Note: Descriptions are shown in the official language in which they were submitted.
Q~
NEW CEPHEM COMPOUNDS AND
PROCESSES FOR PREPARATI ON THEREOF
This invention relates to new cephem compoundO
More particularly, it relates to new 7-substituted~3-
cephem-4-carboxylic acid, its pharmaceu~ically accep~a-
ble salt and pharmaceutically acceptable bioprecursor
thereof, which have antimicrobial activities and are
lS a].so useful as intermediates for preparing other highly
potent antimicrobial cephem compounds, and processes
for preparation thereof, ~nd to pharmaceutical composi-
tlon 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-4-carboxylic acid, its
pharmaceutically acceptable salt and pharmaceutically
~5 acceptable bioprecursor thereof9 which exhibit excel-
lent antimicrobial activi~ies against a wide variety of
~athogeni~ microorganisms including Gram negative and
Gram positive bacteria,
processes for preparation of the same9
.30 pharmaceutical composition comprising one of the
same as an active ingredient9 and
a method of using the same prophylactically and
therapeutically for treatment of infectious diseases
caused by pathogenic microorganisms in human being and
animals.
.
. .
.
'~
,
. .
The cephem compounds provided by this invention
include the ones represented by the formula
N
Rl ~ ~ A-CONH ~ ,S ~ (I)
0~
O ~3
wherein Rl is amino or protected amino,
A is lower alkylene which may be substituted
with oxo, hydroxy, amino, prot~cted h~dxoxy
or protected amino,
R is hydrogen, halogen or hydroxy, and
R3 is carboxy or functionally modified carboxy,
and pharmaceutically acceptable salts and bioprecursors
thereof
It is to be noted *hat the cephem compounds (I) as
illustrated above include a compound useful as an anti-
microbial agent and also a compound useful as an inter-
; mediate for preparing the other antimicrobial agent,
~o particularly as illustrated below.
The terms and definitions desrribed in this spe-
cification are illustrated as follows.
As being wéll known, the 2-amino- or 2-pro*ected
amino- thiazolyl group lies in tautomeric relation with
the corresponding 2-imino- or 2-protected imino-
thiazolinyl group. The tautomerism between the said
~hiazolyl and thiazolinyl groups can be illustrated by
the following equilibrium :
N ~ r--- Rl ~ S ~
wherein Rl is amino or protected amino~ and
Rl is imino or protected imino.
Accordingly, it is to be understood that both of
the said groups are substantially the same, and the
~ . .
" ,: ~
,
'7
-- 3 --
tautomers consisting of such groups are regardecl as
the same compounds, especially in the manufac~uring
chemis~ry. Therefore, both of the ~automeric ~orms
of the compounds having such groups in ~heir molecule
are included in the scope of this invention and de-
signated inclusively with one expression ~'2-amn~no-or
protected amino- thiazolyl" and represented by the
formula: Rl~ ~ (wherein Rl is as defined above)
for the S convenient sake throughout this
specification.
And further, it is well kno~n that the 3-hydroxy-
3-cephem compound having the partial structure of the
formula:
~ ~ ~ OH
' R
lies in a tautomeric rèlation with the 3-oxo-cephem
compound of the formula:
~0 - S
0~
R5
each of which is referred 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 ~he same
scope of the compound, and thereore, the structure
and nomenclature of such tautomers are expressed
inclusively with one expression of ~he stabilized
anol tautomer, i.e. "3-hydroxy-3-cephem" compound,
~hroughout this speciication.
In the abo~e and subsequent descriptions of ~his
.
.
., , . , . ,: ,.",, , ., ,.,:, i;. .. i." . ,
; : -
.
: . . ,
, ' .,
. .
-- 4 --
specifi.cation, suitable examples and illustratlon of
the various definitions which this invention intends
to include within the scope thereof are explained in
detail as follows.
"Lower alkylene" for A may be straight or branched
bivalent hydrocarbon residue such as methylene,
ethylene, trimethylene, propylene, ethylethylene,
tetramethylene, pentamethylene, l-methylpentamethylene,
2-methylpentamethylene, 3-methylpentamethylene,
hexamethylene and the like and more preferable one may
be alkylene of 1 ~o 4 carbon atoms and the most pre-
ferable one is methylene.
"Lower alkylene substituted with oxo" for A may be
referred to as "oxo(lower)alkylene", and particularly
it may be oxome~hylene~ oxoethylene, l-oxotrimethylene,
2-oxotrimethylene, 2-oxo*etramethylene, 3-oxopenta-
methylene, and the like, and more preferabiy the one
having 1 to 4 carbon atoms and the most preferably
oxomethylene (namely, carbonyl).
"Lower alkylene substituted with hydroxy" for
A may be referred to as "hydroxy(lower)alkylene", and
particularly it may be hydroxymethylene, hydroxyethy-
lene,;;l-hydroxytrimethylene, 2-hydroxy~rimethylene~ 2-
hydroxytetramethylene, 3-hydroxypentamethylene, and the
like, more prefsrably the one having 1 to 4 carbon
atoms and the most preferably hydToxymethylene.
"Lower alkylene substituted with amino" for A may
be referred to as "amino(lower~alkylene", and particu-
larly it may be aminomethylene, aminoethylene, 1-
aminotrimethylsne, 2-aminotrimethylene, I-amino-3-
methyltrimethylene, 2-aminotetramethylene, 3-amino-
pentamethylene and the like, more preferably the one
having 1 to 4 carbon atoms and the most preferably
aminomethylene.
Suitable "protected hydroxy" group in the
. .
definition for A may include an acyloxy and hydroxy
substituted with a conventional protective group other
than khe acyl group (e.g. benzyl, tetrahydropyranyl,
etc.). And, suitable acyl moiety in the "acyloxy" as
mentioned above can be referred to the same examples of
~'acyl" as illustraked hereinafter for that of
"acylamino" for Rl, and preferabie examples of
"acyloxy" may be substituted or unsubstituted lower
alkanoyloxy, substituted or unsubs~ituted lower alkoxy-
carbonyloxy, substituted or unsubs~ituted ar~lower~-
- alkanoyloxy, heterocycle~lower)alkanoyloxy and the like.
Suitable "protected amino" group in the defini-
tion for A includes the same ones as the "protected
amino" for Rl as illustrated in the following.
"Protective group" in the "protected amino" for
may be the conventional N-pro~ec~ive group such as
substituted or unsubstituted artlower)alkyl (e.g. b~nzyl,
benzhydryl, trityl, 4-methoxybenzyl, 3,4-dimethoxybenzy~
etc.), halo(lower~alkyl te.g. trichloromethyl, tri-
chloroethyl J tri1uoromethyl, etc.), tetrahydropyranyl 9
substituted phenylthio, substituted alkylidene~ sub-
stituted aralkylidene, substi~u~ed cycloalkylidene,
acyl J or the like.
Suitable acyl for the protective group may be sub-
stituted or unsubstituted lower alkanoyl (e.g. formyl,
acetyl, chloroacetyl~ trifluoroacetyl, etc.j, substi-
tuted or unsubstituted ar(lower)alkanoyl (e.g. phenyl-
acetyl, phenylpropionyl, etc.~, substituted or unsub-
- stituted lower alkoxycarbonyl (e.g. me~hoxycarbonyl,
ethoxycarbonyl, propoxycarbonyl, l-cyclopropylethoxy~
carbonyl, isopropoxycarbonyl, butoxycarbonyl, t-
butoxycarbonyl, pentyloxycarbonyl, t-pentyloxycarbonyl,
hexyloxycarbonyl, trichloroethoxycarbonyl, 2-pyridyl-
methoxycarbonyl J etc.), substituted or unsubstituted
ar(lower)alkoxycarbonyl (e.g. benzyloxycarbonyl,
, .
.
- : .
:; :
- 6 ~ 3 ~
ben~hydryloxycarbonyl, 4-nitrobenzyloxyoarbonyl, etc.),
lower cycloalkoxycarbonyl ~e.g. cyclo~e~tyloxycarbonyl,
cyclohexyloxyca~bonyl, etc.), 8-quinolyloxycarbonyl,
succinyl, phthaloyl, or the like.
And further, the reaction product ~f 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 trlmethyl-
silyl chloride, trimethoxysilyl chloride, boron tri-
chloride, butoxyboron dichloride, aluminum trichloride,
diethoxy aluminum chloride, phosphorus dibromide,
phenylphosphorus dibromide, or ~he like.
