Discovery of novel trans-3,5-disubstituted pyrrolidinylthio-1β-methylcarbapenems

Article abstract of DOI:10.1016/S0968-0896(00)00125-5

Novel trans-3,5-disubstituted pyrrolidinylthio-1β-methylcarbapenems were designed and synthesized to provide J-111,347 (1a) as the first example of an exceptionally broad-spectrum antibiotic, showing activity against methicillin-resistant Staphyloccocus aureus (MRSA) as well as Pseudomonas aeruginosa. Further derivation of 1a afforded J-111,225 (2a), J-114,870 (3a), and J-114,871 (3b), which showed improved safety profiles and retained broad-spectrum antibacterial activities. Copyright (C) 2000 Elsevier Science Ltd.

Full text of DOI:10.1016/S0968-0896(00)00125-5

Bioorganic & Medicinal Chemistry 8 (2000) 1969±1982
Discovery of Novel trans-3,5-Disubstituted Pyrrolidinylthio-
1ꢀ-Methylcarbapenems
Hideaki Imamura,* Norikazu Ohtake, Aya Shimizu, Hiroki Sato, Yuichi Sugimoto,
Shunji Sakuraba, Rie Nagano, Masato Nakano, Shinnosuke Abe, Chihiro Suzuki-Sato,
Ikuko Nishimura, Hisaki Kojima, Yoshimi Tsuchiya, Koji Yamada,
Terutaka Hashizume and Hajime Morishima
Banyu Tsukuba Research Institute, Okubo-3, Tsukuba 300-2611, Ibaraki, Japan
Received 17 February 2000; accepted 26 April 2000
AbstractÐNovel trans-3,5-disubstituted pyrrolidinylthio-1b-methylcarbapenems were designed and synthesized to provide J-
111,347 (1a) as the ®rst example of an exceptionally broad-spectrum antibiotic, showing activity against methicillin-resistant Sta-
phyloccocus aureus (MRSA) as well as Pseudomonas aeruginosa. Further derivation of 1a a€orded J-111,225 (2a), J-114,870 (3a),
and J-114,871 (3b), which showed improved safety pro®les and retained broad-spectrum antibacterial activities. # 2000 Elsevier
Science Ltd. All rights reserved.
Introduction
good ecacy against P. aeruginosa, and with signi®cant
but insucient ecacy against MRSA.
Emergence of multidrug-resistant bacteria, especially
MRSA, has caused serious concern in clinics worldwide.
Since there is no better chemotherapeutic agent against
MRSA, vancomycin has been used in spite of its signi®cant
adverse e€ect pro®le.1 Recently, the emergence of van-
comycin-resistant MRSA has raised an alarm for the
overuse of vancomycin.2 Therefore, development of new
chemotherapeutic agents showing potent activity against
MRSA and good safety pro®les has been urgently
demanded. Carbapenem antibiotics, such as imipenem,
meropenem, panipenem, and biapenem, possess better
safety pro®les and superior activity against a broad range
of pathogens including Pseudomonas aeruginosa, another
important pathogen in the clinic. However, these carba-
penems have no appreciable ecacy against MRSA.
On the other hand, we also reported BO-3482,4 a dithio-
carbamate carbapenem possessing potent anti-MRSA
activity, and demonstrated that increased lipophilicity of
the C-2 side chain enhanced anti-MRSA activity. Simi-
larly, Sumitomo's structure±activity relationship studies
of carbapenems having substituted-thiazol-2-ylthio side
chains at the C-2 position indicated that more lipophilic
phenyl-substituted thiazolthio analogues showed greater
activity against MRSA than simple alkyl-substituted
thiazolthio analogues.5
Based on these observations, we designed and synthesized
new carbapenems with pyrrolidinylthio side chains which
possessed well-balanced hydrophobicity and basicity, in
order to obtain novel carbapenems exhibiting sucient
ecacy against MRSA as well as P. aeruginosa. That is, a
phenyl ring was introduced as a hydrophobic site between
the pyrrolidine ring and an aminoalkyl substituent, a
basic site (Fig. 1).
In the course of our structure±activity relationship studies
of carbapenems having pyrrolidinylthio side chains at the
C-2 position, we found that introduction of an additional
basic function onto the pyrrolidine ring was important for
enhancing anti-pseudomonal activity. Consequently, we
reported the development of the potent carbapenem BO-
27273 which exhibited broad-spectrum activity with
Among the compounds with or without several kinds of
spacers between the pyrrolidine ring and the phenyl ring,
J-111,347 (1a), having a 4-(aminomethyl)phenyl group
directly attached at C-5 of the pyrrolidine ring, was
found to have excellent antibacterial activity against
MRSA and P. aeruginosa. With regard to the structure
*Corresponding author. Fax: +81-298-77-2027; e-mail: imamrahk@-
banyu.co.jp
0968-0896/00/$ - see front matter # 2000 Elsevier Science Ltd. All rights reserved.
PII: S0968-0896(00)00125-5

1970
H. Imamura et al. / Bioorg. Med. Chem. 8 (2000) 1969±1982
Figure 1.
of the pyrrolidine ring part, it should be noted that 1a
had an unusual trans-(3S,5R)-stereochemistry di€ering
from the well-known 1b-methylcarbapenems such as
meropenem,6 S-4661,7 MK-826 (L-749,345),8 and BO-
2727, which have a cis-(3S,5S)-disubstituted pyrrolidine
ring as a C-2 side chain. Since 1a showed epileptogenicity
in the rat head assay (200 mg/rat-head), it should be
modi®ed for clinical use.
biological properties of the new carbapenems, 2a, 3a,
3b, and their related compounds (Fig. 1).
Results and Discussion
Chemistry
We synthesized carbapenems having various spacers,
such as methylene (4a, 4b), thiomethylene (5a, 5b),
aminomethylene (6b), and oxymethylene (7b) functions,
between the pyrrolidine and phenyl ring, by using the
method described in our previous paper10 (Fig. 2) or
that used for FR21818.11
In order to eliminate the undesirable epileptogenic
potency, the aminomethyl moiety of 1a was modi®ed by
introducing various kinds of substituents, 1) onto the
nitrogen of the primary amino function, or 2) onto the
benzilic carbon adjacent to the amino function. These
derivations a€orded three analogues, J-111,225 (2a), J-
114,870 (3a), and J-114,871 (3b), which maintained
excellent antimicrobial activity similar to 1a and showed
no epileptogenicity in the rat-head assay. Following the
preliminary account of this work presented previously,9
we describe herein the details of the creation of 1a, and
the synthesis, structure±activity relationships, and some
J-111,225 (2a), J-114,870 (3a), J-114,871 (3b), and their
related compounds (1a, 1b, 2b±j, 3c±l, 8a, 8b, 9a, 9b, 10a
and 10b) were prepared as described for the preparation
of 1a12 or according to our previous method.13 We
describe herein the synthetic process of J-111,347 (1a) as
a representative example (Scheme 1).
Figure 2.

H. Imamura et al. / Bioorg. Med. Chem. 8 (2000) 1969±1982
1971
Scheme 1. Reagents; (a) 1-bromo-4-(tert-butyldimethylsilyloxy)methylbenzene, n-BuLi, THF, 70 ꢀC, 88%, (b) TPAP, NMO, molecular sieve 4A,
CH2Cl2, 0 ꢀC, 88%, (c) LiAlH4, LiI, THF, 78 ꢀC, 76%, (d) i: DEAD, PPh3, DPPA, THF, 0 ꢀC, 86%; ii: PPh3, THF-H2O, r.t.; iii: TEA, Alloc-Cl,
THF, 0 ꢀC, 90%, (e) i: n-Bu4NF, THF, 0 ꢀC, 98%; ii: TEA, MsCl, CH2Cl2, 0 ꢀC; iii: NaN3, DMF, r.t., 91%, (f) i: PPh3, THF-H2O, r.t.; ii: TEA,
Alloc-Cl, 0 ꢀC; iii: HCl-MeOH, 78%, (g) i: TEA, MsCl, CH2Cl2, 0 ꢀC; ii: t-BuOK, THF, 20 ꢀC, 97%, (h) AcSK, DMF, 65 ꢀC, 86%, (i) i: NaOH,
0 ꢀC; ii: i-Pr2NEt, 18, CH3CN, 0 ꢀC, 65%; iii: (PPh3)2PdCl2, n-Bu3SnH, H2O, CH2Cl2, 72%.
Thioacetate 17, a protected C-2 side chain of 1a, was pre-
pared starting from butanal 11 which was obtained from
commercially available d-malic acid by a conventional
method.14 Condensation of the butanal 11 with sub-
stituted phenyl lithium generated from the corresponding
bromide a€orded a diastereomeric mixture of alcohol 12.
Formation of the chiral alcohol 13 was performed by the
oxidation of 12 with a tetrapropylammonium perruthe-
nate (TPAP)/N-methylmorpholine N-oxide (NMO)
combination and following diastereoselective reduction
with the LAH-LiI system described by Mori and
Suzuki.15 The stereochemistry of the carbinol center of 13
was determined by 1H NMR analysis16 of the correspond-
ing MTPA esters. The secondary hydroxyl group of 13
was transformed to an allyloxycarbonyl(Alloc)-protected
amino group, and after subsequent desilylation, the other
benzilic hydroxyl group was substituted with NaN3 to
a€ord an azide 14. The azide 14 was reduced with tri-
phenylphosphine and H2O to provide an amino group
which was protected with an Alloc group, and then the
acetonide moiety was removed by acid treatment to
furnish diol 15. Mesylation of 15 gave dimesylate which
was subjected to cyclization reaction by potassium tert-
butoxide to provide the desired pyrrolidine 16 in excellent
yield. Conversion to 1a from thioacetate 17, obtained by
the substitution of 16 with AcSK, is as follows: Thioa-
cetate 17 was hydrolized to the corresponding thiol
which was in turn coupled with carbapenem diphenyl-
phosphate 1817 in the presence of diisopropylethyla-
mine in CH3CN. Deprotection of the coupling adduct
was accomplished according to the method of Guibe et
al.,18 and the resulting crude carbapenem was puri®ed
by reversed-phase column chromatography to furnish
the ®nal compound 1a as an amorphous powder after
lyophilization.
The cis isomer 1b corresponding to the trans isomer 1a
was prepared as shown in Scheme 2. Selective mono-
silylation of a mixture of diol 19 gave a separable mixture
of mono-silylated alcohol 20a and 20b (20a:20b=ca.
3:4). Then, the optically active alcohol 20a was converted
to 1b as described above.
Scheme 2. Reagents; (a) i: DEAD, PPh3, DPPA, THF, 0 ꢀC; ii: PPh3, THF-H2O, r.t.; iii: TEA, Alloc-Cl, THF, 0 ꢀC; v: TEA, MsCl, CH2Cl2, 0 ꢀC;
vi: NaN3, DMF, r.t.; vii: p-TsOH, MeOH, r.t., (b) TBS-Cl, imidazole, CH2Cl2, r.t., 20a: 29.8%, 20b: 40.5%.

