Note: Descriptions are shown in the official language in which they were submitted.
134049~
THIS APPLICATION IS A DIVISIONAL OF CANADIAN PATENT
APPLICATION NO. 298,883 FILED ON NARCH 14, 1978
This invention relates to new cephem and cepham
compounds. More particularly, it relates to new 7-
substituted-3-cephem(or cepham)-4-carboxylic acid, its
pharmaceutically acceptable salt and pharmaceutically
acceptable bioprecursor thereof, which have antimicrobial
activities, and processes for preparation thereof, to
intermediate for preparing the same and processes for
preparation thereof, and to pharmaceutical composition
comprising the same and methods of using the same
prophylactically and therapeutically for treatment of
infectious diseases in human being and animals.
Accordingly, the objects of this invention are
to provide:-
New 7-substituted-3-cephem(or cepham)-4-
carboxylic acid, its pharmaceutically acceptable salt and
pharmaceutically acceptable bioprecursor thereof, which
exhibit excellent antimicrobial activities against a wide
variety of pathogenic microorganisms including Gram
negative and Gram positive bacteria,
Processes for preparation of the same,
Pharmaceutical composition comprising one of
the same as an active ingredient, and
A method of using the same prophylactically and
therapeutically for treatment of infectious diseases
caused by pathogenic microorganisms in human being and
animals; and further
Intermediate to be used for preparation of
pharmaceutically active 7-substituted-3-cephem(or cepham)-4-
J'~L
13~0~3
carboxvlic acid, its pharmaceutically acceptable salt or
pharmaceutically acceptable bioprecursor ~hereof, and
methods for preparation of the same.
The cephem ~nd cepllam compoullds provided by this
invention can be represented by the formula ~
Rl-A-CONH ~ ~ R (I)
wherein Rl is thiadiazolyl, thiazolyl of the formula :
R6~ ~ in which RG is amillo or protected amino,
or haloacetyl,
A is methylene or a group of the formula :
-C- in which R2 is hydrogen
N
o-R2
or an aliphatic hydrocarbon residue which may be
substituted with halogell, carboxy or esterified
carboxy,
R is hydrogen or lower alkyl,
R4 is hydrogen, halogen, lower alkyl or a group of
the formula : -o-R7 in which R7 is hydrogen,
lower alkyl or acyl,
R5 is carboxy or functionally modified carboxy,
and
the dotted line represents 3-cephem and cepham
nuclei, inclusively,
: provided that
i) R4 is hydrogell, halogen or a group of the formula :
13~0~ 99
-o-R7 in which R7 is as defined above, when R3 is
hydrogen,
ii) R4 is lower alkyl, when R3 is lower alXyl,
iii) -A is a group of the formula : -C-
o-R2
in which R2 is as defined above, when Rl is
thiadiazolyl or thiazolyl of the formula :
R6~ ~ in which R6 is as defined above, and
iv) the dotted line represents 3-cephem nucleus and R4
is hydrogen, halogen, lower alXyl or -oR7 in which
R7 is lower alkyl, when R is haloacetyl, and
v) when R is halogen or a group of the ~ormula
-o-R7 in which R7 is lower alkyl, R2 is othex
than hydrogen and lower alkyl.
.,; ,. . ............
13~ 19.9
It is to be noted that the cephem and cepham
compounds (I) as illustrated above include a compound useful
as an antimicrobial agent and also a compoulld useful as an
intermediate for preparing the aboYe antimicrobial agent,
particularly as illustrated below.
The.compound useful as an antimicrobial agent can
be represented by the formula (I') :-
Rl-C-CONH S R3
N O ~ ~ R (I')
O-R R5
wherein Ra is thiadiazolyl or thiazolyl of the formula :
R ~ ~ in which R6 is as defined above, and
R2, R3, R4 and R5 are each as defined above.
On the other hand, the compound useful as an
intermediate for preparing the above compound (I') can be
represented by the formula (I") :-
Rb-A-COl~H
.. . .
. ~ ~
- .
13~0~93
~
wherein Rb is haloacetyl,
R4 is hydrogen, halogen, lower alkyl or a group of
the formula : -o-R7 in which R7 is lower alkyl,
and
R3, R5 and A are cach as defined above.
And further, it is to be noted that the compound
(I') where Ra is thiazolyl of the formula :- R6 ~ ~ in
whic]l R6 is protected amino, R4 is a group of the formula :
~; -o-R7 in which R7 is hydrogen or acyl and/or R5 is
functionally modified carboxy is also useful as an
intermediate for preparing the more active compound as
explained below.
Accordingly, the more preferred active compound
can be reprcsented by the formula (I"') :-
Rc-C-CONII ~ ~ 3 (I ~)
o - R2 COOI I
wherein Rc is thiadiazolyl or thiazolyl of the formula :
il2N ~ ~ and
R2, R3 and Ra are each as defined above.
The terms and definitions described in this
specification and claims are illustrated as follows.
a) Partial structure of the formula :
Rl-c-co
N
~ 2
O-R
is intended to mean both of the ge-ometric formulae :
-- 4 -
13~0 -'193
.
Rl - C- CO - Rl - C - CO -
Il 2 and z
N-0-R R -0-N
(S) (A)
The geometry of the formula (S) is referred to as
syn" and another formula (A) is referred to as "anti .
Accordingly, one isomer of the compound having
the partial structure shown by the above formula (S) is
referred to as syn isomer and another isomer of the compound
having the alternative one shown by the above formula (A)
is referred to as "anti isomer , respectively.
From the view poin. of structure-activity
relationship, it is to be noted that a syn isomer of the
compound (I'3 tcnds to be of much higher antimicrobial
activity than the corresponding anti isomer, and accordingly
the syn isomer of the compound (I') is more ~referable
antimicrobial agent than the corresponding anti isomer in
the prophylactic and therapeu~ic value.
b) The thiazolyl group of the formula : R6 ~ ~
(wherein R6 is as defined above) is well known to lie in
tautomeric relation with a thiazolinyl group of the formula:
HN ~ (wherein R6 is imino or protected imino).
The tautomerism between the said thiazolyl and
thiazolinyl groups can be illustrated by the following
equilibrium :
R6 ~ S ~ = R ~ ~
(wherein R6 and RG are each as defined above).
Accordingly, it is to be understood that both of
the said groups are substantially the same and the tautomers
consisting of such groups are regarded as the same compounds,
- 5 --
13404~3
especially in the manufacturillg chemistry. Therefore,
both of the tautomeric forms of tle compounds having such
groups in their molecule are included in the scope of this
invention and designated inclusively witll one expression
"thiazolyl" ~nd represcnted by the formula:
R6 ~ ~ (wllerein R6 is as defined above) only for the
convenient sake throughout this specification and claims.
c) It is well known that the 3-hydroxy-3-cephem
compound havillg the partial structure of tlle formula:
S
lies in a tautomeric relation with the 3-oxo-cepham compoulld
of the formula:
T~S~
N ~ 0
R5
each of whicll is reLerred to as the enol- or keto-tautomer,
and that the enol-tautomer is usually the stabilized one.
Accordingly, both of the compounds having such
tautomeric structures are included within the same scope of
the compound, and therefore, thc structure and nomenclature
of such tautomers are expressed inclusively with one
expression of the stabilized enol tautomer, i.e. 3-hydroxy-3-
cephem compound throughout this specification and claims.
In the above and subsequent descriptiolls of this
....
specification, s,uitable examples and illustratlon of the
various definitions which tllis invention intends to include
within the scope thereof are explained in detail as follows.
. .,.; ~ :.: - .
. _ . ~ . , .
13~0~93
.
The te~m "lower" is used to intend a group having
1 to G carbon atons, unless otherwise provided.
"Tlliadiazolyl" for Rl may be 1,2,3-thiadiazolyl
- ~e.g. 1,2,3-thiadiazol-4-yl or 1,2,3-thiadiazol-5-yl),
1,3,4-thiadiazolyl or 1,2,4-thiadiazolyl, preferably 1,2,3-
thiadiazolyl, and more preferably 1,2,3-thiadiazol-4-yl.
"Aliphatic hydrocarbon residue "for R2 may include
a monovalentradical of a saturated or unsuturated, and
straight, branched or cyclic aliphatic hydrocarbon, and
particularly may include alkyl, alkenyl, alkynyl, cycloalkyl
and the likej the details of which are explained below.
"Alkyl" may include a residue of straight or
branched alkane having 1 to 12 carbon catoms such as methyl,
ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl,
neopentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl,
dodecyl and the like, preferably lower alkyl, and more
preferably the one having 1 to 4 carbon atoms.
"Alkenyl" may include a residue of a straight or
branched alkene having up to 12 carbon atoms, preferably
lower alkenyl such as vinyl, allyl, l-propenyl, isopropenyl,
butenyl, isobutenyl, pentenyl, hexenyl and the like, and
more preferably the ones having up to 4 carbon atoms.
"Alkynyl" may include a residue of a straight
or branched alkyne having up to 12 carbon atoms, preferably
lower alkynyl such as ethyn~l, propargyl, l-propynyl,
3-butynyl, 2-butynyl, 4-pen.ynyl, 3-pentynyl, 2-pentynyl,
l-pentynyl, 5-hexynyl and the like, and more preferably the
ones having up to 4 carbon atoms.
_
13~0 -~93
"Cycloalkyl" may include a residue of a cycloalkane
having up to 8 carbon atoms, preferably lower cycloalkyl SUC]l
as cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl, and ~ re
preferably cyclohexyl.
These aliphatic hydrocarbon residues may be substi-
tuted with halogen atom(s), carboxy or esterified carboxy
group(s). Accordingly, "ali.phatic hydrocarbon residue
substituted with llalogen atom(s), carboxy or esterified
carboxy group(s)" can also be alternatively expressed as
"halogen-substituted aliphatic hydrocarbon residue",
"carboxy-substituted aliphatic hydrocarbon residue" and
"esterified carboxy-substituted aliphatic hydrocarbon
residue", respectively, which may include more particularly
halo-alkyl, alkenyl, alkynyl and cycloalkyl; carboxy-alkyl,
alkenyl, alkynyl and cycloalkyl; and esterified carboxy-
alkyl, alkenyl, alkynyl and cycloalkyl, respectively.
Suitable examples of the "halogen" may include
chlorine, bromine, iodine and fluorine; suitable examples
of the "esterified carboxy" may be alkoxycarbonyl or the
like; and preferred examples o~ the "alkyl", "alkenyl",
"alkynyl", "cycloalkyl" and alkyl moiety of the "alkoxy-
carbonyl" are the corresponding "lower" ones as mentioned
above.
Preferredexamples of the "halo-alkyl, alkenyl,
alkynyl and cycloalkyl" may be chlorometllyl., bromomethyl,
iodomethyl, fluoromethyl, trichloromethyl, trifluoromethyl,
2-chloroethyl, 1,2-dichloroetllyl, 2,2,2-trifluoroetllyl,
3-chloropropyl, 4-iodobutyl, 5-fluoropentyl, 6-bromohexyl,
3-fluoroallyl, 3-chloropropargyl, 4-fluorocyclohexyl, or
the like-
13~0~9~
Preferred examples of the "carboxy-alkyl, alkenyl,
alkynyl and cycloalkyl" may be carboxymethyl, l-carboxyethyl,
2-carboxyethyl~ l-carboxypropyl, 3-carboxypropyl, 4-carboxy-
butyl, 5-carboxypentyl, 6-carboxyhexyl, l-carboxyisopropyl,
l-ethyl-l-carboxyethyl, 2-methyl-2-carboxypropyl,
3-carboxyallyl, 3-carboxypropargyl, 4-carboxycyclohexyl, or
the like.
Preferredexamples of the "esterified carboxyalkyl,
alkenyl, alkynyl and cycloalkyl" may be lower alkoxycarbonyl-
(lower)alkyl (e.g. methoxycarbonylmetllyl, ethoxycarbonylmethyl,
propoxycarbonylmethyl, t-butoxycarbonylmethyl, 2-ethoxycarbollyl-
ethyl, 2-ethoxycarbonylpropyl, 4-ethoxycarbonylbutyl,
l-t-butoxycarbonylisopropyl, l-t-butoxycarbonyl-l-methyl-
propyl, 4-t-butoxycarbonylbutyl, 5-t-butoxycarbonylpentyl,
6-butoxycarbonylhexyl, etc.), lower alkoxycarbonyl(lower)-
alkenyl (e.g. 3-methoxycarbollylallyl, etc.), lower alkoxy-
carbonyl(lower)alkynyl (e.g. 3-methoxycarbonylpropargyl, etc.),
lower alkoxycarbonyl(lower)cycloalkyl (e.g. 4-methoxycarbonyl-
cyclohexyl, etc.) or the like, and more preferably lower
alkoxycarbonylmethyl as exemplified above.
"Lower alkyl" for R3, R4 and R7 is to be referred
to those as exemplified in the term of the aliphatic
hydrocarbon residue for R2, preferably may be the ones having
up to 4 carbon atoms and more preferably methyl.
"Halogen" for R4 may be chlorine, bromine, iodine
or fluorine, and preferredone is chlorine or bromine.
.
"Acyl" for R7 may be lower alkanoyl (e.g. formyl,
g
. ~ :
13~O'~9 3
acetyl, propionyl, butyryl, isobutyryl, isovaleryl, p-ivaroyl,
etc.), aroyl (e.g. benzoyl, etc.),lower alkanesulfonyl (e.g.
mesyl, ethanesulfonyl, l-methylethanesulfonyl, propanesulfonyl,
butanesulfonyl, etc.), arenesulronyl (e.g. benzenesulfonyl,
tosyl, etc.) or the like.
"Protective group" in the "protected amino" for R6
may be the conventional N-protective group such as substituted
or unsubstituted ar(lower)alkyl (e.g. benzyl, benzhydryl,
trityl, 4-methoxybenzyl, 3,4-dimethoxybenzyl, etc.), halo-
(lower)alkyl (e.g. trichloromethyl, trichloroethyl, trifluoro-
methyl, etc.), tetrahydropyranyl, substituted phenylthio,
substituted alkylidene, substituted aralkylidene, substituted
cycloalkylidene, acyl, or the like.
Suitable acyl for the protective group may be sub-
stituted or unsubstituted lower alkanoyl (e.g. formyl, acetyl,
chloroacetyl, trifluoroacetyl, etc.), substituted or unsubstituted
lower alkoxycarbonyl (e.g. methoxycarbonyl, ethoxycarbonyl,
propoxycarbonyl, l-cyclopropylethoxycarbonyl, isopropoxycarbonyl,
butoxycarbonyl, t-butoxycarbonyl, pentyloxycarbonyl, t-pentyloxy-
carbonyl, hexyloxycarbonyl, trichloroethoxycarbonyl, 2-pyridyl-
methoxycarbonyl, etc.), substituted or unsubstituted ar(lower)-
alkoxycarbonyl (e.g. benzyloxycarbonyl, benzhydryloxycarbonyl,
4-nitrobenzyloxycarbonyl, etc.), lower cycloalkoxycarbonyl (e.g.
cyclopentyloxycarbonyl, cyclohexyloxycarbonyl, etc.), 8-quinolyl-
oxycarbonyl, succinyl, phthaloyl, or the like.
And further, the reaction product of a silan,
boron, aluminium or phosphorus compound with the amino group
may also be included in the protective group. Suitable
examples of such compounds may be trimethylsilyl chloride,
- 10 -
- ~340~
trimethoxysilyl chloride, boron trichloride, butoxyboron
dichloride, aluminwll trichloride,dietlloxy aluminum chloride,
phosyhorus dibromide, phenylphosphorus dibromide, or the
like.
"Functionally modified carboxy" for R5 may be an
ester, amide or the like.
Suitable examples o~ the ester may be
alkyl ester (e.g. methyl ester, ethyl ester, propyl
ester, isopropyl ester, butyl ester, isobutyl ester, t-butyl
ester, pentyl ester, t-pentyl ester, hexyl ester, heptyl ester,
octyl ester, l-cyclopropylethyl ester, etc.);
alkenyl ester (e.g. vinyl ester, allyl ester, etc.);
alkynyl ester (e.g. ethynyl ester, propynyl ester,
etc.);
alkoxyalkyl ester (e.g. methoxymethyl ester,
ethoxymethyl ester, isopropoxymethyl ester, l-methoxyethyl
ester, l-ethoxyethyl ester, etc.);
alkylthioalkyl ester (e.g. methylthiolllethyl ester,
ethyl~}liomethyl ester, ethylthioethyl ester,isopropylthio-
methyl ester, etc.);
haloalkyl ester (e.g. 2-iodoethyl ester, 2,2,2-
trichloroethyl ester, etc.);
alkanoyloxyalkyl ester (e.g. acetoxymethyl ester,
propionyloxymethyl ester, bùtyryloxymethyl ester, valeryloxy-
methyl ester, pi~aloyloxymethyl ester, hexanoyloxymethyl
ester, 2-acetoxyethyl ester, 2-propionyloxyethyl ester,
palmitoyloxymethyl ester, etc.);
alkanesulfonylalkyl ester (e.g. mesylmethyl ester,
2-mesylethyl ester, etc.);
- 11 -
. . . ~ .. .
~. .
. . .
13~0~93
.
substituted or unsubstituted aralkyl ester (e.g.
benzyl ester, 4-methoxybenzyl ester, 4-nitrobenzyl ester,
phenetnyl ester, trityl ester, benzhydryl ester, bis~methoxy-
phenyl)methyl ester, 3,4-dimethoxybenzyl ester, 4-hydroxy-3,5-
di-t-butylbenzyl ester, etc.);
substituted or unsubst-ituted aryl ester ~e.g. phenyl
ester, tolyl ester, t-butylphenyl ester, xylyl ester, mesityl
ester, cumenyl ester, salicyl ester, etc.);
an ester with a sily] compound SUC}I as trialkylsilyl
compound, dialkylalkoxysilyl compowld or trialkoxysilyl
compound, for exampleJ trialkylsilyl ester (e.g. trimethyl
silyl ester, triethylsilyl ester, etc.), dialkylalkoxy
silyl ester (e.g. dimethylmethoxysilyl ester, dimethylethoxy-
silyl ester, diethylmethoxysilyl ester, etc.) or trialkoxysilyl
ester (e.g. trimethoxysilyl ester, triethoxysilyl ester, etc.)
or the like.
With regard to the ~erms "protected amino" for R6
and "functionally modified carboxy" for R5, it is to be
understood that these groups bear the meaning not only in
synthetic manufacture of the object compound by chemical
process(es), but also in physiological and pharmaceutical
properties of the object compound per se.
That is, in the meaning of the synthetic manufacture,
free amino group for R and/or free carboxy group for R5 may
be transformed into the "protected amino" and/or "functionally
modified carboxy" as mentioned above before conducting the
process(es) for preventing any possible undesired side
reaction(s), and tlle "protected amino" and/or "functionally
modified carboxy" group in thc resultant compound may be
- 12 -
... .
~ 13~0~9~
transformed into free amino and/or carboxy group after the
reaction is conducted. This will be apparent from the
explanation of the processes in the following.
On the other hand, in the meaning of the physiological
and pharmaceutical properties of the object compound, the
compound bearing the "protected amino"and/or "functionally
modified carboxy" group is optionally used for improving the
properties such as solubility, stability, absorbability,
toxicity of the particularly actiYe object compound bearing
the free amino and/or carboxy group
Suitable "pharmaceutically acceptable salt'l o~
the object compound ~I') may be conventional IlOn-tOXiC salt,
and may include a salt with an inorganic base or acid,
for example, a metal salt such as an alkali metal salt (e.g.
sodium salt, potassium salt, etc.) and an alkaline earth
metal salt te.g. calcium saltJ magnesiulll salt, etc.),
ammonium salt, an inorganic acid salt ~e.g. hydrochloride,
hydrobromide, sulfate, phosphate, carbonate, bicarbonate,
etc.), a salt l~ith an organic base or acid, for example,
an amine salt (e.g. trimethylamine salt, triethylamine salt,
pyridine salt, procaine salt, picoline salt, dicyclohexylamine
salt, N,N'-dibenzylethylenediamine salt, N-methylglucamine
salt, diethanolamine salt, triethanolamine salt, tris-
(hydroxymethylamino)methane salt, phenethylbenzylamine salt,
etc.), an organic carboxylic or sulfonic acid salt (e.g.
acetate, maleate, lactate, tartrate, mesylate, benzene-
sulfonate, tosylate, etc.), a basic or acidic amino acid
salt te.g. arginine salt, aspartic acid salt, glutamic acid
salt, lysine salt,serin¢ salt, etc.) and the li~e.
It is well knowll in the pharmaceutical field that
- 13 -
13~0~'13
the active drug, when it has any undesired physiological or
pharmaceutical property such as solubility, stability,
absorbability, etc., is converted into modified derivative
thereof for improving such undesired properties, and then
said derivative, upon admillistratioll to a patient, exhibits
the active efficacy by being converted in the body to the
parent drug. In this meaning, the term "pharmaceutically
acceptable bioprecursor" used throughout this specification
and claim is intended to fundamentally mean all of the
10 modified derivatives, which have structural ~ormulae
dif~erent from those o~ the active compounds of this
invention, but are converted in the body to the active
compounds of this invention upon administration, and also
to mcan the derivatives whicll are sometimes dcrived
physiologically from the compounds of ~his invention in the
body and exhibit antimicrobial ef~icacy.
- 14 -
13~33
The compounds (I) of this invention can be
prepared by processes as shown in the following scheme.
Process A : N-Acylation
A-coo~ -A-coN~ cR4
R5 R
(~) (I)
Process B : C-Nitrosation
~b-Cl~2C~NIl S~_ R3 Nitrosating Rl-C-CONII ~ S ~ - R3
:. ~N ~ Ra ~ ~ll O ~ ~ a
(IV) R5 (Ib)
, ,
Process C : Etherification
Rl-~ CONH , S R3 Etherifying Rl-~-CONI ~rR
~N ~ N~- ~ R4 agent ~ N ~ ~ ~ R4
OH O R5 o-R2 R5
(V)
(Ic)
Process D : Thiazole ring formation
Rl-C-CONH ~ S R3 R6-C-NH2 (V~) N C CO S
O N ~ a R6 ~ ~ N ~ ~R4
(VI) (Id)
- 15 -
. . ~,
~ 13~0~9~
Process E : Elimination of amino-protective group
R6 ~ t ~N~ R4 7 ~ CON~C ~ R4
(VIII) (Ie)
Process F : Reductive formation o~ 3-hydroxycepham
Rl - IlC-CONHn ~ ~ Rl- IC-CONH~
~ N '~OH 1 o
o R2 .R o R2R5
(IX) (If)
Process G : O-Acylation
Rl-C CON~--08 > N 0~} a
(If) (Ig)
Process H : 3-Cephem formation
Rl-C-CONIl~ L ~ Base Ra-l CoNI~
30(Ig) ' (Ih)
- 16 -
.
13~0~9'~
Process I : Halogenation
Rl C-CON ~ ~ Halogenating agent > Ra- Il CON~ g
o l~2 R5 . o R2 R5
(XI) (Ii)
Process J : Esterification
Rl-A-CONH ~ ~ R4 Esterifying agent> R -A-CONI~ R4
COOH Ra
(XII) (Ij)
Process K : Carboxy formation
Rl-A-CONIi ~ ~ R3 Rl A CONI ~ ~ R4
R5 COOH
b
(XIII) (Ik)
wherein Ra is an aliphatic hydrocarbon residue which may
be substituted with halogen, carboxy or
esterified carboxy,
Rb is halogen,
Ra is esterified carboxy,
Rb is functionally modified carboxy,
R6 is protected amino,
30Ra is acyl, and
- 17 -
- 1340499
Rl, Ra, Rb, RZ, R3, R4, Ra, R5 and A are each as
defined above.
The above yrocesses will be explained in detail in
the following
Process A : N-Acylation
A compound (I) and its salt can be prepared by
reacting a 7-amino-3-cephem (or cepl~am) compound (II), its
reactive derivative at the amino or a salt thereof with a
10 carboxylic acid (III), its reactive derivative at the carboxy
or a salt thereof according to a conventional manner of so-
called amidation reaction well known in ~-lactam chemistry.
The starting compound (III) includes both of known
and new ones, and the new compound (III) can be prepared
according to the methods as explained hereinafter in this
specification.
Suitable reactive derivative at the amino group
of the compound (II) may include a conventional reactive
derivative as used in a wide variety of amidation reaction,
20 for example, isocyanato, isothiocyanato, a derivative formed
by the reaction of a compound (II) with a silyl compound
(e.g. trimethylsilylacetamide, bis(trimethylsilyl)acetamide, etc.),
with an aldehyde compound (e.g. acetaldehyde, isopentaldehyde,
benzaldehyde, salicylaldehyde, phenylacetaldehyde, p-nitro-
benzaldehyde, m-chlorobenzaldehyde, p-chlorobenzaldehyde,
hydroxynaphthoaldehyde, furfural, thiophenecarboaldehyde, etc.,
or the corresponding hydrate, acetal, hemiacetal or enolate thereof),
with a ketone compound (e.g. acetone, methyl ethyl ketone, methyl
isobutyl ketone, acetylacetone,ethyl acetoacetate, etc., or
the corresponding ketal, hemiketal or enolate thereof), with
- 18 -
13~0'~'~3
phosphorus compound (e.g. phosphorus oxycl-loride,
phosphorous chloride, etc.), or with a sulfur compound
(e.~. thionyl chloride, etc.), and the like.
Suitable salt o~ t]lC compound (II) may be
referred to the on~ as exemplified for the compound (I).
Suitable reactive derivative at thc carboxy
groùp of the corlpoulld (III~ may include, for example, an
acid halide, an acid anhydride, an activated amide,
an activated ester, and the like, and preferably acid
halide sucll as acid cllloride, acid bromidc; a mixed acid
anhydride with an acid such as substituted phosp}loric
acid (e.g. dialkylphosplloric acid, phenylp]losphoric
acid, diphenylphosphoric acid, dibenzylphosphoric acid,
halogenated phosphoric acid, etc.), dialkylphosphorous
acid, sulfurous acid, thiosulfuric acid, sulfuric acid,
alkylcarbonic acid, aliphatic carboxylic acid (e.g.
pivalic acid, pentanoic acid, isopentanoic acid, 2-
ethylbutyric acid, trichloroacetic acid, etc.), aromatic
carboxylic acid (e.g. benzoic acid, etc.); a symmetrical
acid anhydride; an activated acid amide with imidazole,
4-substituted imidazole, dimethylpyrazole, triazole or
tetrazole; an activated ester (e.g. cyanomethyl ester,
methoxymethyl ester, dimethylarlinollletllyl ester, vinyl
ester, propargyl ester, p-nitrophenyl ester, Z,4-
dinitrophenyl ester, trichlorophenyl ester, pentachlorophenyl
ester, mesylphenyl ester, phenylazophellyl ester, phenyl
thioester, p-nitrophellyl thioester, p-cresyl thioester,
carboxymethyl thioester, pyranyl ester, pyridyl ester,
piperidyl ester, 8-quinolyl thioester, an ester with a
N-hydroxy compound such as N,N-dimethylhydroxylamine,
- 19 -
.. ~.. ~. . ...
-
13~0 199
, - .
l-hydroxy-2-(111)-pyridone, N-hydroxysucci~ ide, N-
hydroxyphthalimide, 1-hydroxybenzotriazole, 1-
hydroxy-6-chlorobenzotriazole, etc.), and the like.
The suitablc reactive derivativesof the compoullds
(II) and (III) can be optionally selected from the above
according to the kind of the compounds (II) and (III~ to be
used practically, and to the reaction conditions.
Suitable salt of the compound (III) may includc
a salt with an inorganic base such as alkali metal salt
(e.g. sodium salt, potassium salt,etc.) and an alkaline
earth metal salt (e.g. calcium salt, magnesium salt, etc.),
a salt Wit}l an organic base such as tertiar~ amine (e.g
trimethylamine salt, triethylamine salt, N,N-dimethylaniline
salt, pyridine salt, etc.) , a salt with an inorganic acid
(e.g hydrochloride, hydrobromide, etc.) and the like.
The reaction is usually carried out in a
conventional solvent such as water, acetone, dioxane,
acetonitrile, chloroform, benzene, methylene chloride,
ethylene chloride, tetrahydrofuran, ethyl acetate, N,N~
dimethylformamide, pyridine or any other solvent whlcll does
not adversely influence to the reaction, or an optional
mixture thereof.
When the acylating agent (III) is used in a
form of free acid or salt in this reaction, the reaction
is preferably carried out in the presence of a condensing
agent such as a carbodiimide compound (e.g. N,N'-dicyclo-
hexylcarbodiimide, N-cyclohexyl-N'-morpholinoethylcarbodiimide,
N-cyclohexyl-N'-(4-diethylaminocyclohexyl)carbodiimide,
N,N'-diethylcarbodiimide, N,N'-diisopropylcarbodiimide,
N-ethyl-N'-~3-dimetllylamillopropyl)carbodiimide, etc.),
- 20 -
13~0~
a bisimidazolide compound (e.g. N,N'-carbonylbis(2-
metl-ylimidazole), etc.), an imine compound (e.g.
pentamcthyleneketene-N-cyclohexylimine, diphenylketene-N-
cyclohexylimine, etc.), an olefinic or acetylenic ether
compound (e.g. ethoxyacctylclle, ~-chlorovillyletllyl ether,
e~c.), 1-(4-chlorobenzenesulfonyloxy)-6-cllloro-lH-
benzotriazole, N-ethylbenzisoxazolium salt, N-ethyl-5-
phenylisoxazolium-3'-sulfonate, a phosphorus compound
(e.g. polyphosplloric acid, trialkyl phosphite, ethyl
polyphosphate, isopropyl polyphosphate, phosphorus
oxychloride, phosphorus trichloride, dietllylclllorophosphite,
orthopllellylene chlorophosphite, etc.), thionyl chloride,
oxalyl chloride, ~ilsmeier rcagent prepared by the
reaction of dimethylformamide with thionyl chloride,
phosphorus oxychloride, phosgelle or the like.
With regard to the geometry of the compound (I)
wherein A is a group of the formula: -1 [ hereinafter
referred to as "oxy.~,ino compound" ~] S 2
0-R
produced by this process, it is to be noted that there
seems to be stereoselectivity between syn and anti isomers,
as explained as follows.
In case that the reaction is conducted by
reacting a compound (II) or its reactive derivative at
the amino group or a salt thereof with a compound (III)
- wherein A is a group of the formula: -~- [ hereinafter
referred to as "oximino acylating ~ 2
0-R
agent (III) ] in the prescnce of a condensing agent, for
example, phosphorus pentachloride, thionyl chloride, etc.,
an anti isomer of the oximino compound (I) tends to be
produced as the dominant product and the corresponding
- 21 -
. . .
1 3 4 0 ~f~9
.~ .
syn isomer thereof can be hardly isolated ~rom the
reaction product even when a syn isomer of the oximino
acylating agent (III) is used. It may be understood that
~ ~,
the tendency of such a isomerizatioll in the reaction
conducted by the method as explained above is due to the
fact that the less stable syn isomer tends to isomerize
partially or wholly to the corresponding more stable
anti isomer in the course of such reactio~, for example,
in so-called activation step of the oximino acylating
agent (III) so that more stable isomer, i.e. the anti
isomer of the oximino compound (I) may be isolated as the
reaction product.
Accordingly, in order to obtain a syn isomer
of the oximino compound (I) selectively and in high yield,
it is pre~erable to use a syn-isomer of the oximino
acylating agent (III), and to conduct the reaction under
a selected reaction condition. That is, a syn isomer of
the oximino compound (I) can be obtained selectively and
in high yield by conducting the reaction of a compound (II)
with a syn isomer of the oximino acylating agent (III),
for example, in the presence of a Vilsmeier reagent as
mentioned above and under around neutral condition
The object compound (I) and salt thereof are
useful as an antimicrobial agent, and a part thereof can
be also used as a starting material in the following
processes.
.~
Process B : C-Nitrosation
An object compound (Ib) and its salt can be
prepared by reacting a compound (IV) or its salt with a
~340~'33
nitrosating agent.
The starting compound (IV) corresponds to the
3-cephem compoulld (I) whcrein Rl is haloacetyl, R4 is
hydrogen, halogen, lower al~yl or a group of the formula:
-o-R7 in which R7 is lower alkyl and A is methylelle, and
can be prepared by the above Process A, preferably by
reacting a compound (II) with diketene and halogen (e.g.
chlorine, bromine, etc.). Thus prepared starting compound
(IV) can be used in this process without any isolation
10 and/or purification~
Suitable nitrosating agent may include nitrous
acid and its conventional derivatives such as nitrosyl
halide (e.g. nitrosyl chloride, nitrosyl bromide, etc.),
alkali metal nitrite (e.g. sodium nitrite, potassium
nitrite, etc.), alkyl nitrite (e.g. butyl nitrite,
pentyl nitrite, etc.) and the like.
In case that a salt of nitrous acid is used as
a nitrosating agent, the reaction is preferably carried
out in the presence of an acid such as an inorganic or
organic acid (e.g. hydrochloric acid, sulfuric acid,
formic acid, acetic acid, etc.). And also, in case that
an ester of nitrous acid is used, the reaction is pre-
ferably carried out in the presence of a strong base such
as alkali metal alkoxide or the like.
This reaction is usually conducted in a solvent
StlCh as water, acetic acid, benzene, methanol, ethanol,
tetrahydrofuran or any other solvent which does not
adversely influence the reaction. The reaction temperature
is not critical and the reaction is preferably conducted
within the ran~e of cooling to an ambient temperature.
- 23 -
-
~ 3 4 0 ~ .~ '3
Thus prepared compoun(l (Ib) and salt ~hereof
can be used as a starting matcrial in the following
Proces6es C and D.
Process C : Ethcrification
An object compound (Ic) and its salt can be
prepared by reacting a compound (V) or its salt with an
etherifying agent.
The starting compound (V) corrcsponds to the
10 compound (I) wherein A is hydroxyiminolllethylene group,
and can be prepared by the above Process A and B and
also by the following Process D.
Suitable examples of the etherifying agent may
include a conventional alky]ating agent such as dialkyl
sulfate (e.g. dimethyl sulfate, diethyl sulfate, etc.),
diazoalkane (e.g. diazomethane, diazoethane, etc-.), alkyl
halide (e.g. methyl iodide, ethyl iodide, ethyl bromide,
etc.), alkyl sulfonate (e.g methyl tosylate, etc ),
the corresponding alkenylating-, alkynylating- or
20 cycloalkylating agent, in which the aliphatic hydrocarbon
moiety may be substituted with halogen,carboxy or
esterified carboxy, for example, alkenyl halide (e.g.
allyl iodide, etc.), alkynyl halide (e.g. propargyl bromide,
etc.), cycloalkyl halide (e.g. cyclohexyl bromide, etc.),
lower alkoxycarbonylalkyl halide (e.g. ethoxycarbonylmethyl
iodide, etc.)and the like.
In case of using diazoalkane as an etherifying
agent, the reaction is usually conducted in a solvent
such as diethyl ether, dioxane or any other solvent which
30 does not adversely influence tlle reaction,at a temperature
- 24 -
. - . ~"; - - ~ -
i3~049~
... .
within a range of cooling to an ambient temperature.
In case of using tlle other et}lerifyillg agent,
the reaction is usually conductcd in a solvent such as
water, acetone, ethanol, diethyl ether, dimethylformamide
or any other solvent which does not adversely influence
the reaction within a temperature range of cooling to
heatin~, preferably in the presence of a base such as
an inorganic or organic base, suitable examples of which
are referred to the ones used for the basic hydrolysis
in the Process ~ as illustrated below.
Some of the object compound (Ic) and salt
thereof are uscful as an antimicrobial agent, and some
of them, especially the compound where Rl is haloacetyl
can be used as a starting material in the ~ollowing Process
D.
This process is an alternative one for preparing
~ the compound (Ic) where R is haloacetyl group, and
further this process is particularly preferable and
advantageous for preparing the compound (Ic) where R is
haloacetyl and Ra is substituted- or unsubstituted-lower
alkyl, lower alkenyl or lower alkynyl, more preferably
lower alkyl.
Process D : Thiazole ring formation
A compound ~Id) and its salt can be prepared
by reacting a compound (~'J) or its salt with a thiourea
compound (VII).
The starting compound (VI) corresponds.to the
3-cephem compound (I) wherein Rl is haloacetyl, R4 is
hydrogen, halogen, lower alkyl or a group of the formula:-0-R7
:~ - 25 -
13~0~9~
in which R7 is lower alkyl and A is a group of the
formula : -C- in which R2 is as defined above, and can be
N
o-R2
prepared by the above Process(es) A, B and/or C.
The reaction is usually conducted in a solvent
such as water, alcohol (e.g. methanol, ethanol, etc.),
ben~ene, dimethylformamide, tetrahydrofurall or any other
solvent which does not adversely influence the reaction
within a temperature range of an ambient temyerature to
heating.
This process is an alternative and highly
advantageous one for providing the active compound (Id),
especially (a) the conlpound (Id) wherein R2 is hydrogen
and R6 is amino from the compound (IV) via the Proce.ss B,
and (b) the compound (Id) wherein R2 is lower alkyl and
R6 is amino from the compound (IV) via the Processes s and C.
Process E : Elimination of amino-protective group
A compound (Ie) and its salt can be prepared
by subjecting a compound (VIII) or its salt to elimination
reaction of the protective group in the protected amino
group for Ra.
The starting compound (VIII) corresponds to the
compound (I) wherein Rl is ~hiazolyl of the formula :
a-~ ~ in which R6a is protected amino and A is a group
of the formula : -~- in whicll R2 is as defined above, and
can be.prepared N 2 . for example, by the above Process A.
The elimination reaction may be conducted in
accordance with a conventional method SUC}I as hydrolysis,
reduction or the like. These methods may be selected
- 26 -
- ; ~ - , .,
.
_ _
- 13~0~93
according to the kind of the protective group to be
eliminated.
The hydrolysis may include a method using an
acid ~acidic hydrolysis), a base (basic hydrolysis) or
hydrazine, and the like.
Among these met}lods, hydrolysis using an
acid is one of the common and preferable methods for
eliminating the protective group such as an acyl group,
for example, substituted or unsubstituted lower alkanoyl,
substituted or unsubstituted lower alkoxycarbonyl,
substituted or unsubstituted ar~lower)alkoxycarbonyl, lower
cycloalkoxycarbonyl, substituted phellyltllio, substituted
alkylidene, substitu~ed aralkylidene, substituted cyclo-
alkylidene or the like, particulars of which are to be
referred to those as illustrated for thc N-protective group,
respectively.
Suitable acid to be used in this acidic hydrolysis
may include an organic or inorganic acid such as formic
acid, trifluoroacetic acid, benzenesulfonic acid, p-
toluenesulfonic acid, hydrochlolic acid, cation-exchange
resin, and the like. Preferable acid is the one which can
be easily separated out from the reaction product by a
conventional manner such as neutralization or distillation
under reduced pressure, for example, formic acid,
trifluoroacetic acid, hydrochloric acid or the like. The
acid suitable for the reaction can be selected in con-
sideration of the chemical property of the starting cor.lpound
and the product as well as the kind of the protective
group to be eliminated. The acidic hydrolysis can be
conducted in the presence or absence of a solvent.
- 27 -
, . . .~
1340~3
.i . ,
Suitable solvent may be a conventional organic solvent,
water or a mixture thereof, wllich does not adversely
infl~ence this reaction. Particularly, when the
hydrolysis is conducted Wit]l trifluoroacetic acid,
the reaction may be accelerated by addition of anisole.
lhe hydrolysis using a base can be applied
for eliminating the protective group such as an acyl
group, preferably, for example, haloalkanoyl (e.g.
trifluoroacetyl, etc.) and tlle like. Suitable base may
include, for example, an inorganic base such as alkali
metal hydroxide (e.g. sodium hydroxide, yotassium
hydroxide, etc.), alkaline eartll metal hydroxide (e.g.
magnesium hydroxide, calcium hydroxide, etc.), alkali
metal carbonate (e.g. sodium carbonate, potassium
carbonate, etc.), alkaline earth metal carbonate (e.g.
magnesium carbonate, calcium carbonate, etc.), alkali
metal bicarbonate (e.g. sodium bicarbonate, potassium
bicarbonate, etc.), alkaline earth metal phosphate (e.g.
magnesium phosphate, calcium phosphate, etc.), alkali
metal hydrogen phosphate (e.g. disodium hydrogen
phosphate, dipotassium hydrogen phosphate, etc.), or the
like, and an organic basc such as alkali metal acetate
(e.g. sodium acetate, potassium acetate, etc.),
trialkylamine (e.g. trimethylamine, triethylalllille, etc.),
picoline, N-methylpyrrolidine, N-methylmorplloline, 1,5-
diazabicyclo[4,3,0]-5-nonene, 1,4-diazabicyclo[2,2,2]oc~ane,
1,5-diazabicyclo[5,4,0]-7-undecene,anion-exchange resin
; or the like. The hydrolysis using a base is often carried
out in water or a conventional organic solvent or a
mixture thereof.
