, vinyl-H),6.95 (s, 1H, thiazole),
6.80 (d, 1H, Jˆ16.5 Hz, vinyl-H), 6.66 (s, 1H, isothiazole),
5.86 (d, 1H, 4.8 Hz, C7-H), 5.29 (s, 1H, Jˆ4.4 Hz, C6-H),
3.95 (s, 3H, ±OCH3), 3.70 (ABq, 2h, C2-H); HRMS (M1H,
FAB): calcd for C18H17N6O6S3 509.0371, found 509.0372.
Sodium (6R,7R)-7-(E)-[2-(2-amino-5-chlorothiazol-4-yl)-
2-hydroxyiminoacetamido]-3-[(3-methoxyisothiazol-5-
yl)vinyl]-3-cephem-4-carboxylate (3e(E)). IR (KBr) 3415,
2937, 1775, 1666, 1636, 1546, 1381 cm21 1H NMR
;
(300 MHz, D2O) d 7.09 (d, 1H, 16.1 Hz, vinyl-H), 6.73
(d, 1H, Jˆ16.0 Hz, vinyl-H), 6.59 (s, 1H, isothiazole),
5.78 (d, 1H, Jˆ4.7 Hz, C7-H), 5.16 (d, 1H, Jˆ4.74 Hz,
C6-H), 3.84 (s, 3H, ±OCH3), 3.61 (ABq, 2H, Jˆ29.6 Hz,
Jˆ17.4 Hz, C2-H); 13C NMR (300 MHz, D2O) d 171.2,
169.2, 166.9, 166.7, 165.7, 164.3, 148.0, 138.6, 132.3,
117.7, 116.2, 109.5, 60.2, 59.2, 56.0, 24.6; HRMS (M1H
FAB): calcd for C18H16ClN6O6S3 542.9982, found
542.9987.
Sodium (6R,7R)-7-(Z)-[2-(2-aminothiazol-4-yl)-2-hydroxy-
iminoacetamido]-3-[(3-methoxyisothiazol-5-yl)vinyl]-3-
1
cephem-4-carboxylate (4e(Z)). H NMR (300 MHz, D2O)
d 7.00 (s, 1H, thiazole), 6.67 (d, 1H, Jˆ11.5 Hz, vinyl-H),
6.62 (s, 1H, isothiazole), 6.53 (d, 1H, Jˆ11.5 Hz, vinyl-H),
5.92 (d, 1H, Jˆ4.8 Hz, C7-H), 5.43 (d, 1H, Jˆ4.7 Hz,
C6-H), 3.95 (s, 3H, ±OCH3), 3.65 (d, 1H, Jˆ14.8 Hz,
C2-H), 3.38 (d, 1H, Jˆ14.8 Hz, C2-H); HRMS (M1H,
FAB): calcd for C18H17N6O6S3 509.0371, found
509.0372.
Sodium (6R,7R)-7-(Z)-[2-(2-amino-5-chlorothiazol-4-yl)-
2-hydroxyiminoacetamido]-3-[(3-methoxyisothiazol-5-
yl)vinyl]-3-cephem-4-carboxylate (3e(Z)). IR (KBr) 3315,
3106, 2937, 1760, 1735, 1621, 1506, 1451, 1362, 1237,
Sodium (6R,7R)-7-(E)-[2-(2-aminothiazol-4-yl)-2-hydroxy-
iminoacetamido]-3-[(3-hydroxyisothiazol-5-yl)vinyl]-3-
cephem-4-carboxylate (4k(E)). 1H NMR (300 MHz, D2O)
d 7.07 (d, 1H, Jˆ15.5 Hz, vinyl-H), 6.90 (s, 1H, iso-
thiazole), 6.75 (d, 1H, Jˆ15.9 Hz, vinyl-H), 6.30 (s, 1H,
thiazole), 5.80 (d, 1H, Jˆ3.6 Hz, C7-H), 5.22 (d, 1H,
Jˆ3.4 Hz, C6-H), 3.65 (ABq, 2H, Jˆ24.8 Hz, Jˆ17.7 Hz,
C2-H); HRMS (M1H, FAB): calcd for C17H15N6O6S3
495.0215, found 495.0219.