1972
H. Imamura et al. / Bioorg. Med. Chem. 8 (2000) 1969±1982
Biological properties
pyrrolidine side chain, J-111,347 (1a), showed anti-
bacterial activities against both MRSA and P. aeruginosa.
The carbapenems obtained above were evaluated for their
in vitro antibacterial activity against S. aureus, including
an MRSA strain (pMS520/Smith), a methicillin-resistant
Staphylococcus epidermidis strain (MRSE, MB5181), E.
coli, and P. aeruginosa, including a ceftazidime-resistant
Pseudomonas aeruginosa strain (AKR17); their suscept-
ibility to porcine renal dehydropeptidase-I (DHP-I); and
their epileptogenicity (Tables 1±6). Standard serial dilution
techniques were employed for MIC determinations. Both
imipenem and vancomycin were used as reference drugs.
Subsequently, several analogues of 1a were synthesized
to con®rm the e€ect of trans stereochemistry of the
pyrrolidine ring on antibacterial activity (Table 3). All the
trans isomers showed better antibacterial activities than
the corresponding cis isomers; however, the aminoethyl
analogues, 8a and 8b, the naphthalene analogues, 9a and
9b, and the thiophene analogues, 10a and 10b, exhibited
reduced antibacterial activities against both the MRSA
and P. aeruginosa strains.
First, the 4-(aminomethyl)phenyl group was introduced
on the pyrrolidine ring directly or by using several linkers.
As expected, remarkable enhancement of the anti-MRSA
activity was observed in the resulting carbapenems, 4b, 5b,
6b and 7b, compared with BO-2727 (Table 1). A thio-
methylene analogue, 5b, and no-spacer analogue, 1b, in
which an aminomethylphenyl structure was directly
attached to the pyrrolidine ring, exhibited better anti-
bacterial activities than other compounds. It was notable
that only the methylene analogue 4b did not show the
undesirable epileptogenicity in the rat head assay at 200
mg/rat head.
Thus, J-111,347 (1a) was the ®rst example of an extremely
broad-spectrum antibiotic which showed potent activity
against MRSA as well as P. aeruginosa. However, 1a
and its analogues were found to be epileptogenic by the
rat intracerebroventricular (ICV) assay. Such seizure
potential often appeared as an adverse e€ect during the
development of carbapenems. The relation between the
epileptogenic potency of carbapenems and the stereo-
chemical environment of the side chain as well as its
basicity was investigated by Sunagawa et al.19 The
structure±activity relationship of BO-2727 derivatives
also suggested this e€ect of the structure of the side
chain, that is, introduction of substituent on the primary
amino group of the side chain reduced the epileptogenicity.
Based on these ®ndings, we modi®ed 1a by introducing
substituents onto the nitrogen of the primary amino
group a€ording derivatives (2a±j), in order to eliminate
the epileptogenicity of 1a without losing its broad-spec-
trum antibacterial activities (Fig. 3 and Table 4). All of
the N-alkylated analogues (2a, 2b, 2d, 2e and 2h) tested,
showed no epileptogenic potency. J-111,225 (2a), an N-
methyl analogue of 1a, showed potency against MRSA
and MRSE similar to that of vancomycin, and increased
potency against P. aeruginosa compared with imipenem.
The N,N-dimethyl analogue and trimethyl quarternary
ammonium analogue (2b and 2c) exhibited reduced
Next, we evaluated diastereomers of 1b, 4b, and 5b which
consist of trans-con®gurated pyrrolidine side chains
(Table 2). trans Diastereomers (4a, 5a) showed decreased
potency against all strains compared with the potencies
of the corresponding cis isomers (4b, 5b). Interestingly,
a trans diastereomer J-111,347 (1a) was two-fold more
active than the corresponding cis isomer (1b) against the
MRSA, MRSE, and P. aeruginosa strains. Such ambi-
dextrous activity of 1a was not seen in the trans isomers,
4a and 5a, which have methylene and thiomethylene
spacers, respectively, between the pyrrolidine and ami-
nomethylphenyl rings. In these studies, we discovered
that a novel carbapenem consisting of a trans-con®gurated
Table 1. In vitro activitya and biological properties of 1b, 4b, 5b, 6b and 7b
X Organism
-CH2-
(4b)
-CH2S-
(5b)
-CH2N-
(6b)
-CH2N-
(7b)
BO-2727
VCM
IPM
(1b)
S. aureus 209P NlHJ JC1
S. aureus pMS520/Smithb
S. epidermidis MB5181b
E. coli NlHJ JC2
P. aeruginosa AK109
P. aeruginosa AKR17c
ꢁ0.006
1.56
3.13
0.025
0.78
6.25
ꢁ0.006
3.13
3.13
0.025
0.39
6.25
ꢁ0.006
1.56
3.13
0.025
0.78
3.13
ꢁ0.006
3.13
6.25
0.025
1.56
ꢁ0.006
3.13
6.25
0.025
1.56
0.012
12.5
12.5
0.05
0.39
1.56
0.39
0.78
1.56
ꢁ0.006
50
50
0.1
1.56
3.13
>100
>100
>100
12.5
12.5
DHP-I susceptibilityd
0.12
4/5
0.17
0/5
0.22
2/5
0.15
2/5
0.15
3/5
0.11
0/5
Ð
Ð
1.0
5/5
Epileptogenicity
(200 mg/rat head, n=5)
aMIC(mg/ml) determined by agar dilution method.
bMethicillin-resistant.
cCeftazidime-resistant.
dRelative rate of hydrolysis to imipenem, porcine renal DHP-I.

H. Imamura et al. / Bioorg. Med. Chem. 8 (2000) 1969±1982
1973
Table 2. In vitro activitya and biological properties of J-111,347 (1a) and related compounds
R Organism
S. aureus 209P NlHJ JC1
S. aureus pMS520/Smithb
S. epidermidis MB5181b
E. coli NlHJ JC2
P. aeruginosa AK109
P. aeruginosa AKR17c
ꢁ0.006
0.78
1.56
0.025
0.39
3.13
ꢁ0.006
1.56
3.13
0.025
0.78
6.25
0.05
12.5
12.5
0.39
ꢁ0.006
3.13
3.13
0.025
0.39
6.25
25
>100
DHP-I susceptibilityd
0.29
5/5
0.12
4/5
0.28
0.17
0/5
Epiletogenicity (200 mg/rat head, n=5)
3/5
R Organism
VCM
IPM
S. aureus 209P NlHJ JC1
S. aureus pMS520/Smithb
S. epidermidis MB5181b
E. coli NlHJ JC2
P. aeruginosa AK109
P. aeruginosa AKR17c
ꢁ0.006
3.13
3.13
0.05
1.56
ꢁ0.006
1.56
3.13
0.025
0.78
3.13
0.39
0.78
1.56
ꢁ0.006
50
50
0.1
1.56
3.13
>100
>100
>100
12.5
DHP-I susceptibilityd
0.14
NTe
0.22
2/5
NTe
NTe
1.0
5/5
Epiletogenicity (200 mg/rat head, n=5)
aMIC (mg/mL) determined by agar dilution method.
bMethicillin-resistant.
cCeftazidime-resistant.
dRelative rate of hydrolysis to imipenem, porcine renal DHP-I.
eNot tested.
antibacterial activities against both MRSA and P. aer-
uginosa. Introduction of a substituent larger than the
methyl group, such as ethyl (2d), iso-propyl (2e), carba-
moylmethyl (2f), hydroxyethyl (2g) and sulfamoyl (2h),
also reduced the antibacterial activities of 2a against
MRSA and P. aeruginosa. Only J-111,225 (2a) showed
potent antibacterial activities against MRSA and P. aeru-
ginosa comparable to those of 1a with no epileptogenic
potency.
to cause seizures, and DHP-I stability. Preliminary in
vivo studies of the selected compounds, 2a, 3a, and 3b,
demonstrated their signi®cant ecacy against both
MRSA and P. aeruginosa.9 Subsequent evaluation of the
candidates, 2a, 3a and 3b, revealed good safety pro®les
with regard to epileptogenic potency (ED50), acute
toxicity (LD50) in mice, and 48-h rabbit nephrotoxicity
(Table 6).
In summary, the novel carbapenems, J-111,225 (2a), J-
114,870 (3a), and J-114,871 (3b), were designed, synthe-
sized, and evaluated. These compounds were selected to
be developed in view of an ultra-broad antimicrobial
spectrum covering MRSA as well as P. aeruginosa,
excellent safety pro®le including high stability to renal
DHP-I and low epileptogenecity, and good physico-
chemical properties.
Next, several substituents were introduced onto the
benzilic carbon adjacent to the primary amino group of
1a, in order to avoid the epileptogenic adverse e€ect
(Fig. 4). Similar to the N-alkylated analogues described
above (Table 4), all the substituents introduced so far
were e€ective in eliminating epileptogenicity (200 mg/head,
Table 5). Among these analogues, J-114,870 (3a) and J-
114,871 (3b), which had a carbamoylmethyl group,
exhibited excellent antibacterial activities comparable to
those of 1a and good stability to porcine renal DHP-I,
while other analogues showed decreased antibacterial
activities.
Experimental
Antibiotics and strains
Three compounds, J-111,225 (2a), J-114,870 (3a), and J-
114,871 (3b), were selected for further evaluation in view
of their excellent antibacterial activities, reduced potential
Imipenem was a product of Banyu Pharmaceutical Co.,
Ltd., Japan. Vancomycin was purchased from Sigma Che-
mical Co., St. Louis, MO. The antibiotics were dissolved in