- 28 -
.
13~0 1~9
The hydrolysis using hydrazine can be applied
for eliminating the protective group such as dibasic
acyl, for example, succinyl, phthaloyl or the like.
The reduction can be applicd for eliminating
the protective group such as acyl, for example,
halotlower)alkoxycarbonyl (e.g. trichloroethoxycarbonyl,
etc.), substituted or unsubstituted ar(lower)alkoxycarbonyl
(e.g. benzyloxycarbonyl, p-nitrobenzyloxycarbonyl, etc.),
- 2-pyridylmethoxycarbonyl, etc., aralkyl (e.g. benzyl,
benzhydryl, trityl, etc.) and the like. Suitable reduction
may include, for example, reduction using an alkali metal
borohydride (e.g. sodium borohydride, etc.), conventional
catalytic hydrogenolysis and the like.
And further, the protective group such as
halo(lower)alkoxycarbonyl or 8-quinolyloxycarbollyl can be
eliminated by treatment with a heavy metal such as copper,
zinc or the like.
The reaction temperature is not critical and
may be optionally selected in consideration of the
chemical property of the starting compound and reaction
product as well as the kind of the N-protective group
and the method to be applied, and the reaction is preferably
carried out under a mild condition such as under cooling,
at ambient temperature or sligh~ly elevated temperature.
The process includes in its scope the cases
- that the functionally modified carboxy for R5 is
simultaneously transformed into the free carboxy group
in the course of the above reaction or in the post-treatment
As to this process, it is to be understood that
the purpose of this process lies in providing the
z9
: . .
.......
13~0~9 ~
generally more active comyound (I') wherein Ra is amino-
thiazolyl by eliminating the protective group in the
protected amino group of the compound (VIII) prepared
by the other processes as mentioned above or below.
Process ~ : Reductive formation of 3-hydroxycepham
A compound (If) and its salt can be prepared
by reducing a compound (IX) or its salt.
The starting compound ~IX) corresponds to the
3-cephem compound (I) wherein p~l is thiadiazolyl or
thiazolyl of the formula : R ~ ~ in whicll R6 is as
defined above, R3 is hydrogen, R4 is a group o~ the
formula:-0-R7 in which R7 is hydrogen and A is a group
of the formula : -~- in which R2 is as defined above,
and can be N 2 prepared, for example, by the
above Process A.
The method of reduction applied to this process
may include a conventional one wllicll is applicable for
reduction of lcetonic carbonyl group including its
tautomeric enol form into hydroxymethylene group, and
the preferable method may be reduction usin~ an alkali
metal borohydride (e.g. sodium borohydride, etc.) or a
combination of an acid (e.g. hydrochloric acid, sulfuric
acid, formic acid, acetic acid, etc.) and a metal (e.g.
zinc, iron, copper, etc.), catalytic reduction using
a conventional catalyst (e.g. palladium on carbon,
palladium sponge, Raney nickel, platinum, platinum black,
etc.) or the like.
The reaction is usually carried out in a
conventional solvent such as water, alcohol (e.g.
- 30 -
; . .. - .
~ 13~0~9~
methanol, ethanol, etc.), dimetllylformamide, tetrahydro-
furan or any other solvent W]liCh does not adversely
influence the reaction within a temperature range from
cooling to somewhat elevated temperature.
Although thus prepared compound (If) and salt
thereof have antimicrobial activities, they are also
useful mainly as an intermediate, especially as a
starting material in the following Process G and suc-
cessively Process H for preparing the more active 3-
cephem compound (Ih).
Process G : 0-acylation
A compound (Ig) and its salt can be prepared by
reacting a compound (If) or its salt with a compound (X),
its salt or its reactive derivative.
As to the compound (X), sui~able eXamples of
the acyl moiety for Ra are to be referred to those as
exemplified above for the acyl group for R of the
compound (I).
The reactive derivative of the compound (X)
may be an acyl halide, anhydride, azide, activa~ed ester,
activated amide and the like, which are to be referred to
those as exemplified above for thc compound (III) in the
Process A, preferably an acyl halide such as lower
alkanoyl halide (e.g. acetyl chloride, etc.), aroyl
halide (e.g. benzoyl chloride, etc.), lower alkanesulfonyl
halide (e.g. mesyl chloride, mesyl bromide, ethanesulfonyl
chloride, etc.), arenesulfonyl halide te.g. tosyl chloride,
etc.), an acyl azide such as lower alkanesulfonyl azide
(e.g. mesyl azide, etc.), arenesulfonyl azide ~e.g.
... .. ~ .
'~ 13~0~9~
tosyl azide, etc.) or the like, and more preferably
lower alkanesulfonyl halide or arenesulfonyl halide.
The reaction is usually carried out in a
conventional solvent such as dimethylforlllalllide,
chloroforln, methylene chloride or any other solvent
- which does not adversely influence the reaction, under
cooling or at an ambient or somewhat elevated temperature.
In case that the acyl halide is used as an
acylating agent, the reaction is generally conducted
in the presence of a base as exemplified in the above
Process E.
This process is the first activation step for
preparing a more active 3-ccphem compound (Ih) from the
3-hydroxycepham compound (If) via the 3-acyloxycepham
compound (Ig), which is successively t~reated with a base -
in the following Process 11.
Process H : 3-Cephem formation
This process is the final step to transform
the 3-hydroxycephem compound (IX) into the more active
3-cephem compound (Ih) or its salt. That is, a compound
(Ih) or its salt can be prepared by treating a compound
(Ig) as prepared in the above Process G or its salt
with a base.
The preferable base includes an inorganic
base such as metal hydroxide (e.g. sodium hydroxide,
potassium hydroxide, etc.), metal carbonate (e.g. sodium
carbonate, potassium carbonate, magnesium carbonate, etc.),
metal bicarbonate (e.g. sodium bicarbonate, potassium
bicarbonate, etc.), organic base such as tertiary amine
- 3Z -
- - . . ... . . . . .
- 13~0~9~
.~ .
~e.g. trimethyl amine, triethyl amine, pyridine, etc.)
alkali metal alkoxide ~e.g. sodiw~l methoxide, sodiw
ethoxide, etc.) and the like.
The reaction is usually carried out in a
conventional solvent such as an alcohol, dimethylformamide,
chloroform, methylenc chloride or any other solvent which
does not adversely influence the reaction, under cooling
or at an ambient or somewhat elevated temperature.
Process I : Halogenation
A compound (Ii) or its salt can be prepared
by halogenating a compound (XI) or its salt.
The starting compound (XI) corresponds to the
compound (I) wherein Rl is thiadiazolyl or thiazolyl of
the formula : R ~ ~ _ in which R6 is as defined above,
R3 is hydrogen, S R4 is a group of the formula:-0-R7
in which R7 is hydrogen and A is a group of the formula:
-~- in which R2 is as defined above, and can be prepared
- ~ 2 by the processes as explained above.
Suitable halogenating agent may include a
conventional halogen compound such as phosphorus halide
(e.g. phosphorus trichloride, phosphorus pentachloride,
phosphorus tribromide, phosphorus pentabromide, phosphoryl
chloride, etc.), thionyl chloride and the like.
The reaction is usually carried out in a con-
ventional solvent such as chloroform, methylene chloride,
dimethylformamide or any other solvent which does not
adversely influence the reaction and preferably under
cooling or at ambient or somewhat elevatcd temperature.
- 33 -
1340~9~
Process J : Esterification
This process is to provide an ester compound
(Ij) and its salt for improving the chemical, phisiological
and/or pharmaceutical properties of the corresponding
free carboxy compound (XII), W]liCh corresponds to the
3-cephem compound (I) wherein R5 is carboxy, or its salt.
The esterification is conducted by reacting a
free carboxy compound (XII), its reactive derivative at
the carboxy or a salt thereof with an esterifying agent.
The preferred reactive derivative at the carboxy
group of the compound (XII) is to be referrcd to those
of the compound (III) as exemplified in the Process A.
The esterifying agent may include a llydroxy
compound and its reaction e~uivalent.
Suitable examples of the hydroxy compoulld may be
a substituted or unsubstituted alcohol such as alkanol,
aralkanol, arenol or the like, particulars of which may
be substituted alcohol SUC]I as
alkanoyloxy(lower)alkanol (e.g. acetoxymethanol,
propionyloxymethanol, butyryloxymethanol, pentanoyloxy-
methanol, hexanoyloxymethanol, acetoxyethanol, propionyl-
oxyethanol, butyryloxyethanol, pentanoyloxye~hanol,
hexanoyloxyetllanol, acetoxypropanol, propionyloxypropanol,
hexanoyloxypropanol, hexanoyloxyhexanol, palmitoyloxymethanol,
etc.), halo(lower)alkanol (e.g. mono-, di- or tri-
chloroethanol, etc.), lower cycloalkyl(lower)alkanol (e.g.
l-cyclopropylethanol, etc.), substituted ar(lower)alkanol
(e.g. 4-nitrobenzyl alcohol, 4-chlorobenzyl alcohol, 4-
methoxybenzyl alcohol, 3,5-di-tert-butyl-4-hydroxybenzyl
alcohol, bis(methoxyphenyl)methanol, etc.), substituted
- 34 -
1 3 ~
-
arenol (e.g. 4-methoxyphenol, etc.) , the corresponding
unsubstitutcd alcohol or the like.
Suitable reactive equivalent of the hydroxy
compound may include a conventional one such as halide,
alkanesulfonate, arencsulfollatc or salt of the hydroxy
compound, diazoalkane, diazoaralkane, and the like.
Preferable halide of the hydroxy compound
may be chloride, bromide or iodide.
Preferable alkane- or arene-sulfonate of the
hydroxy compound may be methallcsulfollate, ethanesulfonate,
benzenesulfonate, tosylate or the like.
Preferable salt of the hydroxy compound may be
an alkali metal salt such as lithium salt, sodium salt,
potassium salt or the like.
Preferable diazoalkane and diazoaralkane may
be diazomethane, diazoethane, diazoyropane,
diphenyldiazomethane or the like.
The reaction can be carried out in the presence
or absence of a solvent such as N,N-dimethylformamide,
- 20 dimethylsulfoxide or any other solvent which does not
adversely influence tlle reaction, and within a
temperature range of cooling to heating. The liquid
hydroxy compound can be also used as a solvent in this
reaction.
This reaction can be preferably conducted in
the presence of an inorganic or organic base as exemplified
in t]le above Process E.
In case of preparing a substituted- or
unsubstituted-aryl ester (Ij), particularly substituted-
or unsubstituted- phenyl ester, this reaction is to be
~. ,.
~ ~34~9~
conducted by reacting a~ a compound (XII) or its salt
with phenol or its salt in the presencc of a condensing
agent as exemplified in the above Process A, or b) a
reactive derivative of the compound (XII) preferably
a mixed acid anhydridc of the compound (XII) with phenol
or its salt in the presence of a base.
In case that a compound (XII), where A is a
group of the formula: -~- in which R2 is an aliphatic
hydrocarbon residue ~ 2 substituted with carboxy,
0-R
is used as a starting material in this reaction, the said
carboxy group may be also esterified in accordance with
the reagent and the reaction conditions, and this mode
of the reaction is included within the scope of this
process.
And further, in case that the 2-cephem compound
corresponding to the compound (Ij) is produced, the said
2-cephem compound can be transfornied into the 3-cephem
compound (Ij) by oxydizing and then reducing the
resultant S-oxide compound in a conventional manner.
This mode of the reactions is also included within the
scope of this process.
Process K : Carboxy formation
This process is to provide a free carboxy
compound (I~) or its salt, especially the compound (Ik)
wherein Rl is thiadiazolyl or thiazolyl of the formula~
R ~ ~ in which R6 is as defined above and A is a group
of the formula:-~- in which R is as defined above, which
generally N 2 exhibits higher antimicrobial
0-R
activities as compared with the corresponding functionally
- 36 -
... .
13~0~93
~ modified carboxy compound (XIII).
Accordingly, the meaning of thc functionally
modified carboxy in the compound (XIII) iies in mainly
synthetic manufacture by chemical process(es) as
illustrated hereinabove.
This process is conducted by transforming the
functionally modified carboxy group of the starting
compound (XIII) into free carboxy group, and the preferred
functionally modified ca.rboxy for Rb in the compound (XIII)
may be an esterified carboxy group as exemplified for R5
of the compound (I).
The method to be applied to this process includes
~ conventional ones such as hydrolysis, rcduction and the
like.
The method of hydrolysis includes a con.vcntional
one using an acid, base, enzyme or enzymatic preparation,
and the like.
Suitable examples of the acid and base are
to be referred to those as exemplified in the above
~ 20 Process E, and the acidic or basic hydrolysis can be
carried out in a similar manner to that of the Process E.
Suitable enzyme includes an esterase and
~ esterase preparation which exhibits an esterase activity
such as a cultured broth of microorganism or processed
materials of microorganism, the preparation of animal or
plant tissues, or the like, and preferably a cultured
broth of microorganism or processed material thereof.
; An esterase to be used in the enzymatic
: hydrolysis may be used not only in a purified state, but
also in a crude state.
~' ' 1340~ 9
Such an esterase is frequcntly found to exist
widely, for example, in various kind o~ microorganisms,
which can be easily isolated from a soil sample and
other sources by conventional means, and further can be
easily selected from the collected cultures available
in public facilities for culture collection such as ATCC
(American Type Culture Collection, Maryland, USA), IAM
(Institute of Applied Microbiology, University of Tokyo,
Japan), IFO (Institute For Ferlllentation, Osaka, Japan),
IID (The Institute for Infectious Diseases, University
of Tokyo, Tokyo, Japan), CBS (Centraalbureau voor
Schimmelcultures, Bearn, Netherlands), FEI~M (Fermentation
Research Insti~ute, Agency of Industrial Science and
Technology, Chiba, Japan) and NRRL (Northerll Utilization
Research and Development Division, U.S. Depar~ment of
Agriculture, Illinois, U.S.A.) and the like.
As to the microorganism having an esterase
activity, there may be exemplified one belonging to the
genus, Bacillus, Corynebacterium, Micrococcus,
Flavovacterium, Salmonella, Staphylococcus, Vibrio,
Microbacterium, Escherichia, Arthrobacter, Azotobacter,
Alcaligenes, Rhizobium, Brevibacterium, Kluyvera, Proteus,
Sarcina, Pseudomonas, Xanthomollas, Protaminobacter,
Comamonus and the like.
Examples of the above microorganisms may be
Bacillus subtilis IAM-lOG9, lAM-1107, lAM-1214, Bacillus
sphaericus IAM-1286, Corynebacterium equi IAM-1308,
Micrococcus varians IAM-1314, Flavobacterium rigeus
IAM-123~, Salmonella typhimurium IA~1-1406, Staphylococcus
epidermidis IAM-1296, Microbacterium f~avum IAM-1642,
- 38 -
.. ... . . .. .. . .
- '' 13~0~99
Alcaligenes faecalis ATCC-8750, Arthrobacter simplex
ATCC-6946, Azotobacter vinelandii I~M-1078, Escherichia
coli IA~I-llOl, Rhizobium ~aponicum IAM-0001, Vibrio
metchnikovii IAM-1039, Brevibacteriwn helvolum IAM-1637,
Protaminobacter alboflavum IA~1-1040, Comamonas terrigena
I~0-12685, Sarcina lutea IAM-1099, Pseudomonus
schuylkilliensis IA~1-1055, Xanthomonas trifolii ATCC-12287
or the like.
In the enzymatic hydrolysis, the esterase can
be preferably used in a form of a cultured broth obtained
by culturing microorganisms having an esterase activity
in a suitable manner, or of its processed material
Cultivation of microorganisms can be generally
conducted in a conventional manner. As a culture medium
to be used, there may be used a nutrient one containing
sources of assimilable carbon and nitrogen and inorganic
salts. The preferred sources of carbon are, for example,
glucose, sucrose, lactose, sugars, glycerol and starch.
The preferred sources of nitrogen are, for example, meat
extract, peptone, gluten meal, corn meal, cotton-seed meal,
soybean meal, corn steep liquor, yeast extracts, casein
hydrolysate and amino acids, as ~ell as inorganic and
organic nitrogen such as ammoniWIl salts (e.g. anunoniwn
sulfate, ammonium nitrate, ammonium phosphate, etc.),
sodium nitrate or the like. If desired, mineral salts
such as calcium carbonate, sodium or potassium phosphate,
magnesium salts and copper salts, and various vitamines
can be also used.
Suitable pH of the culture medium, suitable
cultivation temperature and suitable cultivation time
- 39 -
~ . . .... _ .
134 0~9 3
vary with the kind of the microorganisms to be used.
A desirable pH usually lies in a range of pH 5 to 8.
The temperature is usually selected from about 20~C
to about 35~C. The cultivation time is usually selected
from 20 hours to 120 hours.
The cultured broth per se thus obtained and its
processed material may be employed for enzymatic hydrolysis
of this process. The "processed material" of cultured
broth means any preparation having esterase activity, which
is processed by conventionally suitable means for increas-
ing said esterase activity.
The esterasc activity of the cultured broth
is present in cells (intracellularly) and/or out of cells
(extracellularly).
When the activity exists mainly in cells, the
following preparation, for example, may be used as a
processed material of the cultured brotll.
(1) raw cells, separated from the cultured broth in
conventional malmers such as filtration and centrifugation,
(23 dried cells; obtained by drying said raw cells in
conventional manners such as lyophilization and vacuum
drying,
(3) a cell-free extract; obtained by destroying said raw
or dried cells in conventional manners (e.g. grinding the
cells with almina, sea sand, etc. or treating the cells
with super sonic waves), or
(4) an enzyme solution; obtained by purification or partial
purification of said cell-free extract in a conventional
manner.
When the activi~y exists mainly out of cells,
- 40 -
.
,
1340~ 3
t]le following preparation, for exaDIllle~ may be used as a
processed material.
(1) a supernatant or a filtrate; obtained from the cultured
broth in a conventional manner, or
(2) an enzyme solution; obtained by purification or partial
purification of said supernatant or filtrate in a conven-
tional manner.
The enzymatic hydrolysis is conducted by
contacting the compound (XIII) l~ith the cultured broth
of the microorganism or its processed material in an
aqueous medium such as water or a buffer solution te.g.
phosphate buffer, etc.), preferably in the presence of
conventional surface-active agent. That is, the reaction
is usually conducted by adding the compound (XIII) to the
cultured broth of the microorganism or its liquid processed
material (e.g. supernatant, filtrate, enzyme solution,
etc.), or to the solution or suspension of the cultured
broth or its processed material in an aqueous medium.
Sometimes, an agitation of the said reaction mixture is
preferable.
Preferred pH of reaction mixture, concentration
of substrates, reaction time and reaction temperature may
vary with characteristics of the cultured broth or its
processed material to be used, or the compound (XIII) to
be used. However, the reaction conditions are preferably
selected from a range of at pH 4 to 10, more preferably
at pH 6 to 8, at 20 to 50~C, more preferably at 25 to 35~C
for 1 to 100 hours. The concentration of the starting
compound (XIII~ to be used as a substrate itl the reaction
mixture may be in a range of 0.1 to 100 mg per mQ,
," 13~0'~!3'3
preferably 1 to 20 mg per mQ.
The method of tle reduction for this process may
be c~rried out in a similar manner to that of the above
Process E.
This process includes within its scope the cases
- that the protective group in the protected amino for R6,
which is a substituent on the thiazolyl group for Rl, is
eliminated and/or the esterified carboxy group, which is
an optional substituent on the aliphatic hydrocarbon
residue for R2 in the group A, is transformed into free
carboxy group in the course of tlle reaction or the post-
treatment.
The compound obtained in accordance with the
processes as explained above can be isolated and purified
in a conventional manner.
In case that the object compound (I) has free
carboxy for RS and/or free amino for R6, it may be trans-
forlned into its pharmaceutically acceptable salt by a
conventional method,
Among the object compound (I), the compound (I'),
its pharmaceutically acceptable salt and bioprecursor
thereof exhibit high antimicrobial activities inhibiting the
growth of a wide variety of pathogenic microorganisms
including Gram-positive and Gram-negative bacteria and are
useful as antimicrobial agents.
And further, the compound (I") and its salt are
novel and useful as an intermediate for preparing the active
compound (I'), its pharmaceutically acceptable salt or
- 42 -
13~0~93
bioprecursor thercof.
Accordi.ng to the aforementioncd urocesses, more
specifically the following compounds can be prepared.
7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-
3-methoxy-3-cephem-4-carboxylic acid (syn isomer)
7-[2-(2-amino-4-thiazolyl)-2-met]loxyimilloacetamido]-
2,3-dimethyl-3-cephem-4-carboxylic acid (syn isomer)
7-[2-(2-amino-4-tlliazolyl)-2-metlloxyiminoacetamido]-
3-chloro-3-cephem-4-carboxylic acid (anti isomer~
7-[2-(1,2,3-thiadiazol-4-yl)-2-metlloxyiminoacetamido]-
3-cephem-4-carboxylic acid (syn isomer)
7-{2-(2-amino-4-thiazolyl)-2-hydroxyiminoacetamido]-
3-cephem-4-carboxylic acid (syn isomer)
7-[2-(2-amino-4-thiazolyl)-2.-methoYyiminoacetamido]-
3-tosyloxy-3-cephem-4-carboxylic acid (syn isomer)
7-[2-(2-amino-4-thiazo].yl)-2-ethoxyimilloacetamido]-
3-cephem-4-carboxylic acid (syn isomer)
7-[2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetamido]-
3-chloro-3-cephem-4-carboxylic acid (Syll isomer)
7-[2-(2-amino-4-thiazolyl)-2-isopropoxyiminoacetamido]-
3-cephem-4-carboxylic acid (syn isomer)
7-[2-(2-amino-4-thiazolyl)-2-propoxyiminoacetamido]-
3-chloro-3-cephem-4-carboxylic acid (syn isomer)
7-[2-(2-amino-4-thiazolyl)-2-propoxyimilloacetamido]-
3-cephem-4-carboxylic acid (syn isomer)
7-[2-(2-amino-4-thiazolyl)-2-isobutyloxyiminoacet2mido~-
3-cephem-4-carboxylic acid (syn isomer)
7-[2-(2-amino-4-tlliazolyl)-2-n-butoxyimilloacetamido]-
3-cephem-4-carboxylic acid (syn isomer)
- 43 -
13~0~
7-[2-(2-amino-4-thiazolyl)-2-1l-hexyloxyiminoacetamido]-
3-cepheln-4-carboxylic acid (syn isomer)
7-[2-(2-amino-4-t]liazolyl)-2-cyclohexyloxyimino-
acetamido]-3-cephem-4-carboxylic acid (syn isomcr)
7-[2-(2-amino-4-thiazolyl)-2-allyloxyilllinoacctamido]-
3-cephem-4-carboxylic acid (syn isomer)
7-[2-(2-amino-4-thiazolyl.)-2-propar~yloxyimino-
acetamido]-3-cephem-4-carboxylic acid (syn isomer)
7-[2-(2-amino-4-thiazolyl)-2-ethoxycarbonylmethoxy-
iminoacetamido]-3-cephem-4-carboxylic acid (Syll isomer)
7-[2-(2-amino-4-thiazolyl)-2-carboxymethoxyimillo-
acetamido]-3-cephem-4-carboxylic acid (syn isomer)
7-.[2-(2-amino-4-tlliazolyl)-2-n-pentyloxyimino-
acetamido]-3-cephem-4-carboxylic acid (syn isomer)
7-[2-(2-amino-4-thiazolyl)-2,-(2,2,Z-trifluoro-
ethoxyimino)acetamido]-3-cephem-4-carboxylic acid (syn isomer)
7-[2-~2-amino-4-tlliazolyl)-2-(2-cllloroetlloxyimino)-
acetamido]-3-cephem-4-carboxylic acid (syn isomer)
7-[2-(2-amino-4-thiazolyl)-2-(2,2,2-trif~uoroethoxy-
1mino)acetamido]-3-chloro-3-cephem-4-carboxylic acid (Sy
somer)
7-[2-(2-formamido-4-thiazolyl)-2-methoxyimino-
acetamido]-3-cephem-4-carboxylic acid (syn isomer)
7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-
3-cephem-4-carboxylic acid (anti isomer)
7-[2-(2-amino-4-thiazolyl)-2-n-octyloxyimino-
acetamido]-3-cephem-4-carboxylic acid (syn isomer)
7-[2-(2-amino-4-thiazolyl)-2-(2,3,3-trifluoro-2-
propenyloxyimino)acetamido]-3-cephem-4-carboxylic acid (Sy
isomer)
- 44 -
1340~99
7-[2-(2-amino-4-tlliazolyl)-2-lauroyloxylnetlloxy-
iminoacetamido]-3-cepllem-4-carboxylic acid (syn isomer)
7-[2-(1,2,3-thiadiazol-4-yl)-2-n-hexyloxyilllino-
acetamido]-3-cephem-4-carboxylic acid ~syn isomer)
7-[2-(2-amino-4-thiazolyl)-2-1l-butoxyiminoacetamido]-
3-chloro-3-cephem-4-carboxylic acid (syn isomer)
7-[2-(2-amino-4-thiazolyl?-2-n-butoxyimino$cetamido]-
3-methoxy-3-cephem-4-carboxylic acid (syn isomcr)
7-[2-(2-amino-4-thiazolyl)-2-propargyloxyimino-
acetamido]-3-methoxy-3-cephem-4-carboxylic acid (syn isomer)
7-[2-(2-amino-4-tlliazolyl)-2-trifluoronletlloxyilllino-
acetamido]-3-cephem-4-carboxylic acid (syn isomcr)
the corresponding functionally modified derivative
such as
hexanoyloxymethyl 7-[2-(2-an!ino-4-thia7Olyl)-2-
methoxyiminoacetamido]-3-cephcm-4-carboxylate (syn isomer)
pivaloyloxymethyl 7-[2-(2-ami.no-4-thiazolyl)-2-
methoxyiminoacetamido]-3-cephem-4-carboxylate (syn isomer)
4-nitrobenzyl 7-[2-(2-amino-4-thiazolyl)-2-
methoxyiminoacetamido]-3-cephem-4-calboxylate (syn isomer)
4-nitrobcnzyl 7-[2-(2-amino-4-thiazolyl)-2-
methoxyiminoacetamido]-3-hydroxy-3-cephem-4-carboxylate
(syn isomer)
4-nitrobenzyl 7-[2-(2-amino-4-thiazolyl)-2-ethoxy-
iminoacetamido]-3-hydroxy-3-cephem-4-carboxylate (syn isomer)
4-nitrobenzyl 7-[2-(2-amino-4-thiazolyl)-2-n-
'' propoxyiminoacetamido]-3-hydroxy-3-cepllem-4-carboxylate
(syn isomer)
4-nitrobenzyl 7-[2-(2-amino-4-thiazolyl)-2-
isobutoxyiminoacetamido]-3-hydroxy-3-cephem-4-carboxylate
(syn isonler) 45
~ ~ 13~0~g9
the corresponding salt such as
sodium 7-[Z-~2-amino-4-tlliazolyl)-2-methoxyimino-
acetamido]-3-cephem-4-carboxylate (syn isomer)
calcium 7-[Z-(2-amino-4-tlliazolyl)-2-metlloxyimino-
acetamido]-3-celllem-4-carboxylate Isyn isomer)
magnesium 7-[2-(2-amino-4-thiazolyl)-2-methoxy-
iminoacetamido]-3-cephem-4-carboxylate (Syll isomer)
arginille salt of 7-[2-(2-amino-4-thiazolyl)-2
methoxyiminoacetamido]-3-cephem-4-carboxylic acid (syn isomer)
lysine salt of 7-[2-(2-amino-4-thiazolyl)-2-
methoxyiminoacetamido]-3-cephem-4-carboxylic acid (syn isomer)
7-[2-(2-amino-4-thia4Olyl)-2-metlloxyimino-
acetamido]-3-cep}lem-4-carboY.ylic acid hydrochloride (syn
' isomer)
In order to show the utility of the active compound
(I'), the test data of some representative compounds (I')
are shown in the following.
1. In vitro antibacterial activity :
(1) Test method:
In vitro antibacterial activity was determined by
the two-fold agar-plate dilution method as described below.
One loopful of the 100-fold dilution of an
overnight culture of each test strain in Trypticase-soy broth
was streaked on heart infusion agar (HI-agar) containing
graded concentrations of the test compound and incubated at
37~C for 20 hours. The minimal inhibitory concentration
(MIC) was expressed in ~g/ml.
- 46 -
....
. .: ~ . - - ~ - - - -
. .._~ _
13~0~99
t2) Test compoulld5 :
No.
l --- 7-[2-(2-Amino-4-thiazolyl)-2-metlloxyilllinoacetamido]-
3-cephen--4-carboxylic acid (syn i.somcr)
2 --- 7-[2-(2-Amillo-4-thi.azolyl)-2-hydro~yilllilloacetamido]-
3-cephem-4-carboxylic acid (syn isomer)
3 --- 7-[2-(Z-Amino-4-thiazolyl)-2-ethoxyiminoacetamido]-
3-cephem-4-carboxylic acid (syn isomer)
4 --- 7-[2-(2-Amino-4-thiazolyl)-2-methoxyimilloacetamido]-
3-chloro-3-cephem-4-carboxylic acid (syn isomer)
5 --- 7-[2-(2-Amino-4-thi.azolyl)-2-n-propoxyiminoacetamido]-
3-cephem-4-carboxylic acid (syn isomer)
6 --- 7-[2-(2-Amillo-4-thiaznlyl)-2-n-butoxyiminoacetamido]-
3-cephem-4-carboxylic acid (syn isomer)
7 --- 7-[2-(2-Amino-4-thiazolyl)-2-allyloxyiminoacetamido]-
3-cephem-4-carboxylic a.cid (syn isomer)
--- 7-[2-(2-Amino-4-thiazolyl)-2-propargyloxyiminoacetamido]-
3-cephem-4-carboxylic acid (syn isomcr)
9 --- 7-[2-(2-Amino-4-thiazolyl)-2-n-pentyloxyiminoacetamido]-
3-cephem-4-carboxylic acid (syn isomer)
10 --- 7-[2-(2-Amino-4-thiazolyl)-2-n-hexyloxyiminoacetamido]-
3-cephem-4-carboxylic acid (syn isomer)
11 --- 7-[2-(2-Amino-4-thiazolyl)-2-cyclohexyloxyiminoacetamido]-
3-cephem-4-carboxylic acid (syn isomer)
12 --- 7-[2-(2-Amino-4-thiazolyl)-2-(2-chloroethoxyimino)-
acetamido]-3-cephem-4-carboxylic acid (syn isomer)
13 --- 7-[2-(2-Amino-4-thiazolyl)-2-(2,2,2-trifluoroethoxy-
imino)acetamido]-3-cephem-4-carboxylic acid (syn
isomer)
- 47 -
(3) Test results :
MIC(~g/ml)
\ Compound No.
~ 1 2 3 4 5 6 7 8
Test Strains
S tâphyl OCOCCUS
aureus 209P JC-l6.25 0.39 3.13 12.5 1.56 0.78 1.56 1.56
Escherichia <0.025 0.1 0.05 0.39 0.2 0.39 0.2 0.1
coli. NIHJ JC-2
Proteus
vulgaris I~-1025 -0 025 0.1 50.025 <0.025_O.OZ5 0.05 _0.02550.025
Klebsiella ~0.025 <0.025 ~0.025 0.1<0.025 0.2 50.02i<0.025
preunoniae 20
Proteus <0.025 <0.025 <0.025 0.0125 0.1 0.2 <0.025 <0.025
mirabilis 18
Pseudomonus 0 3~ 6.25 <1.56 - 6.25<1.56 <1.56 51.56 <1.56
aeruginosa NCTC-10490
Serratia 1.56 12.5 0.78 50 3.13 6.25 1.56 3.13
marcescens 35 ~'
Compound No.
9 10 11 12 13
Test Strains
Staphylococcus 0 39 1.55 0.39 1.56 1.56
aureus 209P JC-l
Escherichia 3.l3 1.56 3.13 0.1 0.2
coli NIHJ JC-2
Proteus 0.39 0 2 0.78<0.025 <0.025
vulgaris IAM-1025
Klebsiella 0.2 0.05 0.39 0.1 0.05
pneumaniae 20
Proteus 1.56 0.78 1.56 0.2 0.2
mirabilis 18
Pseudomonus 3.13 <1.56 <1.56 <1.56 <1.56
aeruginosa NCTC-10490
Serratia 3.13 12.5 12.5 12.5 6.25 c3marcescens 35 ~
c~
- 13~0~9
2. Protecting cffect against experimental
infectiolls in mice :
(1) Test method
Male ICI~ strain mice agcd ~ wceks, each weighing
18.5 - 21.5 g. were uscd in groups Or lo mice. The test
bacteria were cultured overnigllt at 37~C on Trypticase-soy
agar and then suspended in 5Po mucin to obtain the suspension
corresponding to each challenge cclls. ~lice were inoculated
intraperitoneally with 0.5 ml. of tlle suspension. A solution
- 10 containing each test compoulld~ was givcn subcutaneously to
the mice in various dosage one hour after the challenge.
The ED50 values were calculated from the number of surviving
mice for each dosage after four days of observation.
(2) Test compounds
No.
- 1 --- 7-[2-(2-Amino-4-tlliazolyl)-2-llletlloxyiminoacetamido]-
3-cephem-4-carboxylic acid (syn isomer)
reference --- 7-[2-(2-Amino-4-thiazolyl)-2-methoxyimino-
acetamido]cephalospolanic acid (syn-isomer)
- 50 -
13404~
.
(3) Test results :
ED50(s.c.)(lllg/kg) MIC (~g/ml.)
TestInoculated
Test Con!poullds Inoculum Test Compounds
EacteriaCells/mousc
1 reference Size 1 reference
Lschericllia 7 2.8 0 *1 0.78 3.13
10 2 0.05 0.]
Klebsiella8 x 106 <0.98 0.995 10~ 0.39 3.13
10 2 ,c0.025 0.05
Proteus9.9 x 106 0.39 1.171 10~ 1.56 50
rettgeri 24 1o-2 <0.025 0.1
Lo Serratia1.2 x 107 3.562 3 31.427*3 10~ 25 50
Marcesc~ns 58 1o~2 0 39 1.56
* 1 : overnigllt culture
* 2 : 100-fold dilution of the overnight culture
* 3 : treated with two divisional doses at 1 hr. and
3 hrs. after infectio
3. Acute toxicity :
(1) Test method :
Ten male and 10 female rats aged 6 weeks (JCL-SD
strain) were used per group. Test compound dissolved in
distilled water was given subcutaneously and intraveneously
to the animals. These animals were observed for 7 days after
dosing. The LD50 values were calculated from the number of
dead animals by the Litchfield-Wilcoxon method.
; (2) Test compound :
7-[2-(2-amino-4-thiazolyl)-2-methoxyimilloacetamido]-
- 51 -
.~ .
. _ . ~ ... , . _ . .. .
1 3 4 0 1 9 ~
3-cephem-4-carboxylic acid (syn isomcr)
(3) Test results :
Test Sex LD50 (mg./kg.)
animal s . c . i . v .
~lale >8000about 8000
Rat
Female >8000 >8000
4. Absorbability
(1) ~est method :
Test compound was given orally to a group o~
5 rats (JCL-SD strain, 6-week-old, male) which llad been
~asted. Bile and urine samples were c~llected at 0~6 and
6~24 hrs. The concentrations o~ the .tcst compound in the
samples werc determined by bioassay (disk method) using
Batillus su~tilis ATCC-6633 as test organism, and the
recoveries in bile and urine were calculated.
(2) Test compound :
7-[2-(2-amino-4-thiazolyl)-2-n-pentyloxyimino-
acetamido]-3-cephem-4-carboxylic acid (syn isomer)
(3) Test result :
Total recovery in bile and urine in 24 hrs. was
22.8%.
For prophylactic and/or therapeutic administration,
the activ~ compound (I') of the present invention is used in
- 52 -
. . ..
13~0~93
the form of conventional pharmaceutical preyaration which
contains said compound, as an active ingredient, in admixture
with pllarmaceutically acceptable carriers sucl- as an organic
or inorganic solid or licluid excipient W]liC]I is suitable for
oral, parenteral or external administration. The pharma-
ceutical preparations may be in solid form such as capsule,
tablet, dragee, ointment or suppository, or in liquid form
such as solution, suspension, or emulsioll. If needed,
there may be included in the above preparations auxiliary
substances, stabilizing agellts, wetting or emulsifying
agents, buffers and the other commonly used additives.
While the dosage of the compounds may vary from and
also depend UpOII the age and conditions of the patient, a
kind of disease and a degree of t]le infection, and further
a kind of the active compound (I') to be applied, etc., an
average single dose of about 50 mg., 100 mg., 250 mg. and
500 mg. of the active compound (I') is sufficient for
treating infectious diseases caused by pathogenic bacteria.
In general, the active compound ~I') can be administered in
an amount between 1 mg / kg and 100 mg / kg, preferably
- 5 mg/kg and 50 mg/kg.
The starting compound (III) can be prepared as
illustrated below.
- 53 -
.. .. .. . .
.. , . . .. _
1340~93
X-CH2CO-C-COOZ (IIIa)
N~ S (VIIa)
N I C--COOZ (IIIC)
X-CI-12CO-C-COOZ (IIIb) Ra~S~ N
o-R2 ~ Ra ICI Nllz (YIIa) ~ 011
N~ C-COOZ (IIId)
~~ Ra
N ~~- C-COOZ (lIIe) N ---C-COOH
H2NJ~S~ N RaJ~3 N (IIIf)
o- Ra o- Ra
N- l C-Coo~l (IIIg)
H2NJ~s~ ~ Ra-ONH2 (XIV)
O-Ra
~a~C-COOH (lIlh)
NNH - Y
H3CCO-C-COOZ (XV) 2 r 3 1 (XVII)
N N
O RZ /2 ( XV I I I ) o - Ra
N~ ICI - COO Z ( I I I i ) NNI~ ICI - COOH ( I I I j )
o R2 O- Ra
- 54 -
13~0 ~93
-
wherein Ra is an aliphatic hydrocarbon residue wllicll may be
substituted witll halogell, carboxy or esterified
carboxy
R6a is protected amino
X is halogen
Y is lower alkoxycarbollyl, and
Z is lower alkyl
Bach of the above processes are explained in the
following.
Process 1 : Etherification
The compound (IIIb) and (IIId) can be prepared by
reacting a compound (IIla) or (IIIC) witll an etherifying
agent, respectively.
This reaction may be conducted substantially in
the same manner as the aforementioned Process C.
Process 2 : Thiazole ring formation
The compound (IIIC) and ~IIId) can be prepared by
reacting a compound (IIIa) or ~IIIb) with a thiourea compound
(VIIa), respectively, and further the compound (IIIe) can be
prepared by reacting a compound ~IIIb) with thiourea.
This reaction may be conducted substantially in the
same manner as the aforementioned Process D.
Process 3 : Elimination of amino-protective group
The compound (IIIe) and (IIIg) can be prepared by
subjecting a compound (IIId) or (IIIf) to elimination reaction
of the protective group in the protected amino group for Ra,
- 55 -
134~99
" ..
respectively.
This reaction may be conducted substantially in
the same manner as the aforementioned Process E.
Process 4 : Carbo~y formatioll
The compound (IIIf), (IIIg) and (IIIj) can be
prepared by transforming the esterified carboxy group of a
compound (IIId), (IIIe) or (IIli) into free carboxy group,
respectively.
This reaction may be conducted substantially in
the same manner as the aforemelltiolled Process K.
Process 5 : Oximation
The compound (If) can be also prepared by reacting
a compound (IIIh) with a hydroxylamine derivative (XIV) or
its salt.
The hydroxylamine derivative (XIV) may be
hydroxylamine substituted with an aliphatic hydrocarbon
residue which may be substituted with halogen, carboxy or
esterified carboxy, particulars of which are to be referred
to those as exemplified before. Suitable salt of the
hydroxylamine derivative (XIV) may be hydrochloride,
hydrobroinide, sulfate or the like.