"Halogen" for R2 may be chlorine, bromine, iodine
or fluorine, and preferred one is chlorine or bromine.
"Functionally modified carboxy" for R3 may be an
ester amide or ~he like, and preferably an ester.
Suitable examples of 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-cyclopropyl-
ethyl ester, etc.); alkenyl ester (e.g. vinyl ester,
allyl ester, etc.~; alkynyl es~er (e.g. ethynyl ester,
propynyl ester, etc.); alkoxyalkyl ester (e.g.
methoxym~hyl ester, ethoxymethyl ester, isopropoxy-
methyl ester, l-methoxyethyl esb~r, l-ethoxyethyl ester,
etc.); alkylthioalkyl ester (e.g. methylthiomethyl
ester, ethylthiomethyl ester, ethylthioethyl es~er,
isopropylthiomethyl ester, e~c.); haloalkyl ester
(e.g. 2-iodoethyl ester, 2,2,2-trichloroethyl ester,
etc.); alkanoyloxyalkyl ester ~e.g. acetoxymethyl
ester, propionyloxymethyl ester, butyryloxymethyl
ester, valeryloxymethyl ester, pivaloyloxymethyl ester,
hexanoyloxymethyl ester, 2-acetoxyethyl ester, 2-
propionyloxyethyl ester, palmitoyloxymethyl ester,et~
. -- - - . .
:. :
:
': ~ ' : '
:
: ,
- 7 ~ Z~l~
alkanesulfonylalkyl ester (e.g. mesylmethyl ester3
2-mesylethyl ester, etc.); substituted or unsubstituted
aralkyl ester ~e.g. benzyl ester, 4-methoxybenzyl ester,
4-nitrobenzyl ester, phenethyl ester, trityl ester,
benzhydryl ester, bis(methoxyphenyl)methyl ester, 3,4-
dimethoxybenzyl es~er, 4-hydroxy-3,5-di-t-butylbenzyl
ester, etc.); substituted or unsubstituted aryl es~er
(e.g. phenyl ester, tolyl ester, t-butylphenyl ester,
xylyl ester, mesi~yl ester~ cumenyl ester, salicyl
ester, etc.); an ester with a silyl compound such as
trialXylsilyl compound, dialkylalkoxysilyl compound
or trialkoxysilyl compound, for example, trialkylsilyl
ester (e.g. trimethylsilyl ester, triethylsilyl ester,
etc.), dialkylalkoxysilyl ester (e.g. dimethylmethoxy-
silyl ester, dimethylethoxysilyl ester, diethylmethoxy-
silyl ester, etc.) or trialkoxysilyl ester (e.g.
trimethoxysilyl ester, triethoxysilyl ester~ étc.) sr
~he like.
With regard to the terms "protected hydroxy",
'Iprotected amino" and "functionally modified carboxy"
in the above, 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 khe meaning of
the synth~tiG manufacture, free hydroxy group, free
àmino group and/or free carboxy group may be trans-
formed into the 'tpr~te~t~d hydro~y", "protected amino"
and/or 'tfunctionally modified carboxy" as mentioned
3~ above before conducting the process~es) for prevent-
ing any possible undesired side reaction(s), and the
"protected hydroxy", "protected amino" and/or "func-
tionally modified carboxy" group in the resultant
compound may be transformed into free hydroxy, amino
35 and/or carboYy group after the eac-ion is conducted.
; , ~.
.
~ ' .
This will be apparent from the explanation of the
processes in the following.
OrL the other hand, in the meaning of the physio-
logical and pharmaceutical properties of the object
S compound, the compound bearing the "protected hydroxy",
"protected amino" and/or "functionally modified
carboxy" group is optionally used for improving the
properties such as solubility, stability, absorbability,
toxicity of the particularly active object compound
bearing the free hydroxy, free amino and/or carboxy
group.
. Suitable "pharmaceutically acceptable salt" of
the object compound ~I) may be co~ventional non-~oxic
sal~, 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 (e.g. calcium salt, magnesium
sal~, etc.), ammonium salt9 an inorganir acid salt
(e.g. hydrochloride, hydrobromide, sulfa~e, phosphate,
carbonate, bicarbonate, etc.), a salt with an organic
base or acid, for example, an amine salt (e.g. tri-
methylamine salt, triethylamine salt, pyridine salt, -
procaine salt, picoline salt, dicyclohexylamine salt,
- N,N'-idibenzylethylenediamine salt, N-methylglucamine
salt, diethanolamine salt, ~riethanolamine salt, tris-
~hydroxymethylamino)methane salt, phenethylbenzylamine
salt, etc.), an organic carboxylic or sulfonic acid
salt (e.g. acetate, maleate, lac~ate, tartrate, mesyl~
ate, benzenesulfonateJ tosylate, etc.~, a basic or
acidic amino acid salt (e.g. arginine saIt, aspartic
acid salt7 glutamic acid salt, lysine salt, serine sal~
etc.) and the like.
It is well known in the pharmaceutical field that
the active drug, when it has any undesired physio-
logical and~or pharmaceutical property such as
,
,.
- ~ :,
, ~. - ~ ,
~' `
g ~ 7~i7
solubility, stability, absorbability, etc., is con-
verted into modified derivative'thereof for improving
such'undesired properties, and then said derivative,
upon administration to a patient, exhibits ~he active
efficacy by being converted to the parent drug in the
body. In this meaning, the term "pharmaceutically
acceptable bioprecursor" used thPoughout this specifi-
cation is intended to fundamentally mean all of the
modified derivatives, which have structural formulae
different from ~hose of the active compounds of this
invention, but are converted in the body to the a~tive
' comppunds of this invention upon administra~ion, and
also to mean the derivatives which are sometimes
derived physiologically from the compounds of this
invention in the body and exhibit antimicrobial efficacy.
The compound (I) of this' invention can be prepared
by processes as shown in the following scheme.
Process A : N-Acylation
,N
H2 ~ S~ R ~S~ (m) 1 ~ ~ A-CON ~ S~
C~ N ~ R _________~ S ~ ~ N ~ ~'
. R3
(II)
.
Process B: Thiazole ring formation
'X-CH2CO-Al-CON ~ S Rl-e-
30~ N ~ R2
I
n3
o~ ~
., (~ .
- Rl ~ ~ ~ R2 (Ib)
' , ~ .
;
Process C: Reductive ormation o-f amino(lower)alkylene
R ~ ~ A2-oONH~ 2 .. ~ Rl~ ~ A3-~ ]-~ ~ 2
. R R
~ (Ic~
P~rocess D: Reductive formation of hydroxy(lower)-
alkylene
Rl ~ A -CONH ~ ~ ~2 agent g Rl~ ~ A -CON ~ 2
R
(Id',) . (Id)
Process E: Elimination of amino protective group
Rl ~A CONH~ R2 H2N~A-CONH~R2
) R
Process F: Carboxy formation
.
R ~ A CONH ~ ~ R2 - ~ R ~ ~A-CONE ~ ' ~ 2
R3 COOH
tIf') (If)
wherein Rl, R2, R3 and A are each as defined above,
- . ' ':
O X is halogeng
Rl is protec*ed amino,
R3 is functionally modified car~oxy,
A~ is lower alkylene which may be substituted
with hyd~oxy amino, protested hydroxy or
protected amino,
A2 is Iower alkylene substituted with a group
of the formula: = N - oR4 wherein R4 is
hydrogen, lower alkyl, lower alkenyl, lower
alkynyl or cycloalkyl 7
A3 is amino(lower)alkylene,
A4 is oxo(lower)alkylene, and
A5 is hydroxy~lower)alkylene
Process A N-Acylation
A compound (I) and i~s salt can be prepared by
reacting a 7-amino-3-cephem compound ~II), its reactive
derivative at the amino or a salt thereof wi~h a
carboxylic acid (III), its reactive deriva~ive at the
carboxy or a salt thereof according to a conventional
manner of so-called N-acylation reaction well known in
~-lactam chemistry.