1974
H. Imamura et al. / Bioorg. Med. Chem. 8 (2000) 1969±1982
Table 3. In vitro activitya and biological properties of J-111,347 (1a) and related compounds
R Organism
S. aureus 209P NlHJ JC1
S. aureus pMS520/Smithb
S. epidermidis MB5181b
E. coli NlHJ JC2
P. aeruginosa AK109
P. aeruginosa AKR17c
ꢁ0.006
0.78
1.56
0.025
0.39
3.13
ꢁ0.006
1.56
3.13
0.025
0.78
6.25
ꢁ0.006
1.56
3.13
0.025
0.78
6.25
ꢁ0.006
3.13
3.13
0.05
0.78
>25
DHP-I susceptibilityd
0.29
5/5
0.12
4/5
0.26
3/5
0.12
5/5
Epiletogenicity (200 mg/rat head, n=5)
R Organism
S. aureus 209P NlHJ JC1
S. aureus pMS520/Smithb
S. epidermidis MB5181b
E. coli NlHJ JC2
P. aeruginosa AK109
P. aeruginosa AKR17c
ꢁ0.006
1.56
0.78
0.05
6.25
ꢁ0.006
1.56
0.78
0.2
6.25
ꢁ0.006
1.56
1.56
0.025
0.78
3.13
ꢁ0.006
3.13
3.13
0.05
1.56
>25
>25
6.25
DHP-I susceptibilityd
0.16
5/5
<0.05
5/5
0.84
5/5
0.22
NTe
Epiletogenicity (200 mg/rat head, n=5)
aMIC (mg/mL) determined by agar dilution method.
bMethicillin-resistant.
cCeftazidime-resistant.
dRelative rate of hydrolysis to imipenem, porcine renal DHP-I.
eNot tested.
Table 4. In vitro activitya and biological properties of J-111,225 (2a) and related compounds
Organism
(2a)
(2b)
(2c)
(2d)
(2e)
S. aureus 209P NlHJ JC1
S. aureus pMS520/Smithb
S. epidermidis MB5181b
E. coli NlHJ JC2
P. aeruginosa AK109
P. aeruginosa AKR17c
ꢁ0.006
0.78
1.56
0.025
0.39
1.56
ꢁ0.006
1.56
3.13
0.05
0.78
ꢁ0.006
3.13
3.13
0.10
0.78
ꢁ0.006
1.56
1.56
0.025
1.56
6.25
ꢁ0.006
1.56
1.56
0.05
0.78
6.25
6.25
6.25
DHP-I susceptibilityd
0.25
0/5
0.22
0/5
0.27
NTe
0.44
0/5
0.46
0/5
Epiletogenicity (200 mg/rat head, n=5)
R Organism
(2f)
(2g)
(2h)
(2i)
(2j)
S. aureus 209P NlHJ JC1
S. aureus pMS520/Smithb
S. epidermidis MB5181b
E. coli NlHJ JC2
P. aeruginosa AK109
P. aeruginosa AKR17c
ꢁ0.006
3.13
1.56
0.025
1.56
6.25
ꢁ0.006
1.56
1.56
0.05
0.78
ꢁ0.006
1.56
6.25
0.025
1.56
3.13
ꢁ0.006
1.56
0.78
0.05
1.56
ꢁ0.006
1.56
0.78
0.012
0.78
3.13
6.25
6.25
DHP-I susceptibilityd
0.31
NTe
0.31
NTe
0.12
0/5
0.25
NTe
NTe
NTe
Epiletogenicity (200 mg/rat head, n=5)
aMIC (mg/mL) determined by agar dilution method.
bMethicillin-resistant.
cCeftazidime-resistant.
dRelative rate of hydrolysis to imipenem, porcine renal DHP-I.
eNot tested.

H. Imamura et al. / Bioorg. Med. Chem. 8 (2000) 1969±1982
1975
Table 5. In vitro activitya and biological properties of J-114,870 (3a), J-114,871 (3b), and related compounds
Organism
(3a)
(3b)
(3c)
(3d)
(3e)
(3f)
S. aureus 209P NlHJ JC1
S. aureus pMS520/Smithb
S. epidermidis MB5181b
E. coli NlHJ JC2
P. aeruginosa AK109
P. aeruginosa AKR17c
ꢁ0.006
1.56
1.56
0.025
0.78
1.56
ꢁ0.006
1.56
1.56
0.025
1.56
1.56
ꢁ0.006
3.13
3.13
0.025
1.56
6.25
ꢁ0.006
1.56
1.56
0.025
1.56
6.25
ꢁ0.006
3.13
6.25
0.05
1.56
ꢁ0.006
0.39
0.39
0.025
3.13
6.25
12.5
DHP-I susceptibilityd
0.27
0/5
0.27
0/5
0.32
0/5
0.41
NTe
0.46
0/5
0.22
NTe
Epiletogenicity (200 mg/rat head, n=5)
Organism
(3g)
(3h)
(3i)
(3j)
(3k)
(3l)
S. aureus 209P NlHJ JC1
S. aureus pMS520/Smithb
S. epidermidis MB5181b
E. coli NlHJ JC2
P. aeruginosa AK109
P. aeruginosa AKR17c
ꢁ0.006
1.56
0.78
0.025
3.13
ꢁ0.006
1.56
0.78
0.025
0.78
3.13
ꢁ0.006
1.56
1.56
0.10
3.13
25
ꢁ0.006
1.56
1.56
0.05
1.56
ꢁ0.006
1.56
1.56
0.025
3.13
6.25
ꢁ0.006
1.56
3.13
0.025
1.56
12.5
6.25
12.5
DHP-I susceptibilityd
0.27
NTe
0.55
0/5
0.20
0/5
0.37
0/5
0.32
0/5
0.29
0/5
Epiletogenicity (200 mg/rat head, n=5)
aMIC (mg/mL) determined by agar dilution method.
bMethicillin-resistant.
cCeftazidime-resistant.
dRelative rate of hydrolysis to imipenem, porcine renal DHP-I.
eNot tested.
Table 6. Safety pro®le of selected compounds
Compound
(1a)
(2a)
(3a)
>400
ca. 1850
No change
(3b)
Epiletogenicitya ED50 (mg/head, n=5)
Acute toxicity in mice LD50 (mg/Kg, single i.v., n=5)
48-hour nephrotoxicity (225 mg/Kg, single i.v., n=3)
ca. 76
>1500
NTb
>400
600ꢂ700
>200
>1000
No change
No change
aInjected intracerebroventricularly.
bNot tested.
10 mM 3-morpholino-propanesulfonate (MOPS) bu€er/
pH 7.0 on the day of use. The strains used in the study
were from our collections; MRSA pMS520/Smith was a
generous gift from M. Inoue, School of Medicine,
Kitasato University, Japan.
(®nal concentration, 0.3 U/mL) was incubated with 50
mM carbapenem at 35 ꢀC in 50 mM MOPS bu€er/pH
7.0. The initial hydrolysis rate was monitored by the
spectrophotometric method. One unit of activity was
de®ned as the amount of enzyme hydrolyzing 1 mM of
glycyldehydrophenylalanine per minute when the sub-
strate, 50 mM, was incubated at 35 C in 50 mM MOPS
bu€er/pH 7.0.
Determination of MIC
MICs were determined by a two-fold serial agar dilution
method using Mueller-Hinton medium (Difco Labora-
tories, Detroit, MI.). An overnight culture was diluted
to give a ®nal concentration of approximately 106 CFU/
mL. A portion of the dilution was delivered onto a drug-
containing agar surface with an inoculum apparatus
(Microplanter: Sakuma Seisakusho, Co., Ltd, Tokyo,
Japan). The ®nal inoculum size was approximately 104
CFU per spot. The MIC was de®ned as the lowest con-
centration of antibiotics that completely prevented visible
growth was inhibited.
Determination of epileptogenicity
Male SD rats, 7 weeks of age, were cannulated into the
right cerebroventricle 1 week before of drug administra-
tion. The carbapenems were dissolved in saline, adjusted
to pH 7.0, and an aliquot of 10 ml/head was injected
intracerebroventricularly (n=5). Convulsant behavior
was monitored for 30 min after administration.
General methods
Melting points were measured on a Yanaco MP micro-
melting point apparatus and were not corrected. The 1H
NMR spectra were recorded on a Varian VXR-300
spectrometer and a JEOL JNM-A500 spectrometer with
tetramethylsilane (TMS) as an internal standard. 13C
NMR spectra were recorded on a JEOL JNM-A500. IR
Determination of susceptibility to renal
dehydropeptidase-1 (DHP-I)
Relative hydrolysis rates of carbapenems by porcine renal
DHP-I were determined, taking the initial hydrolysis rate
of imipenem as 1.0. Partially puri®ed porcine DHP-I

1976
H. Imamura et al. / Bioorg. Med. Chem. 8 (2000) 1969±1982
Figure 3.
Figure 4.
absorption spectra were recorded with a Horiba FT-200
spectrometer. Speci®c rotations were measured with a
Jasco DIP-370 polarimeter. UV spectra were taken on a
SHIMAZU SPD-10A spectrometer in 0.1 M 3-morpho-
linopropanesulfonate bu€er (pH 7.0). Mass spectra
(MS) were measured on a JEOL JMS-SX102A spectro-
meter. TLC was performed with Merck Kieselgel F254
precoated plates. The silica gel used for column chroma-
tography was WAKO gel C-300. Reversed phase column
chromatography was carried out on YMC-gel ODS-AQ
120-S50. All reactions involving air-sensitive reagents
were performed under nitrogen using syringe-septum cap
techniques.
4.30 (2H, m), 7.43 (4H, m); FAB-HRMS m/z calcd for
C22H30N3O4S (M+H)+: 432.1957, Found: 432.1952.
(1R,5S,6S)-2-[(3S,5S)-5-(4-Aminomethylbenzyl)pyrroli-
din-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-
2-em-3-carboxylic acid hydrochloride 4b. IR nmax (KBr)
3425, 1758, 1587, 1384 cm 1; 1H NMR (300 MHz, D2O)
d 1.19 (3H, d, J=7.2 Hz), 1.28 (3H, d, J=6.5 Hz), 1.78
(1H, m), 2.71 (1H, m), 3.29 (5H, m), 3.83 (3H, m), 4.19
(2H, s), 4.22 (2H, m), 7.42 (4H, m); FAB-HRMS m/z
calcd for C22H30N3O4S (M+H)+: 432.1957, Found:
432.1955; UV lmax298 (e 8410).
(1R,5S,6S)-2-[(3S,5R)-5-[(4-Aminomethylphenyl)thiome-
thyl]pyrrolidin-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-
1-carbapen-2-em-3-carboxylic acid hydrochloride 5a. IR
4a, 4b, 5a, 5b, 6b and 7b were synthesized using a method
described in the literature.10,11
1
nmax (KBr) 1749, 1595, 1394, 1263 cm
;
1H NMR
(1R,5S,6S)-2-[(3S,5R)-5-(4-Aminomethylbenzyl)pyrroli-
din-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-
2-em-3-carboxylic acid hydrochloride 4a. IR nmax (KBr)
3435, 1761, 1593, 1388 cm 1; 1H NMR (300MHz, D2O) d
1.21 (3H, d, J=7.8Hz), 1.37 (3H, d, J=6.4 Hz), 2.15 (1H,
m), 2.44 (1H, m), 3.36 (5H, m), 3.87 (3H, m), 4.22 (2H, s),
(300 MHz, D2O) d 1.18 (3H, d, J=7.3 Hz), 1.27 (3H, d,
J=6.3 Hz), 2.28 (2H, m), 3.35 (5H, m), 3.73 (1H, dd,
J=13.0, 6.0 Hz), 4.03 (2H, m), 4.27 (4H, m), 7.45 (2H,
d, J=8.0 Hz), 7.55 (2H, dd, J=8.0 Hz); FAB-HRMS m/
z calcd for C22H30N3O4S2 (M+H)+: 464.1678, Found:
464.1655; UV lmax 297 (e 7590).