The reaction is usually conducted in a conventional
solvent such as water, alcohl, tetrahydrofuran, acetonitrile,
dimethylsulfoxide, pyridine or any other solvent which does
not adversely influence the reaction, or a mixture thereof,
and the reaction temperature is not critical.
In case that a salt of the hydroxylamine derivative
(XIV) is used as a reagent, the reaction is preferably
- 56 -
1340~9~
..
conducted in the presence of a conventiollal base.
Process 6 : Thiadiazol rillg formation
The compound (II]:i) can bc preparcd by rcactin~ a
compound (XV) with a hydrazj.lle dcrivative (XVI), and then
reacting the resultant compound (XVII) with sulfur dihalide
(XVIII).
Among the starting compound (III), the compound
of the formula :-
Rl- C- COOR8
a 11
N (III')
~ 2'
.. ~~Ra
whereill Ra is thiadiazolyl or thiazolyl of thc formula:
6N~ ill W}liCh R6 is a.mino or protected amino,
~' Ra is alkyl, alkellyl or alkynyl having more'~tnanone
carbon atom or cycloalkyl W]liCh Illay be substituted.
with halogen, carboxy or esterified carboxy,
R8 is hydrogell or lower alkyl,
provided that R6 is amino which may be protected with ~ormyl,
and R8 is hydrogen, when Ra is ethyl, isopropyl or allyl,
is novel and useful as a starting material in tlle
aforementioned Process A.
Particulars of each defillitioll in the above are to
be referred to those as explained before.
Following examples are given only for explaining
this invention in more detail.
~; ~ 30
- 57 -
.. . . .
13~0~9'3
Preparation of the startinq compounds:
Example A
(1) A solution of ethyl 2-methoxyiminoacetoacetate
(a mixture of syn and anti isomers) (34.6 g.) and t-
butoxy-carbonylhydrazine (26.4 g.) in ethanol (200 ml.)
was stirred for 7.5 hours at ambient temperature and
allowed to stand overnight to precipitate crystals. The
crystals were collected by filtration, washed with ethanol
and dried to give ethyl 2-methoxyimino-3-t-butoxycarbonyl-
hydrazonobutyrate (a mixture of syn and anti isomers)
(41.7 g.), mp 144 to 145~C.
I.R. v j : 3200, 1750, 1705, 1600, 1520 cm~
max
N.M.R. ~ppm (CDC/3) : 8.52 (lH, broad s), 4.35
(2H, q, J=7Hz), 4.10 (3H, s),
2.00 (3H, s), 1.50 (9H, s),
1.33 (3H, t, J=7Hz)
(2) Sulfur dichloride (15.9 ml.) was added with
stirring at ambient temperature to a solution of ethyl 2-
methoxyimino-3-t-butoxycarbonylhydrazonobutyrate (a
mixture of syn and anti isomers) (14.36 g.) in methylene
chloride (150 ml.), and the mixture was stirred for 1 hour
at ambient temperature. To the reaction mixture was added
ice-water (300 ml.), and the methylene chloride layer was
washed with water, with a saturated aqueous solution of
sodium bicarbonate and with a saturated aqueous solution
of sodium chloride and dried over magnesium sulfate. The
solvent was distilled off to give an oil. The oil was
purified by column chromatography on silica gel using a
mixture of benzene and n-hexane (19:1) as an eluent to
firstly give ethyl 2-methoxyimino-2-(1,2,3-
- 58 -
*Trademark
' ~
13~0~93
, .
.,
thiadiazol-4-yl)acetate (syn isomcr) (1.8 g.), mp 77 to
79~C.
I.R. v NaUxol : ]720, 1595 cm 1
N-~-R- ~ppm (CDCQ3) : 8.92 (lH, s), 4.46 (2H, q,
J~7Hz), 4.06 (3H, s), 1.38 (3H, t,
J=71~z)
From subsequent fractions, ethyl 2-methoxyimino-
2-(1,2,3-thiadiazol-4-yl)acetate (anti isomer) (0.7 g.) was
obtained as an oil.
I.R. v Faxm : 1730, 1590 cm 1
N-M-R- ~ppm (CDCQ3) : 9.38 (lH, s), 4.47 (2H, q,
J=7Hz), 4.20 (3H, s), 1.40 (3H, t,
J=7Hz)
(3) lN Aqueous solution of sodium hydroxide (6.7 ml.)
was added to a solution of ethyl 2-methoxyimino-2-(1,2,3-
thiadiazol-4-yl)acetate (syn isomer) (1.2 g.j in methanol
(10 ml.) and the mixture was stirred for 1.5 hours at
ambient temperature. Methanol was distilled off from the
reaction mixture and water was added to the residue. The
mixture was washed with ether, adjusted to pH 1 with 10~
hydrochloric acid and extracted with ethyl acetate. The
extract was washed with a saturated aqueous solution of
sodium chloride and dried over magnesium sulfate. The
solvent was distilled off to give prisms of 2-methoxyimino-
2-(1,2,3-thiadiazol-4-yl)acetic acid (syn isomer) (0.7 g.),
m.p. 110 to 113~C.
I.R. v INnuaxol : 2750-2150t 1730, 1595 cm 1
N.M.R. ~ppm (d6-DMSO) : 9.47 (lH, s), 4.01 (3H, s)
- 59 -
- ~
~340~93
Example B
(1) Pulverized potassium carbonate (160 g.) was added
to a solution of etlly] 2-hydrox~iminoacetoacetate ta mixture
of syn and anti isomers) (152 g.) in acetone (500 ml.).
Dimethyl sulfate (130 g.) was dropwise added thereto with
stirring over 1 hour at 45 to 50~C and the mixture was stirred
for 2 hours. An insoluble material was filtered off and
tbc filtrate was concentratcd under reduced pressure. ~he
filtered insoluble material was dissolved in water (500 ml.)
and this solution was added to the residue. The mixture
was extracted twice witll ethyl acetate (300 ml.). The
extract was washed twice with water (200 ml.) and with a
saturated sodium chloride aqueous solution (200 ml.) and
dried over magnesium sulfate. The solvent was distilled
o~f under reduced pressure and the residue was distilled
under reduced pressure to give colorless oil of ethyl 2-
metlloxyiminoacetoacetate (a mixture of syn and anti isomers)
(145.3 g.), bp 55 to 64~C¦0.5 mm Hg.
I.R. v maxm : 1745, 1695, 1600 cm 1
N.M.R. ~ppm (CDCQ3) : 4.33 (4H, q, J=8Hz),
4.08 (3H, s), 3.95 (3H, s),
2.40 (31l, s), 1.63 (3H, s),
1.33 (6H, t, J=8Hz)
(2) Sulfuryl chloride (235 ml.) was dropwise added
over 20 minutes with stirring and ice-cooling to a solution
of ethyl 2-methoxyiminoacetoacetate (syn isomer) (500 g.)
in acetic acid (500 ml.), and the mixture was stirred
overnight under cooling with watcr. Nitrogen gas was
_ 60 -
~34~ ~99
introduced to thc reaction mixture for 2 hours, and the
resulting mixture was poured into watcr (2.5 Q.) After
extrracting with methylene chloride (500 ml.) and twice
with methylene chloride (200 ml.), the extracts were
combined. The combined extract were washed with a saturated
aqueous solution of sodium chloride, and adjusted to pH 6.5
by adding water (800 ml.) and sodium bicarbonate. Methylene
chloride layer was separated, washed with an aqueous solution
of sodium chloride and dried over magnesium sulfate. The
solvent was distilled off to give ethyl 2-methoxyimino-4-
chloroacetoacetate (syn isomer) (559 g.).
I.R. ~ maxm : 1735, 1705 cm 1
(3) Ethyl 2-methoxyimino-4-chloroacetoacetate (syn
isomer) (50 g.) was added over 3 minutes with stirring at
ambient temperature to a solution of thiourea (18.4 g.)
and sodium acetate (19.8 g.) in a mixture of methanol (250
ml.) and water (250 ml.). After stirring for 35 minutes
at 40 to 45~C, the reaction mixture was cooled with ice
and adjusted to pH 6.3 with a saturated aqueous solution of
sodium bicarbonate. After stirring for 30 minutes at the
same temperature, precipitates were collected by filtration,
washed with water (200 ml.) and then with diisopropyl ether
(100 ml.), and dried to give colorless crystals of ethyl
2-methoxyimino-2-(2-amino-1,3-thiazol-4-yl)acetate (syn
isomer) (37.8 g.), m.p. 161 to 162~C.
I.R. v Nlauxol : 3400, 3300, 3150, 1725, 1630
1559 cm 1
N-M-R- ~ppm tCDCQ3) : 6.72 (lH, s), 5.91 ~2H,
broad s), 4.38 (2H, q, J=7Hz),
4.03 (3H, s), 1.38 (3ll, t, J=7Hz)
- 61 -
1340~9
., ~
(4) Ethanol (lO ml.) was added to a suspension of
cthyl 2-methoxyimino-2-(2-amino-l,3-thiazol-4-yl)acetate
tsyn isomer) ~2.2 g.) in a lN aqueous solution of sodium
hydroxide (12 ml.) and the mixture was stirred for 15 hours
at ambient temperature. The reaction mixture was adjusted
to pH 7.0 with lO~ hydrochloric acid and ethanol was distil-
led off under reduced pressure. The residual aqueous
solution was washcd with ethyl acetate, adjustcd to p1~ 2.8
with lO~ hydrochloric acid and stirred under ice-cooling
to precipitate crystals. The crystals were collected by
filtration, washed with acetone and recrystallized from
ethanol to gi~e colorless needles of 2-methoxyimino-2-(2-
amino-1,3-thiazol-4-yl)acetic acid (syn isomcr) (l.l g.)
I.R. v N~aUx~l : 3150, 1670, 1610, 1585 cm l
N.M.R. ~ppm (d6-DMSO) : 7.20 (211, broad s),
6.85 (l11, s), 3.83 (31~, s)
- 62 -
13~0493
Example C
(1) Sulfuryl,cllloride ~35.2 g.) was added all at once
to the stirred solutioll o~ etllyl 2-ethoxyi,mino-3-
- oxobutyrate (syn isomer, 48.9 g.) in acetic acid (49 ml.)
at room temperaturc, and stirrcd ,at the same temperature
for an hour. After addin~ the resu]tallt solution into
water (200 ml.), the solution was extracted with methylene
chloride. The extract was washed with a saturated
aqueous solution of sodium chloride, neutralized with
an aqueous solution of sodium bicarbonate and wished with
water. The solution was dried over magncsium sulfate
and conccntrated under,reduced pressure to give ethyl
2-ethoxyimino-3-oxo-4-chlorobutyrate (syn isomer, 53.8 g.),
pale yellow oil.
(2) A mixture of ethyl 2-ethoxyimin~3-oxo-4-chlorobutyrate
(syn isomer 38.7 g.), thiourea (13.2 g.), sodium acetate
~14.3 g.), methanol (95 ml.) and water (95 ml.) was
stirred at 48~C for 40 minutes. After the resultant
solution was adjusted to pH 6.5 with an aqueous solution
of sodium bicarbonate, the appeared precipitates were
collected by filtration and washed with diisopropyl ether
:.~
to give ethyl 2-~2-amino-4-thiazolyl)-2-ethoxyiminoacetate
(syn isomer, 14.7 g.), mp 130 to 131~C.
I.R.v mUaxol : 3450, 3275, 3125, 1715, 16~0 cm 1
- 63 -
.. .. , . ~ . .
13'~0~93
(3) Ethyl 2-~2-llmino-~-t}lia7Olyl)-2-ethoxyiminoacetate
(syn isomer, 5 g.) was added to a mixture of lN sodium
hydroxidc (45.9 ml.) and cthanol (30 ml.) and stirred at
room temperature ~or 5 hours. After removing othallol
from t}-e resu]tant solution under rcduced pressure, the
residue was dissolved in water (60 ml.) and adjusted to
pH 2.0 with 10~ hydrochloric acid. The solution was
subjccted to salting-out, and the precipitates were collected
by filtration and dried to give 2-(2-amino-4-thiazolyl)-
2-ethoxyiminoacetic acid (syn isomer, 2.9 g.).
I. R. vNmUax~l 3625, 3225 (shoulder), 3100,
1650, 1615 cm 1
N.M.R. ~ppm (DMSO-d6) : 1.20 (3H, t, J=7Hz),
4.09 (2ll, q, J=7Hz), 6.82 (lH, s),
7.24 (211; broad s)
(4) 2-(2-Aminothiazol-4-yl)-2-ethoxyiminoacetic acid
(syn isomer, 100 g.), formic acid (85.5 g.) and acetic
anhydride (190.1 g.) were treated in a similar manner to
that of Example F-(5) to give 2-(2-formamidothiazol-4-yl)-
2-ethoxyiminoacetic acid (syn isomer, 99.1 g.).
I.R. v mUaxol : 3200, 3140, 3050, 1700 cm 1
N.M.R. ~(DMS0-d6, ppm) : 1.18 (3H, t, J=6Hz),
4.22 ~2H, q, J=6Hz), 7.56 (lH, s),
8.56 (lH, s), 12.62 (lH, broad s)
- 64 -
1340~3
EzamPle D
(1) To a suspensionof ethyl 2-hydroxyimino-3-oxobutyrate
(syn isomer, 15 g.) and potassium carbonate (19.3 g.)
in acetone (75 ml.) was added dropwise propyliodide (16.2 g.)
with stirring, and the mixture was stirred at ambient
temperature for 1,5 hours. The insoluble substance was
collected by filtration and washed with acetone. The
washings and the filtrate were combined and evaporated
to dryness under reduced pressure.To t~eresultant residue
10 was added water and the aqueous solution was extracted
twice with chloroform. The extract was washed with an
aqueous solution Of sodium chloride, dried over magnesium
sulfate, and then evaporated to dryness under reduced
pressure to give ethyl 3-oxo-2-propoxyiminobutyrate
(syn isomer, 15.4 g.), oil.
(2) Ethyl 3-oxo-2-propoxyiminobutyrate (syn isomer, 15.4 g.)
and sulfuryl chloride (10.6 g.) were dissolved in acetic
acid (15.4 ml.), warmed at 35 to 40~C for 10 minutes with
20 stirrin~ and then stirred at ambient temperature for
additional 6 hours. The reaction mixture was poured into
ice-water (200 ml.) andtlle resultant mixture was extracted
twice with chloroform. The extract was washed with an
aqueous solution of sodium chloride, twice a saturated
aqueous solution of sodium bicarbonate and once with water
in turn, dried over magnesium sulfate, and then evaporated
~ to dryness under reduced pressure to give ethyl 4-chloro-
;' 3-oxo-2-propoxyiminobutyrate (syn isomer, 15.4 g.), oil.
I.R. ~maxm : 1740, 1710, 1695, 1455 cm 1
- 65 -
1340~93
'~ (3) Ethyl 4-chloro-3-oxo-2-propoxyiminobutyrate (syn isomer,
15.4 g.), thiourea t4.97 g.) and sodium acetate hydrate (8.89 g.)
were dissolved in a mixture of water (40 ml.) and ethanol (50 ml.),
and stirred at 40~C for an hour.
The reaction mixture was adjusted to pH 6.5 with a saturated
aqueous solution of potassium carbonate under cooling and
stirred at the same temperature for half an hour. The precipitating
crystals were collected by filtration, washed with water and
diisopropyl ether, and then dried to give crystalline ethyl
2-(2-amino-4-thiazolyl)-2-propoxyiminoacetate (syn isomer,
10.55 g.), mp 142 - 144~C.
I.R. yNmaUxol : 3460, 3260, 3120, 1720, 1620, 1540 cm 1
N.M.R. ~ ppm (d6-DMSO) : 0.88 (3H, t, J=7Hz),
1.27 (3H, t, J=6Hz), 1.60 (2H, sextet, J=7Hz),
4.04 (2EI, t, J=7Hz), 4.28 (2H, q, J=6Hz),
6.86 (lH, s), 7.23 (2H, s)
(4) A solution of ethyl 2-(2-amino-4-thiazolyl)-2-propoxyimino-
acetate (syn isomer, 10 g.) in a mixture of tetrahydrofuran
(39 ml.), methanol (39 ml.) and lN sodium hydroxide (75.8 ml.)
was stirred at 35 to 40~C for 5 hours.
After the resultant solution was concentrated under reduced
pressure, the aqueous residue was adjusted to pH 2.5 with 10%
hydrochloric acid. The precipitates were collected by filtration
and dried to give 2-(2-amino-4-thiazolyl)-2-propoxyiminoacetic
~cid (synisomer, 6.2 g.), mp 161~C (dec.)
I.R. ~ NmaU~ol : 3380, 3120 (broad), 1630, 1610, 1460 cm 1
N.M.R. ~ ppm (DMSO-d6) : 0.89 (3H, t, J=7Hz),
1.63 (2H, sextet, J=7Hz), 4.05 (211, t, J=7Hz),
6.83 (lH, s), 6.9 - 8.8 (3H, broad)
- 66 -
.. .. . ..
13~93
(5) 2-(2-Aminothiazol-4-yl)-2-n-propoxyiminoacetic acid
(syn isomer, 21.8 g.), acetic anhydride (38.8 g.) and formic
acid (17.5 g.) were treated in a similar manner to that of
Example F -(5), and then the obtained oil was triturated with
diisopropyl ether to give 2-(2-formamidothiazol-4-yl)-2-n-
propoxyiminoacetic acid (syn isomer, 19.2 g.), mp. 164~C (dec.).
I.R. v mUa~ol : 3200, 3120, 3050, 1700, 1550 cm 1
N.M.R. ~ ISO-d6, ppm) : 0.92 (3H, t, J=711z),
1.67 (2H, sextet, J=7Hz), 4.12
(2H, t, J=7Hz), 7.53 (lH, s),
~.54 (11~, s)
_ 67 -
~,.
... ; ~ ~ . ... .
. = . . .
1340~ 3
v
Exam~le E
(1) Ethyl Z-hydroxyimino-3-oxobutyrate (syn isomer, 30 g.),
iso-propyl iodidc (3Z.5 g.), potassium carbonate (39.5 g.)
and acetone (150 ml.) were treated in a similar manner to that
of ExampleD -(1) to give ethyl 2-iso-proyoxyimino-3-oxobutyrate
(syn isomer, 35.4 g.), oil.
I.R. v maxm : 1745, 1690, 1600 cm 1
N.M.R. ~(CCQ4, ppm) : 1.33 (3H, t, J=7Hz), 1.35
(6H, d, J=6Hz), 2.32 (3H, s),
4.1~4.7 (3H, m).
(2) Ethyl 2-iso-propoxyimino-3-oxobutyrate (syn isomer
35.4 g.), sulfuryl chloride (24.5 g.) and acetic acid ~35.4
ml.) were treated in a similar manner to that of Example D (2)
to give ethyl 4-chloro-3-oxo-2-iso-propoxyimi.nobutyrate
(Syll isomer, 41.5 g.), oil.
I.R ~ maxm : 1745, 1715, 1375 cm 1
(3) Ethyl 4-chloro-3-oxo-2-iso-propoxyiminobutyrate (syn
isomer, 41.5 g.), thiourea (13.4 g.), sodium acetate (14.4 g.),
water (110 ml.) and ethanol (110 ml.) were treated in a similar
manrler to that of Example D-(3) to give ethyl 2-(2-aminothiazol-
4-yl)-Z-iso-propoxyiminoacetate (syn isomer, 27.3 g.), mp. 162
to 164~C.
I.R. ~ NnUajxol: 3460, 3430, 3260, 3150, 1725, 1615
1540 cm 1
N.M.R. ~(DMSO-d6, ppm) : 1.17 (6H, d, J=6Hz),
1.24 (3ll, t, J=7Hz), 4~4.7 (3H, m),
6.86 (lH, s), 7.24 (2H, s)
- 68 -
1340~93
.~ J
(4) Ethyl 2-(2-aminothiazol-4-yl)-2-iso-propoxyimino-
acetate (syn isomer, 26.8 g.), lN aqueous solution of sodium
hydroxide, (156 ml.), methanol (156 ml.) and te~rahydrofuran
(100 ml.) were treated in a similar manner to that of Example
D -(4) to give 2-(2-aminothiazol-4-yl)-2-iso-propoxyimino-
acetic acid (syn isomer, 15.3 g.), mp. 151~C ~dec.).
I.R. v INnuajxol: 3610, 3580, 3080, 1650, lC10 cm 1
N.M.R. ~(DMSO-d6, ppm) : 1.22 (6H, d, J=6Hz),
4.33 (lH, quintet, J=6Hz),
6.80 (lH, s), 7.22 (2H, broad s)
2-(2 Aminotlliazol-4-yl)-2-iso-propoxyiminoacetic acid
(syn isomer, 4 g.), acetic anhyddride (7.6 g.) and formic
acid (3.4 g.) were treated in a similar manner to that of
Example F -(5) to give 2-(2-formamidothiazol-4-yl)-2~iso-
propoxyiminoacetic acid (syn isomer, 3.75 g.), mp. 168 to 169~C
(dec.).
I.R. v mUaxol : 3200, 3130, lilO, 1600, 1560 cm 1
N.M.R. ~(DMSO-d6, ppm) : 1.26 (6H, ~), 4.4 (lH, m),
7.54 (lH, s), 8.52 (lH, s),
12.56 (lH, broad s)
- 69 -
13~0~93
~ . .
Example F
(1) n-Butyl iodide (46.9 g.) was added dropwise to a
stirred suspension of ethyl 2-hydroxyimino-3-oxobutyrate
(syn isomer, 40 g.), potassium carbonate (52.7 g.) and
acetone (200 ml.) under ice-cooling over S minutes, and
s~irred at room temperature for 4 hours. The resultant
solution was filtered, and washed with acetone. The
filtrate and washing solution were combined together and
concentrated in vacuo. After adding water (300 ml.) to the
residue, the solution was extracted with methylene chloride
three times. The solution was washed with a saturated
aqueous solution of sodium chloride, dried over magnesium
sulfate and concentrated in vacuo to give ethyl 2-n-
butoxyimino-3-oxobutyrate (syn isomer, 48.8 g.), oil.
I.R. v maxm : 1750, 1700, 1470, 1370, 1320 cm 1
(2) A solution of ethyl 2-n-butoxyimino-3-oxobutyrate
(syn isomer, 48.8 g.), sulfuryl chloride (31.5 g.) and
acetic acid (48.8 ml.) was stirred at 40~C for 10 minutes
and further at room temperature for 5.5 hours. After water
(300 ml.) was added to the resultant solution under ice
cooling, the solution was extracted with methylene chloride
three times. The extract was washed with water, an aqueous
solution of sodium bicarbonate and a saturated aqueous
solution of sodium chloride in turn, and dried over magnesium
sulfate. The solution was concentrated in vacuo to give
ethyl 2-n-butoxyimino-4-chloro-3-oxobutyrate (syn isomer,
52.1 g.), oil.
I. R- ~ max : 1740, 1710, 1470, 1370 cm 1
- 70 -
... . .....
13 ~0 i~3
(3) A solution of ethyl 2-n-butoxyimino-4-chloro-3-
oxobutyrate ~syn isomer, 52.1 g.), thiourea (15.9 g.),
sodium acetate 3 hydrate (28.4 g.), water (130 ml.) and
ethanol (180 ml.) was stirred at 40~C for 1.25 hours.
The resultant solution was adjusted to pH 6.5 with an
aqueous solution of sodium carbonate under ice cooling, and
stirred for 20 minutes under ice cooling. The precipitates
were collected by filtration, and washed with water and
diisopropyl ether in turn to give ethyl 2-(2-aminothiazol-
4-yl)-2-n-butoxyiminoacetate (syn isomer, 36.1 g.)~ mp 126
to 128~C.
I. R. v NUaxol : 3460, 3370, 3230, 1720, 1620,
1550 cm 1
N.M.R. ~(DMSO-d6, ppm) : 0.6 - 2.0 (6H, m),
1.28 (3H, t, J=7Hz), 4.12 (3H, t, Js6Hz),
4.31 (2H, q, J=7Hz), 6.89 (lH, s),
7.24 (2H, s)
~4) A solution of ethyl 2-(2-aminothiazol-4-yl)-2-n-
butoxyiminoacetate (syn isomer, 36 g.), methanol (133 ml.),
tetrahydrofuran (133 ml.) and 2N aqueous solution of sodium
hydroxide (133 ml.) was stirred at 30~C for 5 hours. After
the resultant solution was concentrated in vacuo, the
residue was dissolved in water. The solution was adjusted
to pH 7 with 10% hydrochloric acid and treated with acti-
vated charcoal. The solution was adjusted to pH 2.0 with
10% hydrochloric acid and stirred for 20 minutes under ice
cooling. The precipitates were collected by filtration,
washed with water and acetone in turn, and dried to give
2-(2-aminothiaz~-4-yl)-2-n-butoxyiminoacetic acid (syn
isomer, 25.4 g.).
. ~. ~_.
13~0~93
I. R. v maUxol : 3325, 3190, 1660, 1620 cm 1
N.M.R. ~(DMSO-d6, ppm) : 0.88 (3H, t, J=7Hz),
1.0-1.9 (4H, m), 4.06 (2H, t, J=7Hz),
6.81 (lH, s), 7.21 (2H, broad s)
(5) Formic acid (18.95 g.) was added dropwise to acetic
anhydride (42.0 g.) under stirring at room temperature over
5 minutes, and stirred at 50~C for an hour. 2-(2-Aminothiazol-
4-yl)-2-n-butoxyiminoacetic acid (syn isomer, 25 g.) was
added to the solution under ice cooling, and stirred at room
te~perature for 3 hours and additionally at 30~C for an hour.
After concentrating the resultant solution in vacuo, the
residue was dissolved in diethyl ether. The solution was
washed with water and a saturated aqueous solution of sodium
chloride in turn, dried over magnesium sulfate and concentrated
in vacuo. The obtained oil was triturated with a solution
of n-hexane (1 part) and diisopropyl ether (1 part), and
collected by filtration to give 2-(2-formamidothiazol-4-yl)-
2-n-butoxyiminoacetic acid (syn isomer, 20.1 g.).
I. R. v mUaxol : 3350, 3160, 3050, 1700, 1680,
1570 cm~l
N.M.R. ~(DMSO-d6, ppm) : 0.91 (3H, t, J=6Hz),
1.0 - 2.2 (4H, m), 4.18 (2H, t, J=6Hz),
7.57 (lH, s), 8.59 (1ll, s), 12.66 (lH,
broad s)
Example G
(1) Ethyl 2-hydroxyimino-3-oxobutyrate (syn isomer,
40 g.), N,N-dimethylformamide (200 ml.), potassium carbonate
(52.7 g.) and iso-butyl bromide (34.94 g.) were treated in
a similar manner to that of Example F -(1) to give ethyl
- . . .
; 1340~93
2-iso-butoxyimino-3-oxo-butyrate (syn isomer, 42 g.).
I. R. v mUaxol : 1740, 1670 (broad) cm 1
(2) Ethyl 2-iso-butoxyimino-3-oxobutyrate (syn isomer,
42 g.), acetic acid (42 ml.) and sulfuryl chloride (27.1 g.)
were treated in a similar manner to that of Example F-(2)
to give ethyl 2-iso-butoxyimino-4-chloro-3-oxobutyrate
(syn isomer, 31.9 g.).
I. R. v maxm : 1750, 1720, 1680 cm 1
(3) Ethyl 2-iso-butoxyimino-4-chloro-3-oxobutyrate
(syn isomer, 31.9 g.), thiourea (9.72 g.), sodium acetate
3-hydrate (17.4 g.), ethanol (120 ml.) and water (80 ml.)
were treated in a similar manner to that of Example F-(3)
to give ethyl 2-~2-aminothiazol-4-yl)-2-iso-butoxyimino-
acetate (syn isomer, 17.6 g.), mp 122 to 124~C.
I. R. v mUaxol : 3470, 3260, 3120, 1730, 1620,
1545 cm 1
N.M.R. ~DMSO-d6, ppm) : 0.86 (6H, d, J=7Hz),
1.28 (3H, t, J=7Hz), 1.6-2.2 (lH, m),
3.86 (2H, d, J=7Hz), 4.28 (2H, q, J=7Hz),
- 20 6.86 (lH, s), 7.22 (2H, s)
(4) Ethyl 2-(2-aminothiazol-4-yl)-2-iso-butoxyimino-
acetate (syn isomer, 19.6 g.), 2N aqueous solution of sodium
hydroxide (72.2 ml.), methanol (72.2 ml.) and tetrahydrofuran
(72 2 ml.) were treated in a similar manner to that of
Example F -(4) to give 2-(2-aminothiazol-4-yl)-2-iso-
butoxyiminoacetic acid (syn isomer, 16.1 g.), mp 180~C (dec.).
I. R. v mUaxol : 3375, 3300, 3130, 3050, 1640 cm 1
N.M.R. ~(DMSO-d6, ppm) : 0.91 (6H, d, J=7Hz),
1.5-2.3 (lH, m), 3 90 (2H, d, J=7Hz),
30 - 6.87 (lH, s), 7.26 (2H, broad s)
- 73 -
-- 13~0~9~
.1 .
(5) 2-(2-Aminothiazol-4-yl)-2-iso-butoxyiminoacetic
acid (syn isomer, 11.5 g.), acetic anhydride (19.3 g.) and
formic acid (8.7 g.) were treatcd in a similar manner to
that of Example F-(5) to give 2-(2-formamidothiazol-4-yl)-
2-iso-butoxyiminoacetic acid (syn isomer, 11.15 g.), mp
163~C (dec.).
I. R. v NmUaxol : 3175, 3110, 3050, 1695, 1550 cm 1
; N.M.R. 8(DMSO-d6, ppm) : 0.91 (6H, d, J=7Hz),
; 1.7-2.3 (1ll, m), 3.92 (2H, d, J-7Hz),
7.52 (lH, s), 8.52 (lH, s), 12.58 (lH,
broad s)
Example H
(1) Ethyl 2-hydroxyimino-3-oxobutyrate (syn isomer, 30 g.),
N,N-dimethylformamide (100 ml.), potassium carbonate (39.5 g.)
and cyclohexyl bromide (31.1 g.) were treated in a similar
manner to that of Example F-(l) to give ethyl 2-cyclohexyloxy-
imino-3-oxobutyrate (syn isomer, 41.8 g.), oil.
I.R. v mlxm : 1740, 1680 cm 1
(2) Ethyl 2-cyclohexyloxyimino-3-oxobutyrate (syn isomer,
41.3 g.), acetic acid (41.3 ml.) and sulfuryl chlori~e (23.8
g.) were treated in a similar manner to that of Example F-(2)
to give ethyl 4-chloro-2-cyclohexyloxyimino-3-oxobutyrate
(syn isomer, 27.8 g.), oil.
I.R. v maxm : 1745, 1715, 1680 cm 1
(3) Ethyl 4-chloro-2-cyclohexyloxyimino-3-oxobutyrate
(syn isomer, 27.8 g.), thiourea (7.7 g.), sodium acetate
3-hydrate (13.7 g.), water (70 ml.) and ethanol (140 ml.)
were treated in a similar manner to that of ExampleF -(3) to
give ethyl 2-(2-aminothiazol-4-yl)-2-cyclohexyloxyiminoacetate
(syn isomer, 3.6 g.), mp. 125 to 126~C.
- 74 -
, . ... ..
1340~9~
.~ . ,
I.R. v mUaxol : 3430, 3250, 3160, 3130, 1715,
1635 cm 1
N.M.R. ~(DI~SO-d6, ppm) : l.Z8 (31-l, t, J=7Hz),
1.0~2.Z (10ll, m), 4.22 (lH, m),
4.32 (ZH, q, J=7Hz), 6.88 (lH, s),
7.Z4 (211, broad s)
(4) Ethyl 2-(2-aminothiazol-4-yl)-2-cyclohexyloxyiminoacetate
(syn isomer, 3.5 g.), 2N aqueous solution of sodium hydroxide
(11.8 ml.), methanol (11.8 ml.) and tetrahydrofuran (11.8 ml.)
10 were treated in a similar manner to that of Example F-~4) to
give 2-t2-aminothiazol-4-yl)-2-cyclohexyloxyiminoacetic acid
(syn isomer, 2.1 g.), mp. 148~C (dec.).
I.R. v maUxol : 3110, 1630, 1450 cm 1
N.M.R. ~(DMSO-d6, ppm) : 0.8~2.3 (lOH, m),
4.14 (lH, m), 6.86 (lH, s),
7.5 (2H, broad s)
(5) 2-(2-Aminothiazol-4-yl)-2-cyclohexyloxyiminoacetic
acid (syn isomer, 1.5 g.), acetic anhydride ~2.27 g.) and
formic acid (1.03 g.) were treated in a similar manner to
that of Example F-(5), and the oil obtained was suspended in
an aqueous solution of sodium bicarbonate. The suspension
was adjusted to pH 3.5 with 10% hydrochloric acid. The
precipitates were collected by filtration, washed with water
and dried to give 2-(2-formamidothiazol-4-yl)-2-cyclohexyloxy-
iminoacetic acid (syn isomer, 1.0 g.), mp. above 230~C.
I.R. v maUxol : 3175, 3100, 3060, 1680 cm 1
Exam~le I
(1) Ethyl 2-hydroxyimino-3-oxobutyrate (syn isomer, 56.7 g.),
N,N-dimethylformamide (280 ml.), potassium carbonate (72.3 g.)
and propargyl bromide t43 g.) were treated in a similar manner
:
~ - 1340~93
to that of ExampleF ~1) to give ethyl Z-propargyloxyimino-3-
oxobutyrate (syn isomer, 71.2 g.).
I.R. v maxm : 3280, 3220, 2120, 1735, 1670 cm 1
(2) Ethyl 2-propargyloxyimino-3-oxobutyrate (syn isomer,
71.2 g.), acetic acid (81 ml.) and sulfuryl chloride (50.2 g.)
were treated in a similar manner to that of Example F-(z) to
give ethyl 4-chloro-3-oxo-2-propargyloxyiminobutyrate (syn
isomer, 61.6 g.), oil.
I.R. v Film : 3300, 2130, 1745, 1720, 1675 cm
N.M.R. ~(CCQ4, ppm) : 1.39 (3H, t, J=7Hz~, 2.57
(lH, t,,J=2Hz), 4.36 (211, q, J=7Hz),
4.56 (21l, s), 4.86 (2H, d, J=2Hz)
(3) Ethyl 4-chloro-3-oxo-2-propargyloxyiminobutyrate (syn
isomer, 61 g.), thiourea (20 g.), sodium acetate 3-hydrate
(35.8 g.), water (150 ml.) and ethanol (180 ml.) were treated
in a similar manner to that of Example F-(3) to give ethyl
2-(2-aminothiazol-4-yl)-2-propargyloxyiminoacetate (syn isomer,
35.6 g.).
I.R. v NmaU~ol : 3290, 2220, 1729 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 1.28 (3H, t, J=.7~z),
2~ 3.49 (lH, t, J=3Hz), 4.31 (2H,
q, J= 7Hz)" 4.76 (2H, d, J=3Hz),
6.95 (lH, s), 7.29 (2H, s).
(4) Ethyl 2-(2-aminothiazol-4-yl)-2-propargyloxyimino-
acetate (syn isomer, 2.8 g.), methanol (Z3 ml.), tetrahydrofuran
(20 ml.) and lN aqueous solution of sodium hydroxide (22.17 ml.)
were treated in a similar manner to that of Example F-(4) to
give 2-(2-aminothiazol-4-yl)-2-propargyloxyiminoacetic acid
(syn isomer, 1.924 g.).
I.R. v mUajxol : Zl90, 1740
- 76 -
- - -- - ~ . ~ , ,
. ~, : ~ . , .
1 3 ~ O ~ '3
N.M.R. ô (DMSO-d6, ppm) : 3.47 (lH, t, J=1.51iz),
4 . 74 (ZH, d, J=1 . 511z), 6 . ~0 (lH, s)
-- 77 --
~340~93
. ~ . .
Example J
(1) Ethyl 2-hydroxylmino-3-oxobutyrate (syn l~omer, 40 g.), I~,N-
dlmethylformal~c (200 ml.), potas~lum csrbonate (52 e.) and n-hexyl
; bromide (41.4 g.) were treated ln a similar manner to that of Example
F-(l) to,elve ethyl 2-n-hexyloxylmino-3-oxobucyrate (syn lsomer, 60.7 g.),
oll.
I. R. ~ Fllxm : 1740, 1705, 1700 cm 1
N. M. R. ~(CC14, ppm): 0.6~ 2.1 (14H, m), 2.37 (311, 8), 4.1 ~4.6
(4H, m)
(2) Ethyl 2-n-hexylo~ylmino-3-oxobutyrate (Byn lsomer, 60.7 g.),
acetic acld (61 ml.) and sulfuryl chlorlde (34.7 g.) Nere treated ln
a similar manner to that of E~ample F-(2) to glve ethyl 2-n-hexyloxyimlno-
4-chloro-3-oxobutyrate (syn isomer, 55.6 g.).
I. R. ~JFalm : 1740, 1720, 1470 cm 1
; N. M. R. S(CC14, ppm): 0.6~2.2 (14H, m), 4.1~4.6 (4H, m),
4.47 (2~, 8)
t3) Ethyl 2-n-hexyloxylmino-4-chloro-3-oxobutyrate (syn lsomer,
55.6 g.), thiourea (15.2 g.), sodlum acetate 3-hydr~te (27.2 g~,
ethanol (280 ml.) and water (140 ml.) were treated in ~ similar manner
to that of Example F-(3) to give ethyl 2-(2-aminothiazol-4-yl)-2-n-
hexyloxyiminoacetate (syn i~omer, 29.3 g.), mp 77 to 78 ~C.
I. R. L~maUxol : 3460, 3250, 3140, 1720, 1535 cm 1
N. M. R. ~ (DMS0-d6t : 0.85 (3H, t, J=6Hz), 1,0~1.9 (llH, m),
2.07 (2H, t, J=6Hz), 2,26 (2H, q, J=7Hz), 6.85(1H, e),
7.22(2H, 8)
(4) ~thyl 2-(Z-aminothiazol-4-yl)2-n-hexyloxy1~noacetate (syn
lsomsr, 29.1 g.), mèthanol (97.2 ml.), 2N aqueous solutlon oi sodium
hydroxide (97.2 ml.) and tetrahydrofuran (50 ml.) weru treated in a
- 78 -
- 1340~
8imilar manner to that o~ Example ~-(4) to give 2-(2- aminothlazol-4-
yl)-2-n-'~sxyloxylmlnoacetic acid (9yn i90mer, 24.0 g.), mp. 174~C (dec.).
I. R.~ mai~l : 1660, 1625, 1425 cm 1
N.M.R. ~ (DMSO-d6, ppm) 0.6~ 2.1 OE1H, m), 4.07 (2H, t, J=6Hz),
6.83 (lII, ~), 7.19 (2H, 9)
- 79 -
.__,. .
13~0~99
Example ~
(1) Ethyl 2-hydroxyimino-3-oxobutyrate (syn
isomer, 40 g.), N,N-dimethylformamide (200 ml.),
potassium carbonate (52 g.) and pentyl bromide (37.9
g.) were treated in a similar manner to that of
Example F -(1) to give ethyl 2-pentyloxyimino-3-
oxobutyrate (syn isomer, 57.5 g.), oil.
I.R. v maxm : 1745, 1680, 1470 cm 1
;
N.M.R. ~ (CCQ4, ppm): 0.7 - 2.2 (12H, m),
2.36 (3H, s), 4.1 - 4.6 (4H, m)
(2) Ethyl 2-pentyloxyimino-3-oxobutyrate (syn
isomer, 57.5 g.), acetic acid (58.5 ml.) and sulfuryl
- chloride (20.9 ml.) were treated in a similar manner
to that of Example F -(2) to give ethyl 2-
pentyloxyimino-4-chloro-3-oxobutyrate (syn isomer,
51.1 g.), oil.
I.R. v maxm : 1750, 1715, 1470 cm 1
N-M-R- ~ (CCQ4, ppm) : 0.7 - 2.1 (llH, m),
4.1 - 4.6 (4H, m), 4.48 (2H, s)
(3) Ethyl 2-pentyloxyimino-4-chloro-3-oxobutyrate
(syn isomer, 51.1 g.), thiourea (14.7 g.), sodium
acetate trihydrate (26.4 g.), ethanol (175 ml.) and
water (125 ml.) were treated in a similar manner to
that of Example F -(3) to give ethyl 2-(2-
aminothiazol-4-yl)-2-pentyloxyiminoacetate (syn
isomer, 28.7 g.), mp 86 to 88~C.