Suita~le reactive derivative at the amino group
of the compound (II) may include a conventional react-
ive derivative as used in a wide variety of amidation
~eaction, for example, isocyanato, isothiocyanato, a
derivative formed by the reac*ion of a compound (II~
with a silyl compound (e;g. trimethylsilylacetamidel
~30 bis(trimethylsilyl)acetamide, etc.), with an aldehyde
compound (e.g. acetaldehyde, isopentaldehyde, benzalde-
hyde, salicylaldehyde7 phenylacetaldehyde, p-nitIo-
benzaldehyde, m-chlorobenzaldehyde, p-chlorobenzalde-
hyde, hydroxynaphthoaldehyde, furfural, thiophene-
3~ carboaldehyde; etc.7 OT the corresponding hydrate,
: .. ~ , . .
.::
, . .. . .
-: ' ' ~ , ' .
,:
:
,
.
- 12 -
acetal, hemiacetal or enolate thereof), with a ketone~
compound ~e.g. acetone, methyl ethyl keton~, rnethyl isobutyl
ketone, acetylacetone, ethyl acetoacetate, etc., or the cor-
responding ket~, hemiketal or enolate thereof), wi~h phosphorus
compound ~e~g. phosphorus oxychloride~ phospho ~ chloride, etc.),
or with a sulfur compound ~e.g. thionyl chloride, etc.) and the like.
Suitable salt of the compound ~II) may be refer-
red to the one as exemplified for the compound ~I).
Suitable reactive derivative at the carboxy
group of the compound ~III) may include, for example,
an acid halide, an acid anhydride, an activated amide9
an activated ester, and the like, and preferably acid
halide such as acid chloride or acid bromide; a mixed
acid anhydride with an acid such as substituted phos-
lS phoric acid (e.g. dialkylphosphoric acid, phenylphos-
phoric acid, diphenylphasphoric acid, dibenzylphos-
phoric acid, halogenated phosphoric acid, etc.),
dialkylphosphorus acid, sulfurous acid, thios~lfuric
acid, sul~uric acid, alkylcarbonic acid, aliphakic
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,
dimethylpyr~zole, triazole or tetrazole; an ac~ivated
ester (e.g. cyanomethyl ester, methoxymethyl ester~
dimethylaminomethyl ester, vinyl ester, propargyl ester,
p-nitrophenyl ester, 2,4-dinitrophenyl ester, tTichloro-
phenyl ester, pentachlorophenyl ester, mesylphenyl
ester, phenylazophenyl ester, phenyl thioester, p-
nitrophenyl thioester, p-cresyl thioester, carboxy-
methyl thioester, pyranyl ester, pyridyl ester,
piperidyl ester, 8-quinolyl thioester, an es~er with a
N-hydroxy compound such as N,N-dimethylhydroxylamine,
1-hydroxy-2-~lH)-pyridone, N~hydroxysuccinimide,
,, .
,
. ' , ~ ~'"' ;'
,
.
:: '
-
- 13 ~ ~ ~r
N hydroxyph~halimide~ l-hydroxybenzo~riazole, 1~
hydroxy-6-chIorobenzo~riazole, ~tc.~, and ~he like.
~ Suitable salt of the compound (III) may include
a salt with an inorganic base such as alkali metal salt
te.g. sodium sal~, potassium salt, 0tc.) and an
alkaline earth metal salt (e.g. calcium salt~ magnesium
salt, etc.), a salt with an organic base such as
tertiary amine salt (e.g~ trimethylamine salt, tri-
ethylamine salt, N~N-dime~hylaniline salt, pyridine
salt, etc.), a salt with an inorganic acid ~e.g.
hydrochloride, hydrobromide, etc.) and the like.
The suitable reactive derivatives of the com~
pounds (II) and (III) can optionally be selec~ed from
the above according to th~ kind of the compounds (II)
and ~III) to be used practically, and to the reaction
conditions.
The reaction is usually carried out in a conven-
~ional solvent such as water, acetone, dioxane,
acetonitrile, chloro~orm, benzene9 methylene chloride,
~O ethylene chloride, tetrahydrouran, ethyl acetate, N9N-
dimethylformamide9 pyridine or any other solvent which
does not adversely influence the reaction~ or an
opti~nal mixture thereo~
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 con-
densing agent such as a carbodiimide compound (e.g.
N,NI-dicyclohexylcarbodiimide, N-cyclohexyl-N'-
morpholinoethylcarbodiimide, N-cyclohexyl-N'-(4-
diethylaminocyclohexyl)carbodiimide, N,N'-diethyl-
carbodiimide, N,N'-diisopropylcarbodiimide, N-
ethyl-N'-(3-di.methylaminopropy]3carbodiimide, etc.);
a bisimidazolide compound (e.g. N,N'-carbonylbis(2-
- methylimidazole~, etc.), an imine compound (e.g.
pentamethyleneketene-N-cyclohexylimine,
,.; . ,
,, , ~ .
- 14 - ~ 2~
diphenylketene-N-cyclohexylimine, ~tc.), an olefinic or
acetylenic ether compound (e.g. e~hoxyacetylene, ~-
- chIorovinylethyl ether, etc.~, 1-(4-chlorobenzenesul-
fonyloxy)-6-chloro-lH-benzotriazole, N-ethylbenzis-
oxazolium salt, N-e~hyl-5-phenylisoxazolium-3'-
sulfonate, a phosphorus compound (e.g. polyphosphoric
acid, ~rialkyl phosphite, ethyl polyphosphate, isopropyl
polyphosphate, phosphorus oxychloride, phosphorus
trichloride, diethylchlorophosphite, orthophenylene
chlorophosphite, etc.), thionyl chloride, oxalyl
chloride, Vilsmeier reagent prepared by the reaction o
dimethylformamide with a halogen compoun~ such as
thionyl chloride 9 phosphorus oxychloride, phosgene or
the like.
The protective group in the protected amino, and
protected hydroxy group of the compound (III) and the
functionally modified carboxy group of the compound (II)
may occasionally be transformed into free amino, hydroxy
and carboxy group respectively in the course of the
reaction or post-treatment in this pTocessJ and these
cases are included within the scope of this invention.
The object compound (I) and salt thereqf are useful
as an antimicrobial agent, and a part thereof can also
be used as a s~arting material in the following pro~
cesses.
Process B : Thiazole ring formation
The starting compound (IV) and its salt to be used in this
- process can be prepared by reacting a compound (II),
its reactive derivative at the amino group or a salt
thereof with a compound (III'), its reactive deriva~i-
ve at the carboxy group or reaction equivalent thereof,
as illustrated in the following scheme.
~ r ~ R~
R3
(II) . -
. D 1 X- CH2CO-Al - COOH ~I I I t
X- CH2CO-Al- CONH~L
o
(IV)
wherein R2, R3 and X are each as defined above.
Suitable 7'halogen" f~r X may be chlorine,
bromine, iodine, and suitable reactive equivalent of
the compound (III') may be a combina~ion of diketene
and halogen such as chlorine or bromine. The reaction
of a compound tII) with a compou~d (III') can be con-
ducted substan~ially in the same manner as thP above
Process A.
A compound ~Ib) and its salt can be prepared by
reacting a compound (IV) or its salt wi~h a ~hiourea
sompound (V).
The reaction is usually conducted in a solvent
such as water, alcohol (e.g. methanol, ethanol, etc.~,
benzene) dimethylformamide, te~rahydrofuran or any
other solvent which does not adversely influence the
reaction, within a tempera~ure range of an ambien~
temperature to heating.
Process C : Reduc~ire forma~ion of amino~lower)~
_
alkylene
.
. , .
- "
:: ,, :,
. . ,: , . . .
,. . , ~.
,,
" ' '; ,
- 16 -
A compound (Ic) and its salt can be prepared by
reducing a compound (VI) or its sal~, which can be
prepared by reacting a compound ~ s reactive
derivative at the amino group or a salt thereof with a
S compound of the formula:
N 2
wherein Rl and A2 are each as de-fined above, its react-
ive derivative at the carboxy group or a salt thereof,
- in the same manner as the above Process A.
. .The ~eduction in this process may be conducted by
a conventional method, for example, catalytic reduction
using a conventional catalyst (e.g. Raney nickel)
platinum oxide, palladium on carbon, ruthenium on car-
bon, rhodium on alumina, copper chromium oxide, etcO);
chemical reduction using a combination of an acid (e.
g. hydrochloric acid, sulfuric acid, formic acid,
acetic acid, etc.) and a metal or a metal salt (e.g.
iron, zinc, tin, chromium chloride, chromium acetate,
etc.), a combination of a metal or a~ amalgam (e.g.