H. Imamura et al. / Bioorg. Med. Chem. 8 (2000) 1969±1982
1977
(1R,5S,6S)-2-[(3S,5S)-5-[(4-Aminomethylphenyl)thiome-
thyl]pyrrolidin-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-
1-carbapen-2-em -3-carboxylic acid hydrochloride 5b.
20min at the same temperature. Work-up and puri®cation
in the usual manner gave (3R)-1-(4-tert-butyldimethylsil-
oxymethylphenyl)-3,4-O-isopropylidene-3,4-dihydroxy-
20
1
IR nmax (KBr) 3373, 1749, 1558, 1396, 1089cm 1; H
butanone as a colorless oil (9.9 g, 88.0%): ‰ꢀŠ
29.0 (c
d
1
NMR (300MHz, D2O) d 1.18 (3H, d, J=7.3Hz), 1.28
(3H, d, J=6.3 Hz), 1.65 (1H, m), 2.64 (1H, m), 3.30 (3H,
m), 3.46 (3H, m), 3.81 (2H, m), 4.16 (2H, s), 4.23 (2H, m),
7.43 (2H, d, J=8.6Hz), 7.54 (2H, d, J=8.6 Hz); FAB-
HRMS m/z calcd for C22H30N3O4S2 (M+H)+: 464.1678,
Found: 464.1667; UV lmax286 (e 8290).
1.0, CHCl3); IR nmax (KBr) 1684 cm 1; H NMR (300
MHz, CDCl3) d 0.03 (6H, s), 0.84 (9H, s), 1.29 (3H, s), 1.34
(3H, s), 2.99 (1H, dd, J=17.3, 7.1Hz), 3.44 (1H, dd,
J=17.3, 5.0 Hz), 3.53 (1H, dd, J=8.4, 6.6 Hz), 4.20 (1H,
dd, J=8.4, 6.1 Hz), 4.53 (1H, m), 4.69 (2H, s), 7.31 (2H,
d, J=8.3 Hz), 7.82 (2H, d, J=8.3 Hz); FAB-HRMS m/z
calcd for C20H32O4SiNa (M+Na)+: 387.1968, Found:
387.1967. 13 was obtained by reduction with the
LiAlH4-LiI system described by Mori and Suzuki.15
(1R,5S,6S)-2-[(3S,5S)-5-[(4-Aminomethylphenyl)amino-
methyl]pyrrolidin-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-
methyl-1-carbapen-2-em-3-carboxylic acid hydrochloride
1
6b. IR nmax (KBr) 1749, 1595, 1394, 1263 cm
;
1H
To a solution of the butanone described above (2.80g,
7.69mmol) in Et2O (50 mL) was added LiI (10.29g,
76.9mmol) under a nitrogen atmosphere at 40ꢀC and
the mixture was stirred for 5 min at the same temperature.
The resulting mixture was then cooled to 78 ꢀC and
LiAlH4 (2.91 g, 76.9 mmol) 5w>wasadded.Themixture
wasstirredfor30min.Work-upandpuri®cationintheusual
mannergave13asacolorlessoil(2.14 g,76.2%):[HPLCana-
lysis: column, DAICEL CHIRALPAK AS(250Â4.6 mm);
detection, 254 nM; eluent, n-hexane/isopropanol=90:10;
NMR (300 MHz, D2O) d 1.17 (3H, d, J=7.1 Hz), 1.24
(3H, d, J=6.5 Hz), 1.71 (1H, m), 2.71 (1H, m), 3.30 (3H,
m), 3.60 (3H, m), 4.01 (2H, m), 4.09 (2H, s), 4.18 (2H, m),
6.79 (2H, d, J=8.6 Hz), 7.26 (2H, dd, J=8.6 Hz); FAB-
HRMS m/z calcd for C22H30N4O4SNa (M+Na)+:
469.1885, Found: 469.1874; UV lmax300 (e 10100).
(1R,5S,6S)-2-[(3S,5S)-5-[(4-Aminomethylphenyl)oxyme-
thyl]pyrrolidin-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-
1-carbapen-2-em-3-carboxylic acid hydrochloride 7b. IR
nmax (KBr) 3413, 1753, 1610, 1516, 1392, 1248 cm 1; 1H
NMR (300 MHz, D2O) d 1.19 (3H, d, J=7.2 Hz), 1.25
(3H, d, J=6.5 Hz), 1.91 (1H, m), 2.76 (1H, m), 3.37 (3H,
m), 3.68 (1H, dd, J=12.2, 6.0 Hz), 4.10 (2H, s), 4.15 (5H,
m), 4.38 (1H, m), 7.06 (2H, d, J=8.7 Hz), 7.39 (2H, dd,
J=8.7 Hz); FAB-HRMS m/z calcd for C22H30N3O5S
(M+H)+: 448.1906, Found: 448.1904; UV lmax298 (e
9220).
¯owrate, 0.5 mL/min;tR of13,9.1 min;tR ofcorresponding
20
d
1R isomer, 8.6 min]. 90% de, ‰ꢀŠ
27.4 (c 1.0, CHCl3);
IR nmax (KBr) 3518, 1518, 778 cm 1; 1H NMR (300MHz,
CDCl3) d 0.09 (6H, s), 0.92 (9H, s), 1.37 (3H, s), 1.46 (3H,
s), 1.89 (1H, m), 1.98 (1H, m), 3.53 (1H, dd, J=8.1,
8.1 Hz), 4.03 (1H, dd, J=8.1, 5.9 Hz), 4.23 (1H, m), 4.72
(2H, s), 4.89 (1H, dd, J=8.5, 4.2 Hz), 7.27 (2H, d,
J=8.5Hz), 7.32 (2H, d, J=8.5 Hz); FAB-HRMS m/z
calcd for C20H34O4SiNa (M+Na)+: 389.2124, Found:
389.2112.
General procedure for the preparation of 5-aryl pyrrolidi-
nylthio-1ꢀ-methylcarbapenems. The experimental proce-
dure for 1a was described as a representative example.
The stereochemical structure of 13 is shown below
(ÁdÂ10 3 ppm), which was determined by the advanced
Mosher method with the corresponding MTPA esters.16
(3R)-1-(4-tert-Butyldimethylsiloxymethylphenyl)-3,4-O-
isopropylidene-3,4-dihydroxybutanol 12. To a solution of
4-(tert-butyldimethylsiloxymethyl)bromobenzene (14.6
g, 48.7 mmol) in THF (300 mL) was added n-BuLi (1.6
M in n-hexane, 27.9 mL, 44.7 mmol) dropwise under a
nitrogen atmosphere at 70 ꢀC, and the mixture was
stirred for 20 min at the same temperature. To this mix-
ture was added a solution of 11 (5.86 g, 40.6 mmol) in
THF (25 ml) dropwise under a nitrogen atmosphere at
70 ꢀC. Work-up and puri®cation in the usual manner
gave 12 as a colorless oil (13.2 g, 88.8%): IR nmax (KBr)
(1R,3R)-N-Allyloxycarbonyl-1-(4-azidomethylphenyl)-3,4-
O-isopropylidene-3,4-dihydroxybutylamine 14. To a solu-
tion of 13 (1.31 g, 3.56 mmol) in THF (30 mL) were
added triphenylphosphine (1.97 g, 7.5 mmol), diethyl
azodicarboxylate (1.12 ml, 7.14 mmol) and diphenylpho-
sphoryl azide (1.54 mL, 7.14 mmol) dropwise under a
nitrogen atmosphere at 0 ꢀC, and the mixture was stirred
for 20min at the same temperature. Work-up and pur-
i®cation in the usual manner gave (1R,3R)-1-(4-tert-
butyldimethylsiloxymethylphenyl)-3,4-O-isopropylidene-
1
3518, 1516, 778 cm 1; H NMR (300 MHz, CDCl3) d
0.09 (6H, s), 0.92 (9H, s), 1.37 (3H, s), 1.46 (3H, s), 1.94
(2H, m), 3.57 (1H, m), 4.05 (1H, m), 4.26 (1H, m), 4.73
(2H, s), 4.92 (1H, m), 7.27 (2H, d, J=9.3 Hz), 7.34 (2H,
d, J=9.3 Hz); FAB-HRMS m/z calcd for C20H34O4S-
iNa (M+Na)+: 389.2124, Found: 389.2131.
(1S,3R)-1-(4-tert-Butyldimethylsiloxymethylphenyl)-3,4-
O-isopropylidene-3,4-dihydroxybutanol 13. To a solution
of 12 (11.3 g, 30.8 mmol) and molecular sieve 4A (15.4 g)
in CH2Cl2 (100 mL) were added N-methylmorpholine N-
oxide (5.4l g, 46.2 mmol) and tetrapropylammonium
perruthenate (541 mg, 1.54 mmol) under a nitrogen
atmosphere at 0 ꢀC, and the mixture was stirred for
3,4-dihydroxybutylazide as a colorless oil (1.21 g,
1
86.4%): H NMR (300 MHz, CDCl3) d 0.10 (6H, s),
0.93 (9H, s), 1.31 (1H, s), 1.36 (2H, s), 1.41 (3H, s), 1.93
(2H, m), 3.54 (1H, m), 4.09 (1H, m), 4.34 (1H, m), 4.64
(1H, m), 4.72 (2H, s), 7.34 (4H, m); FAB±MS m/z 392
(M+H)+. To a solution of the azide described above
(1.03 g, 2.62 mmol) in THF (30 mL) and H2O (6 mL)