I.R. v mUax~l : 3450, 3250, 3130, 1715,
1535 cm 1
- 80 -
~ ~ 13~0~9~
..
N.M.R. ~ (DMSO-d6, ppm) : 0.6 - Z.0 (lZH,
m), 4.11 (2H, t, J=6Hz), 4.32 (2H,
q, J=7Hz), 6.90 (lH, s), 7.25 (2H, s)
(4) Ethyl 2-(2-aminothiazol-4-yl)-2-
pentyloxyiminoacetate (syn isomer, 28.6 g.), 2N aqueous
solution of sodium hydroxide (100.2 ml.), methanol
(100 ml.) and tetrahydrofuran (100 ml.) were treated
in a similar manner to that of Example F -(4) to
give 2-(2-aminothiazol-4-yl)-2-pentyloxyiminoacetic
acid (syn isomer, 22.4 g.), mp 176~C (dec.).
I.R. v maUxol : 3160, 1655, 1620, 1460 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 0.6 - 2.2 (9H, m),
4.07 (2H, t, J=6Hz), 6.82 (lH, s),
7.20 (2H, s)
(5) 2-(2-Aminothiazol-4-yl)-2-pentyloxyiminoacetic
acid (syn isomer, 15 g.), acetic anhydride (23.8 g.)
and formic acid (10.7 g.) were treated in a similar
manner to that of Example F -(5) to give 2-(2-
formamidothiazol-4-yl)-2-pentyloxyiminoacetic acid
(syn isomer, 14.7 g.), mp 125~C (dec.).
I.R. v maUxol : 3200, 3140, 1700, 1565 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 0.6 - 2.0 (9H, m),
4.13 (2H, t, J=6Hz), 7.53 (lH, s),
7.54 (lH, s), 12.66 (1ll, s)
- 81 -
. ., . ~ . , .
; 1340~9~
Example I.
(1) Allyl bromide (2.91 g.) was addcd dropwise to a stirred
suspensi,on of ethyl 2-(2-tritylaminothiazol-4-yl)-2-hydroxyimino-
ace~ate (syn isom~er, 10 g.), N,N-dimethylformamide (100 ml.)
and potassium carbonate (4.54 g.) under ice cooling over 5 minutes,
and stirred at the same tennperature for 4 hours. After adding
water (200 ml.) to the resultant solution, the solution was
extracted w1th diethyl ether twice. The extract was washed
with a saturated aqueous solution of sodium chloride and dried
over magnesium sulfate. The solution was concentrated in
vacuo, and the residue was triturated with a solution of
n-hexanc and diethyl ether. The precipitates were collected
by filtration to give ethyl 2-(2-tritylaminothiazol-4-yl)-2-
allyloxyiminoacetate (syn isomer, 9.4 g.), mp.-130 to 132~C.
I.R. v mUaxol : 3380, 1735, 1520, lS00 cm 1
N.M.R. ~(DMSO-d6, ppm) : 1.08 (3H, t, J~7Hz),
3.96 (2H, q, J~7Hz), 4.54 (2H,
broad d, J=SHz), 5.0~5.5 (2H, m),
5.6~6.3 (lH, m), 6.90 (1511,
broad s), 7.74 (lH, s)
(2) A solution of ethyl 2-(2-tritylaminothiazol-4-yl)-2-
allyloxyiminoacetate (syn isomer, 8.7 g.), 50% formic acid
(42.5 ml.) and tetrahydrofuran (42.5 ml.) was stirred at 60~C
for 40 minutes. After concentrating the resultant solution in
vacuo, the residue was dissolved in ethyl acetate, washed with
an aqueous solution of sodium bicarbonate and a saturated
aqueous solution of sodium chloride in turn, and dried over
magnesium sulfate. After concentrating the resultant solution
- 82 -
13404.9.~
in vacuo, the residue was subjected to column chromatography
on silica gel wit1l benzene and ethyl acetate in turn, to give
ethyl 2-(2-aminothiazol-4-yl)-2-allyloxyiminoacetate (syn isomer,
3.7 g.J, mp. 102 to 104~C.
I.R. v NlUajx~l : 3460, 3Z60, 3130, 1725, 1620,
1540, 1460 cm~l
N.M.R. ~ (DMSO-d6, ppm) : 1.25 (3H, t, J=7Hz),
4.30 (2H, q, J=7Hz), 4.61 (2H,
dd, J=5Hz, lHz), 5.0~5.5 (2H, m),
5.6~6.5 (lH, m), 6.95 (lH, s),
7.28 (2~{, s)
(3) A solution of ethyl 2-(2-aminothiazol-4-yl)-2-allyloxy-
iminoacetate (syn isomer, 3.6 g.), 2N aqueous solution of
sodium hydroxide (14.1 ml.), tetrahydrofuran (14.1 ml.) and
methanol (15 ml.) was stirred at 40~C for l.S hours. The
resultant solution was concentrated in vacuo, and the residue
was dissolved in water. After the solution was adjusted to
pH 2.8 with 10% hydrochloric acid under ice cooling, the
precipitates were collected by filtration, washed with water
and acetone in turn and dried to give 2-(2-aminothiazol-4-yl)-
2-allyloxyiminoacetic acid (syn isomer, 1.91 g.), mp. 187~C
(dec.).
I.R. ~ maUxol : 3350, 1630, 1580, 1460 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 4.61 (2H, d, J=6Hz),
5.1~5.5 (2H, m), 5.7~6.2 (lH, m),
6.84 (lH, s), 7.25 (2H, broad s)
Example M
(1) P ropargyl bromide (4.16 g.) was added to a suspension
of ethyl 2-(2-tritylaminothiazol-4-yl)-2-hydroxyiminoacetate
(syn isomer, 10 g.), potassium carbonate (4.84 g.) and
- 83 -
! 1 3 ~ 0 4
N,N-dimethylformamide (22 ml.) ullder atmosphere of nitrogen gas
and stirred at room temperature for 100 minutes. The insoluble
substance was filtered off and washed with a little of N,N-
dimethylformamide. The filtrate and washing solution were
combined together, and water (400 ml.) was added to the solution.
After the suspension was extracted with ethyl acetate (400
ml.), the extract was washed with a saturated aqueous solution
of sodium chloride and dried over magnesium sulfate. After
treating the solution with activated charcoal, the solution
was concentrated in vacuo. The residue was triturated with
diisopropyl ether. The precipitates were collected by
filtration, and washcd with diisopropyl ether to give ethyl
2-(2-tritylaminothiazol-4-yl)-2-propargyloxyiminoacetate (syn
isomer, 8.34 g.).
I.R. v NmaUxol : 3290, 2225, 1735 cm 1
N.M.R. ~(DMSO-d6, ppm) : 1.12 (3H, t, J= 7HZ),
3.47 (lH, t, J= ~Hz), 3.97 (2H,
q, J= 7Hz), 4.67 (2H, d, J= 3Hz)~ -
6.95 (lH, s), 7.26 (15H, s),
8.77 (lH, s)
(2) 50~ Formic acid (41 ml ) was added to a solution of
ethyl 2-(2-tritylaminothiazol-4-yl)-2-propargyloxyiminoacetate
(syn isomer, 8.2 g.) and tetrahydrofuran (41 ml.), and stirred
at 60~C for an hour. The resultant solution was concentrated
to a half of initial volume under reduced pressure, and the
precipitates were collected by filtration and washed with
diisopropyl ether. The filtrate and washing solution we~e
combined together and c~ncentrated in vacuo. The residue
~as added to ethyl acetate (200 ml.) under stirring. The
ihsoluble substance was collected by filtration, and washed
- 84 -
1340~9~
..
with diethyl ether to give ethyl Z-(2-aminotlliazol-4-yl)-2-
propargyloxyiminoacetate (syn isomer, 0.3 g.). The filtrate
and ethyl acetate washin~ solution were combined together,
washed with a saturated aqueous so]ution of sodium bicarbonate
- and a saturated aqueous solution of sodium chloride twice in
turn, and dried over magncsium sulfate. The solution was
trea~ed with activated charcoal and concentrated in vacuo.
The residue was dried in vacuo after adding benzene. The
residue was subjected to column chromatography on silica gel
with bcnzene and ethyl acetate in turn. The eluate was
concentrated in vacuo, and the residue was triturated with
diisopropyl ethcr. The precipitates were collected by
filtration, washed with diisopropyl ether to give the same
compound as mentioned above (syn isom,er, 2.658 g.).
The I.R. spectruol and N.M.R. spectrum are the same as those
of the compound obtained in E~ample I-(3).
- 85 -
,
1 ~ ' 1 3 ~ O
Example N
Sodium bicarbonate ~0.84 g.) was added to a
suspension of 2-(2-formamidothiazol-4-yl)oxalic
acid (2 g.) in water (120 ml.) to prepare a solution.
Ethyl 2-aminoxyacetate hydrochloride (4.56 g.) was
added to the solution and stirred at room temperature
for 3 hours while adjusting to pH 6 with sodium
bicarbonate. The resultant solution was adjusted to
pH 1.5 with hydrochloric acid, salted out and
extracted with ethyl acetate three times. The
extract was dried over magnesium sulfate and concent-
rated in vacuo. The residue was pulverized with
diethyl ether, and the precipitates were collected by
filtration and dried to give 2-(2-formamidothiazol-
4-yl)-2-ethoxycarbonylmethoxyiminoacetic acid (syn
isomer, 1.44 g.), mp 112~C (dec.).
I.R. v mUajxol : 3150, 1740, 1670, 1550 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 1.23 (3H, t,
J=7Hz), 4.16 (2H, q, J=7Hz),
4.77 (2H, s), 7.56 (lH, s), 8.54 (lH, s)
- 86 -
134~493
..
Example O
(1) Ethyl 2-hydroxyimino-3-oxobutyrate (syn isomer,
60 g.), 1-bromo-2-chloroethane (54.1 g.), potassium
carbonate (78 g.) and N,N-dimethylformamide (200 ml.)
were treated in a similar manner to that of Example
F-(l) to give ethyl 2-(2-chloroethoxyimino)-3-
oxobutyrate (syn isomer, 83.6 g.), oil.
I.R. v maxm : 1740, 1680, 1430 cm 1
N.M.R. ~ ~CCQ4, ppm) : 1.34 ~3H, t, J=7Hz),
2.34 ~3H, s), 3.72 ~2H, t, J=6Hz),
4.28 ~2H, q, J=7Hz), 4.46 ~2H, t, J=6Hz)
(2) Ethyl 2-(2-chloroethoxyimino)-3-oxobutyrate
(syn isomer, 83.6 g.), sulfury chloride (52.4 g.) and
acetic acid (83.6 ml.) were treated in a similar manner
to that of Example F-(2) to give Ethyl 2-(2-
chloroethoxyimino)-3-oxo-4-chlorobutyrate ~syn isomer,
68 g.), oil.
I.R. v maxm : 1740, 1720 cm 1
N.M.R. ~ ~CCQ4, ppm) : 1.32 ~3H, t, J=7Hz),
3.70 (2H, t, J=611z), 4.29 (2H, q,
J=7Hz), 4.47 (2H, s), 4.48 (2H, t, J=6Hz)
(3) Ethyl 2-(2-chloroethoxyimino)-3-oxo-4-
chlorobutyrate (syn isomer, 68 g.), thiourea (20.2 g.),
sodium acetate trihydrate (36.2 g.), ethanol (270 ml.)
and water (170 ml.) were treated in a similar manner
to that of Example F-(3) to give ethyl 2-~2-
aminothiazol-4-yl)-2-(2-chloroethoxyimino)acetate (syn
isomer, 33.7 g.), mp 126 to 128~C.
_ 87 -
13~99
I.R. v max~l : 3440, 3260, 3140, 1725,1620, 1540 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 1.30 (3H, t, J=7Hz),
3.78 (2H, t, J=6Hz), 4.1-4.6 (4H, m),
6.96 (lH, s), 7.27 (2H, s)
(4) Ethyl 2-(2-aminothiazol-4-yl)-2-(2-
chloroethoxyimino)acetate (syn isomer, 30.5 g.),
lN aqueous solution of sodium hydroxide (220 ml.),
methanol (110 ml.) and tetrahydrofuran (140 ml.) were
treated in a similar manner to that of Example F-(4)
to give 2-(2-aminothiazol-4-yl)-2-(2-chloroethoxyimino)-
acetic acid (syn isomer, 23.4 g.), mp 201~C (dec.).
I.R. v maUxol : 3210, 3100, 1640, 1620,
1580 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 3.83 (211, t, J=6Hz),
4.36 (2H, t, J=6Hz), 6.92 (lH, s),
7.30 (2H, s)
(5) 2-(2-Aminothiazol-4-yl)-2-(2-chloroethoxyimino)-
acetic acid (syn isomer, 15 g.), acetic anhydride
(24.5 g.), formic acid (11.0 g.) and tetrahydrofuran
(S0 ml.) were treated in a similar manner to that of
Example F-(5) to give 2-(2-formamidothiazol-4-yl)-2-
(2-chloroethoxyimino)acetic acid (syn isomer, 13.4 g.),
mp 155~C (dec.).
I.R. v NmaUxol : 3100, 1740, 1690, 1660 cm 1
N.M.R. ~(DMSO-d6, ppm) : 3.87 (2H, t, J=6Hz),.
4.40 (2H, t, J=6Hz), 7.60 (lH, s),
8.56 (lH, s), 12.62 (lH, broad s)
- 88 -
i .
13~0~ ~ )
Example P
A suspension of 2-(2-formamidothiazol-4-yl)oxalic
acid (3.0 g.) in methanol (60 ml.) and water (60 ml.)
was adjusted to pH 8 witll lN aqueous solution of
sodium hydroxide under stirring. 2,2,2-Trifluoroethoxyamine
hydrochloride (2.24 g.) was added to the solution, and
the solution was adjusted to pH 2.5 to 3 with lN
aqueous solution of sodium hydroxide. After the
solution was stirred at room temperature for 1.5 hours,
methanol was removed from the resultant solution under
reduced pressure. The concentrated aqueous solution
was adjusted to pH 7 with lN aqueous solution of sodium
hydroxide and washed with ethyl acetate. Ethyl acetate
was added to the aqueous solution and adjusted to pH 1.5
with 10~ hydrochloric acid, and then extracted with
ethyl acetate. The aqueous layer was extracted again
with ethyl acetate. The extracts were combined, washed
with a saturated aqueous solution of sodium chloride and
dried over magnesium sulfate. The solution was con-
centrated in vacuo to give 2-(2-formamidothiazol-4-yl)-
2-(2,2,Z-trifluoroethoxyimino)acetic acid (syn isomer,
2.4 g.), mp 162 to 163~C (dec.).
I.R. v NmaUxol : 3200, 1700, 1600, 1560 cm 1
N.M.R. & (DMS0-d6, ppm) : 4.83 (2H, q, J=8.5Hz),
7.65 (lH, s), 8.58 (lH, s), 12.60 (lH,
broad s)
Example Q
2-(2-Formamidothiazol-4-yl)oxalic acid (10 g.),
sodium bicarbonate (4.2 g.) and tert-butyl 2-aminooxyacetate
- 89 -
13'~0~93
.,
(8.1 g.) were treated in a similar manner to that of
Example N to give an oil. The oil was triturated with
n-hexane and the precipitates were collected by
filtration and dried to give 2-(2-formamidothiazol-4-
yl)-2-tert-butoxycarbonylmethoxyiminoacetic acid
(syn isomer, 11.3 g.), mp 117~C (dec.).
I.R. v NmUaxol : 3180, 3140, 1750, 1690,
1630 cm 1
N.M.R. ~ (DMS0-d6, ppm) : 1.46 (9H, s), 4.66
(2H, s), 7.56 (lH, s), 8.56 (lH, s),
12.67 (lH, broad s~
-- 90 --
. - , .
~ 13~0~99
Example 1
(i) N,N-Dimethylformamide (0.16 g.) and phosphorous
oxychloride (0.34 g.) were mixcd to prepare Vilsmeier
reagent in a usual manller, and the resultant Vilsmeier
reagent was suspcnded in dry ethyl acetate. To the sus-
pension was added 2-(2-formamido-4-thiazolyl)-2-
methoxyiminoacetic acid (syn isomer, 0.46 g.) under ice-
cooling with stirring, and then the solution was stirred
at the same temperature for 30 minutes to prepare the activated
acid solution. p-Nitrobenzyl 7-amino-3-chloro-3-
cephem-4-carboxylate hydrochloride (0.81 g.) was dissolved
in a solution of trimethylsilylacetamide (2.10 g.) in ethyl
acetate (200 ml.). To the solution was addcd the activated
acld solution obtained above all at once at -20~C, and
the solution was stirred a~ -20 --5~C for 1.5 hours.
After water and ethyl acetate (100 ml.) were added to the
resultant solution at -20~C, the insoluble product was
separated by filtration, washed with water and acetone in
turn and tllen dried to give p-nitrobenzyl 7-{2-(2-
formamido-4-thiazolyl)-2-metlloxyiminoacetamido}-3-ch1Oro-
3-cephem-4-carboxylate (syn isomer, 0.6 g.).
After ethyl acetate was removed from the above filtrate,
the aqueous layer was extracted with ethyl acetate (50 ml.)
twice. The ethyl acetate layer and the extract were com-
bined together, washed with 10% hydrochloric acid, a satu-
rated aqueous solution of sodium bicarbonate and a satu-
rated aqueous solution of sodium.chloride in turn and then
dried over magnesium sulfste. After removing ethyl acetate
~rom the solution, dietllyl ether was added to the residue.
_ 91 -
13~0493
The insoluble product was collected by filtration to
give the same object compound (0.25 g.), m.p. 226 to
228~C ~dec.), Total yield 0.85 g.
I.R. v NmUaxol : 3250, 1780, 1720, 1685, 1645,
1605, 1550, 1520 cm~l
N.M.R. ~ppm (DMSO-d6) : 3.45 (2H, broad s),
3.93 (3H, s), 5.35 (lH, d, J=5Hz),
5.50 (2H, s), 5.95 (lH, dd, J=5,8Hz),
7.43 (lH, s), 7.72 (2H, d, J=9Hz),
8.28 (211, d, J=9Hz), 8.55 (lH, s),
9.80 (lH, d, ~=8Hz)
' '
(2)p-Nitrobenzyl 7-12-(2-formamido-4-thiazolyl)-
2-methoxyiminoacetamido}-3-chloro-3-cephem-4-carboxylate
(syn isomer, 0.8 g.) ~ae dleeolved
in a mixed solution of methanol (30 ml.) and tetrahydrofuran
(60 ml.). After adding 10% palladium carbon (0.4 g.) to
~he solution, the mixture was subjected to catalytic
reduction at room temperature under atmospheric pressure.
The catalyst was filtered off, and the filtrate was con-
centrated under reduced pressure. Water (30 ml.) was
added to the residue and the mixture was adjusted to pH
7.5 with an aqueous solution of sodium bicarbonate.
After removing the insoluble substance from the mixture
by filtration, the filtrate was washed with ethyl acetate
(50 ml.). Ethyl acetate (70 ml.) was added to the
solution, and the mixture was adjusted to pH 1.5 with 10~
hydrochloric acid and then shaked sufficiently. After the
ethyl acetate layer was removed, the a~ueous layer was
- 92 -
~ 1340~99
extracted with ethyl acetate (30 ml.) twice. The ethyl
acetate layer and the extracts were combined together,
washed with a saturated aqueous solution of sodium chloride,
dried over magnesium sulfate and then concentrated under
reduced pressure to give 7~{2-(Z-formamido-4-thiazolyl)-
2-methoxyiminoacetamido}-3-chloro-3-cephem-4-carboxylic
acid (syn isomer 0.48 g.), m.p. 165 to 174~C (dec.).
I.R. v mUaxol : 3250, 1780, 1730, 1690, 1660,
1550 cm 1
N.M.R. ~ppm (DMSO-d6) : 3.57 (2H, broad s),
3.91 (3H, s), 5.30 (lH, d, J=5Hz),
5.88 (lH, dd, J=5, 8Hz), 7.44 (lH, s),
8.52 (lH, s), 9.78 (lH, d, J=8Hz),
lZ.60 (lH, s)
(~) 7-{2-(2-Formamido-4-thiazolyl)-2-methoxyimino-
acetamido}-3-chloro-3-cephem-4-carboxylic acid (syn isomer,
0.4 g.) wag suspended in methanol
(15 ml.). After adding conc-hydrochloric acid (0.16 g.)
to the suspension, the mixture was stirred at room tem-
perature for 2.5 hours. Methanol was distilled off from
the resultant mixture under reduced pressure, and the
residue was dissolved in water (15 ml.). The solution was
washed with ethyl acetate (30 ml.) and dichloromethane
(30 ml.) in turn. To the acqueous layer was introduced
nitrogen gas to remove the remaining organic solvent comp-
letely, and the solution was lyop~ilized to give 7-{2-(2-
amino-4-thiazolyl)-2-methoxyiminoacctamido}-3-chloro-3-
cephem-4-carboxylic acid hydrochloride (syn isomer 0.35 g.),
'~
- 93 -
. ~ .--. ~..
.:
13~0~93
.. ~
m.p. 170 to 180~C tdec.)
Il~n. v maUxol: 3300- 1780, 1730, 1670, 1630,
1545 cm 1
N.lVI.R. ~ppm (DMSO-d6): 3.88 (Zll, AB-q, J=17Hz),
3.94 (31-1, s), 5.26 ~111, d, J=5Hz),
5.80 (111, dd, J=5, 8Hz), 6.9Z (lH, s),
9.88 (lH, d, J=811z)
(4) 7-[2- (2-Amino-4-thiazolyl)-2-methoxyiminoacetamido]-
10 3-chloro-3-cephem-4-carboxylic acid hydrochloride ~syn-
isomer : 1.5 g.) and sodium bicarbonate (0.56 g.) were dis-
solved in water (50 ml.) at room 1:emperature with stirring and
lyophilized. A solution of iodomethyl hexalloate (0.93 g.) in
dimethylformamide (5 ml.) was added droywise to a solution
of the product obtained above in dimetllyl foTmamide (15 ml.)
at -5~C and stirred at the same tempcrature for 30 minutes.
-- 94 --
13~0~
Ethyl acetate (50 ml.) and water (100 ml.) were added to the
resultant solution and the etl~yl acetate layer was separated.
The aqueous layer was extracted with ethyl acetate (S0 ml.)
twice. The extracts wer~ combined with the ethyl acetate
layer, washed with a saturated aqueous solution of sodium
bicarbonate three times and a saturated aqueous solution of
sodium chloride three times in turn, dried over magnesium sul-
fate, treated with activated charcoal and then concentrated
under reduced pressure. After washing the concentrate with
10 n-hexane ~50 ml.), n-hexane (50 ml.) and diethylether (25 ml.)
were added to the residue and allowed to stand in a refrigerator
overnight. The precipitating powder was collected by filtrat-
ion, washed with n-hexane and dried to give a mixture (1.0 g.)
of n-hexanoyloxymethyl 7-[2-(2-amino-4-thiazolyl)-2-
methoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylate (syn-
isomer) and n-hexanoyloxymethyl 7-[2-(2-amino-4-thiazolyl)-2-
methoxyiminoacetamido~-3-chloro-2-cephem-4-carboxylate (syn-
isomer. )
(5) Thus obtained mixture (1.0 g.) was added to methylene
chloride (10 ml.) To the solution were added acetic acid(7 ml.), a solution of sodium tungstate (Na2W04 2H20) (20 mg.)
in water (0.5 ml.)j methylene chloride (5 ml.) and 35~ hydrogen
peroxide (180 mg.), and then stirred under ice-cooling for 4
hours. Ice-water was added to the resultant solution and
extracted with methylene chloride. The extract was washed with
water, dried over magnesium sulfate and concentrated under
reduced pressure. The residue was pulverized with diethyl ether
(20 ml.), washed with diethyl ether (10 ml.) twice and dried.
The product was purified with column chromatography on silica
gel (eluent : ethyl acetate) to give n-hexanoyloxymethyl 7-[2-
- g5 -
. .
1~0~
~2-amino-4-thiazolyl)-2-Jnethoxyiminoacetamido]-3-chloro-3-
cephem-4-carboxylate-1-oxide (syn-isomer : G00 mg.)
I.R. v mUaxol : 3300, 1790, 1760, 1680, lG30, 1540,
1380 cm 1
N.M.R. ~ppm (DMSO-d6) : 0.67 - 2.5 (llH, m),
3.90 (3H, s), 4.20 (2H, broad s),
5.17 (1ll, d~, 5.83 - 6.17 (3H, m),
6.88 (lH, s), 9.17 (lH, d)
(6) Phosphorus trichloride (210 mg.) was added to a
solution of n-hexanoyloxymethyl 7-[2-(2-amino-4-thiazolyl)-
2-methoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylate-l-
oxide (syn-isomer : 570 mg.) in dry dimethylformamide (10 ml.)
at -30~C, and stirred at -20 to -30~C for 50 minutes.
10~ Aqueous solution (50 ml.) of sodium chloride was added
to the resultant solution, adjusted to pH 8.0 with a saturated
aqueous solution of sodium bicarbonate and extracted with
ethyl acetate. The extract was washed with a saturated
aqueous solution of sodium bicarbgnate and a saturated
aqueous solution of sodium chloride, dried over magnesium
sulfate, and then concentrated under reduced pressure. The'
residue (560 mg.) was purified with column chromatography on
silica gel (20 g.) (eluent:ethyl acetate), and the resultant
residue (180 mg.) was pulverized with n-hexane (10 ml.) and
diethyl ether (5 ml.) to give n-hexanoyloxymethyl 7-[2-(2- .
amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-chloro-3-
cephem-4-carboxylate (syn-isomer : 150 mg.)
- 96 -
- .
' 1340~9~
.! 'I ~ ~ V max : 3400 (broad), 1780 ~broad), 1760 (shoulder),
1~70, 1620, 1530 cm 1
N.M.R. ~ppm (CDCQ3): 0.67 - 2.5 (1111, m), 3.67 (2H, q),
4.00 (311, s), 5.17(111, d), 5;90 (lH, s), 6 00 (1~1, m),
G.77 (lH, s), 7.83 (1~l, d)
Example 2
(1) 2-{2-(2,2,2-Trifluoroacetamido)-4-thiazolyl}-2-
methoxyiminoacetic acid (syn isomer, 0.65 g.) was added at
0~C to Vilsmeier reagent which had been prepared from
dimethylformamide and phosphorus oxychloride in ethyl acetate
(10 ml.), and the mixture was stirred at the same temperature
for 40 minutes to prepare the activsted ecid solution.
Thcscti~rated scid solution was added dropwise to a solution
of 7-amino-2,3-dimethyl-3-cephem-4-carboxylic acid (0.5 g.)
and trimethylsilylacetamide (1.73 g.) in ethyl acetate
(30 ml.) at -20~C, and the mixture was stirred at the same
20 temperature for 40 minutes. To the resultant mixture was
added water (10 ml.), and the ethyl acetate layer was
separated from the mixture and washed with water. Water
(30 ml.) was added to the solution and the mixture was
adjusted to pl~ 7.5 with sodium bicarbonate under ice-cooling.
After shaking the mixture, the aqueous layer was separated.
Ethyl acetate (50 ml.) was added to the aqueous solution,
and the mixture was adjusted to pll 2 with dilute hydrochloric
acid with stirring, and the ethyl acetate layer was separated,
washed with water and a saturated aqueous solution of sodium
30 chloride in turn, treated with acti~ated charcoal, dried
-- 97 --
13~0~9~
over magnesium sulfate and then concentrated under reduced
prcssure. The residue was pulverized with diisopropyl ether
ta give 7-[2-{2-(2,2,2-trifluoroacetamido)-4-thiazo~yl}-2-
methoxyiminoacetamido]-2,3-dimetllyl-3-ccpllem-4-carboxylic
acid ~syn isomer, 0.9 g.).
I.R. v maUxol : 3250, 1780, 1725, 1680, 1650 cm 1
N.~S.R. ~ppm (DMSO-d6) : 1.43 (3H, d, J=8Hz),
; 1,92 (lH, s), 3.82 (3H, s),
3.98 (lH, q, J=8Hz), 5.18 (lH, d,
J-6Hz), 5.73 (lH, AB-q, J=6Hz),
7-43 tll~, s), 9.63 (111, d, J=8Hz)
(2) 7-[2-{2-~2,2,2-Trifluoroacetamido)-4-thiazolyl}-
-2-methoxyiminoacetamido]-2,3-dimethyl-3-cephem-4-carboxylic
acid (syn isomer, 0.86 g.) wae dlo-
solved in an aqueous solution (g ml.) containing sodium
acetate trihydrate (2.3 g.), and the solution was stirred
at room temperature for 19 hours. After removing the
insoluble substance from the resultant mixture by filtration,
the filtrate was adjusted to around pH 2.5 with 10%
hydrochloric acid under ice-cooling. The precipitates
were collected by filtration, washcd with water and dried
to give 7-{2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido}-
2,3-dimethyl-3-cephem-4-carboxylic acid (syn isomer, 0.16 g.).
I.R. v nN~aUx~l : 3320, 3200, 1770 (shoulder),
1670, 1630 cm 1
N.M.R. ~ppm (DMSO-d6) : 1.44 (3H, d, J=7Hz),
1.98 (3H, s), 3.57 (lH, q, J=7Hz),
3.82 (3H, s), 5.18 (lH, d, J=5Hz),
5.73 (lH, dd, J=5, 8Hz), 6.76 (lH, s),
9.63 (lH, d, J=811z)
- 98 -
-. i . ...
1340~.~.9
EY~am~le 3
(1) A mixture of 2-{2-(2,2,2-trifluoroacetamido)-4-
thiazolyl}-2-methoxyiminoacetic acid (syn isomer, 0.8 g.),
dimethylformamide (0.20 g.), phosphorus oxychloride (0.41
g.) and ethyl acetate (10 ml.) was stirred for 30 minutes
under ice-cooling to prepare the activated acid solution
in a similar manner to that of Example 2-(1). On the
other hand, a solution of 7-amino-3-methoxy-3-cephem-4-
carboxylic acid hydrochloride (0.6 g.) and
trimethylsilylacetamide (3 g.) in ethyl acetate (15 ml.)
was stirred at 40~C for 3 hours. To the solution was
added dropwise the activated acid solution at -10 to -20~C
in 2 minutes, and the mixture was stirred at the same
temperature for 1.5 hours. After water (10 ml.) was added
to the resultant mixture, the ethyl acetate layer was
separated and allowed to stand. The precipitates were
collected by filtration to give 7-[2-{2-(2,2,2-trifluoro-
acetamido)-4-thiazolyl}-2-methoxy-iminoacetamido]-3-
methoxy-3-cephem-4-carboxylic acid (syn isomer, 0.3 g.).
The aqueous layer, which was separated from the ethyl
acetate layer, was extracted with ethyl acetate, and the
extract was combined with the mother liquor obtained
above. The ethyl acetate solution was washed with a
saturated aqueous solution of sodium chloride, dried over
magnesiumsulfate and then filtered. The filtrate was
concentrated under reduced pressure, and the residue was
washed with diethyl ether to give the same object compound
(0.25 g.), Total yield 0.55 g.
I.R. v ~ : 3230, 1770, 1715, 1650, 1580 cm
99 _
.
~3~0~9~
N-M-R- ~pplll (DMSO-d6) : 3.62 ~211, AB-q, J=1611z),
3.78 ~3H, s), 3.93 (3H, s),
5.17 (lH, d, J=4Hz), 5.54 ~lH, dd,
J=8, 4Hz), 7.60 ~lH, s),
9.69 (lH, d, J=8Hz)
(2) A solution of 7-[2-{2-(2,2,2-trifluoroacetamido)-
4-thiazolyl}-2-methoxyiminoacetamido]-3-methoxy-3-cephem-
4-carboxylic acid ~syn isomer, 0.55 g.)
and sodium acetate trihydrate (1,76 g.) ln
- ethyl acetate ~3 ml.), tetrahydrofuran ~3 ml.) and water
~5.5 ml.) was stirred at room temperature overnight. The
aqueous layer was separated ~rom the resultant mixture,
washed with dichloromethane, and then evaporated under
reduced pressure to remove the organic solvent. The
aqueous solution was adjusted to pH 4.2 under ice-cooling,
and subjected to column chromatography on Diaion HP-20
resin ~Trade mark : manufactured by Mitsubishi Chemical
Industries Ltd., 15 ml.). After washing the column with
waterJ the object compound was eluted with 20~ aqueous
isopropyl alcohol. The eluate was concentrated under
' reduced pressure and the residue was lyophilized to give
7-{2-~2-amino-4-thiazolyl)-2-methoxyiminoacctamido}-3-
methoxy-3-cephem-4-carboxylic acid ~syn isomer 0.4 g.),
m.p. 185 to 190~C (dec.).
I.R. v maUxol : 3300, 1770, 1660, 1630, 1540 cm 1
N.M.R. ~ppm ~DMS0-d6) : 3.60 (2H, broad s),
3.75 (3H, s), 3.83 (3H, s), 5.12
(lH, d, J-4llz), 5.55 ~lH, dd, J=4,8Hz),
6.82 ~lH, s), 9.52 ~lH, d, J=8Hz)
-- 100 --
13~0 ~9~
Example 4
(1) To a suspension of p-nitrobenzyl 7-phenylacetamido-
3-cephem-4-carboxylate (lP.50 g.) in dry dichloromethane
(100 ml.) was added dry pyridine (Z.14 g.). Phosphorus
pentachloride (5.50 g.) was added to the solu~ion at -10~C,
and the mixture was stirred at -5~C for 45 minutes and
further at 10~C for an hour. After adding mcthanol (520
g.) to the resultant mixture, the mixture was stirred at
-20~C for 1.5 hours. The precipitates were collected by
filtration, washed with dichlorometllane (120 ml.) and
diethyl ether (130 ml.) in turn, and then dried to give
p-nitrobenzyl 7-amino-3-cephem-4-carboxylate (7.90 g.),
m.p. 182~C (dec.).
I.r~ lauxol : 1790, 1730, 1638, 1600 cm 1
N-M-R- ~ppln (DMSO-d6) : 3.78 (2H, d, J=4Hz),
5.27 (2H, dd, J=SHz), 5.44 (2H, s),
6.78 (lH, t, J=4H2), 7.72 (2H, d,
J=9Hz), 8.26 (2H, d, J-9Hz)
-- 101 --
13~0 199
) Vilsmeier reagent prepared from dimethylformamide
(0.43 g.) and phosphorus oxychloride (0.92 g.) was suspended
in dry ethyl acetate (10 ml.). To the suspension was
added 2-(2-formamido-4-thiazolyl)-2-methoxyimino acetic
acid (syn isomer, 1.15 g.) under ice-cooling with stirring,
and the mixture was stirred at the same temperature for 30
j minutes to prepare the actlvated acidsolution. On the
- other hand, p-nitrobenzyl 7-amino-3-cephem-4-carboxylate
hydrochloride (1.79 g.) and trimethylsilylacetamide (5.0 g.)
were dissolved in ethyl acetate (40 ml.). To the solution
was added the actlvated acidsolution at -20~C all at once,
and the mixture was stirred at the same temperature for 2.5
hours. Water (60 ml.) and ethyl acetate (200 ml.) were added
to the resultant solution, and the ethyl acetate layer was
separated, washed with 10% hydrochloric acid (60 ml.), a
saturated aqueous solution of sodium bicarbonate (60 ml.)
and an aqueous solution of sodium chloride (50 ml.) in turn,
dried over magnesium sulfate, treated with activated charcoal,
and then evaporated under reduced pressure. Diethyl ether
was added to the residue, and the precipitates were collected
by filtration to give p-nitrobenzyl 7-{2-(2-formamido-4-
thiazolyl)-2-methoxyimino-acetamido}-3-cephem-4-carboxylate
(syn isomer, 1.30 g.), m.p. 210 to 212~C (dec.).
I.R. v mUax~l : 3240, 1780, 1730, 1690, 1655
1605, 1550, 1520 cm 1
N.M.R. ~ppm (DMSO-d6) : 3.65 (2H, broad s),
3.90 (3H, s), 5.20 (lH, d, J~5Hz),
5.43 (2H, s), 5.95 tlH, q, J~5,8Hz),
- 102 -
,
13~0~99
6.68 (lH, t, J=411z), 7.42 ~ 1, s),
7.72 (2H, d, J=9Hz), 8.28 (2H, d,
J=9Hz), 8.46 (lH, s), 9.72 (lH, d,
J=8Hz)
(~) To a solution of p-nitrobenzyl 7-{2-(2-formamido-4-
thiazolyl)-2-methoxyiminoacetamido}-3-cephem-4-carboxylate
(syn isomer, 1.25 g.) in
methanol (40 ml.) and tetrahydrofuran (50 ml.) was added
10% palladium carbon (0.65 g.), and the mixture was subjected
to catalytic reduction at room temperature under atmospheric
pressure for 3.5 hours. After removing the catalyst from
the reaction mixture, the filtrate was concentrated under
reduced pressure. Water (80 ml.) was added to the residue,
and the mixture was adjusted to pH 7.5 with an aqueous solution
of sodium bicarbonate, and then the insoluble substance
was filtered off. The filtrate was washed with ethyl
acetate (50 ml.), and then ethyl acetate (100 ml.) was
added to the solution. After adjusting to pH 1.5 with 10%
hydrochloric acid, the ethyl acetate layer was separated.
The remaining aqueous layer was extracted with ethyl acetate
(80 ml.) twice, and the extracts were combined with the
ethyl acetate layer obtained above, washed with an aqueous
solution of sodium chloride, dried over magnesium sulfate,
and then concentrated under reduced pressure to give 7-{2-t2-
formamido-4-thiazolyl)-2-methoxyiminoacetamido}-3-cephem-4-
carboxylic acid (syn isomer, 0.60 g.), m.p. 176 to 183~C
(dec.).
- 103 -
..
13~0~99
,
I.R. v max~l: 3250, 1780, 1690, 1660, 1550 cm 1
N.M.R. ~ppm (DMSO-d6) : 3.63 (ZH, d, J=4Hz),
3.93 (3H, s), 5.10 (lH, d, J=5Hz),
5.90 (lH, q, J=5,8Hz), 6.53 (lH, t,
J=4Hz), 7.47 (lH, s), 8.57 (lH, s),
9.70 (lH, d, J=8Hz), 12.63 (lH, s)
(4) 7-{2-(2-Formamido-4-thiazolyl)-2-methoxyiminoacet-
amido}-3-cephem-4-carboxylic acid (syn isomer, 95 mg.)
was euspended ln methanol (4 ml.).
To the suspension was added conc. hydrochloric acid (110 mg.)
and the solution was stirred at room temperature for 4 hours.
After distilling methanol under reduced pressure, the
residue was dissolved in water (30 ml.) and the aqueous
solution was washed with ethyl acetate (10 ml.) and dichloro-
methane (15 ml.) in turn. Nitrogen gas was introduced
into the aqueous solution to exclude the remaining organic
solvent, and the aqueous solution was lyophilized to give
7-{2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido}-3-
cephem-4-carboxylic acid hydrochloride (syn isomer, 83 mg.),
m.p. 180 to 190~C (dec.).
I.R. v maUxol : 3300, 1770, 1710, 1660, 1630 cm 1
N.M.R. ~ppm (DMSO-d6) : 3.64 (2H, broad s), 3.95
(3H, s), 5.14 (lH, d, J=5Hz), 5.82
(lH, t, J=4Hz), 6.95 (lH, s), 9.80
(lH, d, J=8Hz)
- 104 -
13~0~93
(5) The solution of 7-f2-(2-formamido-4-thiazolyl)-2-
methoxyiminoacetamido~-3-cephem-4-carboxylic acid (syn
isomer 10.8 g.), conc. hydrochlorie acid (11 g.) and
methanol (350 ml.) was stirred at room temperature for
4 hours. After concentrating the resultant solution under
reduced pressure, ethyl acetate was added to the residue.
The solution was adjusted to pH 8.0 with a saturated
aqueous solution of sodium bicarbonate and the aqueous
layer was separated and washed with diethyl ether.