- sodium, zinc amalgam, sodium amalgam~ aluminum amalgam,
etc.) and a solvent (e g. water, methanol, ethanol,
etc.), or a reducing agent such as a complex of metal
hydride (e.g. sodium borohydride, alkyl-tin-hydride,
lithium aluminum hydride, diethyl aluminum hydride 9
etc.); electrolytic reduction or the like. The react-
îon conditions such as temperature, pressure, time
and solvent can be selec~ed according to the kinds of
the starting compound (VI) and the reduction method
to be applied.
Process D : Reductive ~ormation of hydroxy(lower)-
= .
alkylene
A compound (Id) and its sal~ can be prepared by
..
- ~ ;
- 17 ~
reducing a compound (Id') or its salt, which can be
preparea by the abo've'Proces's A.
The reduc'tion method of this process may be a
conventional one which can be applied for the reduction
of oxo group into hydroxy group, and particular of
which is substantially the same as t~ose explained in
the above Process C.
Process E : Elimina~ion of amino-protective group
10A compound (Ie) and i.ts salt can be prepared by
subjecting a compound (Ie') or its salt to elimination
reaction of the amino-protective group in the pro-
tected amino group for Ra.
The starting compound (Ie') can be prepared, for
example, by the above Process A.
The elimination reaction may be çonducted in
accordance with a conventional method such as hydro-
lysis, reduction (reductive elimination) or the like.
These methods may be selected according to the kind of
the amino-protective group ~o be eliminated.
The hydrolysis may include a method using an acid
tacidic hydrolysis), a base (basic hydrolysis) or
hydrazine, and the like.
Among these methods, hydrolysis using an acid is
one of the common and preferable methods or eliminat-
ing the amino-protective group such as an acyl group,
~or example, substituted or unsubstituted.lower
alkanoyl, substituted or unsubstituted lower alkoxy-
carbonyl, substituted or unsubstituted ar(lower)-
- 30 alkoxycarbonyl, lower cycloalkoxycarbonyl, substituted
phenylthio J .substituted alkylidene, substituted In I
aralkylideneJ substituted cycloalkylidene or the like.
Suitable acid to be used in this acidic hydrolysis may
include an organic or inorganic acid such as formic
acid, trifluoroacetic acid, benzenesulfonic acid,
, ... . .
18 -
p-toluenesulfonic acidJ hydrochloric acid, cation-
ex`change resin, and the like. Prefera~le acid is the
one which can easily be separated out from the react-
ion product by a conventional manner such as neutrali-
zation or distîllation under reduced pressure, forexample, formic acid, trifluoroacetic acid, hydrochloric
acid or the like. The acid suitable for the reaction
can be selected in consideratîon of the chemical pro-
perty of the starting compound and the product as well
as the kind of the pro~ec~ive group to be eliminated.
The acidic hydrolysis can be conducted in the presence
or absence of a solvent. Suitable solvent may be a
conventional organic solvent, water or a mixture
thereof, which does not adversely influence this react-
ion. Particularly, when the hydrolysis is conductedwith trifluoroacetic acid, the reaction may be acoele-
rated by addition of anisole.
The hydrolysis using a base can be applied for
eliminating the protective group such as an acyl group,
preferably, for example, haloalkanoyl (e.g. trifluoro-
acetyl, etc.) and the like. Suitable base may include,
or example, an inorganic base such as alkali metal
hydroxide (e.g. sodium hydroxide, potassium hydroxide,
etc.),~alkaline earth 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 hydro-
gen phosphate, dipotassium hydrogen phosphate, etc.),
or the like, and an organic base such as alkali metal
acetate (e~g. sodium acetate, potassium acetate, etc.),
.
~ . ~
- -
, .
' - 1 9 ~ ~
trialkylamine ~e.g. trimethylamine, triethylamine, etc.),
picoline, N-methylpyrrolidine, N:methylmorpholine~ 1,5-
- diazabicyclo~4,3,0]-5-nonene, 1,4-diazabicyclo[2,2,2]-
octane, 1,5-diazabicyclo[5,4,0~-7-undeceneanion-
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.
The hydrolysis using hydrazine can be applied for
eliminating the protective group such as dibasic acyl,
for example, succinyl, phthaloyl or the like.
The reductive elimination can be applied for eli-
minating the protective group such as acyl, for example,
halo~lower)alkoxycarbonyl ~e~g. trichloroethoxycarbonyl,
etc.), substituted or unsubstituted ar(lower)alkoxy-
carbonyl (e.g. benzyloxycarbonyl, p-nitrobenzyloxy-
carbonyl, etc.), 2-pyridylmethoxycarbonyl, etc.,
aralkyl ~e.g. benzyl~ benzhydryl 9 trityl, etc.~ and
the like. Suitable reduction may include, for example,
reduction using an alkali metal borohydride (e.g.
sodium borohydride, etc.~, conventional catalytic
hyd~ogenolysis and the like.
And further, the protective group such as halo-
tlower)alkoxycarbonyl or 8-quinolyloxycarbonyl can be
eliminated by treatment with a heavy metal such as
copper, zinc or the like.
The reaction temperature is not critical and may
optionally be selected in consideration o-f 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 cool-
ing~ at ambient temperature or slightly elevated tem-
perature.
The process includes in its scope the cases that
t~e functionally modified carboxy for R3 is
.. .. . .. .... . .. . .
, .. . ..
, ;. , .
,; . ,
- 20 - ~L~
simultaneously txans~ormed into ~he free carboxy group
in the course of the above reaction or in ~he post-
treatment.
As to this process, it is to be understood that
the purpose of this process lies in providing the
generally more active compound (Ie) having an amino-
thiazolyl group by eliminating the protective group in
the protected amino group of the compound (Ie') pre-
pared by the other processes as mentioned ab~ve or
below.
Process F : Carboxy formation .
This process is to provide a free carboxy compound
~If) or its salt, which generally exhibits higher anti-
microbial activity as compared with the corresponding
functîonally modified carboxy compound (If').
Accordingly, the meaning of the unc~ionally
modified carboxy in the compound (If') lies în mainly
synthetic manufacture by chemical process(es) as
~0 illustrated hereinabove.
This process is conducted by transforming the
unctionally modified carboxy group of the star~ing
compound (I~) into free carboxy group, and the prefer-
red functionally modified earboxy for Ra in the eom-
pound (If') may be an esterified carboxy group as exem-
plified for R3 of the compound (I).
The method to be applied to this process includes
conventional ones such as hydrolysis, reduction and
the like.
.The method of hydrolysis includes a conventional
one using an acid~ base, enzyme or enzymatic preparat-
ion, and the like.
Suitable examples of the acid and base are to be
~eferred to those as exemplified ;n the above Process
E, and the acidic or basic hydrolysis can be carried
,
' , :
,
..
;
- 21 ~ `7
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 prefeT-
ably a cultured broth of microorganism or processed
material thereof.
An esterase to be used in the e~zymatic hydro-
lysis may be used not only in a purified state, butalso in a crude state.
. The compound obtained in accordance with the
processes as explained above can be isola*ed and
purified in a conventional manner.
In case that th0 object compound (I) has free
caTboxy and/or ree amino, it may be transformed into
its pharmaceutically acceptable salt by a conven-
~ional method.
The object compound (I), its pharmaceutically
acceptable salt and bioprecursor thersof 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.
In order to show the utility of the compound (I),
the test data of some representative compounds (I)
are shown in the following.
In vitro antibacterial activity:
Test Method
In vitro antibact0rial activity w s determined
by the two-fold agar-pla~e dilution method as descri-
bed below.
One loopful of an overnight culture of each test
strain in Trypticase-soy broth (lO8 viable cells per
1.
.
.~ ~
- I , .
~ '7
- 22 -
ml.) was streaked on heart infusion agar (HI-agar) con-
taining graded concentrations of antibiotics, and the
minimal inhibitory concentration ~MIC) was express~d in
terms of ~g/ml. after incubation at 37C for 20 hours.
Test_com~ounds
No. 1 H N~
COOH
N CH2CONH~S ~
No. 2 H2NIl~ o~LN~oOH
N--CHCONH S
No. 3 H2 J~ SJJ NH2
~00
HCOOH salt
2 5 No . ~ N CHC011H~ S~ .