1978
H. Imamura et al. / Bioorg. Med. Chem. 8 (2000) 1969±1982
was added triphenylphosphine (1.38 g, 5.26 mmol), and
the mixture was stirred for 5 h at 50 ꢀC. After evapora-
tion under reduced pressure, to a solution of the residue
in THF (60 mL) were added triethylamine (1.32 g,
13.1 mmol) and allyl chloroformate (420mL, 3.94mmol)
at 0 ꢀC. Work-up and puri®cation in the usual manner
gave (1R,3R)-N-allyloxycarbonyl-1-(4-tert-butyldimethyl-
room temperature. Work-up and puri®cation in the usual
20
manner gave 15 as a colorless oil (941mg, 78.6%): ‰ꢀŠ
d
+56.0 (c 0.5, CHCl3); IR nmax (Nujol) 3413, 1732, 1695,
1
769 cm 1; H NMR (300MHz, CDCl3) d 1.80 (1H, m),
2,36 (1H, m), 3.46 (1H, m), 3.54 (1H, m), 3.77 (2H, m), 4.32
(2H, s), 4.58 (4H, m), 4.97 (1H, m), 5.22 (4H, m), 5.53 (1H,
m), 5.88 (2H, m), 7.27 (4H, s); FAB-HRMS m/z calcd for
C19H26N2O6Na (M+Na)+: 401.1689, Found: 401.1676.
siloxymethylphenyl)-3,4-O-isopropylidene-3,4-dihydroxy-
20
butylamine as a colorless oil (1.019 g, 90.8%): ‰ꢀŠ
d
1
+44.1 (c 1.0, CHCl3); IR nmax (KBr) 1733, cm 1; H
(2R,4R)-2-[4-(N-Allyloxycarbonylanimomethyl)phenyl]-1-
allyloxycarbonyl-4-mesyloxypyrrolidine 16. To a solution
of 15 (243 mg, 0.64 mmol) in CH2Cl2 (8 mL) were added
triethylamine (259 mg, 2.56 mmol) and methanesulfonyl
chloride (150 mL, 1.93 mmol) under a nitrogen atmo-
sphere at 0 ꢀC. The reaction mixture was poured into
H2O, and the whole was extracted with EtOAc. The
organic layer was washed with brine, dried over MgSO4,
and the mixture was evaporated under reduced pressure.
To a solution of the residue in THF (10 mL) was added
potassium tert-butoxide (160mg, 1.42mmol) under a
nitrogen atmosphere at 20ꢀC. Work-up and puri®cation
NMR (300 MHz, CDCl3) d 0.10 (6H, s), 0.96 (9H, s),
1.33 (3H, s), 1.44 (3H, s), 1.94 (1H, m), 2.08 (1H, m),
3.46 (1H, m), 3.87 (1H, m), 4.50 (2H, m), 4.69 (2H, s),
4.96 (1H, m), 5.22 (2H, m), 5.79 (1H, m), 7.24 (2H, d,
J=7.5 Hz), 7.27 (2H, d, J=7.5 Hz); FAB-HRMS m/z
calcd for C24H39NO5SiNa (M+Na)+: 472.2495, Found:
472.2500. To a solution of above (1.25 g, 2.75 mmol) in
THF (30 mL) was added tetra-n-butylammonium ¯uo-
ride (1 M in THF, 3.75 mL, 3.75 mmol) dropwise under
a nitrogen atmosphere at 0 ꢀC, and the mixture was
stirred for 30 min at the same temperature. Work-up
and puri®cation in the usual manner gave (1R,3R)-N-
allyloxycarbonyl-1-(4-hydroxymethylphenyl)-3,4-O-iso-
propylidene-3,4-dihydroxybutylamine as a colorless oil
in the usual manner gave 16 as a colorless oil (274 mg,
20
d
97.3%): ‰ꢀŠ +29.6 (c 1.0, CHCl3); IR nmax (Nujol)
1
1693, 1248, 769 cm 1; H NMR (300 MHz, CDCl3) d
1
(917 mg, 98.3%): IR nmax (KBr) 3543, 1708, cm 1; H
2.36 (1H, m), 2.69 (3H, s), 2.71 (1H, m), 3.90 (1H, m),
4.02 (1H, m), 4.29 (2H, m), 4.53 (4H, m), 5.02 (3H, m),
5.23 (3H, m), 5.90 (2H, m), 7.22 (4H, s); FAB-HRMS
m/z calcd for C20H26N2O7SNa (M+Na)+: 461.1358,
Found: 461.1367.
NMR (300 MHz, CDCl3) d 1.31 (3H, s), 1.44 (3H, s),
1.95(1H, m), 2.07 (1H, m), 3.50 (1H, m), 3.98 (2H, m),
4.55 (2H, m), 4.64 (2H, s), 4.99 (1H, m), 5.22 (2H, m),
5.90 (1H, m), 7.27 (4H, m); FAB-HRMS m/z calcd for
C18H25NO5Na (M+Na)+: 358.1630, Found: 358.1631.
To a solution of above (835mg, 2.49mmol) in CH2Cl2
(30 mL) were added triethylamine (1.74 mL, 12.5 mmol)
and methanesulfonyl chloride (290mL, 3.75mmol) under
a nitrogen atmosphere at 0 ꢀC. The reaction mixture was
poured into H2O, and the whole was extracted with
EtOAc. The organic layer was washed with brine, dried
over MgSO4, and the mixture was evaporated under
reduced pressure. To a solution of the residue in DMF
(50 mL) was added sodium azide (485 mg, 7.45 mmol),
and the mixture was stirred for 5 h at room temperature.
(2R,4S)-4-Acetylthio-2-[4-(N-allyloxycarbonylanimo-
methyl)phenyl]-1-allyloxycarbonylpyrrolidine 17. To a
solution of 16 (274 mg, 0.62 mmol) in DMF (10 mL) was
added potassium thioacetate (215 mg, 1.88mmol) under a
nitrogen atmosphere at room temperature, and the mix-
ture was sttired for 3 h at 65 ꢀC. Work-up and puri®cation
in the usual manner gave 17 as a colorless oil (227 mg,
86.8%): ‰ꢀŠ20 +36.6 (c 1.0, CHCl3); IR nmax (Nujol) 1699,
d
1249, 769, 727 cm 1; 1H NMR (300MHz, CDCl3) d 2.22
(1H, m), 2.34 (3H, s), 3.60 (1H, m), 4.03 (2H, m), 4.34
(2H, m), 4.56 (4H, m), 5.02 (3H, m), 5.26 (3H, m), 5.90
(2H, m), 719 (4H, m) ; FAB-HRMS m/z calcd for
C22H28N2O5SNa (M+Na)+: 455.1617, Found: 455.1619.
Work-up and puri®cation in the usual manner gave 14
20
d
as a colorless oil (818 mg, 91.3%): ‰ꢀŠ +23.5 (c 1.0,
1
CHCl3); IR nmax (KBr) 1760, 1703, cm
;
1H NMR
(300 MHz, CDCl3) d 1.32 (3H, s), 1.44 (3H, s), 1.87 (1H,
m), 2.02 (1H, m), 3.47 (1H, m), 3.92 (2H, m), 4.32 (2H,
s), 4.56(2H, m), 4.50 (1H, m), 5.22 (2H, m), 5.90 (1H,
m), 7.29 (4H, s); FAB-HRMS m/z calcd for C18H24
N4O4Na (M+Na)+: 383.1695, Found: 383.1683.
(1R,5S,6S)-2-[(3S,5R)-5-(4-Aminomethylphenyl)pyrroli-
din-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-
2-em-3-carboxylic acid hydrochloride 1a (J-111,347). To
an ice-cooled solution of 17 (227mg, 0.54mmol) in
MeOH (8 mL) was added 1 M aqueous NaOH (540mL)
under a nitrogen atmosphere. After being stirred for
20min at 0 ꢀC, the reaction mixture was adjusted to pH
7.0 with 1 M aqueous HCl and concentrated under
reduced pressure. The mixture was poured into H2O, and
the whole was extracted with EtOAc. The organic layer
was washed with brine, dried over MgSO4, and evapo-
rated under reduced pressure. To a stirred solution of the
residue and allyl (1R,5S,6S)-2-diphenylphosphoryloxy-6-
[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-car-
boxylate (18, 271 mg, 0.54mmol) in CH3CN (10 mL) was
added diisopropylethylamine (153 mL, 0.87 mmol) drop-
wise at 0 ꢀC. After being stirred overnight at 4 ꢀC, the
mixture was poured into H2O, and the whole was
extracted with EtOAc. Work-up and puri®cation in the
(1R,3R)-N-Allyloxycarbonyl-1-(4-N-allyloxycarbonylami-
nomethyl)phenyl-3,4-dihydroxybutylamine 15. To a solu-
tion of 14 (1.14 g, 3.17mmol) in THF (40 mL) and H2O
(8mL) was added triphenylphosphine (1.25g, 4.76mmol),
and the mixture was stirred for 2 h at room temperature.
After evaporation under reduced pressure, to a solution
of the residue in THF (70 mL) were added triethylamine
(2.23 mL, 16.0 mmol) and allyl chloroformate (507 mL,
4.76 mmol) at 0 ꢀC. The reaction mixture was poured
into H2O, and the whole was extracted with EtOAc. The
organic layer was washed with brine, dried over MgSO4,
and evaporated under reduced pressure. To a solution
of the residue in MeOH (20 mL) was added 10% HCl/
MeOH (10 mL), and the mixture was stirred for 1 h at