After nitrogen gas was bubbled in the aqueous solution,
the aqueous solution was adjusted to pH 4.0 with 10% hydro-
ehlorie aeid. The preeipitates were eolleeted by filtration
and washed with water to give 7-(2(2-amino-4-thiazolyl)-2-
methoxyiminoaeetamido~-3-cephem-4-earboxylic acid (syn
isomer, 8.2 g.), m.p.~ 290~C.
IR ~ NUaxol : 3470, 3280, 3200, 1780, 1695, 1655,
1622 em
~MR d ppm (DMSO-d6) : 3.60 (2H, broad s), 3.84
(3H, s), 5.12 (lH, dd, J=5Hz), 5.8a
~1~1, dd, J-5,8Hz), 6.52 (lH, broad t)
6.76 (lH, s), 7.26 (2H, broad s),
9.65 (lH, d, J=8Hz)
.
- 105 -
.. .... ~
1340493
.
(6) Sodium bicarbonate (1.04 g.) was added to a solution of
7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-
4-carboxylic acid hydrochloride (syn isomer, 2.6 g.) in water
(100 ml.) under ice-cooling and stirred at room temperature.
The resultant solution was lyophilized to give sodium 7-[~-
(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-
carboxylate (syn isomer).
I.R. ~ max~l : 3100, 1760, 1650, 1590, 1530 cm 1
N.M.R. ~ (D2O, ppm) 3.60 (2H, broad q), 4.00 (3H, s),
5.22 (lH, d), 5.88 (lH, d), 6.35 (lH, q), 7.03 (lH, s)
(7) The product obtained above was dissolved in dry
N,N-dimethylformamide (20 ml.). To the solution was dropwise
added a solution of iodomethyl n-hexanoate (1.33 g.) and dry
l~,N-dimethylformamide (5 ml.) at -40~C over 5 minutes, and
then stirred at the same temperature for 40 minutes and then
under ice-cooling for 45 minutes. The resultant solution
was added to a mixed solution of ethyl acetate (60 ml.) and
water (125 ml.). The ethyl acetate layer was separated, washed
with a saturated sodium bicarbonate aqueous solution and a
staturated sodium chloride aqueous solution in turn dried over
magnesium sulfate, and then treated with an activated charcoal.
After removing ethyl acetate from the solution, the residue was
triturated with diethyl ether to give n-hexanoyloxymethyl 7-
~ -(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-
carboxylate (syn isomer, 750 mg.)
~ maUxol : 3170, 1780, 1750 (shoulder), 1670, 1630, 1530 cm
N.M.R. ~ (CDC13, ppm) 0.68-1.84 (9H, m), 2.20-2.48 (2H, t),
3.20-3.80 (2H, m), 4.02 (3H, s), 5.04 (lH, d),
5.60-6.20 (3H, m), 6.62 (1~, q), 6.80 (lH, s),
7.72 (lH, d)
- 106 -
.. .. ... . .
.
134049~
(8) p-Nitrobenzyl 7-[2-(2-formamido-4-thiazolyl)-2-
methoxyiminoacetamido]-3-cephem-4-carboxylate (syn-isomer :
1.1 g.) was suspended in a mixture of ethanol (10 ml.) and
water (15 ml.) lN Aqueous solution of potassium hydroxide
(6 ml.) was added dropwise to the suspension at 5 to 7~C
over 10 minutes and stirred for 10 minutes. The resultant
solution was adjusted to pH 7.5 with 10% hydrochloric acid,
washed with ethyl acetate and adjusted to pH 2.5 with 10%
hydrochloric acid. The precipitating crystals were
collected by filtration to give the mixture of 7-[2-(2-
formamido-4-thiazolyl)-2-methoxyimino-acetamido]-3-cephem-
4-carboxylic acid (syn-isomer : 0.32 g.) and 7-{2-(2-amino-
4-thiazolyl)-2- methoxyiminoacetamido}-3-cephem-4-
carboxylic acid (syn-isomer : 0.035 g.)
(9) 7-[2-(2-Amino-4-thiazolyl)-2-methoxyimino-
acetamido]-3-cephem-4-carboxylic acid (syn-isomer : 5 g.)
was gradually added to an aqueous solution (30 ml.) of
sodium bicarbonate (1.04 g.) at 35 to 40~C, and stirred at
20 50 to 53~C for 30 minutes. After removing the insoluble
substance from the resultant solution, the filtrate was
treated with activated charcoal (0.3 g.), and filtered.
The filtrated was lyophilized to give sodium 7-[2-(2-
amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-
carboxylate (syn-isomer : 4.2 g.)
max : 3300-3100, 1760, 1670, 1595, 1530 cm~
N.M.R. ~ppm (DMSO-d6) : 3.50 (2H, broad s),
3.83 (3H, s), 5.00 (lH, d, J=5Hz)
5.68 (lH~ dd, J=5Hz~ 8Hz) ~ 6.13 (lH~ broad s),
6.73 (lH, s),7.3 (2H, broad s), 9.60 (lH, d,
J=8Hz)
- 107 -
,~.~ .
A.
. . , ~
134049~
(lO) 7-[2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-
3-cephe~-4-carboxylic acid ~syn isomer, l.lS g.) was
added to an aqueous solution of calcium hydroxide
(0.111 g.) in water (100 ml.), and the solution was
stirred at room temperature for 10 minutes. After the
solution was filtered, the filtrate was lyophilized to
give calcium 7-[2-(2-aminothiazol-4-yl)-2-
methoxyiminoacetamido]-3-cephem-4-carboxylate ~syn
isomer, 1.2 g.).
I.R. v mUaxol : 3350, 1760, 1670, lS90,
1535, 1465cm 1
N.M.R. ~ ~D2O, ppm) : 3.51 ~lH, d, J=SHz),
3.59 (lH, d, J=3Hz), 3.97 (3H, s),
5.15 (lH, d, J=5Hz), 5.82 ~lH, d,
J=5Hz), 6.33 (lH, dd, J=SHz, 3Hz),
6.95 (lH, s)
(11) 7-[2-(2-Aminothiazol-4-yl)-Z-methoxyiminoacetamido]-
3-cephem-4-carboxylic acid ~syn isomer, 1.15 g.) was
added to a suspension of magnesium hydroxide (0.088 g.)
in water (100 ml.), and the mixture was stirred at
70~C for 30 minutes to give a solution. After the
resultant solution was filtered, the filtrate was
lyophilized to give magnesium 7-[2-(2-aminothiazol-4-
yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate
(syn isomer, 1.1 g.).
I.R. v mUaxol : 3350, 1760, 1660, 1610,
1530, 1460cm 1
- 108 -
1340~99
:...
N.M.R. ~ (D2O, ppm) : 3.53 (lH, d, J=SHz),
3.59 (lH, d, J=3Hz), 3.96 (3H, s),
5.16 (lH, d, J=5Hz), 5.84 (lH, d,
J=5Hz), 6.32 (lH, dd, J=5Hz, 3Hz),
7.98 (lH, s)
(12) 7-[2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-
3-cephem-4-carboxylic acid (syn isomer, 1.15 g.) was
added to a solution of arginine (0.523 g.) in water
(50 ml.), and the solution was stirred at room temperature
for 10 minutes.
After the resultant mixture was filtered, the filtrate
was lyophilized to give an arginine salt of 7-[2-(2-
aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-
4-carboxylic acid (syn isomer, 1.35 g.).
I.R. v maUxol : 3350, 3150, 1770 1650 (broad),
1580, 1530, 1460cm 1
N.M.R. ~ (D2O, ppm) : 1.4 - 2.1 (4H, m),
3.22 (2H, t, J=6Hz), 3.55 (lH, d,
J=6Hz), 3.65 (lH, d, J=3Hz),
3.82 (lH, d, J=6Hz), 3.97 (3H, s),
5.18 (lH, d, J=5Hz), 5.85 (lH, d,
J=5Hz), 6.33 (lH, dd, J=6Hz; 3Hz),
7.00 (lH, s)
~13) Sodium 7-[2-(2-aminothiazol-4-yl)-2-
methoxyiminoacetamido]-3-cephem-4-carboxylate (syn
isomer, 1.21 g.) was added to a solution of lysine
hydrochloride (0.55 g.) in water (12 ml.).
-- 109 --
13 1049.~3
The solution was lyophilized to give a lysine salt of 7-
[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-
cephem-4-carboxylic acid (syn isomer, 1.6 g.).
I R Nujol : 3350 3150, I770, 1600 (broad),
max
1530, 1460 cm1
N.M.R. ~(D2O, ppm) : 1.3 - 2.1 (6H, m),
3.03 (2H, t, J=7Hz), 3.54 (lH, d,
J=5Hz), 3.64 (lH, d, J=3Hz), 3.80 (lH,
d, J=6Hz), 3.97 (3H, s), 5.17 (lH, d,
J=5Hz), 5.84 (lH, d, J=5Hz), 6.32 (lH,
dd, J=5Hz, 3Hz), 6.99 (lH, s)
(14) 20% Aqueous solution of sodium hydroxide was
added to a suspension of 7-[2-(2-aminothiazol-4-yl)-2-
methoxyiminoacetamido]-3-cephem-4-carboxylic acid (syn
isomer, 15 g.) in a mixture of ethanol (8 ml.) and water
(8 ml.) at room temperature to make a solution of pH 7.5.
After filtration and washing, the filtrate and washings
were combined (which contained 18.3 ml. of water) and
added dropwise to ethanol (46 ml.) at 20 to 25~C under
stirring and stirred at the same temperature for 30
minutes. Ethanol (28 ml.) was added dropwise to the
mixture over 30 minutes, and stirred at the same
temperature for 2 hours. The precipitates were collected
by filtration, washed with ethanol (20 ml.) and dried in
vacuo at room temperature to give plates of sodium 7-[2-
(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-
carboxylate dihydrate (syn isomer, 13.5 g.) mp 260~C(dec.).
- 110 -
,. ,~
~ t'
. .
1340493
I.R. v maUxol : 3430, 3250, 1760 (shoulder),
1745, 1650, 1630 ~shoulder),
1590, 1540 cm 1
(l5? Sodium 7-[Z-(2-aminothiazol-4-yl)-2-
methoxyiminoacetamido]-3-cephem-4-carboxylate (syn
isomer, 15 g.) was dissolved in water (13 ml.) at 35 to
45~C under stirring. Warmed ethanol (52 ml., 30~C) was
added dropwise to the stirred solution, and stirred at
jthe same temperature for 5 minutes and then at room
temperature for 2 hours. The precipitates were collected
by filtration, washed with ethanol and dried under
reduced pressure to give plates of sodium 7-[2-(2-
aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-
carboxylate dihydrate (syn isomer, 13.45 g.).
(16) 4N Aqueous solution of sodium hydroxide was
carefully added dropwise to a stirred suspension of
7-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-
3-cephem-4-carboxylic acid ~syn isomer, 52 g.) in water
(100 ml.) below 5~C to make a solution of pH 7.0 to 7.5.
After filtration and washing, the combined filtrate and
the washings (200 ml.) was added dropwise to ethanol
~2 Q.) under stirring over 30 minutes, and stirred at
room temperature for 15 minutes and then at 5 to 10~C
for an hour. The precipitates were collected by
filtration, washed with ethanol (200 ml.) and dried in
vacuo at 30~C to give amorphous sodium 7-[2-(2-
aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-
4-carboxylate (syn isomer, 46.3 g.).
I.R. ~ maUxol : 3400, 3300, 3170, 1750,
1650, 1580 cm 1
~ ... .
1340~93
(17) A suspension of sodium 7-[2-(2-aminothiazol-4-
yl)-2-methoxyiminoacetamdio]-3-cephem-4-carboxylate (syn
isomer, 10 g.) in methanol (250 ml.) was treated with a
supersonic apparatus to make a clear solution. The
solution was allowed to stand at room temperature, and
then stirred at the same temperature for 3 hours. The
precipitates were collected by filtration and washed with
methanol to give amorphous sodium 7-[2-(2-aminothiazol-4-
yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate (syn
isomer).
(18) The crystals obtained in the above Example
4-(14) were dried over P2Os in vacuo for one day at room
temperature to give another plates of sodium 7-[2-(2-
aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-
carboxylate (syn isomer).
- 112 -
13~0~93
~xample 5
(l) Vilsmeier reagent prepared from dimethylformamide
(0.22 g.) and phosphorus oxychloride (0.46 g.) was suspended
in dry ethyl acetate (20 ml.). 2-(2-Formamido-4-thiazolyl)-
2-methoxyiminoacetic acid (anti isomer, 0.62 g.) was added
to the suspension under ice-cooling with stirring, and the
mixture was stirred at the same temperature for 30 minutes
- ito prepare the activated acidsolution. The solution was
added all at once to a solution of p-nitrobenzyl 7-amino-3-
chloro-3-cephem-4-carboxylate (1 g.) and trimethylsilyl-
acetamide (2.58 g.) in ethyl acetate (20 ml.) at -20~C
witll stirring, and the mixed solution was stirred at -10
- to -20~C for 1.5 hours. To the resultant solution was
- added water (20 ml.), and the solution was stirred at -20~C.
After separating the ethyl acetate layer, the aqueous layer
was extracted with ethyl acetate (20 ml.). The ethyl
acetate layer and the extract were combined togetller,
washed with 10% hydrochlorlc acid (20 ml.) twice, water
~20 ml.) once, a 5~ aqueous solution of sodium bicarbonate
(20 ml.) three times, and an aqueous solution of sodium
chloride (20 ml.) once in turn, dried, and then concentrated
under reduced pressure. The residue was washed with
diethyl ether (50 ml.) to give p-nitrobenzyl 7-{2-(2-
formamido-4-thiazolyl)-2-methoxyiminoacetamido} - 3-chloro-3-
cephem-4-carboxylate (anti isomer, 1.27 g.), m.p. 135 to
145~C. (dec.).
I.R. v NmUaiXol : 3150 to 3300 tbroad), 1780, 1730,
1670 to 1690 (broad) cm 1
- 113 -
. ~ , ~ - . . . .... . .
13~0~99
N.M.R. ~ppm (DMSO-d6) : 3.83 ~2H, AB-q, J=17Hz),
3.97 (3H, s), 5.23 (lH, d, J=5Hz),
5.41 (2H, s), 5.9 (lH, dd J=5,8Hz),
7.62 (2H, d, J=8Hz), 8.0 (lH, s)
8.2 (2H, d, J=8tlz), 8.42 (lH, s),
9.55 (lH, d, J~8Hz), 12.43 (lH, s)
'(2) 10% Palladium carbon (0.6 g.) was added to a
solution of p-nitrobenzyl 7-{2-(2-formamido-4-thiazolyl)-2-
methoxyiminoacetamido}-3-chloro-3-cephem-4-carboxylate
(anti isomer, 1.16 g.)
in methanol (20 ml.) and tetrahydrofuran (40 ml.), and the
mixture was subjected to catalytic reduction at room
tcmperature under atmospheric pressure for 5 hours.
After removing the catalyst from the reaction mixture, the
filtrate was concentrated under reduced pressure. Water
(30 ml.) and ethyl acetate (60 ml.) were added to the
residue, and the mixture was adjusted to pH 7.S with an
aqueous solution of sodium bicarbonate and shaked sufficiently.
The aqueous layer was separated and ethyl acetate (90 ml.)
was added to the aqueous solution. The aqueous layer was
adjusted to pH 2.5 with 10% hydrochloric acid with stirring
under ice-cooling, and the ethyl acetate layer was separated.
The remaining aqueous layer was extracted with ethyl acetate
(30 ml.), and the extract and the ethyl acetate layer were
combined together, washed with an aqueous solution of sQdium
chloride, dried and then concentr~ted under reduced pressure.
The residue was washed with diisopropyl ether to give 7-{2-
-- (2-formamido-4-thiazolyl)-2-methoxyiminoacetamido}-3-chloro-
- 114 -
'~ :"'':' - -
13~0499
.
3-cephem-4-carboxylic acid (anti isomer, 0.47 g.). The
compound was colored at 210~C and decomposed at above than
250~G.
I.R. v mUaxol: 3250, 1780, 1720 (shoulder), 1670
to 1690 cm 1
N-M-R. ~ppm (DMSO-d6) : 3.8 (2H, AB-q, J=17Hz),
4.0 (3H, s), 5.21 (lH, d, Js5Hz),
5.83 (lH, dd, J=5,8Hz), 8.05 (lH, s),
8.47 (lH, s), 9.55 (lH, d, J=8Hz),
12.55 (lH, broad s)
(3) 7-{2-(2-Formamido-4-thiazolyl)-2-methoxyimino-
acetamido}-3-chloro-3-cephem-4-carboxylic acid (anti isomer,
0.4 g.) was suspended in methanol
(15 ml.). Conc. hydrochloric acid (0.16 g.) was added to
the suspension and the mixture was stirred at room temperature
for 5 hours. The precipitates were collected by filtration,
washed with a mixed solvent of methanol and diethyl ether
(1:1), and dried to give 7-{2-(2-amino-4-thiazolyl)-2-
methoxyiminoacetamido}-3-chloro-3-cephem-4-carboxylic acid
hydrochloride (anti isomer, 0.31 g.), m.p. above than 250~C.
I.R. v NmaUxol : 3250, 3200, 1788, 1720, 1680,
1640 cm 1
N.M.R. ~ppm (DMS0-d6) : 3.81 (2H, AB-q, J=17Hz),
4.08 (3H, s), 5.22 (lH, d, J=5Hz),
5.7 (lH, dd, J=5,8 Hz), 7.59 (lH, s),
9.5 (lH, d, J=8Hz)
- 115 -
1340493
~xample 6
(1) A solution of Vilsmeier reagent was prepared from
dry dimethylformamide (0.39 g.), dry ethyl acetate (1.2 ml.)
and phosphorus oxychloride (0.84 g.) in a usual manner.
To the solution was added a solution of 2-~1,2,3-thiadiazol-
4-yl)-2-methoxyiminoacetic acid ~syn isomer, 0.93 g.) in
ethyl acetate ~10 ml.) at -15~C to prepare the nctlvated
acid solution. On the other hand, a p-nitrobenzyl
7-amino-3-eephem -4-carboxylate ~1.5 g.), trimethylsilyl-
acetamide ~4.6 g.) and bis~trimethylsilyl)acetamide ~1 ml.)
in dry ethyl acetate ~50 ml.) was stirred at 45~C for 5
hours to give a solution. To the solution was added
all at once the activated acid solution obtained above at
-10~C with stirring, and the mixed solution was stirred at
-5~C for l.S hours. Water was added to the reaction
mixture, and the insoluble product was separated by
filtration, washed with ethyl acetate and water in turn,
and then dried to give pale yellow powder of p-nitrobenzyl
7-{2-(1,2,3-thiadiazol-4-yl)-2-methoxyiminoacetamido}-3-
cephem-4-carboxylate ~syn isomer, 1.9 g.), m.p. 243 to 245~C
(dec.).
I.R. ~ mUajxol : 3250, 1782, 1725, 1655, 1630,
1600, 1520, 1345 cm 1
N.M.R. ~ppnl ~DMSO-d6) : 3.69 ~2H, AB-q, J=14Hz),
4.00 ~3H, s), 5.24 ~lH, d, J=SHz),
5.46 ~2H, s), 6.00 ~lli, dd, J=5,8Hz),
6.68 ~lH, ~, J=41iz), 7.7 to 8.4 ~4H,
m), 9.44 ~lH, s), 9.88 ~lH, d, J=8Hz)
- 116 -
~' 1340~3
(2) 10% Palladium carbon (0.85 g.) was added to a
solution of p-nitrobenzyl 7-{2-(1,2,3-thiadiazol-4-yl)-2-
methoxyiminoacetamido}-3-cephem-4-carboxylate (srn lsomer, 1.65 g.)
in methanol (70 ml.) and
tetrahydrofuran (90 ml.), and the mixture was subjected to
catalytic reduction at room temperature under atmospheric
pressure for 3.5 hours. After removing the catalyst from
the reaction mixture by filtration, the filtrate was
concentrated under reduced pressure. Water was added to
the residue, and the mixture was adjusted to pH 7 to 8 with
sodium bicarbonate, washed with ethyl acetate, adjusted to
pH 1.5 with 10% hydrochloric acid, and then extracted with
ethyl acetate. The extract was washed with a saturated
aqueous solution of sodium chloride, dried over magnesium
sulfate, and then filtered. The filtrate was concentrated
under reduced pressure, and the residue was pulverized with
diethyl ether. The precipitates were collected by ~iltration
and the dried to give yellow powder of 7-{2-(1,2,3-thiadiazol-
4-yl)-2-methoxyiminoacetamido}-3-cephem-4-carboxylic acid
(syn isomer, 0.3 g.), m.p. 200 to 210~C (dec.).
I.R. v NmaUxol : 3250, 2550 to 2600, 1785, 1715,
1655, 1630, 1600 cm l
N M R ~ (DMSO-d6) : 3.58 (2H, AB-q, J=14Hz),
4.00 (3H, s), 5;15 (lH, d, J=5Hz),
5.90 (lH, dd, J=5,8Hz), 6.52 (lH,
t, J~5Hz), 9.38 (lH, s), 9.84 (lH,
d, J=8Hz)
- 117 -
134~93
r;xample 7
7-Amino-3-cephem-4-carboxylic acid (1.7 g.) and
sodium bicarbonate (2.84 g.) were dissolved in a mixture of
water ~35 ml.) and acetone (35 ml.). On the other hand,
phosporus oxychloride (1.95 ml.) was added dropwise to a
suspension of Z-(2-amino-4-thiazolyl) 2-methoxyiminoacetic acid
- (syn-isomer: 3.42 g.) in dry ethyl acetate t34 ml.) over 10
minutes at 0 to 6~C, and the mixture was stirred at the same
temperature for 30 minutes. To the solution was added dropwise
lO a solution of trimethylsilylacetamide (2.39 g.) in ethyl acetate
(5 ml.) at 0 to 6~C over 20 minutes, and the mixture was stirred
for 20 minutes. After phosphorus oxychloride (1.95 ml.) was
added dropwise to the mixture at the above temperature over
10 minutes, the mixture obtained thus was stirred for 30 minutes.
And further, dimcthylformamide (1.29 ml.) was added dropwise to
the mixture over 10 minutes at the same temperature and stirred
for one hour to give a clear solution. The solution was added
dropwise to the solution of 7-amino-3-cephem-4-carboxylic acid
at -5 to 5~C, over 30 minutes, at pH 6.5 to 7.5, and the react-
ion mixture was stirred for one hour at the same temperature.
Ethyl acetate (200 ml.) was added to the resultant solution,
and the aqueous layer was separated, washed Wit]l methylene
chloride, bubbled with nitrogen gas and adjusted to pH 4 with
acetic acid. The solution was subjected to column chromato-
graphy on macroporous, non-ionic adsorption resin "Diaion
HP-20" (Trade mark: manufactured by Mitsubishi Chemical
Industries Ltd.) and eluted with 20~ aqueous solution of iso-
propyl alcohol. The eluate was concentrated under reduced
pressure and lyophilized to give 7-[Z-(2-amino-4-thiazolyl)-2-
methoxyiminoacetamido]-3-cephem-4-carboxylic acid (syn-isomer:
- 118 -
~ - . . .~ .
... ... .. .
1340 ~93
':
2.0 g.) The product was identified with the authentic
sample by IR and NMR spectrum.
Exampl~8
(1) Phosphorus oxychloride (1.2 g.) was added all at
once to a suspension of 2-(2-amino-4 thiazolyl)-2-
methoxyiminoacetic acid (syn-isomer: 1.23 g.) in ethyl acetate
(12 ml.) at 5~C and stirred at 4 to 6~C for 30 minutes.
Trimethylsilylacetamide (1.0 g.) was added to the solution
lO and stirred at 4 to 6~C for 30 minutes. Phosphorus oxychloride
(1.2 g.) was added again to the solution and stirred for 15
minu~es. And further, dimethylformamide (0.5 g.) was added
all at once to the solution at 4 to 6~C and stirred for 40
minutes to give a clear solution. On the other hand, p-
nitrobenzyl 7-amino-3-cephem-4-carboxylate hydrochloride
(1.9 g.~ was added to a mixture of tetrahydrofuran (30 ml.)
and acetone (10 ml.), and an aqueous solution (20 ml.) of
sodium bicarbonate (0.6 g.) was added to the mixture. To the
solution was added dropwise the solution obtained above at -0
20 to 5~C, pH 8Ø After stirring the mixture at -2 to 2~C, at
pH 8.0 for 30 minutes, the insoluble substance was filtered
out. The filtrate was extracted with ethyl acetate, and the
extract was washed with a saturated aqueous solution of sodium
chloride, dried over magnesium sulfate and then concentrated
under reduced pressure. The residue was pulverized with di-
isopropyl ether to give p-nitrobenzyl 7-[2-(2-amino-4-thiazolyl)-
2-methoxyiminoacetamido]-3-cephem-4-carboxylate tsyn-isomer:
1.6 g.)
I.R. ~ NmUa~ol : 3300, 1780, 1730, 1670, 1520 cm 1
-- 119 --
' 1340~99
N.M.ll. ~ppns (DMS0-d6) : 3.60 (2ll, m), 3.81 (311, s~,
5.12 (1l-l, d, J=5l-lz), 5.85 (lH, dd, J=SHz, lOHz),
6.64 (111, m), 6.70 (1~l, s), 7.20 (2H, s),
7.65 (2H, d, J~-lOHz), 8.19 (2H, d, J=lOHz),
9.60 (lH, d, J=lOHz)
(2~
p-Nitrobenzyl 7-~2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-
3-cephem-4-carboxylate (syn-isomer : 7.8 g.) was suspen~ed
in a mixture of ethanol (60 ml.) and water (60 ml.) lN
Aqueous solution of potassium hydroxide (45 ml.) was added
dropwise to the stirred suspension under ice-cooling over
10 minutes and stirred at 5~C for 15 minutes. The resultant
solution was adjusted to pH 7.0 with conc. hydrochloric acid,
was]led with ethyl acetate and the concentrated under reduced
pressure to half of its initial volume. The concentrated
solution was adjusted to pH 5.0 and subjected to column
chromatography on macroporous, non-ionic adsorption resin
"Diaion HP-20" (Trade mark; manufactured by Mitsubishi Chemical
Industries Ltd.; 80 ml.), and eluted with 5~ aqueous solution
of isopropyl alcohol. The fractions containing the object
compound were collected and adjusted to pH 3.2 with lO~
hydrochloric acid. The precipitating crystals were collected
by filtration, and dried to give 7-[2-~2-amino-4-thiazolyl)-
2-methoxyiminoacetamido]-3-cephem-4-carboxylic acid (syn-
isomer : 2.3 g.)
- 120 -
13~0~93
Example 9
p-Nitrobenzyl 7-amino-3-cephem-4-carboxylate (3.4 g.)
was suspended in tetrahydrofuran (60 ml.) and an aqueous
solution (20 ml . ) .of sodium bicarbonate (1.2 g.) was added
to the suspension. lN Aqueous sodium hydroxide (30 ml.) was
added dropwise to the solution at 3 to 4~C and stirred for 20
minutes. The resultant solution was adjusted to pH 7.0 with
; 10~ hydrochloric acid and concentrated under reduced pressure.
The insoluble substance was filtered out and the filtrate was
~ washed with ethyl acetate. Acetone (30 ml.) wa,s added to the
filtrate and cooled to -5~C. A solution prepared from
phospllorus oxychloridc, dime~hylformamide, trimethylsilylacetamide
and 2-(2-amino-4-thiazolyl)-Z-methoxyiminoacetic acid (syn-
isomer : 2.2 g.) in a similar manner to Example 7 was added
to the solution obtained above at -5 to 0~C, at pH 7.5 to 8,5.
The mixture was stirred at 3 to 7~C, at p}l 7.5 to 8.5 for 2
hours, and the insoluble substance was filtered out. The
aqucous layer was separated from the filtrate, washed with
ethyl acetate and adjusted to pH 3.0 to give 7-~2-(2-amino-
4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylic
acid tsyn-isomer : 1.1 g.)
- 121 -
: '
1340493
Example lO
(1) Phosphoryl chloride (1.764 g.) was added to a sus-
pension of 2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetic
acid (syn isomer, 1.0 g.) in tetrahydrofuran (10 ml.)
below 5~C and stirred at the same temperature for 20
minutes. To the solution were added trimethylsilylacetamide
~0.4 g.) and N,N-dimethylformamide ~0.4 g.), and the
solution was stirred below 5~C for 40 minutes[Solution A].
On the other hand, trimethylsilylacetamide ~3.5 g.) was
added to a suspension of 4-nitrobenzyl 7-amino-3-cephem-
4-carboxylate ~1.5 g.) in tetrahydrofuran ~15 ml.), and
stirred at room temyerature for 1.5 hours. l'o the solution
was added all at once the above Solution A at -20~C, and
the solution was stirred at -5 to 0~C for an hour. 1~1ater
- 122 -
.
... ,; - , . . :
1340~9~
(20 ml.) was addcd to the resultant solution at -20~C,
and the solution was adjusted to pH 7.5 with an aqueous
solution of sodium bicarbonate.
Tetrahydrofuran (70 ml.) and a saturated aqueous solution
of sodium chloride (50 ml.) were added to the solution,
and the solution was shaken sufficiently. The aqueous
layer was separated and extracted with tetrahydrofuran.
The tetrahydrofuran layer and extract were combined and
washed with a saturated aqueous solution of sodium chloride.
The solution was dried over magnesium sulfate and con-
centrated under reduced pressure. The residue was
triturated with diisopropyl ether to give 4-nitrobenzyl
7-[2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetamido]-3-
- cephem-4-carboxylate (syn isomer, 2.5 g.).
I. R.v NmaUxol : 3330, 1780, 1730, 1680, 1640,
1610 cm 1
N.M.R. ~ppm(DMSO-d6) : 1.17 (3H, t, J=7Hz),
3.50 (2H, m), 4.05 (2H, q, J=7Hz),
5,10 (lH, d, J=5Hz), 5.85 (lH, dd,
J=5Hz, 8Hz), 6.67 (lH, s), 7.17 (2H, m),
7.63 (2H, d, J=8Hz), 8.18 (2H, d, J=8Hz),
10.13 (lH, d, J=8Hz).
(2) Palladium on carbon (1.0 g.) moistened with water
(3 ml.) was added to a solution of 4-nitrobenzyl 7-[Z-
(2-amino-4-thiazolyl)-2-ethoxyiminoacetamido]-3-cephem-4-
carboxylate (syn isomer, 2.3 g.) in a mixture of tetra-
hydrofuran (30 ml.), methanol (15 ml.) and acetic acid
(0.3 ml.), and the suspension was subjected to catalytic
- 123 -
1340~93
reductioll at room temperature wldcr ordinary pressure for
2 llours. Artcr rcmovillg the catalyst from the resultallt
mixture by filtration, the filtrate was concentrated
under reduced pressure. Ethyl acetate was added-to the
residue, and the solution was adjusted to pH 7.5 with an
aqueous solution of sodium bicarbonate. After removing the
- insoluble substance by filtration, the aqueous solution
was separated, washed with ethyl acetate, adjusted to pH 5.5
and then treated with activated charcoal. The aqueous
solution was adjusted to pH 3.2, and the precipitates were
collected by filtration and dried to give 7-[2-~2-amino-
4-thlazolyl) 2-ethoxyiminoacetamido]-3-cephem-4-carboxylic
acid (syn isomer, 0.6 g.).
I.R. v~NIlaux~l : 3500, 3300, 3200, 1785, 1625,
16Q0 cm 1
N.M.R. ~ppm(DMSO-d6): 1.20 (3H, t, Jc711z),
3.57 (2H, m), 4.08 ~2H, q, J-7Hz),
5.08 (lH, d, J=5Hz), 5.83 ~lH, dd, Jc5Hz,
8Hz), 6.47 (lH, m), 6.73(1H, s), 7.20~2H, m),
9.58 (lH, d, J=8Hz)
Exflmple 11
(1) Triethylamille (2.37 g.), dimethylaniline (7.12 g.)
and trimethylsilyl chloride (3.93 g.) were addcd to a stirred
suspensiDn of 4-nitrobenzyl 7-(2-phenylacetamido)-3-
hydroxy-3-cephem-4-carboxylate (10 g.) in methylene chloride
- 124 -
.
- ~340~9~
(200 ml.) i.n turn, aIld the solution was stirred at
room tempcraturc for an hour. Phosphor~ls pentachloride
(4.88 g.) was addcd to the solution at -30 to -25~C and
stirred at -25 to -20~C for 3 hours. Methanol (42 ml.)
was added to the solution at -25 to -20~C, and stirred
for an hour. To the solution was added water (35 ml.)
at -25 to -20~C, and the solution was stirred at room
temperature. The precipitates were collected by filtration,
washed with methylene chloride and diethyl ether in turn,
and dried to give 4-nitrobenzyl 7-amino-3-hydroxy-3-cephem-
4-carboxylate (5.2 g.), mp 148~C (dec.).
I.R. v nNIauxol : 3440, 3300, 1760, 1740 cm 1
N.M.R. ~ ppm (DMSO-d6) : 2.8 - 3.7 (2H, m),
4.90 (11{, t, J=411z), 5.29 (lH, d, J-4Hz),
5.38 (211, s), 7.71 ~21{, d, J-8~Iz),
8.26 ~2H, d, J=8Hz).
~2) Phosphoryl chloride (2.87 g.) was dropwise added to
a solution of N,N-dimethylformamide ~l.37 g.) in ethyl
acetate (lO ml.) at 5 to 10~C. Ethyl acetate (40 ml.) was
added to the solution, and stirred under ice cooling for
40 minutes. To the solution was added 2-(2-formamido-4-
thiazolyl)-2-methoxyiminoacetic acid (syn isomer, 3.58 g.),
and the solution was stirred at 0 to 5~C for 40 minutes.
The resultant solution was added all at once to a mixture
of 4-nitrobenzyl 7-amino-3-hydroxy-3-cephem-4-carboxylate
(5 g.?, ethyl acetate (50 ml.), trimethylsilylacetamide
(14.3 g.) and bis(trimethylsilyl)acetamide (5.8 g.) at -15~C,
and stirred at -20 to -15~C for 1.2 hours.
- 125 -
,. 13~0493
IYater (S0 ml.) was added to tlle resultant sol~r6~nl~b
-25 to -20~C, and stirred until the temperature rise
5~C. 1he aqueous layer was separated and extracted
with ethyl acetate. The cthyl acetate layer and
extract wcre combined, washed with a saturated aqueous
solution of sodium cllloride and dried over ~nagnesium
sulfate. After thc solution was concentrated to a volume
o~ 50 ml. under reduced yressure, the precipitates were
collected by filtration and washed with ethyl acetate to
give 4-nitrobenzyl 7-[2-(2-formamido-4-thiazolyl)-2-
methoxyiminoacetamido]-3-11ydroxy-3-cepllem-4-carboxylate
(syn isomer, 3.5 g.), mp 163~C (dec.).
I.R. v NmUaxol : 3210, 3160, 3050, 1780,
1665 cm~l
N.M.R. ~ppm (D~lS0-d6): 3 0 - 4.2 ~2H, m), 3.95
(3H, s), 5.28 (lH, d, J=4Hz), 5.41 (2H, s),
5.64 (11-l, dd, J=4Hz, 9Hz), 7.49 (lH, s)
7.67 (2H, d, J=811z), 8.21 (ZH, d, J=8Hz),
8.50 (lH, t, J=9Hz).
- 126 -
~ 13~0~.93
; (3) A solution of the compound obtained above (1 g.) in
methanol (15 ml.), tetrahydrofuran (5 ml.) and conc. hydrochloric
acid (0.72 g.) was stirred at room temperature for an hour.
Diethyl ether (lOOml.) ~as added to the resultant solution and
then triturated. The crystals were collected by filtration to
give 4-nitrobenzyl 7-[2-(2-aminoth~ol-4-yl)-2-methoxyimino-
acetamido]-3-hydroxy-3-cephem-4-carboxyl~tu hydrochloride
(syn isomer, 0.65 g.).
I.R. ~ NUa~ol : 3180, 1780, 1680, 1670, 1640 cm 1
i N.M.R. ~ ppm (DM~O-d6) : 3.2 - 4.0 (2H, m), 3.97
(3H, e), 5.27 (lH, d, J=4Hz), 5.41 (2H, 8),
5.60 (lH, dd, J=4Hz, 8Hz), 7.10 (IH, 8),
7.66 (2H, d, J=9Hz), 8.25 (2H, d, J=9Hz),
9.73 (lH, d, J=8Hz)
Example 12
(1) Phosphoryl chloride ~1.76 g.) and trimethylsilylacetamide
(U,4 g.) were added to a stirred suspension of 2-~2-
amino-4-thiazolyl)-2-ethoxyiminoacetic acid (syn isomcr, 1.0 g~
in tetrahydrofuran (10 ml.) below 5~C, and stirred at the
same temperature for 30 minutes. N,N-Dimethylformamide (0.4 g.)
was added to the solution and stirred below 5~C for 20
~; minutes [Solution A]. Trimethylsilylacetamide (4.8 g.) was
added to a stirred suspension of 4-nitrobenzyl 7-amino-3-
chloro-3-cephem-4-carboxylate hydrochloride ~1.9 g.) in
tetrahydrofuran (15 ml.), and the solution was stirred at
room temperature for an hour. To the solution was added
the above Solution A all at once at -20~C, and the so~ution
- 127 -
13~0~99
was stirred at 0~C for an hour. Water (50 ml.) was added
to the resultant solution at -20~C, and adjusted to pH 8.0
with an aqueous solution of sodium bicarbonate.
Tetrahydrofuran (50 ml.) and a saturated aqueous solution
of sodium chloride (50 ml.) were added to the solution.
The organic layer was separated, washed with a saturated
aqueous solution of sodium chloride and dried over magnesium
sulfate. After concentrating the solution under reduced
pressure, the residue was triturated with diisopropyl ether
and the precipitates wcre collected by filtration to give
4-nitrobenzyl 7-[2-(2-amino-4-thiazolyl)-2-ethoxyimino-
acetamido]-3-chloro-3-cephem-4-carboxylate (syn isomer,
2.0 g.).
I. R. v maUxol: 3200, 1780, 1730, 1670 cm 1
N.M.R. ~ppm (DMSO-d6): 1.23 (3H, t, J=?Hz)~
3.96 (2H, s), 4.13 (2H, q, J=7Hz),
5.31 (lH, d, J=5Hz), 5.88 ~lH, dd,
J=5Hz, 8Hz), 6.77 (lH, s), 7.67 ~2H,
d, J=8Hz), 8.25 ~2H, d, J-8Hz), 10.30
(lH, d, J=8Hz).
- (2) A suspension of palladium on carbon (0.8 g.) in
water (5 ml.) was added to a mixture of 4-nitrobenzyl 7-
[2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetamido]-3-
chloro-3-cephem-4-carboxylate (syn isomer, 2.0 g.), acetic
acid (0.6 ml.) and tetrahy~rofurar, (60 ml.), and the
suspension was subjected to catalytic reduction under
ordinary pressure at room temperature for 3 hours.
- 128 -
~ 13~0~3
After removing the catalyst by filtration, the filtrate
was concentrated under reduced pressure. After adding
ethyl acetat:e (50 ml.~ to the residue, the solution was
adjusted to pH 7 5 with an aqucous solution of sodium
-~ bicarbonate, and the insoluble substance was filtered out.
The aqeuous layer was separated and adj usted to pll ~.0
with lO~ hydrochloric acid, and then the organic solvent
was removed under reduced pressure. The aqueous solution
was subjected to column chromatography on macroporous,
lO nonionic adsorption resin "Diaion HP-20" (Trademark,
manufactured by Mitsubishi Chemical Industries Ltd.) (30 ml.).
The column was washed with water and eluted with 5% aqueous
isopropyl alcohol. The eluate was lyophili~ed to give
sodium 7-[2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetamido]-
3-chloro-3-cephem-4-carboxylate (syn isomer, 0.3 g.).
I. R. v NUaxol : 3350, 3200, 1770, 1675, 1620 cm 1
N.M.R. ~sppm (D2O): 1.33 (3H, t, J=7Hz), 3.76
(211, q, J=18Hz, 30Hz), 4.30 (2H, q, J=7Hz),
5.33 (lH, d, J=511z), 5.83 (lH, d, J=5Hz),
7.06 (111, s).