H2NI~bH o~ N~J .
COOH .;~
.
..
:,
f3;~J
- 23 -
Test ~esults
~IC (~g/mQ)
5 ¦ Compound No. 1 Z 3 4
Salmonella
enteritidis 1.56 1.56 3 f 133.13
_. .. _ _ .. .. ._
! Escherichia
coli 3~ 1.56 1.56 0.78 1.56
..... - - _ .- ~
Klebsiella
aerogenes 417 0.1 0.2 0.39 0.78
Proteus
mirabilis 520 0.78 0.3g 0.78. 1.56
_
Proteus .
vulgaris 616 12.5 6.25 1.56 12.5
.
For prophylactic and/or therapeutic administrat-
ion, the active compound (I) of the present invention
is used in the orm of a conventional pharmaceutical
preparation which contains said compound, as an
active ingredient, in admixture with pharmaceutically
acceptable carriers such as an organic or inorganic
solid or liquid excipient which 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, suspension7 or emul-
sion. If needed, there may be included in the above
preparations auxiliary substances, stabilizing agents,
wetting or emulsifying agents, buffers and the other
commonly used additives.
While the dosage of the compounds may vary from
: ,,
- '. -: , t ;
,:" ' ; '
:: :: : :
- 24 -
and also depe~d upon the age and conditions of the
patient, a kind of disease and a degreé o~ the in-
fection, and further a kind of the active compound (I)
to be applied, etc., an average single dose of about
50 mg. 9 100 mg., 250 mg. and 500 mg. of the active
compound tI) ;s sufficient for treating infectious
diseases caused by pathogenic bacteria. In general,
the active compound (I) can be adminis~ered in an
amount between 1 mg/kg and 100 mg/kg, pref~rably
5 mg/kg and 50 mg/kg.
And urther, it is ~o be noted tha~, among the
objeGt compound (I) 5 the compound (Id') and its salt
are us~ful as an intermediate or preparing the more
active cephalosporin compound o the formula (VI),
its nontoxic, pharmaceutically acceptable salt or a
bioprecursor thereof.
A compound (VI) and its salt can be prepared by
~eacting a compound (Id') or i~s salt with a compound
~VII) or its salk as illustrated by the ollowing
schema:
Rl~ ~ A4-coNH ~ ~ 2 (Id'~
.r
R4 - ONH2 ~VII)
Rl~ ~ A2-CONH ~ ~ ~VI~
R3
h i Rl R2 R3 R4 A2 ~nd A4 are each as defined above.
.
. . ;. "
22.~7 1~
- 2S -
o The suitable example of the compound (VII) may be
hydroxylamine, lower alkoxyamine (e.g. methoxyamine,
ethoxyamine, propoxyamine, butoxyamine, etc.), lower
alkenyloxyamine (e.g. vinyloxyamine, allyloxyamine,
propenyloxyamine, butenyloxyamine, etc.), lower
alkynyloxyamine (e.g. ethynyloxyamine, propynyloxy-
amine, butynyloxyamine, etc.) or cycloalkoxyamine
(e~g. cyclobutoxyamine, cyclopentyloxyamine, cyclo-
hexyloxyamine, etc.).
The salt of the compound (VII) may be acid salt
such as hydrochloride, hydrobromide, sulfate or the
like.
This reaction is usually conducted in a solvent
such as water, alcohol or any other solvent which does
not adversely influence the reaction~ within a tem-
perature range from cooling to heating.
When a salt of the compound (VII) is used, the
reaction is preferably conducted in the presence of a
base as exemplified in the Process E.
Still further, it is to be noted that, among the
object compounds (I), the compounds (Ic) and (Id) and
their salts are also useful as intermediates ~or pre-
paring the other cephalosporin compounds, which may be
prepared by acylating the amino group in the symbol
A of the compound (Ic) oresterifying the hydroxy
group in the symbol A5 of the compound ~Id), respec-
tively. .
The partial structure of the formula~ -CO-
of the compound (VI) is intended to mean N
both of the geometric ormula: o-R4
-c- co- -c-co-
~-o-R4 R4-~-~
(5) (A)
I
,
- 26 -
The geomet~y of the form~la (S) is referrea to as "syn"
and another formula (A) is referred to as "anti".
From the view point of struc~ure-activity reIationship,
i~ is to be noted that the syn isomer of the compound
(VI) tends to be of much higher antimicrobial ac~ivity
than the corresponding anti isomer,.and accordingly
the syn isomer of the compound (VI) is more preferable
antimicrobial agent than the corresponding ~nti isomer
in the prophylactic and therapeutic value.
1~ Following examples are given only for explaining
this invention in more detail.
.
~ . .
:'
: ,
~ 30
,
~ 35
, . . .. . .
'' . : ' ., ;
- 27 -
Example l
(1) 4-Nitrobenzyl 7-amino-3-cephem-4-carboxylate
hydrochloride ~9 g), ~rimethylsilylacetamide (24.81 g3
and bis(trimethylsilyl)acetamide (9 ml) were added to
dry ethyl acetate (100 ml) and stirred at 45C for an
hour.
On the other hand, phosphoryl chloride (8.4 ml;
was added dropwise to a stirred mixture of dimethyl-
formamide ~4.0 ml) and dry ethyl acetate (16.0 ml)
under ice cooling, and stirred for a while. To the
solution were added ethyl ace~ate (240 ml) and 2-(2-
formamido-4-thiazolyl)glyoxylic acid ~5.35 g) gradual-
ly at -3C~ and ~he mixture was stirred at the same
temperature for 15 minutes. The solution was added
dropwise to the solution containing the cephalosporin
compound prepared above at -15C and stirred at ~he
same temperature for 30 minutes. Water (50 ml) was
added to the resultant solution, and the precipitates
wsre collected by filtration, washed with water and
~20 dried over phosphorus pentoxide under reduced pressure
to give 4-nitrobenzyl 7-E2-formamido-4-thiazolyl)-
glyoxyloylaminoJ-3-cephem-4-carboxyla~e ~7.124 g).
The ethyl acetate layer was separated from the filt-
rate, washed with a saturated aqueous solution of
sodium chloride, dried over magnesium sulfate9 and
concentrated under reduced pressure to give the same
object compound as abov~ ~1.03 g). Total yield 8.154
I.R. ~mU~oi . 1775, 1725, 1650 cm 1
N.M.R. ~(DMSO-d6, ppm) : 3.66 ~2H, m), 5.17 (lH,
d, J=5.2Hz), 5.42 ~2H, s), 5~90 (lH,
dd, J=5 2Hz, 7.8Hz), 6.66 (lH, t, J=
5.0Hz), 7.67 (2H, d, J-9Hz), 8.22 (2H,
d~ J=9Hz), 8.39 (lH, s), 8.55 (lH, s),
.
de ~7 q y k
.
,
- 28 --
9.87 (1H, d7 J-7.8Hz)
(2) A mixture of 4-nitrobellzyl 7-~2-(2-formamido-~-
thiazolyl)glyoxyloylamino]-3-cephem-4-carboxyla~e (3.0
g) methanol (60 ml) and tetrahydrofuran (gQ ml) was
5added to a mixture of 10% palladium-carbon (1.5 g),
ace~ic acid ~10 ml) a~d methanol (10 ml) in an a~mos-
phere of nitrogen gas, and then subjected to ca~alytic
reduction at room temperature under ordinary pressure
for 4 hours. Th~ resultant mixture was filtered, and
10the ~iltrate was concentrated under reduced pressure.
The precipitates were collected by filtration, washed
with~diisopropyl ether (50 ml) and dried to give powder
(I.34 g). A mixture of water (100 ml) and ethyl acetate
(100 ml) was added to the powder adjusted to pH 6.0 with
15sodium bicarbonate and the aqueous layer was separated,
washed with ethyl acetate and diethyl ether. The re-
maining ether was removed of-~ by bubbling with nitrogen
gas, and the aqueous solu~ion was adjusted to pH 2.0
with 10% hydrochloric acid. The resultant precipi~ates r
20were collected by filtration and dried over phosphorus
pentoxide ~o give 7-[2-(2-formamido-4-thiazolyl)glyoxy-
loylamino]-3-cephem~4-carboxylic acid (0.47 g~. The
above filtrate was concentrated under reduced pressure,
and the residue was pulverized with a mixture of diethyl
~5ether and pekroleum ether. The precipitates were col-
lected by filtrationa washed with diethyl ether and
petroleum ether and dried under reduced pressure to
give the same objective compound tl.l g). Total yi~ld
1.57 g.