H. Imamura et al. / Bioorg. Med. Chem. 8 (2000) 1969±1982
1979
usual manner gave the adduct, allyl (1R,5S,6S) - 2 -
[(3S,5R)-1-allyloxycarbonyl-5-[4-(N-allyloxycarbonyl-
aminomethyl)phenyl]pyrrolidin-3-ylthio]-6-[(R)-1-hy-
droxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate as a
foam (222 mg, 65.4%): IR nmax (KBr) 3373, 2966, 1751,
(1H, m), 4.14 (1H, m), 4.20 (2H, s), 4.24 (1H, m), 7.52
(2H, d, J=8.0 Hz), 7.55 (2H, d, J=8.0 Hz); FAB-HRMS
m/z calcd for C21H28N3O4S (M+H)+: 418.1801, Found:
418.1793; UV lmax298 (e 9910).
1
1587, 1392, 1086 cm 1; H NMR (300MHz, CDCl3) d
(1R,5S,6S)-6-[(R)-1-Hydroxyethyl]-2-[(3S,5R)-5-[4-(N-
methylaminomethyl)phenyl]pyrrolidin-3-ylthio]-1-methyl-1-
carbapen-2-em-3-carboxylic acid dihydrochloride 2a (J-
1.22 (3H, d, J=7.0Hz), 1.36 (3H, d, J=6.2 Hz), 2.24 (1H,
m), 2.41 (1H, m), 3.21 (1H, dd, J=7.2, 2.4 Hz), 3.30 (1H,
m), 3.78 (2H, m), 4.03 (1H, m), 4.24 (2H, m), 4.49 (5H, m),
4.63 (2H, m), 4.64 (1H, m), 4.75 (1H, m), 5.04 (2H, m),
5.26 (4H, m), 5.43 (1H, m), 5.94 (2H, m), 7.20 (4H, m);
FAB-HRMS m/z calcd for C32H40N3O8S (M+H)+:
626.2536, Found: 626.2534. To an ice-cooled solution of
above (210mg, 0.34mmol) in CH2Cl2 (10 mL) were suc-
cessively added H2O (30 mL), bis(triphenylphosphine)
palladium(II) dichloride (11.8 mg, 0.017 mmol), and tri-
butyltin hydride (329 mL, 1.22 mmol) under a nitrogen
atmosphere. Work-up and puri®cation in the usual
manner gave 1a as an amorphous powder (110 mg,
72.9%): IR nmax (KBr) 3421, 1749, 1646, 1558 cm 1; 1H
NMR (300 MHz, D2O) d 1.22 (3H, d, J=7.0 Hz), 1.27
(3H, d, J=6.5 Hz), 2.51 (1H, m), 2.73 (1H, m), 3.40 (3H,
m), 3.86 (1H, dd, J=12.5, 6.0 Hz), 4.25 (5H, m), 5.03
(1H, dd, J=10.5, 7.0 Hz), 7.20 (4H, m); FAB-HRMS
m/z calcd for C21H28N3O4S (M+H)+: 418.1801, Found:
418.1800; UV lmax298 (e 9520).
20
111,225). ‰ꢀŠ +9.0 (c 1.0, H2O); IR (KBr) nmax 3373,
d
1751, 1587, 1392, 1086 cm 1; 1H NMR (500 MHz, D2O,
as a hydrochloride) d 1.02 (3H, d, J=7.3 Hz), 1.08 (3H,
d, J=6.4 Hz), 2.33 (1H, dd, J=14.0, 6.7 Hz), 2.52 (3H,
s), 2.57 (1H, m), 3.17 (1H, dq, J=9.1, 7.3 Hz), 3.27 (2H,
m), 3.70 (1H, dd, J=12.8, 5.8 Hz), 4.04 (5H, m), 4.88 (1H,
dd, J=11.0, 6.7 Hz), 7.35 (4H, m); 13C NMR (125MHz,
D2O, as a hydrochloride) d 15.4, 19.7, 31.9, 35.9, 40.7,
42.1, 51.4, 51.9, 55.6, 58.5, 61.2, 64.7, 128.1, 130.2, 131.8,
134.2, 135.2, 136.7, 167.3, 176.3; FAB-HRMS m/z calcd
for C22H30N3O4S (M+H)+: 432.1957, Found 432.1950.
.
.
Anal. calcd for C22H29N3O4S 2HCl H2O: C,50.57; H,
6.37; N, 8.04; S, 6.14; Found: C, 50.87; H, 6.45; N, 7.83;
S, 6.02.
(1R,5S,6S)-2-[(3S,5R)-5-[4-(N,N-Dimethylaminomethyl)-
phenyl]pyrrolidin-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-
methyl-1-carbapen-2-em-3-carboxylic acid hydrochloride
2b. IR nmax (KBr) 3371, 2968, 1755, 1589, 1390, 1286,
1263, 1074cm 1; 1H NMR (300MHz, D2O) d 1.22 (3H, d,
J=7.3Hz), 1.27 (3H, d, J=6.6 Hz), 2.55 (1H, m), 2.74
(1H, m), 2.83 (6H, s), 3.37 (1H, m), 3.50 (2H, m), 3.88 (1H,
m), 4.22 (3H, m), 4.32 (2H, s), 5.08 (1H, m), 7.56 (4H, m);
FAB-HRMS m/z calcd for C23H32N3O4S (M+H)+:
446.2114, Found: 446.2109; UV lmax298 (e 10940).
(1S,3R)-N-Allyloxycarbonyl-1-(4-azidomethylphenyl)-4-
tert - butyldimethylsiloxy - 3-hydroxybutylamine 20a and
(1R,3R)-N-Allyloxycarbonyl-1-(4-azidomethylphenyl)-4-
tert-butyldimethylsiloxy-3-hydroxybutylamine 20b. To a
solution of 19 (650 mg, 2.03 mmol) in CH2Cl2 (15 mL)
were added imidazole (208 mg, 3.05 mmol) and tert-
butyldimethylchlorosilane (368 mg, 2.44 mmol) and the
mixture was stirred overnight at room temperature.
Work-up and puri®cation in the usual manner gave 20a
(1R,5S,6S)-6-[(R)-1-Hydroxyethyl]-2-[(3S,5R)-5-[4-(N,N,
N-trimethylammoniomethyl)phenyl]pyrrolidin-3-ylthio]-1-
methyl-1-carbapen-2-em-3-carboxylate hydrochloride 2c.
IR nmax (KBr) 3402, 2968, 1751, 1589, 1389, 1288, 1263,
as a colorless oil (263mg, 29.8%) and 20b as colorless oil
20
d
(358mg, 40.5%). 20a: ‰ꢀŠ
21.6 (c 1.0, CHCl3); IR nmax
(KBr) 3524, 1703, 788 cm 1; 1H NMR (300MHz, CDCl3)
d 0.06 (6H, s), 0.89 (9H, s), 1.86 (1H, m), 2.49 (1H, m),
3.39 (1H, dd, J=9.8, 5.7 Hz), 3.54 (1H, dd, J=9.8,
3.5 Hz), 3.60 (1H, m), 4.22 (2H, s), 4.59 (2H, m), 4.76 (1H,
m), 5.22 (2H, m), 5.89 (1H, m), 7.27 (4H, s); FAB-HRMS
1074cm 1; H NMR (300MHz, D2O) d 1.22 (3H, d,
1
J=7.3Hz), 1.27 (3H, d, J=6.3 Hz), 2.55 (1H, m), 2.78
(1H, m), 3.08 (9H, s), 3.38 (1H, m), 3.52 (2H, m), 3.85 (1H,
m), 4.22 (3H, m), 4.51 (2H, s), 5.14 (1H, m), 7.63 (4H, m);
FAB-HRMS m/z calcd for C24H34N3O4S (M+H)+:
460.2270, Found: 460.2266; UV lmax298 (e 8930).
m/z calcd for C21H34N4O4SiNa (M+Na)+: 457.2247,
20
d
Found: 457.2247. 20b; ‰ꢀŠ +39.4 (c 1.0, CHCl3); IR
nmax (KBr) 3525, 1709, 788 cm 1; H NMR (300 MHz,
(1R,5S,6S)-2-[(3S,5R)-5-[4-(N-Ethylaminomethyl)phenyl]-
pyrrolidin-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-car-
bapen-2-em-3-carboxylic acid hydrochloride 2d. IR nmax
(KBr) 1749, 1558, 1394 cm 1; 1H NMR (300 MHz, D2O)
d 1.24 (9H, m), 2.52 (1H, m), 2.75 (1H, m), 3.11 (2H, q,
J=7.3 Hz), 3.36 (1H, m), 3.47 (2H, m), 3.88 (1H, m), 4.24
(5H, m), 5.08 (1H, m), 7.55 (4H, m); FAB-HRMS m/z
calcd for C23H32N3O4S (M+H)+: 446.2114, Found:
446.2113; UV lmax298 (e 9190).
1
CDCl3) d 0.06 (6H, s), 0.84 (9H, s), 1.78 (1H, m), 2.92
(1H, m), 3.41 (2H, m), 3.74 (1H, m), 4.38 (2H, s), 4.53
(2H, m), 5.03 (2H, m), 5.21 (1H, m), 5.79 (1H, m), 7.27
(4H, s); FAB-HRMS m/z calcd for C21H34N4O4SiNa
(M+Na)+: 457.2247, Found: 457.2233.
The following compounds (1b, 2a±j, 3a±l, 8a, 8b, 9a, 9b,
10a and 10b ) were prepared as described for the pre-
paration of 1a or according to the method described in
the literature.13
(1R,5S,6S)-6-[(R)-1-Hydroxyethyl]-2-[(3S,5R)-5-[4-(N-iso-
propylaminomethyl)phenyl]pyrrolidin-3-ylthio]-1-methyl-1-
(1R,5S,6S)-2-[(3S,5S)-5-(4-Aminomethylphenyl)pyrroli-
din-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-
2-em-3-carboxylic acid hydrochloride 1b. IR nmax (KBr)
3421, 1749, 1652, 1558 cm 1; 1H NMR (300 MHz, D2O)
d 1.22 (3H, d, J=7.0 Hz), 1.27 (3H, d, J=6.0 Hz), 2.14
(1H, m), 3.00 (1H, m), 3.37 (1H, m), 3.46 (2H, m), 3.80
carbapen- 2-em-3- carboxylic acid hydrochloride 2e. IR
;
1H NMR
(300 MHz, D2O) d 1.22 (3H, d, J=7.3 Hz), 1.26 (3H, d,
J=6.3 Hz), 1.35 (6H, d, J=6.6 Hz), 2.52 (1H, m), 2.75
(1H, m), 3.38 (4H, m), 3.88 (1H, m), 4.20 (5H, m), 5.08
(1H, m), 7.54 (4H, m); FAB-HRMS m/z calcd for
1
nmax (KBr) 1751,1650, 1618, 1394 cm