1
1177, 1053, 699 cm21; H NMR (300 MHz, D2O) d 6.60
(d, 1H, Jˆ11.4 Hz, vinyl-H), 6.56 (s, 1H, isothiazole) 6.43
(d, 1H, Jˆ11.3 Hz, vinyl-H), 5.87 (d, 1H, Jˆ4.8 Hz, C7-H),
5.35 (d, 1H, Jˆ4.8 Hz, C6-H), 3.88 (s, 3H, ±OCH3), 3.61 (d,
1H, Jˆ18.2 Hz, C2-H), 3.32 (d, 1H, Jˆ18.3 Hz, C2-H); 13C
NMR (300 MHz, D2O) d 169.3, 168.3, 166.9, 164.8, 161.9,
146.9, 136.3, 132.5, 131.1, 121.6, 120.69, 114.9, 112.5,
58.8, 57.3, 26.7, 20.9; HRMS (M1H, FAB): calcd for
C18H16ClN6O6S3 542.9982, found 542.9987.
Sodium (6R,7R)-7-(Z)-[2-(2-aminothiazol-4-yl)-2-hydroxy-
iminoacetamido]-3-[(3-hydroxyisothiazol-5-yl)vinyl]-3-
cephem-4-carboxylate (4k(Z)). 1H NMR (300 MHz, D2O)
d 6.94 (s, 1H, isothiazole), 6.58 (d, 1H, Jˆ11.4 Hz, vinyl-
H), 6.41 (d, 1H, Jˆ11.3 Hz, vinyl-H), 6.23 (s, 1H, thiazole),
5.85 (d, 1H, Jˆ4.7 Hz, C7-H), 5.36 (d, 1H, Jˆ4.8 Hz,
C6-H), 3.63 (d, 1H, Jˆ18.2 Hz, C2-H), 3.33 (d, 1H,
Jˆ18.2 Hz, C2-H); HRMS (M1H, FAB): calcd for
C17H15N6O6S3 495.0215, found 495.0219.
Sodium (6R,7R)-7-(Z)-[2-(2-amino-5-chlorothiazol-4-yl)-
2-hydroxyiminoacetamido]-3-[(3-hydroxyisothiazol-5-
yl)vinyl]-3-cephem-4-carboxylate (3k(Z)). IR (KBr) 3375,
1
3226, 2927, 1785, 1656, 1606, 1546, 1347, 1267 cm21; H
NMR (300 MHz, D2O) d 6.87 (d, 1H, Jˆ11.2 Hz, vinyl-H),
6.54 (d, 1H, Jˆ11.5 Hz, vinyl-H), 6.33 (s, 1H, isothiazole),
5.97 (d, 1H, Jˆ4.7 Hz, C7-H), 5.43 (d, 1H, Jˆ4.7 Hz, C6-H),
3.69 (d, 1H, Jˆ18.2 Hz, C2-H), 3.40 (d, 1H, Jˆ18.1 Hz,
C2-H); 13C NMR (300 MHz, D2O) d 172.3, 168.4, 167.2,
165.2, 164.7, 158.2, 147.1, 136.0, 133.4, 131.4, 122.1, 120.2,
115.3, 114.1, 58.9, 57.3, 26.4; HRMS (M1H FAB): calcd for
C17H14ClN6O6S3 528.9825, found 528.9826.
Sodium (6R,7R)-7-(Z)-[2-(2-amino-5-chlorothiazol-4-yl)-
2-hydroxyiminoacetamido]-3-[(1,2,5-thiadiazol-5-yl)-
vinyl]-3-cephem-4-carboxylate (5a(Z)). 1H NMR (300
MHz, D2O) d 8.54 (s, 1H, thiadiazole), 6.65 (d, 1H,
Jˆ12.9 Hz, vinyl-H), 6.57 (d, 1H, Jˆ12.0 Hz, vinyl-H)
5.78 (d, 1H, Jˆ4.8 Hz, C7-H), 5.24 (d, 1H, Jˆ4.8 Hz,
C6-H), 3.60 (d, 1H, Jˆ17.9 Hz, C2-H), 3.28 (d, 1H,
17.8 Hz, C2-H); 13C NMR (300 MHz, D2O) d 168.9,
163.1, 161.1, 159.2, 151.2, 149.0, 133.5, 130.6, 122.1,
119.6, 115.2, 59.8, 58.9, 57.9, 27.2, 23.2.