I.R. v mUa~ol : 1780, 1670 cm 1
N.M.R- ~(DMS0-d6, ppm) : 3.63 (2H, m, J=4Hz),
5.17 (lH, d, J=5.2Hz), 5.87 (lH, dd9
J=5.2Hz, 8.2Hz)~ 6.53 ~lH, t, J=4Hz),
8.42 (lH, s), 8.59 (lH, s), 9.83 (lH,
., ~. , . . '- .
,
. ' ' .
- 29 -
d, J=8.2Hz)
~3) A mixture of conc.hydrochloric ac;d (2.44 g~ and
- methanol ~10 ml) was added to a mixture of 7-[~-(2-
formamido-4-thiazolyl)glyoxyloylamino]-3-cephem-4-
S carboxylic acid (2,44 g) in methanol (40 ml) under
ice-cooling, stirred at 20 to 22C for 5 hours and fil-
tered, The filtrate was concentra~ed under reduced
pressure, and water (100 ml) was added to the residue
adjusted to pH 6.5 with sodium bicarbonate with stirring
and filtered. The filtrate was washed with ethylacetate) and adjusted to pH 3.5 with 10% hydrochloric
acid. The resultant precipitates were collected by
filtration, washed with water and dried over phosphorus
pentoxide under reduced pressure to give 7-[2-(Z-amino-
4-thiazolyl)glyoxyloylamino]-3-cephem-4-carboxylic acid
(0.492 g). The filtrate and wash~ngs were subjected to
column chromatography on macroporous, non-ionic adsorp-
tion resin "Diaion HP-20" tTrademar~, manufactured by
Mitsubishi Chemical Industries Ltd.3, washed with water
2Q and eluted with 15~ isopropyl alcohol. The isopropyl
alcohol was distilled o~f under reduced pressure and
~he remaining solutio~ was lyophilized to gl~e the same
object compound (1.561 g). Total yield 2.053 g.
I.R. v NUa~ol : 1780, 1668 cm 1
N.M.R. ~(D~O, ppm) : 3.57 (2H, m), 5.17 (lH, d,
J=4.8Hz), 5.78 ~lH,d, J=4.8Hz), 6.33
(lH, m), 8.26 (lH, s)
Example 2
(1) Phosphoryl chloride (0.7 g~ was added to N~N-
dimethylformamide (10 ml) at 20C and stirred at 40C
for 30 minutes. 2-(2-Formamidothiazol-4-yl)glyoxylic
acid ~0.4 g) was added to the solution at 0 to 5C and
stirred for 40 minutes. The solution was added to a
solution of 4-nitrobenzyl 7-amino-3-hydroxy-3-cephem-
-': , ", ,.
,
.. , ~ ' '
:
4-carboxyla~e ~0.7 g), trimethylsilylace~amid~ (1.85 g)
and bis(trimethylsilyl)acetamide (1.62 g) in ethyl
acetate (20 ml) at -20C and stirred at -Z0C for an
hour. After adding water (20 ml) to the resultant
solution~ the ethyl acetate layer was separated, washed
with an aqueous solution of sodium bicarbonate, dried
over magnesium sulfate and concentTated in vacuo.
The residue was triturated with diethyl ether to give
4-nitrobenzyl-7-~2-~2-formamidothiazol-4-yl~glyoxyloyl-
amino]-3-hydroxy-3-cephem-4-carboxylate ~0.85 g).
I.R. v NaU~ol : 3150, 1770, 1660, 1600 cm l
N.M.R. ~(DMSO-d6, ppm) : 3;;58 (2H~As-q, J=18Hz),
5.48 (2H, s), 5.25-5.83 (3H, m), 7.75 ;
(2H, d, J=9Hz), 8.32 (2H, d, ~=9Hz~,
8.58 (lH, s~, 8.63 (lH, s), 9.92 (lH,
t, J=8Hz)
(2) A mixture o-f 4-nitrobenzyl 7-[2-(2-formamido-
thiazol-4-yl)glyoxyloylamino]-3-hydroxy-3-cephem-4-
carboxylate (0.7 g) and conc.hydrochloric acid (0.23
g) in methanol (10 ml~ was stirred at room temperature
for 3 hours. The sol~ent was evapora~ed in vacuo and
the residue was triturated with diisopropyl e~her.
The precipitates wer~ collected by filtra~ion and
washed with diisopropyl ether to give 4-nitrobenzyl 1-
[2-(2-aminothiazol-4-yl)glyoxyloylamino]-3-hydroxy-
3-cephem-4-carboxylate hydrochloride (0.6 g).
I.R. VmaJl : 3300, 1770, 1660, 1630, 1600,
1510 cm ;
N.M.R. ~(DMS0-d6, ppm) : 3.52 (2H, broad s),
5.40 (2H, s), 5.18-5.57 (3H, m), 7.70
(2H,- d, J=9Hz), 8.25 (2H, d, J=9Hz),
8.27 (lH, s), 9.97 (lH, t, J=9Hz)
. ~ , . . .
~;
,
.~
'1
.
s~3~ 1~
i
31 l
Example 3 I
N,N-Dimethylformamide (88 mg) and phosphorus oxy-
chloride (184 mg) were mixed to prepare Vilsmeier re- I
agent in a conventional manner, and the resul~ant
Vilsmeier reagent was suspended in dry tetrahydrofuran
~10 ml). To the suspension was added 2-(2-formamido-
thiazol-4-yl)glyoxylic acid (0.2 g) under ice-cooling
with stirring, and the solution was stirred at the .
same temperature for 30 minutes. to prepare the acti- .
vated acid solution. 4-Nitrobenzyl 7-amino-3-chloro- .
3-cephem-4-carboxylate (0.4 g) was dissolved in a
solution of trimethylsilylacetamide (1.0 g) in ethyl
acetate (100 ml). To the solution was added the
: activated acid solution obtained above all at once at
-20~C, and the solution was stirred a~ -20 to. -5C for ,
1.5 hours. After water and ethyl acetate (50 ml~ were .
`` . added to the resuitan~ solu~ion at -20C, the insoluble 1substance was separated by filtration, washed with !
water and acetone in turn and then dried to give 4-
nitrobenzyl 7-[2-(2-formamidothiazol-4-yl)glyoxyloyl-
` amino]-3-chloro-3-cephem-4-carboxylate (0.1 g).
I R v Nu~ol : 3350, 1780, 1730, 1650, 16009
1520 cm
N.M.R. ~(~MS0-d6~ ppm) : 3.96 (2H, q, J=18Hz),
5.50 ~2H, s), 5.92 (lH, dd, J=5.8Hz),
7.74 (2H, d, J=9Hz), 8.28 ~2H, d, J=9Hz),
8.48 (lH, s)~ 8.~0 (lH, s), 10.00 (lH, :
d, J=8Hz), 12.63 (lH, broad s)
-30 Example 4
(l~ 7-Amino-3-cephem-4-carboxylic acid (2.54 g) was
dissolved in a solution of trimethylsilylace~amide
(11.7 g) and bis(trimethylsilyl)acetamide (15 ml) in
dried ethyl acetate (50 ml). A solu$ion of bromine
~2.43 g) in dried methylene chloride (10 ml) was added
.. . .. . .
.
.. .. . .
:.
...
- 32 '~ '7
dropwise to a solution of dike~ene (1.28 g) ;n dried
methylene chloride (25 ml~ at -30C over 10 minutes
and stirred at the same tempera~ure for 1.5 hours. The
solution was added to the above solution containing 7-
amino-3-cephem-4-carboxylic acid at -15C over 10 minu- .
tes, and stirred at -15 to-10C for 1.5 hours.
Water (50 ml) was added to the resultant solution. The
ethyl acetate layer was separated, and extracted with
aqueous solution o sodium bicarbonate. The aqueous
extract was adjusted to pH 2.0 wi~h 10% hydrochloric
acid and extracted with ethyl acetate. The etbyl
acetate extract was washed with water, dried o~er mag-
nesium sulfate and concentrated under reduced pressure
to give 7-[2-(2-bromoacetyl)acetamido]-3-cephem-4-
carboxylic acid ~2.82 g).