-- 1~9
- i 13~0~.9
Example 13
(1) Phosphoryl chloride (4.6 g.), trimethylsilylacetamide
(0.95 g.) and N,N-dimethylforlllamidetl.2 9.) were added to a stirred
suspension of 2-(2-amino-4-thiazolyl)-2-isopropoxyiminoacetic
acid (syn isomer, 2.8 g.) in tetrahydrofuran (25 ml.) below
5 C for 30 minutes [Solution A]. On the other hand, trimethyl-
silylacetamide (10.5 g.) was added to a suspension of 4-nitro-
benzyl 7-amino-3-cephem-4-carboxylate (3.9 g.) in tetrahydrofuran
(50 ml.), and stirred at room temperature for 1.5 hours.
To the solution was added the above solution A at -20 C all at
once, and the solution was stirred at -5 to 0~C for 40 minutes-.
Water (70 ml.) and tetrahydrofuran (100 ml.) were added to the
resultant solution at -20~C. l'he solution was adjusted to p~l 7.5
with an aqueous solution of sodium bicarbonate and stirred for
an hour. After a saturated aqueous solution of sodium chloride
(200 ml.) was added, the organic layer was separated. The
rc ~;n;ng aqueous layer was extracted with tetrahydrofuran,
and the extract and the above organic layer were combined, washed
with a saturated aqueous solution of sodium chloride, dried
over magnesium sulfate and then concentrated under reduced
pressure. The residue was triturated with diisopropyl ether
and the precipitates were collected by filtration to give 4-
nitrobenzyl 7-[2-(2-amino-4-thiazolyl)-2-isopropoxyiminoacetamido]-
3-cephem-4-carboxylate (syn isomer, 6.0 g.).
I.R. ~maUxol : 3320, 3270, 1775, 1730, 1670, 1630 cm 1
N.M.R. d ppm (DMSO-d6) : 1.17 (6H, d, J=6Hz),
3.63 (211, m), 4.33 (111, q, J=6Hz), 5.17 (111, d, J=5Hz),
5.42 (211, s), 5.92 (lH, dd, J=5Hz, 8ilz), 6.G7 (lH, m),
- 130 -
13~0~93
6.70 (11l, s), 7.22 (211, m), 7.70 (211, d, J=811z),
8.25 (2H, d, J=811z), 10.13 (111, d, J=8Hz)
(2) ~cetic acid (1 ml.) and a suspension of 10% palladiwn on
carbon (2.0 g.) in water (8 ml) were added to a solution of
4-nitrobenzyl 7-[2-(2-amino-4-th1azolyl)-2-isopropoxyimino-
acetamido]-3-cephem-4-carboxylate (syn isomer, 5.0 g.) in
tetrahydrofuran (150 ml.), and the suspension was subjected
to catalytic reduction at room temperature under ordinary
pressure. After removing the catalyst by filtration, the
filtrate was concentrated under reduced pressure. Ethylacetate
(80 ml.) was added to the residue, and adjusted to pH 7.5
with an aqueous solution of sodium bicarbonate. The organic
layer was separated and extracted with an aqueous solution of
sodium bicarbonate. The extract and the aqueous layer obtained
above were eombined, adjusted to pH 3.0 with conc. hydrochloric
acid and extracted with tetrahydrofuran. The extract was
washed with a saturated aqueous solution of sodium chloride,
dried over magnesium sulfate and concentrated under reduced
pressure.
The precipitating crystalls were eollected by filtration and
dried to give 7-[2-(2-amino-4-thiazolyl)-2-isopropoxyimino-
- aeetamido]-3-eephem-4-earboxylie aeid (syn isomer, 0.8 g.).
I.R. I~maj : 3320, 1780, 1670, 1635 em 1
N.M.R. dppm (DMSO-d6): 1.20 (6H, d, J=6Hz), 3.55 (2H, m),
4.30 (lH, q, J=6Hz), 5.08 (lH, d, J=5Hz),
5.82 (lH, dd, J=5Hz, 8Hz), 6.45 (1ll, m), 6.68 (lH, s),
7.10 (2H, m), 10.08 (lH, d, J=8Hz)
-- 131 --
.. ~ ,.
~ 13~0493
E~ample 14
(1) Phosphoryl chloride (4.6 g.), trimethylsilJ~lacetamide
(0.95 g,) and N,N-di~ethylformamide (1.2 ~.) were added to
a stirred suspension of 2-(2-amino-4-thiazolyl)-2-propoxyimino-
acetic acid (syn isomer, 2.8 g.) in tetrahydrofuran (25 ml.)
below 5~C, and stirred for 20 minutes. The solution was
dropwise added to a suspension of 4-nitrobenzyl 7-amino-3-
cephem-4-carboxylate (3.9 g.) in a mixture of tetrahydrofuran
(20 ml.), water (20 ml.) and acetone (20 ml.) at -5 to 5~C
while keeping the pH value at 6.9 to 7.1 with 20% aqueous
solution of sodium carbonate. The solution was stirred at
-5 to 5~C for 30 minutes and further at 10~C for an hour, and
adjusted to pH 7.5. After tetrahydrofuran (100 ml.) and a
saturated aqueous solution of sodium chloride (200 ml.) were
added to the resultant solution, the insoluble substance was
filtered out. The organic layer was separated from the filtrate,
washed with a saturated aqueous solution of sodium chloride,
dried over magnesium sulfate and then concentrated under reduced
pressure. The residue was triturated with diisopropyl ether and
the precipitates were collected by filtration to give 4-nitrobenzyl
7-[2-(2-amino-4-thiazolyl)-2-propoxyiminoacetamido]-3-cephem-
4-carboxylate (syn isomer, 5.8 g.).
I-R-IJmaUx 1 3300, 1780, 1730, 1670, 1640 cm 1
N.M.R. ~ ppm (DMSO-d6): 0.93 (3H, t, J=6Hz), 1.70 (2H, m),
3.70 (2H, m), 4.08 (2H, t, J=6Hz), 4.5 (2H, m),
5.23 (lH, d, J=51iz), 5.50 (2H, s), 5.97 (lH, dd,
J=5~z, 8Hz), 6.73 (lH, m), 6.80 (lH, s), 7.75 (2H, d,
J=9Hz), 8.30 (2H, d, J=9Hz), 9.65 (lH, d, J=8Hz).
(2) 4-Nitrobenzyl 7- [2-(2-amino-4-thiazolyl)-2-propoxyimino-
acetamido]-3-cephem-4-carboxylate (syn isomer, 5.0 g.) was
-- 132
... .
1340493
,
treated in a similar manner to that of Example 13 (2) to give
7-[2-(2-amino-4-thiazolyl)-2-propoxyiminoacetamido]-3-cephem-4-
carboxylic acid (syn isomer, 0.9 g.)
I.R. ~ max : 3250, 1770, 1650, 1660, 1620 cm 1
N.M.R. d ppm (DMSO-d6) : 0.93 (3H, t, J=7Hz), 1.67 t2H,
sextet, J=7Hz), 3.60 (2~1, m), 4.03 (2H, t, J=7Hz),
5.13 (lH, d, J=5Hz), 5.83 (lH, dd, J=5Hz, 8Hz),
6.48 (2H, t, J=4Hz), 6.70 (lH, s), 7.18 (2H, m),
9.53 (lH, d, J=8Hz)
- 133 -
13~0493
... . .
~xam~ le 15
(1) Phosphoryl cl-loride tl3.2 g.) was added dropwise to a
stirred solution of N,N-dimethylformamide (6.3 g.) and tetra-
hydrofuran (24.7 ml.) at -5~C, and stirred at the same
temperature for 30 minutes. Tetrahydrofuran (120 ml.) and
2-(2-formamidothiazol-4-yl)-2-n-butoxyiminoacetic acid (syn
isomer, 19.5 g.) were added to the solution at -5~C, and
stirred at the same temperature for 30 minutes. The solution
was added dropwise a stirred suspension of 4-nitrobenzyl 7-
amino-3-cephem-4-carboxylate (24.7 g.), tetrahydrofuran
(120 ml.), acetone (60 ml.) and water (60 ml.) at -5 to 5~C over
15 minutes while adjusting to pH 7 to 7.5 with 20~ aqueous
solution of sodium carbonate, and then the solution was stirred
for 30 minutes. The insoluble substance was filtered off, and
a saturated aqueous solution of sodium chloride was added to
the filtrate. The solution was extracted with tetrahydrof~ran
twice. The extract was washed with a saturated aqueous solution
of sodium chloride, dried over magnesium sulfate and concentra~ed
in vacuo. The residue was triturated with diisopropyl ether
to give 4-nitrobenzyl 7-[2-(2-formamidothiazol-4-yl)-2-n-
butoxyiminoacetamido]-3-cephem-4-carboxylate (syn isomer,
34.6 g.)-
I.R. v maUxol: 3240, 3050, 1780, 1730, 1695,
1660 cm 1
N.M.R. ~(DMSO-d6, ppm) : 0.92 (3H, t, J=7Hz),
0.8-2.2 (4H, m), 3.67 (2H, d,
J=4Hz), 4.16 (2H, t, J=7Hz),
5.23 (lH, d, J=SHz), 5.46 (2H, s),
S.99 (lH, dd, J=5Hz, 8Hz), 6.71
(lH, t, J=5Hz), 7.43 (lH, s),
- 134 -
. .
~ .
1340~93
7.76 (21-l, d, J=9llz), 8.30 (2H, d,
J=9Hz), 8.58 (1ll, s), 9.72 (lH, d,
J=~l~z), 12.66 (1l~, s)
(2) A mixture of 4-nitrobenzyl 7- [2- (2-formamidothiazol-4-
yl)-2-n-butoxyiminoacetamido]-3-cephcm-4-carboxylate (syn isomer,
34.5 g.), tetrahydrofuran (345 ml.), 10% palladium carbon
(14 g.), methanol (140 ml.), acetic acid (2.5 ml.) and water
(50 ml.) was subjectcd to catalytic reduction under ordinary
pressure at room temperature for 3 hours. The resultant
lO mixture was filtered, and washed with tetrahydrofuran. The
fiitrate was concentrated in vacuo, and the residue was
dissolved in a mixture of ethyl acetate and an aqueous solution
of sodium bicarbonate. The insoluble substance was removed
by filtration. After the ethyl acetate layer was separated
and extracted with an aqueous solution of sodium bicarbonate,
the aqueous layer and the aquenus extract were combined.
After the aqueous solution was washed with ethyl acetate and
diethyl ether in turn, the solution was adjusted to pH 2.0
with 10~ hydrochloric acid and stirred for 30 minutes. The
20 precipitates were collected by filtration, washed with water
and dried over magnesium sulfate to give 7- [2- (2-formamidothiazol-
4-yl)-2-butoxyiminoacetamido~- 3-cephem-4-carboxylic acid
(syn isomer, 18.3 g.).
I.R. v mUaxol: 3330. 3040, 1780, 1725, 1695,
1655 cm 1
~J.M.R. ô(D~SO-d6, ppm) : 0.90 (3H, t, J=7Hz),
1.9 (4l~, m), 3.58 (2H, d, J-5Hz),
4.12 (2H, t, J=7Hz), 5.13 (lH, d,
J=5Hz), 5.86 (lH, dd, J=5Hz,
8l1z), 6.46 (lH, t, J=4Hz), 7.40 (lH, s),
-- 135 --
, . - ~ .~.,- - . -: -
1340~.q3
8.50 (lH, s), 9.63 (lH, d, J=~!lz),
12.57 (lH, broad s)
(3) A mixture of 7-[2-(2-formamidothiazol-4-yl)-2-n-
butoxyiminoacetamido]-3-cephem-4-carboxylic acid (syn isomer,
12.7 g.), conc. hydrochloric acid (9.6 ml.), methanol (9.5 ml.)
and tetrahydrofuran (9.5 ml.) was stirred at room temperature
for 3 hours. The resultant solution was concentrated in vacuo,
and the residue was suspended in water. The suspension was
adjusted to pH 3.5 with sodium bicarbonate under ice cooling,
and stirred at same temperature for 30 minutes. The precipitates
- were collected by filtration and dried over magnesium sulfate
to give the powder (10 g.). The powder was suspended in water
(300 ml.) and adjusted to pH 7.0 with sodium bicarbonate.
The solution was adjusted to pH 6.0 with 10% hydrochloric acid
and subjected to colunm chromatography on nonionic adsorption
resin (Diaion HP-20 : trademark, manufactured by Mitsubishi
Chemical Industries Ltd.) (300 ml.) with 10% aqueous solution
of isopropyl alcohol. The eluate was adjusted to pH 3.5
with 10% hy~rorhlnr;c acid under ice cooling, and the
precipitates were collected by filtration, washed with water
and dried to give 7-[2-(2-aminothia2O1-4-yl)-2-n-butoxyimino-
acetamido]-3-cephem-4-carboxylic acid (syn isomer, 7.2 g.).
I.R. v NmUaxol : 3320, 1775, 1660 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 0.88 (311, t, J=7Hz),
1.1~1.9 (4H~ m), 3.58 (2H, broad s),
4.05 (2H, t, J=7Hz), 5.08 (111, d,
J=5Hz), 5.80 (lH, dd, J=5Hz, 8Hz),
6.44 (lH, braod s), 7.18 (2H, s),
9.51 (lH, d, J=8Hz)
- 136 -
13~0~9.~
~xample 16
(1) 2-(2-Formalnidothiazol-4-yl)-2-iso-butoxyiminoacetic
acid (syn isomer, 6.48 g.), N,N-dimethylformamide (2.10 g.),
phosphoryl chloride (4.40 g.), tetrahydrofuran (110 ml.),
4-nitrobenzyl 7-amino-3-cephem-4-carboxylate (8.23 g.), acetone
(16 ml.) and water (16 ml.) were troated in a similar manner
to that of Example 15-(1) to give 4-nitrobenzyl 7-[2-(2-form-
amidothiazol-4-yl)-2-iso-butoxyiminoacetamido]-3-cephem-4-
carboxylate (syn isomer, 12.8 g.).
I.R. v maUjxol : 3240, 3050, 1780, 1720j 1700,
1655 cm 1
N.M.R. ~(DMSO-d6, ppm) : 0.92 (611, d, J=7Hz),
1.7~2.2 (lH, m), 3.67 (2H, broad s),
3.91 (2H, d, J=7Hz), 5.21 (lH,
d, J=5Hz), 5.95 (lH, dd, J=5Hz,
9Hz), 6.67 (lH, t~ J=4Hz),
7.37 (lH, s), 7.72 (2H, d, J~8Hz),
8.24 (2H, d, Jz8Hz), 8.52 (lH, s),
9.68 (lH, d, J=9Hz), 12.58 (lH,
broad s)
(2) 4-Nitrobenzyl 7-[2-(2-formamidothiazol-4-yl)-2-iso-
butoxyiminoacetamido]-3-cephem-4-carboxylate (syn isomer,
14.2 g.), 10% palladium carbon (5.7 g.), methanol (57 ml.),
tetrahydrofuran (142 ml.), acetic acid (1 ml.) and water
~10 ml.) were treated in a similar manner to that of Example
15-(2) to give 7-[2-(2-formamidothiazol-4-yl)-2-iso~butoxyimino-
acetamido]-3-cephem-4-carboxylic acid (syn isomer, 4.25 g.).
I.R. v nNIuaxol: 3260, 1790, 1725, 1670 cm
- 137 -
1340~93
N.M.R. ~(DMSO-d6, ppm) : 0.92 t6H, d, J=6Hz),
1.6~2.3 (lH, m), 3.61 (2H, d, J=4Hz),
3.91 (2H, d, J=6Hz), 5.14 (11l, d,
J=5Hz), 5.88 (lH, dd, J=SHz, 8Hz),
6.50 (lH, t, J=5Hz), 7.40 (lH, s),
8.56 (lH, s), 9.64 (lH, d, J=8Hz)
(3) 7-[2-(2-formamidothiazol-4-yl)-2-iso-butoxyiminoacet-
amido]-3-cephem-4-carboxylic acid (syn isomer, 4.1 g.), conc.
hydrochloric acid (3.65 g.) and methanol (61.5 ml.) were
treated in a similar manner to that of Example 15-(33 to give
7-[2-(2-aminothiazol-4-yl)-2-iso-butoxyiminoacetamido]-3-cephem-
4-carboxylic acid (syn isomer, 2.4 g.).
I.R. v mUa~ol : 3330, 1780, 1665, 1630, 1545 cm 1
N.M.R. ~(D~ISO-d6, ppm) : 0.89 (6H, d, J=7Hz),
1.6~2.2 (lH, m), 3.58 (2H, broad s),
3.84 (2H, d, J=7Hz,, 5.10 (lH, d,
J=5Hz), 5.82 (lH, dd, J=5Hz, 9Hz),
6.46 (lH, braod s), 6.68 (lH, s),
7.20 (2H, s), 9.53 (lH, d, J=9Hz)
Example 17
(1) 2-(2-'Formamidothiazol-4-yl)-2-cyclohexyloxyiminoacetic
acid (syn isomer, 0.9 g.), N,N-dimethylformamide (266 mg.),
phosphoryl chloride (557 mg.), tetrahydrofuran (20 ml.),
4-nitrobenzyl 7-amino-3-cephem-4-carboxylate (1.05 g.),
acetone (3 ml.) and water (3 ml.) were treated in a similar
manner to that of Example 15-(1) to give 4-nitrobenzyl 7-[2-
(2-formamidothiazol-4-yl)-2-cyclohexyloxyiminoacetamido] 3-
cephem-4-carboxylate (syn isomer, 1.69 g.).
I.R. v mUaxol : 3260, 3170,3070 , 1785, 1725, 1700,
1655 cm 1
- 138 -
.: - ' ': ' ''
.
~- 1340~9~
N.M.R. ~(DMS0-d6, ppm) : 0.8~2.2 (10l{, m), 3.66 (2H,
broad s), 4.10 (lH, m), 5.16 (lH,
d, J=5Hz), 5.42 (2H, s), 5.95 (lH,
dd, J=5Hz, 9Hz), 6.66 (lH, broad s),
7.37 (1ll, s), 7.70 (2H, d, J=8Hz),
8.22 (2H, d, J=8Hz), 8.50 (lH, s),
9.63 (lH, d, J=9Hz), 12.60 (1ll,
broad s)
(2)4-Nitrobenzyl 7-[2-(2-formamidothiazol-4-yl)-2-cyclo-
hexyloxyiminoacetamido]-3-cephem-4-carboxylate (syn isomer,
2.0 g.), 10% palladium carbon (0.8 g.), methanol (8 ml.),
tetrahydrofuran (20 ml.), acetic acid (0.14 ml.) and water
(1.4 ml.) were treated in a similar manner to that of Example
15-(2) to give 7-[2-(2-formamidothiazol-4-yl)-2-cyclohexyloxyimino-
acetamido]-3-cephem-4-carboxylic acid (syn isomer, 0.77 g.).
I.R. v NmaUjxol : 3275, 3070, 1780,.1675 cm 1
N.M.R. ~(DMSO-d6, ppm~ : 0.8~2.2 (lOH, m)~ 3.62
t2H, broad s), 4.12 (lH, m),
5.13 (lH, d, J=5Hz), 5.87 (lH,
dd, J=5Hz, 9Hz), 6.47 (lH, broad s),
7.37 (lH, s), 8.50 (lH, s),
9.58 (lH, d, J=9Hz), 12.61 (lH,
broad s)
(3) 7-[2-(2-Formamidothiazol-4-yl)-2-cyclohexyloxyimino-
acetamido]-3-cephem-4-carboxylic acid (syn isomer, 0.72 g.),
methanol (10.8 ml.) and conc. hydroc]lloric acid (0.61 g.)
were treated in a similar manner to that of Example 15(3) to
give 7-[2-(2-aminothiazol-4-yl)-2-cyclohexyloxyiminoacetamido]-
3-cephem-4-carboxylic acid (syn isomer, 0.28 g.).
- 139 -
1 3 4 0 ~ .9 3
I.R. IJ NUaj~l ; 3~50, 1775, 1665, 1620,1540 cm 1
N.M.R. ~(DMSO-d6, ppm): 0.8~2.2 (l0H, m)~ 3.60 (2H,
broad s), 4.04 (l~1, m), 5.09 (111,
d, J=511z), 5. 83 (l11, dd, J=5Hz, 9Hz),
6.45 (lH, t, J=411z), 6.67 (lH, s),
7.19 (211, s), 9.48 (lH, .d, J=9Hz)
Example 18
Phosphoryl chloride (0.84 g.) was added dropwise to a
stirred suspension of 2- (2-aminothiazol-4-yl)-2-allyloxyimino-
acetic acid (syn isomer, 1.0 g.), tetrahydrofuran (10 ml.) and
water (0.05 ml.) at 5~C, and stirred at the same temperature
for 20 minutes. Trimethylsilylacetamide (0.66 g.), phosphoryl
chloride (0.84 g.) and N,N-dimethylformamide (0.45 g.) were
added to a solution, and stirred at 5~C for an hour to prepare the
activated acid solution. On the othcr hand, trimethylsilyl-
acetamide (4.0 g.) was added to a suspension of 7-amino-3-
cephem-4-carboxylic acid (0.88 g.) in tetrahydrofuran (10 ml.)
at 40~C, and stirred for 30 minutes. To the solution was
added all at once the activated acidsolution obtained above
at -20~C, and stirred at 0~C for an hour. After water (20
ml.) was added to the resultant solution at -20~C, the solution
was adjusted to pH 7.5 with an aqueous solution of sodium
bicarbonate. Ethyl acetate was added to the solution, and
the aqueous layer was separcted. The solution was washed with
ethyl acetate and diisopropyl ether in turn, adjusted to pH 5.0
and treated with activated charcoal. After the solution was
adjusted to pH 3.0, the precipitates were collected by filtration,
washed with water and dried over phosphorus pentoxide to give
7-[2- (2-aminothiazol-4-yl)-2-allyloxyiminoacetamido]-3-cephem-
4-carboxylic acid (syn isomer, 0.8 g.).
I.R. v mUa~~l : 3300, 1780, 1660, 1630 cm 1
-- 140 --
1340~9'3
N.M.R. ~(DMSO-d6, ppm): 3.67 (2H, d, J=411z),
4.67 (211, m), 5.17 (lH, d, J=5Hz),
5.25 (lll, m), 5.50 (lH, m), 5.90
(lH, dd, J=51-1z, 8Hz), 6.03 (lH, m),
6.55 (lH, m), 6.80 (lH, s),
7.50 (2H, m), 9.68 (lH, d, J=8Hz)
Example l9
Phosphoryl chloride (l.4 g.) was added dropwise to a
suspension of 2- (2-aminothiazol-4-yl)-2-propargyloxyiminoacetic
ac1d (syn isomer, l.7 g.) in tetrahydrofuran (15 ml~) below
7~C, and stirred at the same temperature for l0 minutes.
Phosphoryl chloride (l.4 g.), trimethylsilylacetamide (l.3 g.)
and N,N-dimethylformamide (0.76 g.) were added to a solution,
and stirred for 20 minutes to prepare the activated acidsolution.
On the other hand, trimethylsilylacetamide (7.8 g.) was added
to a suspension of 7-amino-3-cephem-4-carboxylic acid (1.5 g.)
in tetrahydrofuran (20 ml.), and stirred at 40~C for 30 minutes.
To the solution was added all at once the activated acid solution
obtained above at -20~C, and stirred for 30 minutes at 0~C.
After adding water (20 ml.) to the resultant solution at -20~C,
the solution was adj usted to pH 7.5 with an aqueous solution
of sodium bicarbonate. The aqueous layer was separated,
washed with ethyl acetate and diisopropyl ether in turn, and
treated with activated charcoal at p11 5.5. The solution was
adj usted to pH 3.0, and the precipitates were collected by
filtration and dried over phosphorus pentoxide under reduced
pressure to give 7-[2- (2-aminothiazol-4-yl)-2-propargyloxy-
iminoacetamido]-3-cephem-4-carboxylic acid (syn isomer, 1.47 g.).
I.R. v NmUaixol: 3500, 3300, 1780, 1720, 1660,
1630 cm l
-- 141 --
13404~3
. .
N.M.R. ~(DMSO-d6, ppm) : 3.48 (lH, m), 3.67 (2H, m),
4.80 (2H, d, J=2Hz), 5.17 (lH, d,
J=5Hz), 5.88 (lH, dd, J=5Hz, 8Hz),
6.55 (lH, m), 6.85 (lH, s),
7.33 (2H, m), 9.73 (lH, d, J=8Hz)
Example 20
(1) N,N-Dimethylformamide (3 drops) was added to a
suspension of 2-(2-formamidothiazol-4-yl)-2-methoxyimino-
acetic acid (syn isomer, 23 g.) in thionyl chloride (230
ml.), and stirred at 60~C for 5 minutes. After
concentrating the solution in vacuo, benzene was added to
the residue. The precipitates were collected by
filtration, washed with benzene (30 ml.) three times and
diethyl ether in turn to give 2-(2-formamidothiazol-4-yl)-
2-methoxyiminoacetyl chloride (anti isomer, 18 g.). On
the other hand, trimethylsilylacetamide (46 g.) was added
to a suspension of 4-nitrobenzyl 7-amino-3-cephem-4-
carboxylate (16.8 g.) in methylene chloride (168 ml.), and
stirred at 40~C for an hour. To the suspension was
gradually added 2-(2-formamidothiazol-4-yl)-2-methoxy-
iminoacetyl chloride (anti isomer, 13.6 g.) at -5 to 0~C,
and stirred at the same temperature for an hour. Water
(150 ml.) was added to the resultant solution and stirred
for 15 minutes. The precipitates were collected by
filtration, washed with water and dried over phosphorus
pentoxide under reduced pressure to give 4-nitrobenzyl 7-
[2-(2-formamidothiazol-4-yl)-2-methoxyimino-acetamido]-3-
cephem-4-carboxylate (anti isomer, 25.7 g.).
Nujol
I.R. max : 3300 (broad), 1780, 1730, 1680,
1520 cm~1
N.M.R. ~(DMSO-d6, ppm) : 3.70 (2H, broad s), 4.07
(3H, s), 5.19 (lH, d, J=5Hz), 9.57
(lH, d, J=8Hz), 6.00 (lH, dd, J=5Hz, 8Hz)
5.30 (2H, s),
- 142 -
~-
.. . .. . ... . ....
1340499
. .
6.70 (111, t, J-4~1z), 7.71, 8.25
(4~ 2B2~ J=9Hz), 8.07 (111, s),
8.50 (lH, s), 12.55 (lH, broad s)
(2) A suspension of 4-nitrobenzyl 7-[2-(2-formamidothiazol-
4-yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate (anti
isomer, 4.2 g.), 10~ palladium carbon (1.7 g.), acetic acid
(0.63 ml.), water (6.3 ml.), methanol (42 ml.), and tetra-
hydrofuran (84 ml.) was subjected to catalytic reduction in a
hydrogen atmosphere at room temperature for 2 hours. After
10 removing the catalyst by filtration, the filtrate was concentrated
to a volume of about 15 ml. under reduced pressure. Water
(30 ml.) and ethyl acetate (50 ml.) were added to the concentrated
solution, and the solution was adjusted to pH 8.0 with sodium
bicarbonate under stirring. The insoluble substance was
removed by filtration, and the aqueous layer was separated and
washed with ethyl acetate (50 ml.). The solution was treated
with activated charcoal, and adjusted to pH 2.2 with 10%
hydrochloric acid under ice cooling. The precipitates were
collected by filtrationandwashed with water to give 7-[2-(2-
20 formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-
carboxylic acid (anti isomer, 2.52 g.).
I.R. v NmUaxol: 3300 (broad), 1780, 1680, 1670,
1550 cm 1
N.M.R. ~(DMSO-d~j, ppm) : 3.63 (2H, broad s),
4.08 (311, s), 5.15 (lH, d, J-5Hz),
5.87 (lH, dd, J=5Hz, 8Hz),
6.55 (11-1, t, J=411z), 8.09 (lH, s),
8.52 (lH, s), 9.46 (lH, d, J=8H~)
(3) A suspension of 7-[2-(2-formamidothiazol-4-yl)-2-
30 methoxyiminoacetamido]-3-cephem-4-carboxylic acid (anti isomer,
-- 143 --
, ~ ~ . . . .
1340~9.~
. . .
2.5 g.), conc. hydroc}lloric acid (2.5 ml.) and methanol (38 ml.)
was stirred at room temperature for two hours. After treating
the resultant solution with activated charcoal, the solution
was eoncentrated in vacuo. The residue was crystallized out
with diisopropyl ether (100 ml.), and the precipitates were
collected by filtration, and washed with diisopropyl ether
(30 ml.) to give 7-[2-~2-aminothiazol-4-yl)-2-methoxyimino-
acetamido]-3-cephem-4-carboxylic acid hydrochloride (anti
isomer, 2.1 g.). The crystals were added to water (20 ml.)
and adjusted to pH 6.0 with sodium bicarbonate. T~e solution
was subjected to column chromatography on nonionic adsorption
resin "Diaion IIP-20" [Trademark: manufactured by Mitsubishi
Chemical Industries Ltd.] (75 ml.) with 10% diisopropyl ether.
The eluate was adjusted to pH 3.5 with 10% hydrochloric acid,
and the precipitates were collected by filtration and dried to
give 7-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-
cephem-4-carboxylic acid (anti isomer, 0.7 g.).
I.R. v Nluaxol : 3400~3200 (broad), 1770, 1680,
1640, 1520 cm 1
N.M.R. ~(DMSO-d6, ppm) : 3.60 (211, d, J=5Hi),
4.00 (3H, s), 5.10 (lH, d, J=5Hz),
5.82 (lH, dd, J=5Hz, 8Hz),
6.48 (lH, t, J=4Hz), 7.13 (2H,
broad), 7.47 (lH, s), 9.42 (lH, d)
Example 21
(1) The Vilsmeier reagent was prepared from N,N-dimethyl-
formamide (0.4 g.) and phosphoryl chloride (0.86 g.) in a usual
manner. After the reagent was suspended in ethyl acetate
(10 ml.), 2-(2-formamidothiazol-4-yl)-2-n-propoxyiminoacetic
acid (syn isomer, 1.3 g.) was added to the stirred suspensi~n
- 144 -
~ ~ 1340~9 ~
under ice cooling and st:irred at the same temperature for
30 minute. The solution was added to a solution of 4-nitrobenzyl
7-amino-3-chloro-3-cephem-4-carboxylate hydrochloride (2.0 g.),
trimethylsilylacetamide (5.2 g.) and ethyl acetate (40 ml.) at
-25~C, and stirred at -20 to -10~C for 1.5 hours. After
adding water into the resultant solution, tlle solution was
extracted with ethyl acetate (60 ml.). The aqueous layer was
extracted with ethyl acetate (50 ml.). The extracts were
combined together, washed with a saturated aqueous solution
of sodium bicarbonate and a saturated aqueous solution of
sodium chloride in turn, and dried over magnesium sulfate.
The solution was concentrated in vacuo, and the residue was
triturated with diethyl ether. The precipitates were
collected by filtratation, washed and dried to give 4-nitrobenzyl
7-[2-(2-formamidothiazol-4-yl)-2-n-propoxyiminoacetamido]-3-chloro-
- 3-cephem-4-carboxylate (syn i~somer, 2.55 g.).
I.R. v maUxol: 3250~3150, 1780, 1730, 1690,
1660, 1610, 1550, 1520 cm 1
N.M.R. ~(DMS0-d6, ppm) : 0.87 (3H, t, J=7Hz),
1.63 (lH, m), 3.88 (2H, q, J=17Hz),
3.97 (2H, q, J=7Hz), 5.30 (lH, d,
J=5Hz), 5.40 (2H, s), 5.92 (lH, dd,
J=SHz, 81-lz), 7.33 (lH, s),
7.65 (2H, d, J=9Hz), 8.20 (2H, d,
J=9Hz), 8.47 (lH, s), 9.70 (lH, d,
J=8Hz), 12.40 (lH, s)
(2) A suspension of 4-nitrobenzyl 7-[2-(2-formamidothiazol-
4-yl)-2-n-propoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylate
tsyn isomer, 2.4 g.), 10~ palladium carbon (1.0 g.), methanol
(24 ml.), water (3.6 ml.) and tetrahydrofuran (48 ml.) was
- 145 -
; .
1340~93
subjected to catalytic reduction under ordinary pressure at room
temperature. After removillg insoluble substance by filtration,
the filtrate was concentrated in vacuo. Water and ethyl acetate
were added to the residue, and adjusted to pl3 8 with a saturated
aqueous solution of sodium bicarbonate. The insoluble substance
was removed by filtration, and the aqueous layer was separated.
Ethyl acetate was added to the solution, adjusted to pH 2.0
with hydrochloric acid and extracted with ethyl acetate. The
ethyl acetate solution and extract were combined together,
washed with a saturated aqueous solution of sodium cllloride
- and dried over magnesium sulfate. The solution was concentrated
in vacuo, and the residue was triturated with diethyl ether
and collected by filtration to give 7-[2-(2-formamidothiazol-
4-yl)-2-n-propoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylic
acid (syn isomer, 1.6 g.).
I.R. v NmUaxol : 3300~3150, 1780, 1720, 1685,
1650, 1540 cm 1
N.M.R. ~(DMSO-d6, ppm) : 0.93 (311, t, J=7Hz),
1.72 (lH, m), 3.88 (2H, q, J=18Hz),
4.08 (2H, q, J=7Hz), 5.33 (lH, d,
J=5Hz), 5.92 (lH, dd, J=5Hz, 8Hz),
7.43 (lH, s), 8.57 (lH, s),
9.73 (lH, d, J=813z)
(3) A suspension of 7-[2-(2-formamidothiazol-4-yl)-2-n-
propoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylic acid
(syn isomer, 1.5 g.), conc. hydrochloric acid (0.7 ml.) and
methanol (30 ml.) was stirred at room temperature for 1.5 hours.
After removing methanol from the resultant solution in vacuo,
water (30 ml.) was added to the residue. After the solution
was adjusted to pH 7.5 with a saturated aqueous solution of
'
- 146 -
. ~ ....
13a~0~93
sodium bicarbonate, the insoluble substance was removed by
filtratiol~. The filtrate was adjusted to pH 3 with 10%
hydrochloric acid. The precipitates were collected by filtration
and dried over phosphorus pentoxide to give 7-[~-(2-aminothiazol-
4-yl)-2-n-propoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylic
acid (syn isomer, 0.6 g.).
I.R.v Nu~ol : 3300, 1780, 1670, 1630, 1530 cm 1
N.M.R. ~(DMS0-d6, ppm) : 0.92 (3H, t, J=7Hz),
1.67 (lH, m), 3.70 (2H, q, J=18Hz),
4.00 (2H, q, J=7Hz), 5.25 (lH, d,
J~5Hz), 5.83 (lH, dd, J=5Hz, 8Hz),
6.75 (lH, s), 9.63 (lH, d, J=8Hz)
Example 22
2-(2-Aminothiazol-4-yl)-2-n-hexyloxyiminoacetic acid (syn isomer,
3 g.), water (0.15 g.), pbcsphoryl chl oride (3.8 g.), trimethylsily-
ace'amide (10.7 g.), N,N-dimethylformamide (1.0 g.), tetrahydroiuran
(50 ml.) and 7-amino-3-cephem-4-carboxylic acid (2.0 e.) were trested
in a 8imilar manner to that oi Examplel8 to give 7-[2-(2-aminothiazol-
4-yl)-2-n-hexyloxyiminoacetamido~-3-cephem-4-carbo~ylic acid (syn isomer,
1.1 g.).
I.R. VmU8~~l 3250, 1760, 1640, 1600 cm 1
N.M.R. d (DM~0-d6, ppm) : 1.88 (3H, m), 1.1 ~ 1.9 (8H, m), 3.60
(2H, m), 4.06 (2H, t, J=6Hz), 5.10 (IH, d, J=5Hz), 5.82
(lH, dd, J= 5Hz, 8Hz), 6.46 (lH, m), 6.70 (~, 8),
7.26 (2H, m), 9.56 (lH, d, J=8Hz)
- 147 -
. . .
1340 i~
Example 23
2-(2-formamidothiazol-4-yl)-2-propargyloxyiminoacetic
acid (syn isomer, 0.506 g.), N,N-dimethylformamide (0.161
g.), phosphoryl chloride (0.337 g.) and ethyl acetate (7.9
ml.) were treated in a similar manner to that of Example
15-(1) to give the activated acid solution.
On the other hand, trimethylsilylacetamide (1.85 g.) and
bis(trimethylsilyl)acetamide (1.60 g.) were added to a
suspension of 4-nitrobenzyl 7-amino-3-hydroxy-3-cephem-4-
l0 carboxylate (0.703 g.) in ethyl acetate (10 ml.) and
stirred at room temperature for an hour.
To the solution was added the activated acid solution
obtained above at -10~C all at once, and stirred at the
same temperature for an hour. Water (20 ml.) and ethyl
acetate (20 ml.) were added to the solution.
The organic layer was separated, washed with a saturated
aqueous solution of sodium bicarbonate (8 ml.) and an
aqueous solution of sodium chloride (15 ml.), and dried
over magnesium sulfate.
After the solution was concentrated in vacuo, the residue
was pulverized with diethyl ether and the precipitates
were collected by filtration to give 4-nitrobenzyl 7-[2-
(2-formamidothiazol-4-yl)-2-propargyloxyiminoacetamido]-3-
hydroxy-3-cephem-4-carboxylate (syn isomer, 0.71 g.)
max : 3270, 1770, 1740, 1670 cm~
N.M.R. ~(DMSO-d6, ppm) : 3.39 (lH, m),
3.63 (2H, broad s), 4.90 (2H, broad s),
5.23 - 5.90 (4H, m), 7.57 (lH, s),
- 148 -
~X
13'1~ 493
7.83 (2H, d, J=9Hz), 8.40 (2H, d,
J=9Hz), 8.67 (lH, s), 9.80 (lH, d,
J=8Hz), 12.83 (lH, broad s)
Example 24
2-(2-Formamidothiazol-4-yl)-2-propoxyiminoacetic acid
(syn isomer, 0.515 g.) N,N-dimethylformamide (0.161
g.),phosphoryl chloride (0.337 g.) and ethyl acetate (7.9
ml.) were treated in a similar manner to that of Example
15-(1) to give the activated acid solution.
I0 On the other hand, trimethylsilylacetamide (1.85 g.) and
bis(trimethylsilyl)acetamide (1.60 g.) were added to a
suspension of 4-nitrobenzyl 7-amino-3-hydroxy-3-cephem-4-
carboxylate (0.703 g.) in ethyl acetate (10 ml.), and
stirred at room temperature for an hour.
To the solution was added the activated acid solution
obtained above at -10~C all at once, and stirred at the
same temperature for an hour. To the resultant solution
were added water (20 ml.) and ethyl acetate (20 ml.). The
organic layer was separated, washed with a saturated
aqueous solution of sodium bicarbonate (8 ml.) and water
(14 ml.) and dried over magnesium sulfate. After the
solution was concentrated in vacuo, the residue was
pulverized with diethyl ether, and the precipitates were
collected by filtration to give 4-nitrobenzyl 7-[2-(2-
formamidothiazol-4-yl)-2-propoxyiminoacetamido]-3-hydroxy-
3-cephem-4-carboxylate (syn isomer, 0.75 g.).
max : 3250, 1765, 1740, 1670 cm
N.M.R. ~(DMSO-d6, ppm) : 0.90 (3H, t, J=8Hz),
1.67 (2H, m), 3.40 (2H, AB-q, J=20Hz),
4.08 (2H, q, J=8Hz),
- 149 -
t ~,
13404'33
- 5.03 - 5.83 (4H, m), 7.40 (lH, s),
7.70 (2H, d, J=9Hz), 8.27 (2H, d,
J=9Hz), 8.53 (lH, s), 9.50 (lH, d,
J=8Hz), 12.60 (lH, broad s).
Example 25
2-(2-Formamidothiazol-4-yl)-2-isobutoxyiminoacetic acid
(syn isomer, 0.54 g.), N,N-dimethylformamide (0.16 g.),
phosphoryl chloride (0.34 g.) and ethyl acetate (10 ml.)
were treated in a similar manner to that of Example 15-(1)
to give the activated acid solution. On the other hand,
trimethylsilylacetamide (1.85 g.) and bis(trimethylsilyl)-
acetamide (1.62 g.) were added to a suspension of 4-
nitrobenzyl 7-amino-3-hydroxy-3-cephem-4-carboxylate (0.7
g.) in ethyl acetate (10 ml.) and stirred at 40~C for 30
minutes. The activated acid solution obtained above was
added to the solution at -20~C all at once, and stirred at
the same temperature for an hour. Water (10 ml.) was
added to the resultant solution, and the organic layer was
separated, washed with water, an aqueous solution of
sodium bicarbonate and a saturated aqueous solution of
sodium chloride, and then dried. After the solution was
concentrated in vacuo, the residue was pulverized with
diisopropyl ether. The precipitates were collected by
filtration, washed with diisopropyl ether and dried to
give 4-nitrobenzyl 7-[2-(2-formamidothiazol-4-yl)-2-2-
isobutoxyiminoacetamido]-3-hydroxy-3-cephem-4-carboxylate
(syn isomer, 1.09 g.)