Sodium (6R,7R)-7-[2-(2-aminothiazol-4-yl)-2-hydroxy-
iminoacetamido]-3-[(isothiazol-5-yl)vinyl]-3-cephem-4-
carboxylate (4a(E,Z)). IR (KBr) 3320, 2365, 1758, 1656,
1538, 1387, 1312, 1264, 1055, 1028 cm21 1H NMR
;
(300 MHz, D2O) E: d 8.26 (s, 1H, isothiazole), 7.19 (s,
1H), 7.14 (d, 1H), 7.14 (d, 1H, Jˆ4.9 Hz, vinyl-H), 6.88

A. N. Pae et al. / Tetrahedron 56 (2000) 5657±5665
5665
Sodium (6R,7R)-7-[2-(2-amino-5-chlorothiazol-4-yl)-2-
hydroxyiminoacetamido]-3-[(3-methylisothiazol-4-yl)-
vinyl]-3-cephem-4-carboxylate (5d(E,Z)). IR (KBr) 3385,
3315, 3196, 2847, 1735, 1641, 1621, 1491, 1367, 1272,
C6-H), 3.40 (d, 1H, Jˆ17.7 Hz, C2-H), 3.15 (d, 1H,
Jˆ17.8 Hz, C2-H), 2.34 (s, 3H), E: d 8.85 (s, 1H, iso-
thiazole), 7.18 (d, 1H, Jˆ16.3 Hz, vinyl-H), 6.67 (d, 1H,
Jˆ16.5 Hz, vinyl-H), 6.47 (s, 1H, thiazole), 5.86 (d, 1H,
C7-H), 5.29 (d, 1H, C6-H), 3.77 (ABq, 2H, C2-H); 13C
NMR (300 MHz, D2O) d 171.0, 169.0, 167.3, 165.5,
164.3, 148.5, 146.8, 141.4, 133.7, 130.9, 129.7, 123.0,
121.7, 112.6, 59.2, 58.4, 27.2, 17.6, 17.2; HRMS (M1H,
FAB): calcd for C18H17N6O5S3 493.0422, found 493.0425.
1
1013, 918 cm21; H NMR (300 MHz, D2O) E; d 8.85 (s,
1H, isothiazole), 7.18 (d, 1H, Jˆ15.4 Hz, vinyl-H), 6.68 (d,
1H, Jˆ16.6 Hz, vinyl-H), 6.47 (s, 1H), 5.86 (d, 1H,
Jˆ4.53 Hz, C7-H), 5.29 (d, 1H, Jˆ4.6 Hz, C6-H), 3.86 (d,
1H, Jˆ17.6 Hz, C2-H), 3.72 (d, 1H, Jˆ17.6 Hz, C2-H) Z; d
8.62 (s, 1H, isothiazole), 6.95 (d, 2H, Jˆ15.3 Hz, vinyl-H),
6.47 (s, 1H), 6.47 (s, 1H), 5.81 (d, 1H, Jˆ4.6 Hz, C7-H),
5.26 (d, 1H, Jˆ4.7 Hz, C6-H), 3.46 (d, 1H, Jˆ17.6 Hz,
C2-H), 3.23 (d, 1H, Jˆ17.6 Hz, C2-H); 13C NMR
(300 MHz, D2O) d 169.1, 167.4, 167.1, 164.4, 146.9,
133.7, 130.6, 129.7, 123.1, 120.7, 120.3, 59.1, 58.9, 58.2,
58.0, 26.9, 17.1; HRMS (M1H, FAB): calcd for
C18H16CIN6O5S3 527.0033, found 527.0037.
Acknowledgements
We are grateful to the Korean Ministry of Science and
Technology for ®nancial support.
Sodium (6R,7R)-7-(E)-[2-(2-aminothiazol-4-yl)-2-hydroxy-
iminoacetamido]-3-[(1,2,5-thiadiazol-5-yl)vinyl]-3-cephem-
4-carboxylate (6a(E)). IR (KBr) 3395, 2971, 1745, 1666,
1606, 1536, 1381, 1207 cm21; 1H NMR (300 MHz, D2O) d
8.76 (s, 1H, thiadiazole), 7.60 (d, 1H, Jˆ11.3 Hz, vinyl-H),
6.98 (d, 1H, Jˆ11.3 Hz, vinyl-H), 6.92 (s, 1H, thiazole).
5.81 (m, 1H, C7-H), 5.28 (m, 1H, C6-H), 3.74 (q, 1H,
C2-H); 13C NMR (300 MHz, D2O) d 163.9, 163.0, 161.1,
149.7, 149.0, 136.5, 133.2, 132.6, 119.4, 117.3, 113.3,
110.9, 103.3, 59.3, 59.2, 23.7; HRMS (M1H, FAB): calcd
for C16H14N7O5S3 480.0218, found 480.0221.