1980
H. Imamura et al. / Bioorg. Med. Chem. 8 (2000) 1969±1982
C24H34N3O4S (M+H)+: 460.2270, Found: 460.2265;
UV lmax298 (e 8640).
(3H, d, J=6.4 Hz), 2.36 (1H, dd, J=14.0, 7.0 Hz), 2.61
(1H, m), 2.81 (1H, dd, J=15.6, 7.3 Hz), 2.88 (1H, dd,
J=15.6, 7.3), 3.18 (1H, dq, J=8.8, 7.0 Hz), 3.28 (1H, dd,
J=5.8, 2.8 Hz), 3.31 (1H, br d, J=12.8 Hz), 3.72 (1H, dd,
J=12.8, 5.8 Hz), 4.05 (3H, m), 4.59 (1H, t, J=7.3 Hz),
4.92 (1H, dd, J=11.0, 7.0 Hz), 7.36 (4H, m); 13C NMR
(125 MHz, D2O) d 15.3, 19.7, 35.7, 37.8, 40.7, 42.0, 51.2,
51.8, 55.6, 58.5, 61.2, 64.7, 127.6, 128.2, 134.3, 134.7,
136.3, 136.5, 167.3, 173.3, 176.3; FAB-HRMS m/z calcd
for C23H31N4O5S (M+H)+: 475.2015, Found 475.2011.
(1R,5S,6S)-2-[(3S,5R)-5-[4-[N-(Carbamoylmethyl)amino-
methyl]phenyl]pyrrolidin-3-ylthio]-6-[(R)-1-hydroxyethyl]-1
-methyl-1-carbapen-2-em-3-carboxylic acid hydrochloride
1
2f. IR nmax (KBr) 1751, 1677, 1587, 1392 cm
;
1H
NMR (300 MHz, D2O) d 1.21 (3H, d, J=7.3 Hz), 1.27
(3H, d, J=6.5 Hz), 2.47 (1H, m), 2.74 (1H, m), 3.39
(3H, m), 3.52 (2H, s), 3.86 (1H, dd, J=12.5, 5.9 Hz),
3.96 (2H, s), 4.22 (3H, m), 5.00 (1H, m), 7.47 (4H, s);
FAB-HRMS m/z calcd for C23H31N4O5S (M+H)+:
475.2015, Found: 475.2033; UV lmax298 (e 10700).
.
.
Anal. calcd for C22H30N4O5S HCl 2H2O: C, 50.50; H,
6.45; N, 10.24; S, 5.86; Found: C, 50.61; H, 6.55; N,
10.32; S, 5.60.
(1R,5S,6S)-2-[(3S,5R)-5-[4-[N-(2-Hydroxyethyl)aminome-
thyl]phenyl]pyrrolidin-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-
methyl-1-carbapen-2-em-3-carboxylic acid hydrochloride
(1R,5S,6S)-2-[(3S,5R)-5-[4-[(R)-1-Amino-2-carbamoyle-
thyl]phenyl]pyrrolidin-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-
methyl-1-carbapen-2-em-3-carboxylic acid hydrochloride
20
1
2g. IR nmax (KBr) 1749, 1558, 1394, 1081 cm
;
1H
3b (J-114,871). ‰ꢀŠ
12.6 (c 1.0, H2O); IR (KBr) nmax
d
1
NMR (300 MHz, D2O) d 1.22 (3H, d, J=7.0 Hz), 1.27
(3H, d, J=6.3 Hz), 2.51 (1H, m), 2.72 (1H, m), 3.17
(2H, m), 3.40 (3H, m), 3.84 (3H, m), 4.24 (5H, m), 5.03
(1H, m), 7.53 (4H, m); FAB-HRMS m/z calcd for
C23H32N3O5S (M+H)+: 462.2063, Found: 462.2048;
UV lmax298 (e 9600).
3417, 1751, 1673, 1583, 1390, 1261, 1147, 572cm 1; H
NMR (500 MHz, D2O) d 1.04 (3H, d, J=7.3 Hz), 1.08 (3H,
d, J=6.4 Hz), 2.36 (1H, dd, J=14.0, 6.7 Hz), 2.60 (1H, m),
2.81 (1H, dd, J=15.8, 7.3 Hz), 2.88 (1H, dd, J=15.8, 7.3),
3.18 (1H, dq, J=9.4, 7.3 Hz), 3.28 (1H, dd, J=6.1, 2.8 Hz),
3.31 (1H, br d, J=12.8 Hz), 3.72 (1H, dd, J=12.8, 5.8 Hz),
4.05 (3H, m), 4.59 (1H, t, J=7.3 Hz), 4.92(1H, dd, J=11.0,
6.7 Hz), 7.36 (4H, m); 13C NMR (125 MHz, D2O) d 15.3,
19.7, 35.7, 37.8, 40.7, 42.0, 51.2, 51.8, 55.6, 58.5, 61.2, 64.7,
127.6, 128.2, 134.3, 134.7, 136.3, 136.5, 167.3, 173.3, 176.3;
FAB-HRMS m/z calcd for C23H31N4O5S (M+H)+:
(1R,5S,6S)-6-[(R)-1-Hydroxyethyl]-2-[(3S,5R)-5-[4-(sul-
famoylaminomethyl)phenyl]pyrrolidin-3-ylthio]-1-methyl-
1-carbapen-2-em-3-carboxylic acid hydrochloride 2h. IR
1
nmax (KBr) 1753, 1589, 1394, 1152 cm
;
1H NMR
.
(300 MHz, D2O) d 1.24 (3H, d, J=7.0 Hz), 1.29 (3H, d,
J=6.5 Hz), 2.49 (1H, m), 2.74 (1H, m), 3.40 (3H, m),
3.76 (1H, m), 4.22 (3H, m), 5.02 (1H, m), 7.48 (4H, m);
FAB±MS m/z 497 (M+H)+; UV lmax298 (e 10200).
475.2015, Found 475.2042. Anal. calcd for C22H30N4O5S
.
HCl 2H2O: C, 50.50; H, 6.45; N, 10.24; S, 5.86; Found: C,
50.80; H, 6.66; N, 10.18; S, 5.65.
(1R,5S,6S)-2-[(3S,5R)-5-[4-(1-Amino-1-carbamoylmeth-
yl)-phenyl]pyrrolidin-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-
methyl-1-carbapen-2-em-3-carboxylic acid hydrochloride
(1R,5S,6S)-2-[(3S,5R)-5-[4-[N-(Acetimidoyl)aminome-
thyl]phenyl]pyrrolidin-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-
methyl-1-carbapen-2-em-3-carboxylic acid hydrochloride
1
3c. IRnmax (KBr) 1749, 1697, 1394 cm
;
1H NMR
1
2i. IR nmax (KBr) 1749, 1681, 1648, 1560, 1394 cm
;
(300 MHz, D2O) d 1.22 (3H, d, J=7.3 Hz), 1.26 (3H, d,
J=6.3 Hz), 2.55 (1H, m), 2.78 (1H, m), 3.36 (1H, m), 3.50
(2H, m), 3.90 (1H, m), 4.23 (3H, m), 5.14 (2H, m), 7.57 (4H,
m); FAB-HRMS m/z calcd for C22H29N4O5S (M+H)+:
461.1859, Found: 461.1847; UV lmax299 (e 8940).
1H NMR (300 MHz, D2O) d 1.21 (3H, d, J=7.1 Hz),
1.26 (3H, d, J=6.3 Hz), 2.26 (3H, s), 2.48 (1H, m), 2.78
(1H, m), 3.36 (1H, m), 3.48 (2H, m), 3.87 (1H, dd,
J=12.5, 5.9 Hz), 4.21 (3H, m), 4.51 (2H, s), 5.05 (1H,
m), 7.42 (2H, d, J=8.3 Hz), 7.49 (2H, d, J=8.3 Hz);
FAB-HRMS m/z calcd for C23H31N4O4S (M+H)+:
459.2066, Found: 459.2094; UV lmax298 (e 7590).
(1R,5S,6S)-2-[(3S,5R)-5-[4-(1-Amino-2-hydroxyethyl)phe-
nyl]pyrrolidin-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-
carbapen-2-em-3-carboxylic acid hydrochloride 3d. IR
1
(1R,5S,6S)-2-[(3S,5R)-5-[4-(Guanidinoaminomethyl)phe-
nyl]pyrrolidin-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-
1-carbapen-2-em-3-carboxylic acid hydrochloride 2j. IR
nmax (KBr) 1745, 1650, 1540, 1396 cm
;
1H NMR
(300 MHz, D2O) d 1.23 (3H, d, J=7.3 Hz), 1.27 (3H, d,
J=7.0 Hz), 2.54 (1H, m), 2.78 (1H, m), 3.38 (1H, m),
3.44 (2H, m), 3.90 (3H, m), 4.23 (3H, m), 4.51 (1H, m),
5.10 (1H, m), 7.53 (4H, m); FAB-HRMS m/z calcd for
C22H30N3O5S (M+H)+: 448.1906, Found: 448.1902;
UV lmax299 (e 9420).
1
nmax (KBr) 1749, 1653, 1558, 1396, 804 cm 1; H NMR
(300 MHz, D2O) d 1.22 (3H, d, J=7.3 Hz), 1.26 (3H, d,
J=6.6 Hz), 2.48 (1H, m), 2.79 (1H, m), 3.38 (1H, m),
3.43 (2H, m), 3.87 (1H, m), 4.21 (3H, m), 4.51 (2H, s),
5.02 (1H, m), 7.46 (4H. m); FAB-HRMS m/z calcd for
C22H30N5O4S (M+H)+: 460.2019, Found: 460.2045;
UV lmax298 (e 8640).
(1R,5S,6S)-2-[(3S,5R)-5-[4-(1-Amino-3-hydroxypropyl)-
phenyl]pyrrolidin-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-meth-
yl-1-carbapen-2-em-3-carboxylic acid hydrochloride 3e.
1
(1R,5S,6S)-2-[(3S,5R)-5-[4-[(S)-1-Amino-2-carbamoyle-
thyl]phenyl]pyrrolidin-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-
methyl-1-carbapen-2-em-3-carboxylic acid hydrochloride
IR nmax (KBr) 1743, 1650, 1560, 1540 cm 1; H NMR
(300 MHz, D2O) d 1.23 (3H, d, J=7.3 Hz), 1.27 (3H, d,
J=6.5 Hz), 2.20 (2H, m), 2.52 (1H, m), 2.73 (1H, m), 3.40
(4H, m), 3.60 (1H, m), 3.85 (1H, m), 4.24 (3H, m), 4.52 (1H,
m), 5.03 (1H, m), 7.51 (2H, d, J=8.6 Hz), 7.55 (2H, d,
J=8.6 Hz); FAB-HRMS m/z calcd for C23H32N3O5S
20
3a (J-114,870). ‰ꢀŠ +1.2 (c 1.0, H2O); IR (KBr) nmax
d
1
1751, 1672, 1585, 1388, 1259, 1147, 773, 665 cm 1; H
NMR (500 MHz, D2O) d 1.04 (3H, d, J=7.0 Hz), 1.08