I.R. v NaUJol : 17~0, 1660 cm 1
N.M.R. ~(DMSO-d6, ppm) : 3.58 (2H, d, J=4H~),
3.65 t2H, s)~ 4.40 (2H, s), 5.06 (lH, d,
J=5Hz), 5.73 (lH, dd, J=8Hz, 5Hz~,
6.50 (lH, t, J~4Hz), 9.08 (lH, d, J=8Hz)
~2) Thiourea (663 mg), sodium bicarbonate (732 mg)
and water (20 ml) were added to a stirred solution of
7-[2-(2-bromoacetyl)acetamido]-3-cephem-4-carboxylic
acid (2.11 g) in tetrahydrofuran (20 ml) under ice-
cooling and stirred at the same temperature for an hour.
Af~er ~he resultant solution was adjusted to pH 400
with dilute hydrochloric acid, the precipita~es were
30 . collected by filtration, washed with water and dried
over phosphorus pentoxide under reduced pressure to
give ~?-[2-(2-amino-4-thiazolyl~acetamido]-3-cephem-4-
carboxylic acid (1.01 g). .
I.R, v mUJl : 3550, 3330, 1750, 1670, 16~0 cm 1
N.M.R. ~(DMS0-d6, ppm) : 3.42 ~2H, s), 3.60 (2H,
d, J=4Hz), 5.08 (lH, d, J=5Hz), 5.77 ~1
dd, J=8Hzf 5Hz), 6.30 (lH, s), 6.52 (lH,
t, J-4Hz), 8.87 (lH, d, J=8Hz~
Example 5 .
(l) 4-Nitrobenzyl 7-amino-3-cephem-4-carboxylate
(5 g) was dissolved in a solution of trimethylsilyl- ,
acetamide (13.8 g) and bis(~rimethylsilyl)acetamide .
(10 ml~ in dry ethyl acetate (50 ml) and stirred at
45C for l.S hours. A solution of bromine (2.88 g~ in .
methylene chloride (7 ml) was added dropwise to a solu-
tion of diketene (1.5 g) in methylene chloride (7 ml~
at -40C over 20 minutes and stirred at -30C for 1
hour. The solution obtained thus was added to drop-
wi~ to the above solu~ion of 4-ni~robenzyl 7-amino-3-
cephem-4-carboxyla~e under cooling at -15~C and then
stirred at the same temperature for 30 minutes. Water
(50 ml~ was added to ~he resultant solution and ex-
tracted with ethyl ace~a~e. 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 NUaJol : 17~09 1740, 1630 cm 1
N.M.R. ~(DMS0-d6, ppm) : 3.62 (2H, broad s),.
4,37 (2H, s), 5.08 ~lH, d, J=5Hz),
5.40 (2H, s), 5.77-6.05 (m), 6.67 (lH,
t, J-5Hz), 7.68, 8.04 (4H, m), J=9Hz)~
9.07 (lH, d, Jz8Hz)
(2) Thiourea tl.l3 g), sodium bicarbonate (1.24 g~
,. I
,:
, ` ' ~ ` ' `
` !
--34
and water ~20 ml) were added to a solukion o 4-
nitrobenzyl 7-~2-(2-bromoacetyl)acetamido~-'3-'cephem-
4-carboxylate ~6.15 g) in te~rahydrofuran (60 ml~ ~ and
stirred at room tempcrature for an hour. After con-
centratin~ the resultant solution under reduced pres-
sure~ the residue was extracted with ethyl ace~ate.
The extract was washed with water, dried over magnesium
sulfate and concentrated under reduced pressure. The
oily residue was subjected to column chromatography on
sili'ca gel, and eluted with benzene, a mixture of
benzene (1 part) and ethyl acetate (1 part) and then
ethyl acetate in turn. The ethyl acetate fractions
were collected and concentrated under reduced pressure
to give 4-nitrobenzyl 7-[2-~2-amino-4-thiazolyl)-
acetamido]-3-cephem-4-carboxylate (1.5 g).
I.R. ~ NuJol : 3350, 1780, 1740, 16809 1610 cm 1
N.M.R. ~(DMSO-d6, ppm) : 3.40 (2H, broad s),
~0 3.6~ (ZH, broad s), 5.12 (lH, d, J=5Hz),
5,43 (2H, s), 5.84 (lH, dd, J=8Hz, 5Hz)~
6.30 (lH, s), 6.70 (lH, broad s),
7.72 (2H, d, J=9Hz), 8.27 (2H, d, J=9Hz),
8.93 (lH, d, J=8Hz)
(3) A solution of 4-nitrobenzyl 7-[2-(2-amino-4-
thiazolyl)acetamido]-3-cephem-~-carboxylate (1.4 g) in
a mixture of methanol ~25 m1) and tetrahydrofuran (60 l
ml) was added to a mixture of 10% palladium -on-carbon ¦
(1.7 g), glacial acetic acid (7 ml) and methanol (20 ¦
'30 ml) in an atmosphere of nitrogen. Hydrogen gas was
introduced into the mixture and shaken for 8.5 hours
at room temperature. After removing the insoluble
substance by filtratio~, the iltrate was concen~rated
under reduced pressure. The residue was pulverized
with diethyl ether, and the resultant substance was
.
. .
,
35 -
dissolved in an aqueous solu~ion of sodium bicarbonate,
washed with ethyl acetate and diethyl ether in turn and
then the remaining ether was removed by bubbling with
nitrogen gas. The resultant solution was adjusted to
! 5 pH 4.5 with dilute hydrochloric acid, subjected to
column chromatography on macroporous non-ionic adsorpt-
ion resin "Diaion HP-20" (Trademark, manufactured by
Mitsubishi Chemical Industries Ltd.), and eluted with
2Q~ isopropyl aicohol. Isopropyl alcohol was distilled
of under reduced pressure and the r~maining solu~ion
was lyophilized to give 7-[2-(2-amino-4-thiazolyl)
acetamido]-3-cephem-4-carboxylic acid (lB5 mg).
I.R. ~Na~l : 3550, 3330, 1750, 1670, 1620 cm 1
N.M.R. l~DMSO-d6, ppm) 3.42 (2H, s), 3.60 ~2H,
d, J=4Hz), 5.Q8 (lH, d, J=5Hz), 5.77 (lH,
dd, J-8Hz, 5Hz), 6.30 ~lH, s), 6.52 (lH,
t, J=4Hz), 8.87 (lH, d, J=8Hz)
Example 6
~1) 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
handJ phosphorus oxychloride (1.95 ml) was added drop~
wise to a suspension of 2-~Z-amino-4-thiazolyl)-2-
methoxyiminoacetic acid (syn-isomer: 3.42 g) in dry
3thyl acetate (34 ml) over lD minutes at 0 to 6C, and
the mixture was stirred at the same temperature for 30
minutes. To the solution was added dropwise a solution
of trimethylsilylacetamide (2.39 g) in ethyl aceta~e
(5 ml) at 0 to 6C over 20 minutes, and the mixture was
stirred for 20 minutes. After phosphorus oxychloride
(1.95 ml) was added dropwise to ~he mixture at the
above temperature over 10 minutes, the mixture obtained
thus was stirred for 30 minutes. And further,
- 36 - I .-
I
dimethylformamide (1.2~ ml) was added dropwise *o the
mixture over 10 minutes at the same temperature and
stirred for one hour to gi~e a clear solution. The
solution was added dropwise to the solution of 7-amino-
3-cephem-4-carboxylic acid at -S to 5C, over 30 i
minutes, at pH 6.5 to 7.5, and the reaction mixture i
was stirred for one hour a~ the same temperature.