- 150 -
,~,~
'~ X ,. ..
1340493
, .
I. R. v Na~~l : 3250, 3050, 1750, 1650,
1610 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 0.97 (6H, d, J=6Hz),
2.0 (lH, m), 3.60 (ZH, AB-q, J=18Hz),
3.95 (2H, d, J=6Hz), 5.1 - 5.8 (4H, m),
7.53 (lH, s), 7.63 (2H, d, Js8Hz),
8.18 (2H, d, J~8Hz), 8.58 (lH, s),
9.47 (lH, d, J=9Hz), 12.77 (lH, broad s)
Example 26
N,N-Dimethylformamide (0.114 g.), phosphoryl
chloride (0.240 g.) and ethyl acetate (0.5 ml.) were
- reacted in a conventional manner to give a Vilsmeier
reagent. Ethyl acetate (5 ml.) and 2-(2-formamido-
thiazol-4-yl)-2-ethoxyiminoacetic acid (syn isomer,
0.35 g.) were added at the same temperature for 30
minutes to give the activated acid solution.
On the other hand, trimethylsilylacetamide (1.31 g.)
and bis(trimethylsilyl)acetamide (1.14 g.) were added
to a suspension of 4-nitrobenzyl 7-amino-3-hydroxy-3-
cephem-4-carboxylate (0.50 g.) in ethyl acetate
(7.5 ml.) and stirred at room temperature for an hour.
To the solution was added the activated acid solution
obtained above all at once at -10~C, and stirred at
the same temperature for 30 minutes. Water (15 ml.)
- and ethyl acetate (15 ml.) were added to the resultant
solution. The organic layer was separated, washed
with a saturated aqueous solution of sodium bicarbonate
(15 ml.) and water (10 ml.), dried over magnesium
sulfate and concentrated in vacuo. The residue was
pulverized with diethyl ether and the precipitates were
- 151 -
,
1340~ 93
collected by filtration to give 4-nitrobenzyl 7-[Z-
(2-formamidothiazol-4-yl)-2-ethoxyiminoacetamido]-
3-hydroxy-3-cephem-4-carboxylate (syn isomer, 0.634 g.).
I.R. ~ maU~ol : 3220, 1760, 1740, 1670cm 1
N.M.R. ~ (DMSO-d6, ppm) : 1.24 (3H, t,
J=8Hz), 3.36 (2H, AB-q, J=20Hz),
4.15 (2H, q, J=8Hz), 5.10 - 5.60 (4H,
m), 7.37 (lH, s), 7.65 (2H, d, J=9Hz),
8.22 (2H, d, J=9Hz), 8.48 (lH, s),
9.52 (lH, d, J=8Hz), 12.58 (lH, broad s)
- 152 -
.. . .. , . . ~
1340493
Example 27
(1) Z-(2-Formamidothiazol-4-yl)-2-
pentyloxyiminoacetic acid (syn isomer, 4.14 g.),
4-nitrobenzyl 7-amino-3-cephem-4-carboxylate (4.5 g.),
N,N-dimethylformamide (1.41 g.), phosphoryl chloride
(Z.96 g.), tetrahydrofuran (7Z ml.), acetone (15 ml.)
and water (15 ml.) were treated in a similar manner
to that of Example 15 -(1) to give 4-nitrobenzyl
7-[2-(2-formamidothiazol-4-yl)-2-pentyloxyiminoacetamido]-
3-cephem-4-carboxylate (syn isomer, 8.1 g.).
I. R. v maUxol : 3240, 3050, 1780, 1730,
1655 cm 1
N.~l.R. ~ (DMSO-d6, ppm) : 0.6 - 2.0 (9H, m),
3.66 (2H, s), 4.10 (2H, t, J=6Hz),
5.19 (lH, d, J=5Hz), 5.42 (2H, s),
5.95 (lH, dd, J=5Hz, 8Hz), 6.16 (lH,
broad s), 7.38 (lH, s), 7.72 (ZH, d,
J=9Hz), 8.Z6 (2H, d, J=9Hz), 8.54 (lH, s),
9.69 (lH, d, J=8Hz), lZ.69 (lH, broad s)
(2) 4-Nitrobenzyl 7-[Z-(Z-formamidothiazol-4-yl)-
2-pentyloxyiminoacetamidol-3-cephem-4-carboxylate (syn
isomer, 8 g.), 10% palladium carbon (3.6 g.), methanol
(36 ml.), tetrahydrofuran (90 ml.), acetic acid (0.63 g.)
and water (6.3 ml.) were treated in a similar manner
to that of Example 15 -(2) to give 7-[2-(2-
formamidothiazol-4-yl)-2-pentyloxyiminoacetamido]-3-
cephem-4-carboxylic acid (syn isomer, 3.4 g.).
I.R. v mUx~l : 3275, 3075, 1795, 1700,
1660, 1630 cm 1
- 153 -
,
134~93
N.M.R. ô (DMSO-d6, ppm) : 0.6 - 2.0 (9H, m),
3.60 (2H, d, J=41tz), 4.12 (2H, t,
J=6Hz), 5.14 (1ll, d, J=5Hz),
5.87 (llt, dd, J=5l-lz, 9Hz), 6.49 (lH,
t, J=3Hz), 7.40 (lH, s), 8.53 (lH, s),
9.64 (lH, d, J=9l-lz), 12.68 (lH, s)
(3) 7-[2-(2-Formamidothiazol-4-yl)-2-
pentyloxyiminoacetamido]-3-cephem-4-carboxylic acid
(syn isomer, 3.3 g.), conc.hydrochloric acid (2.80 g.),
10 tetrahydrofuran (20 ml.) and methanol( 50 ml.) were
treated in a similar manner to that of Example 15~
to give 7-[2-(2-aminothiazol-4-yl)-2-pentyloxyimino-
acetamido]-3-cephem-4-carboxylic acid (syn isomer, 2.3 g.)
I.R. v maUxol : 3300, 1775, 1650, 1540cm 1
N.M.R. ~ (DMSO-d6, ppm) : 0.6-2.0 (9H, m),
3.56 (2H, d, J=2Hz), 4.03 (2H, t,
J=6Hz), 5.08 (lH, d, J=5Hz), 5.81 (lH,
dd, J=5Hz, 8Itz), 6.46 (lH, t, J=4Hz),
6.69 (lH, s), 7.20 (2H, s), 9.15 (lH,
d, J=8Hz)
- Example 28
(1) Triethylamine (3.41 g.), N,N-dimethylphenylamine
(10.3 g.) and trimethylchlorosilan (5.64 g.) were added
to a stirred solution of 7-phenylacetamido-3-tosyloxy-
3-cephem-4-carboxylic acid (15 g.) in methylene chloride
(150 ml.) at room temperature in turn, and the solution
was stirred at room temperature for an hour.
Phosphorus pentachloride (7.03 g.) was added to the
solution at -35~C, and stirred at -25 to -20~C ~or
- 154 -
.
- i 1340~'-)3
1.5 hours. Methanol (61 ml . ) was added to the solution
and stirred at the same temperature for 40 minutes.
To the resultant solution was added water (50 ml.)
at -20 to -10~C. The organic layer was separated and
washed with water twice. The aqueous layer and the
washings were combined, and washed with methylene
chloride twice and diethyl ether in turn.
After the solution was adjusted to pH 4.7 with 10%
aqueous sodium hydroxide under cooling, the precipitates
were collected by filtration, washed with water,
acetone and diethyl ether in turn, and then dried
over phosphorus pentoxide to give 7-amino-3-
tosyloxy-3-cephem-4-carboxylic acid (5.01 g.), mp
172~C (dec.).
I.R. v mUaxol : 3210, 1800, 1653, 1620cm 1
N.M.R. ~ (D2O+NaHCO3, ppm): 2.45 (3H, s),
3.51 (2H, q, J=18Hz), 5.08 (lH, d,
Js5Hz), 5.51 (lH, d, J=5Hz), 7.48
(2H, d, J=9Hz), 7.84 (2H, d, J=9Hz)
(2) 2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetic
acid (syn isomer, 0.76 g.), N,N-dimethylformamide
(0.33 g.), phosphoryl chloride (1.46 g.),
trimethylsilylacetamide (0.5 g.) and ethyl acetate
(10 ml.) were treated to give the activated-acid
solution in a conventional manner.
On the other hand, trimethylsilylacetamide (2.7 g.)
was added to a suspension of 7-amino-3-tosyloxy-3-
cephem-4-carboxylic acid (1.0 g.) in ethyl acetate
(15 ml.) and stirred at room temperature.
-- 155 --
1340~93
To the solution was added the activated acid solution
obtained above at -15~C all at once, and stirred at
-5 - 5~C for an hour. After the resultant solution
was chilled to -20~C, water (30 ml.) was added to the
chilled solution and adjusted to pH 6.5 with an aqueous
solution of sodium bicarbonate. The insoluble substance
was removed by filtration. The aqueous layer was
separated and adjusted to pH 3.0 with hydrochloric
acid. The precipitates were collected by filtration
and dried to give 7-[2-~2-aminothiazol-4-yl)-2-
methoxyiminoacetamido]-3-tosyloxy-3-cephem-4-carboxylic
acid (syn isomer, 1.0 g.).
I.R. v maUxol : 3350, 1780, 1670, 1630cm 1
N.M.R. ~ (DMSO-d6, ppm) : 2.17 (3H, s),
3.71 (2H, m), 3.92 (3H, s), 5.32 (lH,
- d, J=5Hz), 5.87 (lH, dd, J=5Hz, 8Hz),
6.8Z (lH, s), 7.50 (2H, d, J=8Hz),
7.92 (2H, d, J=8Hz), 9.73 (lH, d, J=8Hz)
- 156 -
,.
~ .-, ~ ~- - -
134049t~
.
Example 29
' (1) 2-(2-Formamidothiazol-4-yl)-2-
ethoxycarbonylmethoxyiminoacetic acid (syn isomer,
1.35 g.), 4-nitrobenzyl 7-amino-3-cephem-4-carboxylate
(1.54 g.), N,N-dimethylformamide (393 mg.), phosphoryl
chloride (8Z5 mg.), tetrahydrofuran (21.2 ml.),
acetone (3.9 ml.) and water (3.9 ml.) were treated in a
similar manner to that of Example 15- (1) to give 4-
nitrobenzyl 7-[2-(2-formamidothiazol-4-yl)-2-
,, 10 ethoxycarbonylmethoxyiminoacetamido]-3-cephem-4-
carboxylate (syn isomer 2.52 g.).
I. R. vmaUx~l : 3250, 1790, 1730, 1690,
1640 cm 1
N.M.R. ~(DMSO-d6, ppm):1.23 (3H, t, J=7Hz),
3.66 (2H, s), 4.13 (2H, q, J=7Hz),
4.74 (2H, s), 5.22 (lH, d, J=5Hz),
5.42 (2H, s), 5.98 (lH, dd, J=5Hz,
9Hz), 6.49 (lH, broad s), 7.43 (lH, s),
7.71 (2H, d, J=9Hz), 8.23 (2H, d,
J=9Hz), 8.5Z (lH, s), 9.68 (lH, d,
J=9Hz), 12.66 (lH, s)
(2) 4-Nitrobenzyl 7-[2-(2-formamidothiazol-4-yl)-2-
; ethoxycarbonylmethoxyiminoacetamido]-3-cephem-4-
carboxylate (syn isomer, 2.52 g.), 10% palladium carbon
(1.3 g.), ethanol (13 ml.), tetrahydrofuran (25 ml.),
acetic acid (0.22 ml.) and water (2.2 ml.) were treated
in a similar manner to that of Example 15-(2) to give
7-[2-(2-formamidothiazol-4-yl)-2-ethoxycarbonylmethoxyimino-
acetamido]-3-cephem-4-carboxylic acid (syn isomer, 0.4 g.).
- 157 -
~ .
1340493
I R. v Nujol 3250, 3060, 1780, 1750,
max
1690, 1660 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 1.23 (3H, t, J=7Hz),
3.61 (2H, broad s), 4.15 (2H, q,
J=7Hz), 4.73 (2H, s), 5.13 (lH, d,
J=5Hz), 5.87 (lH, dd, J=5Hz, 9Hz),
6.48 (lH, broad sj, 7.43 (lH, s),
8.50 (lH, s), 9.62 (lH, d, J=9Hz),
12.58 (1~, s)
(3) A solution of 7-[2-(2-formamidothiazol-4-yl)-
2-ethoxycarbonylmethoxyiminoacetamido]-3-cephem-4-
carboxylic acid (syn isomer, 0.35 g.), conc.hydrochloric
acid (0.39 g.), ethanol (5.3 ml.) and tetrahydrofuran
(8 ml.) was stirred at room temperature for 4.5 hours.
After the resultant solution was concentrated in vacuo,
the residue was dissolved in an aqueous solution of
sodium bicarbonate, treated with activated charcoal
and filtered. The filtrate was adjusted to pH 3.5 with
10% hydrochloric acid under ice cooling.
The precipitates were collected by filtration, washed
with water and dried to give 7-[2-(2-aminothiazol-4-yl)-
2-ethoxycarbonylmethoxyiminoacetamido]-3-cephem-4-
carboxylic acid (syn isomer, 0.1 g.).
I. R. v maUxol : 3250, 3050, 1775, 1720,
1660, 1630, 1550 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 1.21 (3H, t, J-7Hz3,
3.59 (2H, s), 4.14 (2H, q, J=7Hz),
4.66 (2H, s), 5.10 (lH, d, J=5Hz),
5.83 (lH, dd, J=5Hz, 8Hz), 6.47 (lH,
broad s), 6.78 (lH, s), 7.23 (2H, s),
9.52 (lH, d, J=8Hz)
- 158 -
1340~93
Example 30
(1) The Vilsmeier reagent was prepared from
N,N-dimethylformamide ~0.32 g.) and phosphoryl chloride
(0.67 g.) in a conventional manner. After the reagent
was suspended in ethyl acetate (10 ml.), 2-~2-
formamidothiazol-4-yl)-2-(2,2,2-trifluoroethoxy~iminoacetic
acid (syn isomer, 1.2 g.) was added to the stirred
suspension under ice cooling and stirred at the same
temperature for 30 minutes. The solution was added
to a solution of 7-amino-3-cephem-4-carboxylic acid
(0.8 g.), and trimethylsilylacetamide ~4.2 g.) in
ethyl acetate (20 ml.) at -25~C, and stirred at -20
to -10~C for an hour. Water and ethyl acetate were
added to the resultant solution, and ethyl acetate
layer was separated. The aqueous layer was extracted
again with ethyl acetate. Water was added to the
combined extract and adjusted to pH 7.5 with a
saturated aqueous solution of sodium bicarbonate, and
then the aqueous layer was separated. Ethyl acetate
was added to the aqueous layer, adjusted to pH 1.5
with hydrochloric acid and the ethyl acetate layer was
separated. The aqueous layer was extracted again with
ethyl acetate. The extracts were combined, washed
with a saturated aqueous solution of sodium chloride
and dried over magnesium sulfate. After concentrating
the solution in vacuo, the residue was triturated with
diethyl ether, and the precipitates were collectbd
by filtration ~nd dried to give 7-[2-(2-formamidothiazol-
4-yl)-2-(2,2,2-trifluoroethoxyimino)acetamido]-3-
cephem-4-carboxylic acid (syn isomer, 1.55 g.).
- 159 -
'- 1340~gg
I.R. ~ maUxol : 3250, 1790, 1690, 1660,
1630, 1605, 1580, 1550 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 3.67 (2H, broad s),
4.78 (2H, q, J=8.5Hz), 5.17 (lH, d,
J=5Hz), 5.92 (lH, dd, J=5Hz, 8Hz),
6.53 (lH, t, J=4Hz), 7.52 (lH, s),
8.57 (lH, s), 9.83 (lH, d, J=8Hz),
12.67 (lH, broad s)
(2) A suspension of 7-[2-(2-fo~mamidothiazol-4-yl)-
2-(2,2,2-trifluoroethoxyimino)acetamido]-3-cephem-4-
carboxylic acid (syn isomer, l.S g.), conc. hydrochloric
acid (1.3 ml.), tetrahydrofuran (10 ml.) and methanol
(30 ml.) was treated in a similar manner to that of
Example 21-(3) to give 7-[2-(2-aminothiazol-4-yl)-2-
(2,2,2-trifluoroethoxyimino)acetamido]-3-cephem-4-
carboxylic acid (syn isomer, 1.1 g.).
I.R. v maUdol : 3450, 3300, 1780, 1660,
1625, 1590, 1550 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 3.60 (2H, broad s),
4.70 (2H, q, J=8.5Hz), 5.13 (lH, d,
J=5Hz), 5.87 (lH, dd, J=5Hz, 8Hz),
6.52 (lH, t, J=4Hz), 6.87 (lH, s),
9.80 (lH, d, J=8Hz)
Example 31
(1) A solution of 2-(2-formamidothiazol-4-yl~-
2-(2-chloroethoxyimino)acetic acid (syn isomer, 3.47 g.),
N,N-dimethylformamide (1.1 g.) and phosphoryl chloride
(2.3 g.) in ethyl acetate (35 ml.), and a solution
- 160 -
134049~
of 7-amino-3-cephem-4-carboxylic acid (2.5 g.) and
bis(trimethylsilyl)acetamide (12.7 g.) in ethyl acetate
t25 ml.) were treated in a similar manner to that of
Example 15-(1) to give 7-[2-(2-formamidothiazol-4-yl)-
2-(2-chloroethoxyimino)acetamido]-3-cephem-4-
carboxylic acid (syn isomer, 1.85 g.).
I.R. v maUxol : 3250, 3050, 1780, 1695,
1685, 1655, 1625 cm~l
N.M.R. ~ (DMSO-d6, ppm) : 3.62 (2H, d, J=4Hz),
3.86 (2H, t, J=6Hz), 4.37 (2H, t,
J=6Hz), 5.16 (lH, d, J=5Hz), 5.90
(lH, dd, J=5Hz, 9Hz), 6.52 (lH, t,
J=4Hz), 7.50 (lH, s), 8.53 (lH, s),
9.68 (lH, d, J=9Hz), 12.72 (lH, broad s)
(2) 7-[2-(2-Formamidothiazol-4-yl)-2-(Z-
chloroethoxyimino)acetamido]-3-cephem-4-carboxylic acid
(syn isomer, 1.8 g.), conc. hydrochloric acid (1.6 g.),
methanol (27 ml.) and tetrahydrofuran (40 ml.) were
treated in a similar manner to that of Example 15-(3)
to give 7-[Z-(2-aminothiazol-4-yl)-2-(2-chloroethoxyimino)-
acetamido]-3-cephem-4-carboxylic acid (syn isomer, 1.4 g.).
I.R. v max~l : 3440, 3300, 3070, 1780,
1660, 1625, 1555 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 3.60 (2H, s),
3.80 (2H, t, J=6Hz), 4.30 (2H, t, J=6Hz),
5.10 (lH, d, J=5Hz), 5.83 (lH, dd,
J=5Hz, 9Hz), 6.47 (lH, s), 6.78 (lH, s),
7.24 (2H, s), 9.58 (lH, d, J=9Hz)
- 161 -
,~ ~ .
1340~9.~
Example 32
(1) A solution of 2-(2-formamidothiazol-4-yl)-
2-(tert-butoxycarbonylmethoxyimino)acetic acid tsyn
isomer, 3.2 g.), N,N-dimethylformamide (0.852 g.) and
phosphoryl chloride (1.79 g.) in ethyl acetate (34 ml.)
and a solution of 7-amino-3-cephem-4-carboxylic acid
(1.95 g.) and bis(trimethylsilyl)acetamide (9.9 g.)
in ethyl acetate (19.5 ml.) were treated in a similar
manner to that of Example 15-(1) to give 7-[2-(2-
; lO formamidothiazol-4-yl)-2-(tert-butoxycarbonylmethoxyimino)-
acetamido]-3-cephem-4-carboxylic acid (syn isomer, 2.9 g.).
I.R. ~ mUaxol : 3260, 3180, 3060, 1785,
1730, 1690, 1640 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 1.44 (9H, s),
3.63 (2H, s), 4.62 (2H, s), 5.12
(lH, d, J=5Hz), 5.87 (lH, dd,
J=5Hz, 9Hz), 6.48 (lH, broad s),
7.42 (lH, s), 8.50 (lH, s), 9.57
(lH, d, J=9Hz), 12.62 (lH, broad s)
. .
(2) A mixture of 7-[2-(2-formamidothiazol-4-yl)-
2-(tert-butoxycarbonylmethoxyimino)acetamido]-3-cephem-
4-carboxylic acid (syn isomer, 2.8 g.), anisole (2.8 ml.)
and trifluoroacetic acid (11.2 ml.) was stirred at
room temperature for an hour. Ethyl acetate and water
were added to the resultant solution and adjusted to
pH 7.0 with sodium bicarbonate. The aqueous layer was
separated, and the ethyl acetate layer was extracted
with water. The aqueous extracts were combined,
- 162 -
~ , ,
....
13~0499
washed with ethyl acetate and diethyl ether in turn,
and then adjusted to pH 2.0 with 10% hydrochloric
acid under ice cooling. The precipitates were col-
lected by filtration, washed with water and dried to
give 7-[2-(2-formamidothiazol-4-yl)-2-
carboxymethoxyiminoacetamido]-3-cephem-4-carboxylic
acid (syn isomer, 1.43 g.).
- I.R. v mUaxol : 3270, 3120, 3070, 1760,
1720, 1690, 1660, 1620 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 3.60 (2H, s),
4.63 (2H, s), 5.11 (lH, d, J=5Hz),
5.88 (lH, dd, J=5Hz, 9Hz), 6.48 (lH,
t, J=4Hz), 7.44 (lH, s), 8.52 (lH, s),
9.59 (lH, d, J=9Hz), 12.64 (lH, broad
s)
(3) A mixture of 7-[2-(2-formamidothiazol-4-yl)-2-
carboxymethoxyiminoacetamido]-3-cephem-4-carboxylic
acid (syn isomer, 1.35 g.), conc. hydrochloric acid
(3.926 g.), methanol (20 ml.), water (10 ml.) and
tetrahydrofuran (40 ml.) was stirred at 30CC for 6 hours.
The resultant solution was concentrated in vacuo in
order to evaporate the methanol, and the aqueous
solution obtained was adjusted to pH 4.2 with 10%
aqueous solution of sodium hydroxide. The solution was
adjusted to pH 3.0 with 10~ hydrochloric acid. The
precipitates were collected by filtration and dried
to give 7-[2-(2-aminothiazol-4-yl)-2-
carboxymethoxyiminoacetamido]-3-cephem-4-carboxylic
acid (syn isomer, 0.8 g.).
- 163 -
.. .. ...
j 1340~93
I.R. v mUxol : 3300 ~broad), 3200 (broad),
1775, 1670, 1635 cm 1
N.M.R. ~(DMSO-d6, ppm) : 3.64 (2H, s),
4.64 (2H, s), 5.13 ~lH, d, J=5Hz),
5.86 (lH, dd, J=5Hz, 7Hz), 6.49
(lH, t, J=4Hz), 6.82 (lH, s), 7.33
(2H, s), 9.57 (lH, d, J=9Hz)
Example 33
(1) A solution of 2-(2-formamidothiazol-4-yl)-2-
(2,2,2-trifluoroethoxyimino)acetic acid (syn isomer,
0.9 g.), N,N-dimethylformamide (0.24 g.) and phosphoryl
chloride (0.5 g.) in ethyl acetate (10 ml.) and a sol-
ution of 4-nitrobenzyl 7-amino-3-chloro-3-cephem-4-
carboxylate hydrochloride (1.23 g.) and trimethylsilylacetamide
(2.8 g.) in ethyl acetate (20 ml.) were treated in a
similar manner to that of Example 21-(1) to give 4-
nitrobenzyl 7-[2-(2-formamidothiazol-4-yl)-2-(2,2,2-
trifluoroethoxyimino)acetamido]-3-chloro-3-cephem-4-
carboxylate (syn isomer, 1.9 g.).
I.R. v mUaxol : 3250, 1790, 1740, 1700,
1660, 1610, 1530 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 3.92 (2H, q, J=17Hz),
4.77 (2H, q, J=8.5Hz), 5.35 (lH, d,
J=5Hz), 5.48 (2H, s), 5.95 (lH, dd~
J=5Hz, 8Hz), 7.50 (lH, s), 7.70 (2H,
d, J=9Hz), 8.27 (2H, d, J=9Hz), 8.53
(lH, s), 9.92 (lH, d, J-8Hz), 12.67
(lH, broad s).
- 164 -
.
~ 1340~9.9
(2) A suspension of 4-nitrobenzyl 7-[2-(2-
formamidothiazol-4-yl)-2-(2,2,2-trifluoroethoxyimino)-
acetamido]-3-chloro-3-cephem-4-carboxylate (syn isomer,
1.8 g.) and 10% palladium carbon (0.9 g.) in methanol
(20 ml.) and tetrahydrofuran (20 ml.) was treated in
a similar manner to that of Example 21-(2) to give
7-[2-(2-formamidothiazol-4-yl)-2-(2,2,2-
trifluoroethoxyimino)acetamido]-3-chloro-3-cephem-4-
carboxylic acid (syn isomer, 1.0 g.).
I.R. v maUxol : 3250, 1780, 1690, 1655,
1530 cm 1
N.M.R. ~(DMSO-d6, ppm) : 3.86 (2H, q, J=17Hz),
4.80 (2H, q, J=8.5Hz), 5.33 (lH, d,
J=5Hz), 5.92 (lH, dd, J=5Hz, 8Hz),
7.53 (lH, s), 8.53 (lH, s), 9.93 (lH,
d, J=8Hz), 12.70 (lH, broad s)
(3) 7-[2-(2-Formamidothiazol-4-yl)-2-(2,2,2-
trifluoroethoxyimino)acetamido]-3-chloro-3-cephem-
4-carboxylic acid (syn isomer, 0.7 g.), conc.hydrochloric
acid (0.43 ml.) and methanol (16 ml.) were treated in a
similar manner to that of Example 21-(3) to give 7-[2-
(2-aminothiazol-4-yl)-2-(2,2,2-trifluoroethoxyimino)-
acetamido]-3-chloro-3-cephem-4-carboxylic acid (syn
isomer, 0.65 g.)
I.R. v maUxol : 3320, 3150, 1775, 1720,
1660, 1645, 1600, 1545 cm 1
N.M.R. ~(DMSO-d6, ppm) : 3.87 (2H, q, J=18Hz),
4.80 (2H, q, J=8.5Hz), 5.30 (lH, d, J=5Hz),
- 165 -
1340493
5.83 ~lH, dd, Js5Hz~ 8Hz),
7.05 (lH, s), 10.00 (lH, d, J=8Hz)
Example 34
(1) A solution of 2-(2-formamidothiazol-4-yl)-
2-butoxyiminoacetic acid (syn isomer, 1.5 g.), N,N-
dimethylformamide (440 mg.) and phosphoryl chloride
(~20 mg.) in ethyl acetate (12 ml.) and a solution of
4-nitroben~yl 7-amino-3-chloro-3-cephem-4-carboxylate
(2.03 g.), trimethylsilylacetamide (7 g.) and
bis(trimethylsilyl)acetamide (2 ml.) in ethyl acetate
(25 ml.) were treated in a similar manner to that of
Example 21-(1) to give 4-nitrobenzyl 7-[2-(2-
formamidothiazol-4-yl)-2-butoxyiminoacetamido]-3-
chloro-3-cephem-4-carboxylate (syn isomer, 2.8 g.),
yellow powder.
I.R. ~ maUxol : 3200-3250, 1780, 1730, 1690,
1655, 1605, 1530, 1350 cm~
N.M.R. ~ (DMSO-d6, ppm) : 0.90 (3H, m),
1.2-1.6 (4H, m), 3.88 (2H, AB-q,
J=19Hz), 4.0-4.2 (2H, m), 5.32 (lH,
d, J=4Hz), 5.44 (2H, s?, 5.92 (lH, d,d,
J=4Hz, 8Hz), 7.36 (lH, s), 7.68 (2H,
d, J=8Hz), 8.22 (2H, d, J=8Hz),
8.50 (lH, s), 9.72 (lH, d, J=8Hz),
12.56 (lH, s)
(2) A mixture of 4-nitrobenzyl 7-[2-(2-
formamidothiazol-4-yl)-2-butoxyiminoacetamido]-3-
chloro-3-cephem-4-carboxylate (syn isomer, 2.7 g.),
- 166 -
. .
.. . . . . .
. .
. .
- 13404~3
10~ palladium carbon (1.3 g.), water ~4 ml.),
acetic acid (0.4 ml.), methanol ~Z7 ml.) and
tetrahydrofuran (54 ml.) was treated in a similar
manner to that of Example 21-(Z) to give 7-[2-~2-
formamidothiazol-4-yl)-2-butoxyiminoacetamido]-3-
chloro-3-cephem-4-carboxylic acid ~syn isomer, 1.4 g.),
pale yellow powder.
I.R. v mUaxol : 3Z50, 2400-Z600, 1780, 1700,
1690, 1650, 1610 cm 1
N.M.R. ~ ~DMSO-d6, ppm) : 0.90 ~3H, m),
l.Z-1.70 ~4H, m), 3.88 ~2H, ABq,
Jel9Hz), 4.0-4.25 ~2lt, 1ll), 5.32
~lH, d, J-5Hz), 5.90 ~lH,d,d,
J-5Hz, 9Hz), 7.42 ~lH, s), 8.50
~lH, s), 9.73 ~lH, d, J-8Hz),
12.60 ~lH, s)
~3) A mixture of 7-[Z-(Z-formamidothiazol-4-yl)-
Z-butoxyiminoacetamido]-3-chloro-3-cephem-4-
carboxylic acid (syn isomer, 1.3 g.), conc. hydrochloric
acid (1.3 ml.) and methanol (Z0 ml.) was treated in a
similar manner to that of Example Zl-~3) to give
7-[Z-~Z-aminothiazol-4-yl)-Z-butoxyiminoacetamido]-3-chloro-3-
cephem-4-carboxylic acid ~syn isomer, l.Z g.), pale
yellow powder.
I.R. ~ NmaUxol : 3300, Z500-Z600, 1785,
1730, 1655, 1630 cm 1
. .
- 167 -
-
1340~93
N.M.R. ~ (DMSO-d6, ppm) : 0.90 (3H, m),
1.2-1.75 (4H, m), 3.88 (2H, AB-q,
J-19Hz), 5.17 (2H, m), 5.33 (lH,
d, J=511z), 5.83 (lH, d,d, J=SHz, 8Hz),
6.93 (lH, s), 9.50 (211, broad s),
9.85 (lH, d, J=8Hz)
Example 35
(1) A solution of Z-(2-formamidothiazol-4-yl)-2-
butoxyiminoacetic acid ~syn isomer, 1.09 g.), N,N-
dimethylformamide (322 mg.) and phosphoryl chloride
(675 mg.) in ethyl acetate (9.2 ml.) and a solution
of 4-nitrobenzyl 7-amino-3-methoxy-3-cephem-4-
carboxylate hydrochloride (1.5 g.), trimethylsilylacetamide
(5 g.) and bis(trimethylsilyl)acetamide (2 ml.) in
ethyl acetate (30 ml.) were treated in a similar manner
to that of Example 3-(1) to give 4-nitrobenzyl 7-[2-
(2-formamidothiazol-4-yl)-2-butoxyiminoacetamido]-3-
methoxy-3-cephem-4-carboxylate (syn isomer,l.8 g.).
I.R. v maUxol : 3300, 3220, 1770, 1715,
1690, 1650, 1610, 1540,
1350 cm 1
N.M.R. ô (DMSO-d6, ppm) : 0.90 (3H, m),
1.2-1.7 (4H, m), 3.72 (2H, broad s),
3.96 (3H, s), 4.10 (2H, m), 5.22 (lH,
d, J=4Hz), 5.32 (2H, s), 5.75 (lH,
d,d, J=4Hz, 8Hz), 7.43 (lH, s),
7.64 (2H, d, J=9Hz), 8.20 (2H, d, J=9Hz),
8~,48 (lH, s), 9.56 (lH, d, J=8Hz),
12.59 (lH, s)
.
-- 168 --
13AL049
r
(2) A mixture of 4-nitrobenzyl 7-[2-~2-
formamidothiazol-4-yl)-2-butoxyiminoacetamido]-3-
methoxy-3-cephem-4-carboxylate (syn isomer, 1.7 g.),
10~ palladium carbon (1 g.), water (3 ml.), acetic
acid l0.3 ml.), methanol (20 ml.) and tetrahydrofuran
(35 ml.) was treated in a similar manner to that of
Example 15-(2~ to give 7-[2-(2-formamidothiazol-4-yl)-
2-butoxyiminoacetamido]-3-methoxy-3-cephem-4-
carboxylic acid (syn isomer, 1 g.), yellow powder.
I. R. ~ mUaxol : 3200-3250, 2600, 1775,
1700, 1690, 1650 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 1.0 (3H, m),
1.2-1.75 (4H, m), 3.67 (2H, broad s),
3.86 (3H, s), 4.0-4.3 (2H, m),
5.23 (lH, d, J=4Hz), 5.68 ~lH, d,d,
J=4Hz, 8Hz), 7.50 (lH, s), 8.58 (lH,
s), 9.63 (lH, d, J=8Hz)
(3) A mixture of 7-[2-(2-formamidothiazol-4-yl)-
2-butoxyiminoacetamido]-3-methoxy-3-cephem-4-
carboxylic acid (syn isomer, 0.9 g.), conc. hydrochloric
acid ~0.8 ml.) and methanol (13.5 ml.) was treated in
a similar manner to that of Example 15-t3) to give
7-[2-(2-aminothiazol-4-yl)-2-butoxyiminoacetamido]-3-
methoxy-3-cephem-4-carboxylic acid (syn isomer, 0.4 g.),
yellowish white powder.
I.R. v NmUaxol: 3200-3300, 2600, 1770,
1705, 1670, 1620 cm 1
- 169 -
,- - , - ~ -
..."~
1340~93
N.M.R. ~ (DMSO-d6, ppm) : 0.90 (3H, m),
l.Z-1.65 (4H, m), 3.60 (2H, s),
3.96 (311, s), 4.0-4.16 (2H, m),
5.12 (lH, d, J=411z), 5.55 (lH,
d,d, J=4Hz, 8Hz), 6.80 ~lH, s),
7.Z-7.6 (ZH, broad s), 9.50 (lH,
d, J=8Hz)
- 170 -
1340~93
.
P~xample 36
(1) 4-Nitrobenzyl 7-amino-3-cephem-4-carboxylate ~5 g.)
was dissolved in a solution of trimetIlylsilylacetamide (13.8 g.)
and bis(trimethylsilyl)acetamide (10 ml.) in dry ethyl acetate
(50 ml.) and stirred at 45~C for 1.5 hours. A solution of
brominc (2.88 g.) in methylene chloride ~7 ml.) was added
dropwise to a solution of diketene (1.5 g.) in methylene
chloride ~7 ml.) at -40~C over 20 minutes and stirred at -30~C
for 1 hour. The solution obtained thus was added to dropwise
to the above solution of 4-nitrobenzyl 7-amino-3-cephem-4-
carboxylate under cooling at -15~C and then stirred at the
same temperature for 30 minutes. Water (50 ml.) was added
to the resultant solution and extractcd with ethyl acetate.
The ethyl acetate extract was washed with water, dried over
magnesium sulfate and concentrated under reduced pressure to
give oily 4-nitrobenzyl 7-[2-(2-bromoacetyl)acetamido]-3-
cephem-4-carboxylate (6.15 g.~
I.R. v mUaxol : 1780, 1740, 1630 cm 1
N.M.R. ~ppm ~DMSO-d6) : 3.62 ~2H, broad 51,
4.37 (2H, s), 5.08 (lH, d, J=5Ilz),
5.40 (2H, s), 5.77 - 6.05 (m),
- 171 -
. ~ . -
13~0~9
G.67 (1ll, t, J=5Hz), 7.68, 8.04 (411, m,
J='3llz), 9.07 (11l, d, J=81-lz)
(Z) 4-Ni~ro~cllzyl 7-[2-(2-~romoacetyl)acetalllido]-3-
cephem-4-carboxyla~e ~8.40 g.) was suspended in a mixture of
tetrahydrofuran (150 ml.) and water (30 ml.) To the sus-
pension were ad~ed acetic acid (50 ml.~ and a solution of
sodium nitrite (1.20 g.~ in water (15 ml.~ under ice-cooling,
and stirred at 20 to 22~C for 1.5 hours. The resultant
solution was poured into ice-water (300 ml.) and stirred for
20 minutes. lhe precipitating substance was collected by
- filtration, washed with water, dried and then recrystallized
from ethyl acetate to give 4-nitrobenzyl 7-[2-(2-bromoacetyl)-
2-hydroxyiminoacetamido]-3-cephem-4-carboxylate (syn-isomer :
3.1 g.), mp 153 to 162~C.
I.R. v NmaUxol : 3250, 1780, 1720, 1705, 1650, 1610,
1600 (shoulder), 1550, 1520 cm 1
N.M.R. ~ppm (DMSO-d6) : 3.67 (2H, d, J~4Hz),
4.63 (1.5H, 9), 4.88 (0.5H, 8), 5.18 (I~,
d, J=5Hz), 5.45 (2H, e), 5.93 (I~, dd, J=5Hz, 8Hz),
6.72 (lH, t, J=4Hz), 7.73 (2H, d, J=9Hz),
8.28 (2H, d, J=9Hz), 9.38 (111, d, J~8Hz),
11.27 (lH, s)
'
(3) A solution of diazomethane in diethyl ether was
added little by little to a solution of 4-nitrobenzyl 7-[2-
(2-bromoacetyl)-2-hydroxyiminoacetamido)-3-cephem-4-
carboxylate (0.9 g.) in tetrahydrofuran (30 ml.) under ice-
cooling until the reaction terminated, and then acetic acid
- i72 -
1 3 ~ .9 3
was added ~o the resultall~ so]utioll to dccompose excess
diazoolc~l-alle. Tlle resultal-t solu~ion was collcen~rated wlder
reduced pressure to give the foallly product of 4-nitrobenzyl
7-[2-(2rbromoacety~-2-metllox~inlinoacctanlido]-3-cephem-4
carboxylate (syn-isomcr : 0.9 g.)
(4) 1hiourea (0.14 g.) was added to a solution of 4-
nitrobenzyl 7-[2-(2-bromoacety~-2-methoxyiminoacetamido]-3-
cephem-4-carboxylate (syn-isomer : 0.8 g.) in ethanol (20 ml.)
and water (S ml.), and stirred at room temperature for 3.5
hours. The resultant solution was concentrated under reduced
pressure, and to the residue were added water and ethyl
acetate. The ethyl acetate layer was separated, washed with
water, dried over magnesium sulfate and concentrated under
reduced pressure to give the crude product (0.6 g.)
The product was purified by column chromatography on silica
gel (eluent : benzene and ethyl acetate (8:2) ) to give 4-
nitrobenzyl 7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido~-
3-cephem-4-carboxylate (syn-isomer : 0.21 g.), mp 165 to 170~C
(dec.)
I.R. ~ NmUaxol : 3350-3200, 1770, 1720, 1665,
1615, 1515 cm 1
N.M.R. ~ppm (DMSO-d6) : 3.60 (211, broad s),
; 3.81 (3H, s), 5.12 (lH, d, J=5Hz),
5.36 (2H, s), 5.83 (1l1, dd, J~5Hz, 8Hz),
6.64 (lH, t, J-4Hz), 6.70 (lH, s), 7.20 (2H, s),
7.65 (2H, d, J=9Hz.), 8.19 ~2H, d, Je9Hz),
9.60 (lH, d, J=8Hz)
- 173 -
, .... ... .