H. Imamura et al. / Bioorg. Med. Chem. 8 (2000) 1969±1982
1981
(M+H)+: 462.2063, Found: 462.2064; UV lmax298 (e
9750).
(300 MHz, D2O) d 1.27 (8H, m), 1.36 (2H, m), 2.49 (1H,
m), 2.72 (1H, m), 3.46 (3H, m), 3.84 (1H, dd, J=13.0,
6.0 Hz), 4.20 (3H, m), 5.03 (1H, dd, J=11.0, 7.0 Hz), 7.49
(4H, s); FAB-HRMS m/z calcd for C23H30N3O4S (M+
H)+: 444.1957, Found: 444.1954; UV lmax298 (e 9650).
(1R,5S,6S)-2-[(3S,5R)-5-[4-(1-Amino-1-cyanomethyl)phe-
nyl]pyrrolidin-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-
carbapen-2-em-3-carboxylic acid hydrochloride 3f. IR
n
max (KBr) 2240, 1749, 1648, 1560, 1394 cm 1; 1H NMR
(1R,5S,6S)-2-[(3S,5R)-5-[4-(1-Amino-3-butenyl)phenyl]-
pyrrolidin-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-
carbapen-2-em-3-carboxylic acid hydrochloride 3l. IR
(300 MHz, D2O) d 1.23 (3H, d, J=7.3 Hz), 1.27 (3H, d,
J=6.3 Hz), 2.52 (1H, m), 2.73 (1H, m), 3.40 (3H, m),
3.78 (1H, m), 4.24 (2H, m), 5.08 (2H, m), 7.57 (4H, m);
FAB±MS m/z 443 (M+H)+; UV lmax298 (e 10300).
1
nmax (KBr) 1749, 1616, 1396, 1288 cm
;
1H NMR
(300 MHz, D2O) d 1.22 (3H, d, J=7.3 Hz), 1.27 (3H, d,
J=6.3 Hz), 2.54 (1H, m), 2.75 (2H, dd, J=7.1, 7.1 Hz),
2.80 (1H, m), 3.37 (1H, m), 3.49 (2H, m), 3.91 (1H, m),
4.23 (3H, m), 4.48 (1H, t, J=7.4 Hz), 5.10 (1H, m), 5.18
(2H, m), 5.69 (1H, m), 7.53 (4H, m); FAB-HRMS m/z
calcd for C24H32N3O4S (M+H)+: 458.2114, Found:
458.2104; UV lmax298 (e 9030).
(1R,5S,6S)-2-[(3S,5R)-5-[4-(1-Amino-2-ethylthioethyl)phe-
nyl]pyrrolidin-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-
carbapen-2-em-3-carboxylic acid hydrochloride 3g. IR
1
nmax (KBr) 1743, 1650, 1558, 1540cm
;
1H NMR
(300MHz, D2O) d 1.17 (3H, d, J=7.4Hz), 1.23 (3H, d,
J=7.2Hz), 1.28 (3H, d, J=6.3Hz), 2.51 (2H, q,
J=7.3Hz), 2.55 (1H, m), 2.78 (1H, m), 3.13 (2H, m), 3.36
(1H, m), 3.47 (2H, m), 3.88 (1H, m), 4.24 (3H, m), 4.55
(1H, m), 5.06 (1H, m), 7.55 (4H, m); FAB-HRMS m/z
calcd for C24H34N3O4S2 (M+H)+: 492.1991, Found:
492.1973; UV lmax300 (e 9810).
(1R,5S,6S)-2-[(3S,5R)-5-(4-Aminoethylphenyl)pyrrolidin-
3-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-
em-3-carboxylic acid hydrochloride 8a. IR nmax (KBr)
1747, 1583, 721 cm 1; 1H NMR (300 MHz, D2O) d 1.21
(3H, d, J=7.1 Hz), 1.25 (3H, d, J=6.3 Hz), 2.47 (1H,
m), 2.74 (1H, m), 3.01 (2H, t, J=7.2 Hz), 3.26 (2H, t,
J=7.2 Hz), 3.35 (1H, m), 3.46 (2H, m), 3.83 (1H, m),
4.22 (3H, m), 5.03 (1H, m), 7.37 (2H, d, J=8.3 Hz), 7.45
(2H, d, J=8.3 Hz); FAB-HRMS m/z calcd for
C22H30N3O4S (M+H)+: 432.1957, Found: 432.1947;
UV lmax298 (e 9190).
(1R,5S,6S)-2-[(3S,5R)-5-[4-(1-Aminoethyl)phenyl]pyrroli-
din-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-
2-em-3-carboxylic acid hydrochloride 3h. IR nmax (KBr)
1751, 1576, 1456, 1389, 1286, 1261, 1146cm 1; 1H NMR
(300MHz, D2O) d 1.22 (3H, d, J=7.3Hz), 1.26 (3H, d,
J=6.2Hz), 1.62 (3H, d, J=6.9 Hz), 2.51 (1H, dd, J=14.1,
6.1 Hz), 2.75 (1H, m), 3.36 (1H, m), 3.46 (2H, m), 3.87
(1H, dd, J=12.5, 6.0 Hz), 4.22 (3H, m), 4.55 (1H, q,
J=6.9Hz), 5.05 (1H, dd, J=11.1, 6.7 Hz), 7.52 (4H, s);
FAB-HRMS m/z calcd for C22H30N3O4S (M+H)+:
432.1957, Found: 432.1955; UV lmax298 (e 9580).
(1R,5S,6S)-2-[(3S,5S)-5-(4-Aminoethylphenyl)pyrrolidin-
3-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-
em-3-carboxylic acid hydrochloride 8b. IR nmax (KBr)
1747, 1621, 1153, 721 cm 1; 1H NMR (300MHz, D2O) d
1.22 (3H, d, J=6.9 Hz), 1.27 (3H, d, J=6.6 Hz), 2.19 (1H,
m), 3.00 (3H, m), 3.27 (2H, t, J=7.7Hz), 3.46 (1H, m),
3.49 (2H, m), 3.77 (1H, m), 4.13 (1H, m), 4.22 (2H, m), 4.81
(1H, m), 7.39 (2H, d, J=7.7 Hz), 7.50 (2H, d, J=7.7 Hz);
FAB-HRMS m/z calcd for C22H30N3O4S (M+H)+:
432.1957, Found: 432.1967; UV lmax298 (e 9440).
(1R,5S,6S)-2-[(3S,5R)-5-[4-(1-Aminopropyl)phenyl]pyrroli-
din-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-
2-em-3-carboxylic acid hydrochloride 3i. IR nmax (KBr)
1751, 1626, 1568, 1392, 1267 cm 1; 1H NMR (300 MHz,
D2O) d 0.83 (3H, t, J=7.4 Hz), 1.21 (3H, d, J=7.3 Hz),
1.26 (3H, d, J=6.3 Hz), 1.99 (2H, m), 2.47 (1H, m), 2.71
(1H, m), 3.38 (3H, m), 3.84 (1H, dd, J=12.6, 5.9 Hz),
4.22 (4H, m), 5.00 (1H, dd, J=10.8, 6.7 Hz), 7.48 (2H,
d, J=8.6 Hz), 7.53 (2H, d, J=8.6 Hz); FAB-HRMS m/z
calcd for C23H32N3O4S (M+H)+: 446.2114, Found:
446.2114; UV lmax298 (e 8210).
(1R,5S,6S)-2-[(3S,5R)-5-(4-Aminomethyl-1-naphtyl)pyrro-
lidin-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carba-
pen-2-em-3-carboxylic acid hydrochloride 9a. IR nmax
(KBr) 1754, 1575, 721 cm 1; 1H NMR (300MHz, D2O) d
1.23 (3H, d, J=7.0 Hz), 1.26 (3H, d, J=6.3 Hz), 2.61 (1H,
m), 3.02 (1H, m), 3.40 (1H, m), 3.47 (1H, m), 3.53 (1H, m),
3.87 (1H, m), 4.23 (2H, m), 4.29 (1H, m), 4.70 (2H, m),
5.86 (1H, m), 7.64 (1H, m), 7.73 (3H, m), 8.16 (1H, m),
8.22 (1H, m); FAB-HRMS m/z calcd for C25H30N3O4S
(M+H)+: 468.1957, Found: 468.1961; UV lmax297 (e
10900).
(1R,5S,6S)-2-[(3S,5R)-5-[4-(1-Amino-1-methylethyl)phe-
nyl]pyrrolidin-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-
1-carbapen-2-em-3-carboxylic acid hydrochloride 3j. IR
nmax (KBr) 2972, 1753, 1626, 1583, 1394, 1302,
579 cm
1
;
1H NMR (300 MHz, D2O) d 1.23 (3H, d,
J=7.3 Hz), 1.27 (3H, d, J=6.5 Hz), 1.74 (6H, s), 2.54
(1H, m), 2.80 (1H, m), 3.39 (1H, m), 3.50 (2H, m), 3.91
(1H, m), 4.24 (3H, m), 5.11 (1H, m), 7.58 (4H, m); FAB-
HRMS m/z calcd for C23H32N3O4S (M+H)+:
446.2114, Found: 446.2089; UV lmax298 (e 8170).
(1R,5S,6S)-2-[(3S,5S)-5-(4-Aminomethyl-1-naphtyl)pyrro-
lidin-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carba-
pen-2-em-3-carboxylic acid hydrochloride 9b. IR nmax
1
(KBr) 1747, 1583, 721 cm 1; H NMR (300 MHz, D2O)
d 1.20 (3H, d, J=7.3 Hz), 1.26 (3H, d, J=6.5 Hz), 2.33
(1H, m), 3.12 (1H, m), 3.41 (2H, m), 3.52 (1H, m), 3.89
(1H, m), 4.20 (3H, m), 4.70 (2H, m), 5.68 (1H, m), 7.68
(1H, m), 7.73 (2H, m), 7.80 (1H, m), 8.18 (2H, m); FAB-
HRMS m/z calcd for C25H30N3O4S (M+H)+:
468.1957, Found: 468.1961; UV lmax296 (e 16100).
(1R,5S,6S)-2-[(3S,5R)-5-[4-(1-Aminocyclopropyl)phenyl]-
pyrrolidin-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-
carbapen-2-em-3-carboxylic acid hydrochloride 3k. IR
1
nmax (KBr) 1749, 1558, 1394, 1261 cm
;
1H NMR

1982
H. Imamura et al. / Bioorg. Med. Chem. 8 (2000) 1969±1982
(1R,5S,6S)-2-[(3S,5R)-5-(5-Aminomethyl-2-thienyl)pyrro-
lidin-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carba-
pen-2-em-3-carboxylic acid hydrochloride 10a. IR nmax
H.; Nakano, M.; Ushijima, R.; Nakagawa, S. Bioorg. Med.
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1
(KBr) 1754, 1579, 721 cm 1; H NMR (300 MHz, D2O)
d 1.21 (3H, d, J=7.2 Hz), 1.25 (3H, d, J=6.3 Hz), 2.51
(1H, m), 2.73 (1H, m), 3.32 (2H, m), 3.42 (1H, m), 3.74
(1H, m), 4.19 (3H, m), 4.34 (2H, s), 5.20 (1H, m), 7.16
(2H, m); FAB-HRMS m/z calcd for C19H26N3O4S2
(M+H)+: 424.1365, Found: 424.1356; UV lmax298 (e
10300).
8. Gill, C. J.; Jackson, J. J.; Gerckens, L. S.; Pelak, B. A.;
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lidin-3-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carba-
pen-2-em-3-carboxylic acid hydrochloride 10b. IR nmax
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1
(KBr) 1754, 1575 cm 1; H NMR (300 MHz, D2O) d
1.20 (3H, d, J=7.0 Hz), 1.26 (3H, d, J=6.2 Hz), 2.04
(1H, m), 2.96 (1H, m), 3.37 (3H, m), 3.58 (1H, m), 4.03
(1H, m), 4.20 (2H, m), 4.34 (2H, s), 7.12 (2H, m); FAB-
HRMS m/z calcd for C19H26N3O4S2 (M+H)+:
424.1365, Found: 424.1362; UV lmax300 (e 10500).
Acknowledgements
We are grateful to Mr. Dan Johnson, Merck & Co., for
his critical reading of this manuscript.
References and Notes
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