References
1. (a) Choi, K. I.; Cha, J. H.; Pae, A. N.; Cho, Y. S.; Kang, H. Y.;
Koh, H. Y.; Chang, M. H. J. Antibiot. 1995, 48, 1371. (b) Choi,
K. I.; Cha, J. H.; Pae, A. N.; Cho, Y. S.; Kang, H. Y.; Koh, H. Y.;
Chang, M. H. J Antibiot. 1995, 48, 1375. (c) Choi, K. I.; Cha, J. H.;
Pae, A. N.; Cho, Y. S.; Kang, H. Y.; Koh, H. Y.; Chang, M. H.
J. Antibiot. 1997, 50, 279. (d) Choi, K. I.; Cha, J. H.; Pae, A. N.;
Cho, Y. S.; Kang, H. Y.; Chang, M. H.; Park, S. H.; Park, S. Y.;
Kim, D. Y.; Jeong, D. Y.; Kim, Y. H.; Kong, J. Y.; Koh, H. Y.
J. Antibiot. 1998, 51, 1122. (e) Choi, K. I.; Cha, J. H.; Pae, A. N.;
Cho, Y. S.; Kim, Y. S.; Chang, M. H.; Koh, H. Y. J. Antibiot. 1998,
51, 1117. (f) Choi, K. I. Drugs Future 1999, 24 (3), 287.
2. Glinka, T.; Cho, I.; Zhang, Z.; Price, M.; Case, L.; Crase, J.;
Frith, R.; Liu, N.; Ludwikow, M.; Rea, D.; Chamberland, S.; Lee,
V.; Hecker, S. Abstract No. F176, 37th Intresci. Antimicrob.
Agents Chemother. Toronto, Ontario, Sept. 28, 1997.
Sodium (6R,7R)-7-(Z)-[2-(2-aminothiazol-4-yl)-2-hydroxy-
iminoacetamido]-3-[(1,2,5-thiadiazol-5-yl)vinyl]-3-cephem-
4-carboxylate (6a(Z)). 1H NMR (300 MHz, D2O) d 8.55 (s,
1H, thiadiazole), 6.90 (s, 1H, thiazole), 6.66 (d, 1H,
Jˆ11.9 Hz, vinyl-H), 6.58 (d, 1H, Jˆ11.9 Hz, vinyl-H),
5.79 (d, 1H, Jˆ4.7 Hz, C7-H), 5.28 (d, 1H, Jˆ4.7 Hz,
C6-H), 3.63 (d, 1H, Jˆ17.7 Hz, C2-H), 3.29 (d, 1H,
Jˆ17.8 Hz, C2-H); 13C NMR (300 MHz, D2O) d 171.2,
168.8, 165.7, 164.2, 159.2, 151.2, 148.7, 141.3, 133.5,
130.6, 122.1, 119.8, 112.8, 59.0, 58.0, 27.2; HRMS
(M1H, FAB): calcd for C16H14N7O5S3 480.0218, found
480.0221.
3. (a) Taylor, E. C.; Ray P. S. J. Org. Chem. 1991, 56, 1812.
(b) Chiarino, D.; Napoletano, M.; Sala, A. Synth. Commun.
1988, 18, 1171.
4. (a) Woodward, R. B.; Reed, W. A. J. Am. Chem. Soc. 1943, 65,
1569. (b) Lykkeberg, J.; Krogsgaard-Larsen P. Acta Chem. Scand.
B 1976, 30 (8), 781. (c) Matzen, L.; Engesgaard A.; Ebert B.;
Didriken, M.; Frolund, B.; Krogsgaard-Larsen, P.; Jaroszewski,
J. W. J. Med. Chem. 1997, 40 (4), 520. (d) Howe R. K.; Gruner,
T. A.; Carter, L. G.; Franz, J. E. J. Heterocycl. Chem. 1978, 15,
1001. (e) Kobori T.; Fujita, M.; Hiyama, T.; Kondo, Kiyosi. Synlett
1992, 95.
Sodium (6R,7R)-7-[2-(2-aminothiazol-4-yl)-2-hydroxy-
iminoacetamido]-3-[(3-methylisothiazol-4-yl)vinyl]-3-
cephem-4-carboxylate (6d(E,Z)). IR (KBr) 3465, 3226,
1
2897, 1775, 1666, 1586, 1391 cm21; H NMR (300 MHz,
5. Leitner, F.; Misiek, M.; Pursiano, T. A.; Buck, R. E.; Chisholm,
D. R.; Deregis, R. G.; Tsai, Y. H.; Price, K. E. Antimicrob. Agents
Chemother. 1976, 10, 426.
D2O) Z: d 8.55 (s, 1H, isothiazole), 6.41 (s, 2H, vinyl-H),
5.79 (d, 1H, Jˆ4.72 Hz, C7-H), 5.18 (d, 1H, Jˆ4.7 Hz,