1340~99
~xample 37
(1) A solution of bromille (43.0 g.) in metllylene chloride
(30 ml.) was dropwise added to a solution of diketene
(22.6 g.) in mctllylene chloride (30 ml.) at -30~C over 35
minutcs, and stirrcd at tl-c same temperature for 30 minutes.
The solution was dropwise added to a stirred solution of
4-nitrobenzyl 7-amino-3-cephem-4-carboxylate (75.1 g.) ànd
bis(trimethylsilyl)acetamide (G8.4 g,) in tetrahydrofuran
(l.S Q) at -15~C over 10 minutes, and the solution was
stirred at the same temperature for 50 minutes. Water
(35 ml.) and an aqueous solution (35 ml.) of sodium nitrite
(18.6 g.) were addcd to the resultant solution while keeping
at p~l 2.0, and the solution was stirred at 10 to 15~C for
15 minutes. Aft.er the solution was adjusted to pH 4.5
with a saturated aqueous solution of sodium bicarbonate,
an aqueous solution (150 ml.) of thiourea (17.1 g.) was
added to the solution, adjusted to p~l 6.0 with a saturated
aqueous solution of sodium bicarbonate, and stirred for 20
minutes. The organic layer was separated and concentrated
under reduced pressure. The residuc was dissolved in e~hyl
acetate (1.5 Q), and washed with water three times. The
solution was dried over magllesium sulfate, treated with
activated charcoal and concelltrated under reduced pressure.
After the residue was triturated with diethyl ether (200 ml.),
~he precipitates were collected by decalltation andwasllcd with ether
acetate ~300 ml.), a mixture of tetrahydrofuran (500 ml.)
and ethyl acetate (1 ~) at fiO~C and ther. witll cthyl acetate
(100 ml.) three times, and dried to give 4-nitro~ellzyl
7-[2-(2-amino-4-thiazolyl)-2-hydroxyiminoacetamido]-3-.
cephem-4-car~oxylate (syn isomer, 55.5 g.).
- 174 -
.. ..
1340493
Nu~ol 1760, 1710, 1660,1630 cm
N.M.R. ~ppm (DMSO-d6) : 3.60 (2ll, d, J=5Hz),
5.12 (1ll, d, J=5Hz), 5.39(2tl, s),
5.88 (lH, dd, J=8Hz, 5tlz), 6.63 (lH, s),
6.53 - 6.77 (lH, m), 7.08 (2H, broad s),
7.68 (ZH, d, J=9Hz), 8.Z2 (2H, d, J~9Hz),
9 47 (ltl, d, J=8Hz), 11.33 (lH, s)
- 175 -
1340 ~19.~
(2) 10% Palladium carbon (0.35 g.) was added to a solut-
ion of 4~nitrobenzyl 7-[2-(2-amino-4-tlliazolyl)-2-
hydroxyiminoacetamido]-3-cephem-4-carboxylate (syn-isomer :
0.7 g.) in methanol (70 ml.), and the mixture was subjected
to catalytic reduction at room temperature under atmospheric
pressurc for 1.5 hours. The resultant mixture was filtered,
and the filtrate was concentrated under reduced pressure.
To the residue was added an aqueous solution of sodium
bicarbonate and the insoluble substance was filtered out.
lO The filtrate was washed with ethyl acetate and methylene
chloride in turn, bubbled with nitrogen gas and then lyophi-
lized. The residue was dissolved in water (30 ml.) and
adjusted to pH 3.8 with 10% hydrochloric acid. The solution
was subjected to column chromatography on macroporous, non-
ionic adsorption resin "Diaion IIP-20" (Trade mark; manufactured
by Mitsubishi Chemical Industries Ltd., 20 ml.), washed with
water, and then eluted with 40% aqueous acetone. After acetone
was removed from the eluate under reduced pressure, the residue
was lyophilized to give 7-[2-(2-amino-4-thiazolyl)-2-
20 hydroxyiminoacetamido]-3-cephem-4-carboxylic acid (syn-isomer:
0.25 g.)
l,R. ~ maUxol : 3350 to 3200, 1770, 1670, 1630 cm 1
N.M.R, ~ ppm (D~ISO-d6) : 3.60 (2H, broad s),
S.10 (lH, d, J=SHz), 5.83 (lH, dd, J-5Hz,
8Hz), 6.47 (lH, t, J=4Hz), 6.67 (lH, s) r
9.47 (lH, d, J=8Hz)
- 176 -
,
13~0~93
Example 38
(1) Thiourea ~0.18 ~.) was added to a suspension of 4-
nitrobenzyl 7-[2-(2-bromoacety~-2-hydroxyiminoacetamido]-3-
cephem-4-carboxylate (syn-isomer : 1.05 g.) in ethanol ( 25
ml.), tetrahydrofuran (25 ml.) and water (5 ml.), and stirred
at roolD temperature for 4 hours. The resultant solution
was concentrated under reduced pressure and cooled. The
residue was crystallized by treating with a mixture of tetra-
hydrofuran and ethyl acetate, and collected by filtration to
give 4-nitrobenzyl 7-[2-(2-amino-4-thiazolyl)-2-hydroxyimino-
acetamido]-3-cephem-4-carboxylate (syn-isomer : O.9S g.),
colorless crystals, mp 172 to 175~C (dec.)
I.R. v nNaUxol : 3350 - 3200, 1770, 1725, 1670,
1625, 1520 cm 1
N.M.R. ~ppm (DMSO-d6) : 3.68 (2H, d, J=4Hz),
5.20 (lH, d, J=511z), 5.43 (2H, s),
5.90 (lH, dd, J=8Hz, 5Hz), 6.70 (lH, t,
J~4Hz), 6.88 (lEI, s), 7.70 ~2H, d, J=9Hz),
8.23 (2EI, d, J=9Hz), 9.68 (lH, d, J=8Hz)
(2) A solution of diazomethane in diethyl ether was
. added little by little to a solution of 4-nitrobenzyl 7-[2-
(2-amino-4-thiazolyl)-2-hydroxyiminoacetamido]-3-cephem-4-
carboxylate (syn-isomer : 0.3 g.) in methanol (30 ml.) until
the reaction terminated. The resultant solution was con-
centrated under reduced pressure, and the residue was pul-
verized with diethyl ether, collected by filtration and
dried to give 4-nitrobenzyl 7-[2-(2-amino-4-thiazolyl)-2-
methoxyiminoacetamido]-3-cephem-4-carboxylate (syn-isomer : 0.26 g.)
- 177 -
, --. ~;.; , .. .
.
1~049~
This product was identified with the authentic sample.
Example 39
7-Amino-3-cephem-4-carboxylic acid (2.54 g.) was
dissolvcd in a solution of trimethylsilylacetamide ~11.7 g.)
and bis(trimethy]silyl)acctamide (15 ml.) in dried ethyl
acetate (50 ml.) A solution of broniine (2.43 g.) in dried
methylcne chloride (10 ml.) was added dropwise to a solution
of diketene (l.Z8 g.) in dried methylene chloride (25 ml.)
at -30~C over 10 minutes and stirred at the same temperature
for 1.5 hours. The solution was added to the above solution
containing 7-amino-3-cephem-4-carboxylic acid at -15~C over
10 minutes, and stirred at -15 to -10~C for 1.5 hours.
Water (50 ml.) was added to the resultant solution. The
ethyl acetate layer was separated, and extracted with aqueous
solutioll of sodium bicarbonate. The aqueous extract was
adjusted to pH 2.0 with 10% hydrochloric acid and extracted
with ethyl acetate. The ethyl acetate extract was washed with
water, dried ovcr magnesium sulfate and concentrated under
reduced pressure to give 7-[2-(2-bromoacetyl)acetamido]-3-
cephem-4-carboxylic acid (2.82 g.)
I.l~. v mUaxol : 1760, 1660 cm 1
N.M.R. ~ppm ~DMSO-d6) : 3.58 (2H, d, J-4Hz),
3.65 ~2H, s), 4.40 (2H, s), 5.06 ~lH, d, J-5Hz),
5.73 (lH, dd, Jc8Hz, 5Hz), 6.50 ~lH, t, J=4Hz),
9.08 (lH, d, J-81-lz)
- 178 -
13~0~
Example 40
The following compounds were prepared in a
similar manner to that of Example 36.
(1) 4-Nitrobenzyl 7-[2-(2-aminothiazol-4-yl)-2-
ethoxyiminoacetamido]-3-cephem-4-carboxylate
(syn isomer)
(2) 4-Nitrobenzyl 7-[2-(2-aminothiazol-4-yl)-2-
propoxyiminoacetamido]-3-cephem-4-carboxylate
(syn isomer)
(3) 4-Nitrobenzyl 7-[2-(2-aminothiazol-4-yl)-2-
isopropoxyiminoacetamido]-3-cephem-4-
carboxylate (syn isomer)
Example 41
The following compounds were prepared in a similar manner
to that of Example 38-(2).
(1) 4-Nitrobenzyl 7-[2-(2-aminothiazol-4-yl)-2-
ethoxyiminoacetamido]-3-cephem-4-carboxylate
(syn isomer)
(2) 4-Nitrobenzyl 7-[2-(2-aminothiazol-4-yl)-2-
propoxyiminoacetamido]-3-cephem-4-carboxylate
(syn isomer)
(3) 4-Nitrobenzyl 7-[2-(2-aminothiazol-4-yl)-2-
isopropoxyiminoacetamido]-3-cephem-4-
carboxylate (syn isomer)
- 179 -
.
... . . ..
13404~9
Ex~mple 42
(1) Sodium boron hydride (160 mg.) was added to a
suspension of 4-nitrobenzyl 7-[2-(2-formamido-4-
thiazolyl)-2-methoxyiminoacetamido]-3-hydroxy-3-cephem-4-
carboxylate (syn isomer, 1 g.) in tetrahydrofuran (10
ml.), acetic acid (3 ml.) and water (1 ml.) at 0~C over 10
minutes, and stirred at 0 to 3~C for 55 minutes. After
water was added to the resultant solution, the solution
was extracted with ethyl acetate. The extract was washed
with a saturated aqueous solution of sodium chloride, a
saturated aqueous solution of sodium bicarbonate and a
saturated aqueous solution of sodium chloride in turn, and
dried over magnesium sulfate. The solution was
concentrated under reduced pressure, and the residue was
pulverized with diethyl ether to give 4-nitrobenzyl 7-[2-
(2-formamido-4-thiazolyl)-2-methoxyiminoacetamido]-3-
hydroxycepham-4-carboxylate (syn isomer, 0.77 g.), mp 172
to 175~C (dec.).
I.R. v j : 3250, 1775, 1745, 1660 cm~
max
N.M.R. ~(DMSO-d6, ppm) : 2.76 (lH, dd, J=14Hz,
3Hz), 3.17 (lH, dd, J=14Hz, 13Hz),
3.92 (3H, s), 4.03 (lH, m), 4.72 (lH, d,
J=6Hz), 5.24 (lH, d, J=4Hz), 5.37 (2H, s),
5.56 (lH, dd, J=9Hz, 4Hz), 6.07 (lH, d,
J=4Hz), 7.44 (lH, s), 7.72 (2H, d, J=8Hz),
8.27 (2H, d, J=8Hz), 8.54 (lH, s),
9.67 (lH, d, J=9Hz).
- 180 -
r
,~~, . .. . ...
134~93
(2) ~esyl chloridc (0.406 g.) was dropwise added to a
stirred mixture of 4-nitrobenzyl 7-[2-(2-formamido-4-
thiazolyl)-2-methoxyiminoacetamido]-3-11ydroxycepllam-4-
carboxylatc (syn isomer, 1 g.), N,N-dimethylformamide ~10
ml.) and potassium carbonate (0 732 g.) at 0 to 5~C over
2 minutes, and the solution was stirred at room temperature
for 2.5 hours. After ethyl acetate and water were added
to the resultant solution, the solution was extracted with
ethyl acetate. The remaining aqueous layer was extracted
again with ethyl acetate. The ethyl acetate extract
solution was washed with a saturated aqueous solution of
sodium chloride,dried over magnesium sulfate, and
concentrated under reduced pressure. The residue was
subjected to column chromatography on silica gel ~30 g.)
and eluted Witll a mixture of chloroform and ethyl ace~ate.
The eluate was concentrated under reduced pressure to give
4-nitrobenzyl 7-l2-~2-formamido-4-thiazolyl)-2-methoxyimino-
acetamido~-3-cephem-4-carboxylate (syn isomer, 0.12 g.),
mp 224~C ~dec.).
- 181 -
.;
13~.19.~
Example 4~
Phosphoryl chloride (704 mg.) was added dropwise to a
solution of N,N-dimethylformamide (336 mg.) in ethyl acetate
(8 ml.) below 5~C and stirred at the same temperature for
30 minutes. 2-(2-Formamidothiazol-4-yl)-2-methoxyiminoacetic
acid (syn isomer, 1 g.) was added to the solution and stirred
at 5 to 10~C for an hour. The solution was added dropwise
to a solution of 7-amino-3-hydroxycepham-4-carboxylic acid
(872 mg.) and trimethylsilylacetamide (1.05 g.) in ethyl
acetate (20 ml.) at -20~C over 5 minutes, and stirred at
-20 to -25~C for an hour. Water (50 ml.) was added to the
resultant solution and adjusted to pH 7.0 with sodium
bicarbonate. The aqueous layer was separated, and the ethyl
acetate layer was extracted again with water (10 ml.).
The aqueous extracts were combined, adjusted to pH 6 and
absorbed on macroporous nonionic adsorption resin Diaion
HP-20 (50 ml., Trademark, manufactured by Mitsubishi Chemical
Industries). The column was washed with water (50 ml.) and
eluted with 30% aqueous isopropyl alcohol. The eluate
containing the object compound was adjusted to pH 6.5 and
concentrated under reduced pressure. The residue was
lyophilized to give sodium 7-[2-(2-formamidothiazol-4-yl)-2-
methoxyiminoacetamido]-3-hydroxycepham-4-carboxylate (syn
isomer, 1.1 g.)
N.M.R. ~ (D2O, ppm): 2.72 - 3.18 (2H, m), 4.02 (3H, s),
4.02 - 4.28 (lH, m), 4.54 (lH, d, J=4Hz), 5.28 (lH, d,
J=4Hz), 5.53 (lH, d, J=4Hz), 7.50 (lH, s), 8.53 (lH, s)
- 182 -
.. . .
1340~93
Example 44
l'hiollyl chl~ridc (0.423 ~.) was dro~wise added to a
stirred solution of 4-nitrobenzyl 7-[Z-~2-formamido-4-
t:hiazolyl)-2-methoxyilllinoacctamido]-3-hydroxy-3-cephem-4-
carboxylate (syn isomer, 1 g.) in N,N-dimethylformamide
~10 ml.) under ice cooling over Z minutes, and the solution
was stirred at room temperature for 1.1 hours. Ethyl
acetate (40 ml.) and water (30 ml.) were added to the
resultant solution and shaken suf~iciently. The aqueous
layer was extracted with ethyl acetate, and the extract
was combined Wit]l the ethyl acetate layer separated above.
The ethyl acetate solution was washed with a saturated
aqueous solution of sodium chloride, dried over magnesium
sulfate and then concentrated under reduced pressure.
The residue was subjected to column chromatography on silica
gel (30 g.) and eluted with chloroform and then a mixture
of chloroform and ethyl acetate (7:3). The latter eluate
was concentrated under reduced pressure to give 4-
nitrobenzyl 7-[2-(2-formalllido-4-thiazolyl)-2-methoxyimino-
acetamido]-3-chloro-3-cephem-4-carboxylate (syn isomer, 0.2 g.),
mp 216~C (dec.).
I.R. ~mUaxol : 3230 (shoulder), 3110, 3050, 1785,
1725, 1690, 1655 cm 1
N.M.R. ~ppm (DMSO-d6) : 3.93 (3H, s), 3.93 (2H,
q, J=18l1z), 5.36 (lH, d, J=51~z), 5.50 (2H,
s), 5.97 (lH, dd, J=5Hz, 9llz), 7.45 (lH, z),
7.73 (2H, d, J=9Hz), 8.29 (ZH, d, J=9Hz),
8.56 (lH, s), 9.78 (lH, d, J=9Hz).
- 183 -
~ ~34049~
E~ample 45
The iollowing compounds were prepared in a similar
manner to that oi E~ample 44.
(1) 4-Nitrobenzyl 7-[2-(2-formamidothiazol-4-yl)-2-propo~y-
iminoacetamido]-3-chloro-3-ceph- q-carbo~ylate
(~yn isomer)
(2) 4-Nitrobenzyl 7-[2-(2-formamidoth~AzQl-4-yl)-2-
(2,2,2-trifluoroetho~yimino)acetamido]-3-chloro-3-
cephem-4-carbo~ylate (syn isomer)
(3) 4-Nitrobenzyl 7-[2-(2-iormamidothiazol-4-yl)-2-
buto~yiminoacetamido]-3-chloro-3-cephem-4-
carbo~ylate (syn isomer)
.
- 184 -
. . .
o493
Example 46
Fermentation:-
Pre-culture mediw~: Trypticase so~ broth (BBL)
Main culture medium:
glycerin 3 g
peptone 1 g
corn steep liquor 1 g
dry yeast 2 g
sodium carbonate 0.1 g
KH2PO4 g
Na2HPO4 2 2.15 g
(The above components were dissolved ir. water
so as to become 100 ml in total, and the
medium was adjusted to pH 7.2.)
A main culture broth (100 ml.) was placed in Sakaguchi-flask
~500 ml) and sterilized at 120~C for 20 minutes. Into this
medium, there was inoculated a culture broth (1 ml.) of each
of microorganisms as given below, which were cultured at 30 C
degree for 18 hours in a pre-culture medium, respectively and
then a shaking culture was conducted at 30 C degree for 48 hours.
Reaction:-
Into above cultured broth, (1 ml)there was added the Substrate (0.1 g) as given below suspended
in 0.1 M phosphate buffer (pH 7.2, 1 ml.), and then the mixture
was shaked at 30~C for48 hours.
Identification and assay:-
After the reaction, in order to identify the generated
product the reaction mixture as obtained above was chromatographed
- 185 -
13~049~
on Eastman chromatogram 6065 cellulose at room temperature.
As a developping agent, there was used (A) the upper layer
of a mixture of n-butanol, ethanol and water (4:1:5 by volume)
and (B) a mixture of n-propanol and water (7:3 by volume).
Rf value was determined by index of antimicrobial activity
against a sensitive strain of Escherichia coli ES 111, and
as the result only one spot showing each of Product I and II
was observed on the Eastman chromatogram G065 cellulose without
showing any spot of each of Substrate I and II. Rf value are
10 shown in the following table.
Developping Solvent
A B
Reaction Mixture 0.85 0.90
(Substrate I) 0.60
(Product II) 0-90 0.92
Reference 0.36 0.54
Note:
Substrate I: 4-nitrobenzyl 7-[2-formamido-4-thiazolyl)-
2-methoxyiminoacetamido]-3-cephem-4-
carboxylate (syn isomer)
Product I: 7-[2-(2-formamido-4-thiazolyl)-2-methoxyimino-
acetamido~-3-cephem-4-carboxyllc acid (syn isomer)
Substrate II: 4-nitrobenzyl 7-[2-amino-4-thiazolyl)-2-
methoxyiminoacetamido]-3-cephem-4-carboxylate
(syn isomer)
Product II: 7-[2-(2-amino-4-thiazolyl)-2-methoxyimino-
acetamido]-3-cephem-4-carboxylic acid (syn isomer)
- 186 -
13~0~
..
The product generated in the reaction mixture as obtained
above was assayed by paper disk-plate method using a sensitive
strain of Escherichia coli ES 111 (culture: 37~C, 16 hours)
and the yield thereof was calculated. The results are shown
in the following.
Microorganism Yield (%)
used for enzymatic
hydrolysis Product I Product II
Bacillus subtilis IAM 1069 75 60
~ sphaericus IAM 1286 75 20
" subtilis IAM 1107 75 95
" subtilis I~l 1214 85 20
Corynebacterium equi IAM 1038 95 95
Micrococcus varians IAM 1314 70 20
Flavobacterium rigens IAM 1238 85 90
Salmonella typhimurium IAM 1406 90 20
Staphylococcus epidermidis IAM 1296 90 95
Microbacterium flavum IAM 1642 90 95
The following are examples of pharmaceutical compositions
prepared in accordance with this invention and containing
7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-
4-carboxylic acid (syn isomer, Compound A) as the active
substance.
- 187 -
- :.: -. -
.
1340 i'~
Example 47. (lyophlized preparation for injection)
Sodium salt of compound A (20 9. po~ency) was dissolved
in water (200 ml.), and the solution (5 ml.) was filled in
each 10 ml. vial. These vials are frozen and dried in a vacuum
(lyophilization).
Example 48. (suspension for injection)
Compound A 2 5 g .
Methyl cellurose 0. 5 g.
Methyl 4-oxobenzoate0.1 g.
Polysolvate 80 0.1 y.
Lidocaine hydrochloride 0.5 g.
Water for injectiosl to make 100 ml.
This aqueous suspension is suitable for intramuscular
injection.
Example 49. (tablets for oral use)
Compound A 5 0 0 mg.
Lactose 375.5 mg.
Hydroxypropylcellurose 2 mg.
Magnesium stearate 22. 5 mg.
This mixture provides a tablet for oral use in the
treatment of infectious deseases caused by pathogenic ~acteria.
Example 50. (capsule for oral use)
Cosnpound A 500 mg.
~agnesium stearate 10 mg.
This mixture provides a capsule for oral use in the
treatment of infectious deseases caused by pathogenic bacteria.
- 188 -
13~0~!3.~
Example 51
(l) A solution of 2-(2-formamidotl1iazol-4-yl)-2-n-
hexyloxyiminoacetic acid (syn isomer, 3.24 g.), N,N-
dimethylformamide (0.951 g.), and phosplloryl chloride
(2.00 g.) in ethyl acetate (20 ml.), and a solution of
4-nitrobenzyl 7-amino-3-chloro-3-cephem-4-carboxylate
hydrochloride (4 g.) in a mixture of acetone (20 ml.),
water (20 ml.) and tetrahydrofuran (40 ml.) were treated
in a similar manner to that: of Example 21-(l) to give
4-nitrobenzyl 7-[2-(2-formamidotl1iazol-4-yl)-2-n-11exyloxy-
iminoacetamido]-3-chloro-3-cephem-4-carl)oxylate (syn
isomer, 5.78 g.).
I.R. v mNUaxol : 3240, 3200, 3050, 1780, 1730,
1695, 1660 cm l
N.M R. ô (DMSO-d6, ppm) : 0.6~2.1 (llH, m),
3.96 (211, q, J=l911z), 4.15 (211, t, J=6Hz),
5.37 (lH, d, J=5Hz), 5.50 (2H, s),
5.97 (lH, d, d, J=SHz, 81-1z), 7.42 (lH, s),
7.72 (2H, d, J=911z), 8.28 (2H, d, J=9Hz),
8.54 (lH, s), 9.74 (111, d, J=811z),
12.73 (lH, broad s)
(2) A mixture of 4-nitrobenzyl 7-[2-(2-formamido-
- thiazol-4-yl)-2-n-hexyloxyiminoacetamido]-3-chloro-3-
cephem-4-carboxylate (syn isomer, 5.6 g.), acetic acid
(0.4 ml.), 10% palladium carbon (2.24 g.), water (4 ml.),
methanol (23 ml.) and tetrahydro~uran (56 ml.) was treated
in a similar manner to that of Example 21-(2) to give
7-[2-(2-formamidothiazol-4-yl)-2-n-11exyloxyiminoacetamido]-
3-chloro-3-cephem-4-carboxylic acid (Syl1 isomer, 2.5 g.).
-- 189 --
1340~9.~
-I.R. ~ IN~axol : 3225, 1785, 1690, 1650 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 0.6~2.0 (1111, m),
3.86 (2H, q, J=18Hz), 4.13 (2H, t, J=6Hz),
5.30 (1l-1, d, J=5Hz), 5 88 (lH, d, d,
J=5Hz, 8Hz), 7;41 (lH, s), 8.54 (]H, s),
9.70 (11-1, d, J=8Hz), 12.68 (1l-1, s)
(3) A mixture of 7-[2-(2-formamidothiazol-4-yl)-2-
n-hexyloxyiminoacetamido]-3-cllloro-3-cepllem-4-carboxylic
acid (syn isomer, 2.4 g.), conc,hydrochloric acid (1.84 g.),
methanol (36 ml.) and tetrahydrofurall (30 ml.) was stirred
at 30~C for 2 hours. The resultant solutiun was concen-
trated in vacuo. Water (60 ml.) was added to the residue
and the precipitates were collectc~ by filtration, washed
with water and dried over phosphorus pentoxidc to give
7-[2-(2-aminothiazol-4-yl)-2-n-hexyloxyiminoacetamido]-3-
chloro-3-cepllem-4-carboxylic acid (syn isomer, 1.86 g.).
I.R. v mUa~ol : 3300, 1780, 1665, 1535 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 0.6~2.0 (11ll, m),
3.84 (2H, q, J=1811z), 4.08 (21-1, t, J=7Hz),
5.28 (1ll, d, J=SHz), 5.82 (11-1, d, d,
J=SHz, 8Hz), 6.77 (1ll, s), 9.66 (lH, d,
J=8Hz), 6.0~8.0 (2H, broad s)
Example 52
(1) 2-(2-Formamidothiazol-4-yl)-2-propar~yloxyimino-
acetic acid (syn isomer, 1.27 g.), N,N-dimethylformamide
(402 mg.), phosphoryl chloride (843 mg.) and ethyl acetate
(11.2 ml.) were treated in a conventional manner to prepare
an activated acid solution. On the other hand, a mixture
of 7-amino-3-methoxy-3-cephcm-4-carboxylic acid
-- 190 --
13~0~3
hydrochloride (1.33 g.), trimethylsilylacetamide (4 g.),
bis(trimethylsilyl)acetamide (2 ml.) and ethyl acetate
(20 ml.) was stirred at 40 to 45~C for an hour. To the
solution was added the activated acid solution obtained
above all at once at -15~C, and then stirred at -5 to
-10~C for 1.5 hours. ~at-er (30 ml.) was added to the
resultant solution, filtered and the organic layer was
separated. The insoluble substance filtered out was
dissolved in a saturated aqueous solution of sodium
bicarbonate, and the solution was added to the organic
layer. The solution was adjusted to pll 7.5 and the
aqueous solution was separated, and tllell extracted with
ethyl acetate at pH 2Ø
The extract was washed with water, dried over magnesium
sulfate and filtered. The filtrate was concentrated in
vacuo to give 7-[2-(2-formamidothiazol-4-ylj-2-
propargyloxyiminoacetamido]-3-methoxy-3-cephem-4-carboxylic
acid (syn isomer, 1.0 g.), yellow powder.
I.R. v mUaxol : 3200~3300, 2500~2600, 2120,
1770, 1710, 1690, 1670 cm~
N.M.R. ~ (DMSO-d6, ppm) : 3.50 (lH, m), 3.65
(2H, s), 3.82 (3H, s), 4.80 (2H, d,
J=21{z), 5.20 (lH, d, J=41{z), 5.62 (lH,
d,d, J=4Hz, 8Hz), 7.52 (lH, s), 8.55
(lH, s), 9.68 (1ll, d, J=8Hz), 12.65
(lH, broad s)
(2) A mixture of 7-[2-(2-formamidothiazol-4-yl)-2-
propargyloxyiminoacetamido]-3-methoxy-3-cepllem-4-carboxylic
acid (syn isomer, 0.9 g.), conc. hydrochloric acid (0.9 ml.)
and methanol (13.5 ml.) was stirred at room temperature
-- 191 --
.
1340~9~
for 4 hours. After concentrating the resultant solution
in vacuo at 35~C, the residue was dissolved in water
and washed with ethyl acetate. The aqueous solution was
adjusted to pH 7.0 with sodium bicarbonate and washed with
ethyl acetate and diethyl ether. After removing the
organic solvent by bubbling nitrogen gas, the solution was
adjusted to pl-l 3.0 with 10% hydrochloric acid and stirred
under ice cooling. The precipitates were collected by
filtration, washed with water and dried to give 7-[2-(2-
aminothiazol-4-yl)-2-proyargyloxyiminoacetamido]-3-
methoxy-3-cephem-4-carboxylic acid (syn isomer, 0.25 g.),
whitish yellow powder.
I.R. v maUxol : 3300, 2500~2600, 2120, 1775,
1710, lG70, 1620 cm 1
N.M.R. ~ (DMSO-dG, ppm) : 3.52 (1ll, m), 3.82
~3H, s), 4.77 (21-1, d, J=21-1z), 5.17
(lH, d, J-4Hz), 5.58 (lH, d,d, J=4Hz,
8Hz), 6.93 (lH, s), 7.1~7.3 (2H, broad s),
9.G7 (lH, d, J=8Hz)
Example 53
(1) 2-(2-Formamidotlliazol-4-yl)-2-propargyloxyimino-
acetic acid (syn isomer~ ].27 g.), N?N-dimethylformamide
(400 mg.), phosphoryl chloride (850 mg.) and ethyl acetate
(11.2 ml.) were treated in a conventional manner to give
the activated acid solution. The activated acid solution
was added to a solution of 7-amino-3-chloro-3-cephem-4-
carboxylic acid hydrochloride (2 g.), trimethylsilyl-
acetamide (6 g.) and bis(trimethylsilyl)acetamide (3 ml.)
in ethyl acetate (40 ml.) at -15~C all at once, and stirred
- 192 -
1340~93
at -5 to -10~C for 1.5 hours. After adding water (30 ml.)
to the resultant solution, the ethyl acetate layer was
separated and extracted with a saturated aqueous solution
of sodium bicarbonate. Ethyl acetate was added to the
aqueous extract, adjustecl to pH 2.0 with 10% hydrochloric
acid. The ethyl acetate layer was separatcd, washed
- with wa1:er, dried over magnesium sulfate and filtered.
The filtrate was concentlated in vacuo, and tlle residue
was crystallized with a mixture of ethyl acetate and
diisopropyl ether. The precipita~es were collected by
filtration and dried to givc 7-[2-(2-formamidot}liazol-4-
yl)-2-propargyloxyiminoacetamido]-3-chloro-3-cepllem-4-
carboxylic acid (syn isome1, 1.5 g.), yellow powder.
I.R. v mUaxol: 3250~3300, 2500~2600, 2120j
1780, 1725, 1690, 1670 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 3.45 (lH, m), 3.57
(2H, AB-q, J=2011z~, 4.77 (21-1, d, J=211z),
5.28 (lH, d, J=411z), 5.80 (lH, d,d>
J=411z, 8Hz), 8.42 (11-1, s), 8.52 (lH, s),
9.78 (111, d, J=811z), 12.72 (lH, broad s)
t2) A mixture of 7-~2-~2-formamidothiazol-4-yl)-2-
propargyloxyiminoacetamido]-3-chloro-3-ceyhem-4-carboxylic
acid (syn isomer, 1.4 g.), conc. hydrochloric acid (1.4 ml.)
and methanol (20 ml.) was treated in a similar manner to
that of Example 51-(2) to give 7-[2-(2-aminothiazol-4-yl)-
2-propargyloxyiminoacetamido]-3-chloro-3-cephem-4-carboxylic
acid (syn isomer, 0.7 g.), yellowish white powder.
I.R. v NUjol 3350, 2500~2600, 2130, 1775,
1710, 1670, 1630 cm 1
-- 193 --
,, ., 13~0493
N.M.R. ~ ~DMSO-d6, ppm) : 4.38 (lll, m), 4.48
(2H, AB-~, J=19Hz), 4.72 (2H, d, J=2Hz),
5.28 (1ll, d, J=4Hz), 5.80 (lH, d,d,
J=4Hz, 8Hz), 6.78 (lH, s), 9.73 (lH, d,
J=8Hz)
Example 54
Thiourea (11 mg.) and a solution of 7-[2-(2-
bromoacetyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylic
acid (syn isomer, 30 mg.) in ethanol (2 ml.) were treated
in a similar manner to that of Example 36-(4) to give
7-[2-(2-amino-4-thiazolyl)-2-methoxyimilloacetamido]-3-
cephem-4-carboxylic acid (syn isomer). The product was
identified with an authentic sample by thin layer
chromatography.
Example 55
(l) A solution of 4-nitrobenzyl 7-amino-3-chloro-3-
cephem-4-carboxylate hydrochloride (15.0 g.), bis(trimethyl-
silyl)acetamide (11.3 g.) and trimethylsilylacetamide (9.7
g.) in tetrahydrofuran (300 ml.), a solution of di~etene
(3.41 ml.) in methylene chloride (4 ml.), a solution of
bromine (2.27 ml.) in methylene chloride (4 ml.), a solution
of sodium nitrite (3.1 g.) in water (20 ml.) and a solution
of thiourea (4.0 g.) in water (40 ml.) were treated in a
similar manner to that of Example 37-(1) to give 4-nitro-
benzyl 7-[2-(2-aminothiazol-4-yl)-2-hydroxyiminoacetamido]-
3-chloro-3-cephem-4-carboxylate (syn isomer, 10.4 g.).
I.R. v maUjxol : 3300, 3180, 1777, 1730, 1670,
1603 cm 1
- 194 -
,, ~
13~0~93
N.M.R. ~ (DMSO-d6, ppm) : 3.93 (2H, d, J=5Hz),
5.33 (lH, d, J=5Hz), 5.49 (~H, s),
5.90 (l11, d,d, J=5Hz, 8.ZHz), 6.68 (lH,
s), 7.14 (lH, broad s), 7.72 (2H, d,
J=9.2Hz), 8.27 (211, d, J=9.2Hz), 9.54
(lH, d, J=8.211z)
(2) 4-Nitrobenzyl 7- [2-(2-aminothiazol-4-yl) -2-
hydroxyiminoacetamido]-3-chloro-3-cephem-4-carboxylate
(syn isomer, 5.0 g.), 10% palladium carbon (3.0 g.),
methanol (lO0 ml.), water (lO ml.) and tetrahydrofuran
(150 ml.) were treated in a similar manner to that of
Example 37-(2), to give 7-[2- (2-aminothiazol-4-yl) -2-
hydroxyiminoacetamido]-3-chloro-3-cephcm-4-carboxylic acid
(syn isomer, l.28 g.) .
I.R. ~ mUax~l : 3330, 1775, 1675, 1630 cm 1
N.M.R. ~ (VMSO-d6, ppm) : 3.72 (21-1, m), 5.24
(lH, d, J=5Hz), 5.80 (lH, d,d, J=5.0Hz,
8.2Hz), 6.66 (lH, s), 9.50 (lH, d)
.
-- 195 --
.. ~.
1~4~'g
Example R
(1) Ethyl 2-hydroxyimino-3-oxobutyrate (syn isomer,
100 g.), N,N-dimethylformamide (300 ml.), potassium
carbonate (130 g.) and bromooctanc (121 g.) were treated
in a similar manner to that of Example F-(l) to give ethyl
Z-n-octyloxyimino-3-oxobutyratc (syn isomer, 165.5 g.),
oil.
I.R. v maxm: 1745, 1695, 1470 cm 1
N.M.II. ~ (CCQ4, ppm ) : 0.6~2.1 (18H, m), 2.35
(3H, s), 4.0~4.6 (4H, m)
(2) Ethyl 2-n-octyloxyimino-3-oxobutyrate (syn isomer,
165.5 g.), sulfuryl chloride (84.7 g.) and acetic acid
(165 ml.) were treated in a similar manner to that of
Example F- t2) to give ethyl 2-n-octyloxyimino-4-chloro-3-
oxobutyrate (syn isomer, 169.6 g.), oil.
I.R. v maxm: 1745, 1710, 1465 cm 1
N.M.R. ~ (CCQ4, ppm) : ().6~Z.1 (1811, m),
4.0~4.6 (4H, m), 4.43 (211, s).
(3) Ethyl 2-n-octyloxyimino-4-chloro-3-oxobutyrate
(syn isomer, 169.6 a.). thiourea (42.3 g.), sodium
acetate trihydrate (75.5 g.), water (420 ml.) and ethanol
(1020 ml.) were treated in a similar manner to that of
Example F-(3) to give ethyl 2-(2-aminothiazol-4-yl) -2-n-
octyloxyiminoacetate (s~n i~omer,65 g.), mp. 77 to 78~C.
I.R v mUax~l: 3470~ 3250, 3125, 1735, 1545,
1465 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 0.81 (3H, t, J=6Hz),
0 6~1.9 (15H, m), 4.07 (2H, t, J=6Hz),
4.28 (ZH, q, J=7Hz), 6.86 (lH, s), 7.02
(ZH, broad s)
-- 196 --
... ...
1~40 ~9~
(4) Ethyl 2-(2-aminothiazol-4-yl)-2-n-octyloxy-
iminoacetate (syn isomer, 64 g.), 2N-aqueous solution of
sodium hydroxide (196 ml.), methanol (196 ml.) and
tetrahydrofuran (300 ml.) were treated in a similar manner
to that of Exan-ple F-(4) to give Z-(2-aminothiazol-4-yl)-
2-n-octyloxyiminoacetic acid (syn isomer, 52.5 g.), mp.
146~C (dec.).
I.R. v mUaxol : 3170, 1635, 1565, 1460 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 0.86 (31-l, t, J=6Hz),
0.6~1.9 (121-l, m), 4.06 (2H, t, J=6Hz),
6.81 (lH, s), 7.2Z (2H, s)
(5) 2-(2-Aminothiazol-4-yl)-2-ll-octyloxyimilloacetic
acid (syn isomer, 20 g.), acetic anhydride (27.3 g.) and
formic acid (12.3 g.) were treated in a similar manner to
that of Example ~-(5) to give 2-(2-formamidothiazol-4-yl?-
2-n-octyloxyiminoacetic acid (syn isomer, 21.3 g.) t mp.
122~C (dec.).
I.R. v mUaxol : 3350, 3150, 3050, 1700,
1675, 1560 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 0.6~2.0 (15H, m),
4.16 (2H, t, J=6Hz), 7.56 (1ll, s),
8.57 (lH, s), 12.67 (]H, s)
- 197 -
13404~3
. ~
Example S6
(1) A solution of 2-(2-formamidothiazol-4-yl)-2-
n-octyloxyiminoacetic acid (syn isomer, 7.52 g.),
phosphoryl chloride (5.4 g.) and N,N-dimethylformamide
(2.58 g.) in tetrahydrofuran (16 ~nl.), wllicll was prepared
in a similar manner to tha~ o~ Example 30-(1), ~nd a
solution of 7-amino-3-cephem-4-carboxylic acid (4 g.) in
a mixture of acetone (20 ml.) water (20 ml.) and tetra-
hydrofuran (20 ml.) were treated in a similar manner to
; lO that of Example 30-(1) to give 7-[2-(2-formamidothiazol-4-
yl)-2-n-octyloxyiminoacetamido]~3-cephem-4-carboxylic acid
(syn isomer, 8.1 g )
I.R. v mUa~ol : 3280, 3200, 3060, 1795, 1705,
1660, 1630 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 0.6~2.1 (15H, m),
3.62 (2H, d, J=4Hz), 4.14 (2H, t, J=6Hz),
5.16 (lH, d, J=5Hz), 5.88 (lH, d,d, J=5Hz,
8Hz), 6.51 (lH, t, J=4Hz), 7.42 (lH, s),
8.54 (lH, s), 9.63 (lH, d, J=8Hz),
12;G6 (lH, s)
(2) 7-[2-(2-Formamidothiazol-4-yl)-4-n-octyloxyimino-
acetamido]-3-cephem-4-carboxylic acid (syn isomer, 8.0 g.),
conc. hydrochloric acid (6.23 g.), tetrahydrofuran (15 ml.)
and methanol (120 ml.) were treated in a similar manner to
that of Example 21-(3) to give 7-[2-(2-aminothiazol-4-yl)-
2-n-octyloxyiminoacetamido]-3-cephem-4-carboxylic acia
(syn isomer, 6.95 g.).
I.R. ~ mUaxol : 3320 (shoulder), 1785, 1660,
1630, 1535
- 198 -
13~0~93
N.M.R. ~ (DMSO-d6, ppm) : 0.6~2.0 (15H, m),
3.62 (21-1, broad s), 4.07 ~211, t,
J=611z), 5.12 (lH, d, J=511z), 5.83 (lH,
d,d, J=5tlz, 9Hz), 6.48 (lH, broad s),
6.72 (lH, s), 7.22 (2H, s), 9.53 (lH,
d, J=9l-lZ)
-- 199 --
.