~thyl acetate ~200 ml) was added to the resultan~
solution, and the aqueous layer was separated, washed
with methylene chloride, bubbled with nitrogen gas and
adjusted topH 4 with acetic acid. The solution was sub-
jected to column chromatography on macroporous, non-
ionic adsorption resin "Diaion HP-20" (Trade mark:
manufactured by Mitsubishi Chemical Industries Ltd.)
and eluted wi~h 20~ aqueous solution of isopropyl
alcohol. The eluate was concentrated under reduced
pressure and lyophilized to give 7-[2-(2-amino-4-
thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-
carboxylic acid (syn isomer: 2.0 g)
I. R. ~ maxl : 3470, 3280, 3200, 17809 1695
1655, 1622 cm 1
N.M.R. ~(DMSO-d6, ppm) : 3.60 (2H, broad s),
3.84 (3H, s), 5.12 (lH, dd, J=5Hz),
5.84 (lH, dd, J=5.8Hz), 6.52 (lH, broad
t), 6.76 (lH, s), 7.26 (2Hj broad s),
9.65 (lH, d, J=8Hz)
(2) Zinc powder (4.5 g) was added to a`stirred
solution of 7-[2-(2-amino-4-thiazolyl~-2-methoxyimino-
acetamido]-3-cephem-4~carboxylic acid (syn isomer :
3 g) in 90~ formic acid ~150 ml) under ice-cooling
over 5 minutes, and stirred at the same temperature
for 15 minutes. Af~er the resul~an~ solu~ion was
filtered and washed with formic acidJ the filtrate
. .
,
~, ' ~ , ,
. .
Z~
37
and the washings were combined and concentrated under
reduced pressure ~o a volume of about 20 ml. The
concentrate w~s dissolved in water ~150 ml) and bubbled
with hydrogen sulfide gas for 20 minutes under ice-
S cooling. The precipitating zinc sulfide was filtered
off, and ~he filtra~e was treated with activated
- charcoal and lyophilized to give 7-~2-(2-amino-4-thiazolyl)glycinamido]-3-cephem-4-carboxylic acid
formate t2.9 g), pale yellow powdera mp~240C.
I.R. v mUaxol : 3330, 32009 3100, 1770, 1690 cm 1
N.M.R. ~(D20; ppm) : 3.42-3.61 (2H, m), 5.03-
5.16 (lH, m), 5.19 (lH, s), 5.60 (1/2H,
d, J=5Hz), 5.79 (1~2H, d, J=SHz), 6.36 -
6.50 tlH, m), 8.32 (lH, s)
Example 7
To a stirred solution of 7-E2-(2-amino-4-
thiazolyl)glyoxyloylamino]-3-cephem-4-carboxylic aacid
(520 mg) in methanol (15 ml) was added sodium boro
hydride (100 mg) under ice-cooling, and stirred at the
same temperature for 3 hours. Methanol was distilled
off under reduced pressu~e from the reaction mixture,
and the residue was dissolved in water (3 ml) and
adjusted to pH 3 with 6N-hydrochloric acid. The pre-
cipitates were collected by iltration, washed with
water and dried over phosphorus pentoxide to give 7-~2-
(2-amino-4-thiazolyl)-2-hydroxyaretamido~-3-cephem-4-
carboxylic acid (290 mg).
I. R. v NmUJol : 1775, 1630 cm 1
N.M.R. ~ (D~O+DMS0-d6, ppm) : 5.03 (lH, s), 5.07 .
~lH, d, J=4.6Hz~, 5.72 (lH, d, J-4.6Hz),
6.49 ~lH, m, J=3.2Hz), 6.67 ~lH, s)
.~ .
, .
- :
~ 3~ - ;
:
A solution of 7-L2-~-amino-4-~hiazolyl)glyoxyloyl-
amino]-3-cephem-4-carboxylic acid hydrochIoride (I.78 g)
in water ~100 ml) was adjusted to pH 6.0 with sodium
bicarbonate under ice-cooling with stirring. Sodium
acetatc (0.38 g) and methoxylamine hydrochloride (1.37
g) were added to the solution adjusted to pH 7.0 with
sQdium bicarbonate and then stirred at 48C for an hour.
The resultant solution was washed with ethyl acetate
(200 ml) and diethyl ether ~100 ml) in turn, and then
nitrogen gas was bubbled into the solution to remove
diethyl ether. The aqueous solution was adjusted to
pH 3.5 with 10% hydrochloric acid under ice-cooling
with stirring. The precipitates were collected by fil-
tration, washed with water and dried over phosphorus
pentoxide under reduced pressure to give 7-[2-(2-
amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-
4-carboxylic acid ~syn isomer, 120 mg). The mQth~r
liquor was subjected to column chromatography on
macroporous, nonionic adsorption resin"Diaion HP-20"
(Trademark: manufactured by Mitsubishi Chemical Indust-
ries Ltd.) and eluted with 40% aqueous acetone. The
eluate was concentrated under reduced pressure, lyo-
philized, and then dried over phosphorus pentoxide to
give the same object compound (950 mg). Total ~eld
1070 mg.
I R ~ Nu~ol : 3460, 3290, 3150, 17809 1655
1623 cm~l
3Q N.M.R. ~(DMSO-d6, ppm) : 3.60 (2H, broad s),
3.84 (3H, s), 5.12 (lH, d, J=5Hz),
5.84 (lH,d,d, J=5~z`~ 8Hz), 6.52 ~lH, t~,
6.76 (lH, s), 7.26 (2H, broad s),
9.65 ~lH, d~ J=8Hz)
.
,
~ .
-39
Example 9
The following compounds may be prepared .in a
similar manner to that of Example 8.
(1) 7-[2-~2-a.minothiazol-4-yl~-2-hydroxyimino-
5 acetamido]-3-cephem-4-carboxylic acid (syn isomer)
I. R~ v NUx~l : 3350-3200; 1770, 1670, 1630 cm 1
N.M.R. ~(DMS0-d6, ppm3 : 3,60 (2H, broad s),
5.10 (lH, d, J=5Hz), 5.83 (lH, dd,
J=5Hz, 8Hz)g 6.47 tlH, t, J=4Hz), 6.67
(lH, s), 9.47 (lH, d, J=8Hz)
~2) 7-[2-(2-aminothiazol-4-yl)-2-ethoxyiminoacet-.
amido]-3-cephem-4-carboxylic acid (syn isomer)
I. R. v Naxl : 3500, 3300, 3200, 1785, 1625,
1600 cm~
N.M.~. ~(DMS0-d6, ppm) : 1.20 (3H9 t, J=7Hz).
3 57 t2H, m), 4.08 (2H, q, J=7Hz),
5.08 (lH, d, J=5Hz), 5.83. (lH, dd9
J-5Hz, 8Hz), 6.47 (lH, m), 6.73 (lH, s),
7.20 (2H9 m), 9.58 (lH, d, J=8Hz~
(3) 7-[2-(2-aminothiazol-4-yl)-2-propoxyimino-
acetamido]-3-cephem-4-carboxylic aeid (syn isomer)
~5
I. R. v NUxl : 3250, 1770, 1650, 1660, 1620 cm 1
N.M.R. ~ (DMS0-d6~ ppm) : 0.93 (3H, t, J~7Hz),
1.67 (2H, sextet, J=7Hz), 3.60 ~2H, m),
4.03 (2H5 t, J-7Hz), 5.13 (lH, diJ~J=5Hz),
5.83 (lH, dd, J-5Hz, 8Hz), 6.48 (2H, t,
J=4Hz), 6.70 (lH, s), 7.18 (2H, m),
~.53 ~lH, d, J=8Hz)
(4) 7-[2-(2-aminothiazol-4~yl)-2-butoxyiminoacet-
35 amido~-3-cephem-4-carboxylic acid (syn isomer)
:
.,. . , :
. , . ~
:: .
~ 40 ~
I R ~ NU]ol 3320, 1775~ 1660 cm 1
N.M.R. ~ (DMSO-d6, ppm) ; 0.8~ (3H, ~, J-7Hz),
1.1-1.9 (4H, m~, 3.58 (2H, broad s),
4 05 ~2H, t, J=7Hz), 5.08 (lH, d, J=5Hz),
5.80 (lH, dd, J=5Hz~ 8Hz), 6.44 (lH,
o broad s) 7 7.1~ (2H, s), 9 51 (lH, d,
J=8Hz)
~5) 7-[2-(2-aminothiazol-4-yl)-2-pentyloxyimino-
acetamido]-3-cephem-4-caTboxylic acid (syn isomer).
I. R. v NU]l : 3300, 1775, 1650~ 1540 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 0.6-2.0 (9H9 m), 3.56
(2H, d, J-2Hz), 4.03 (2H, t, J=6Hz),
5.08 ~lH, d, J=5Hz), 5.81 (lH, dd,
J35Hz, 8Hz~, 6.46 (lH, t, J=4Hz~,
6069 (lH, s~, 7.20 (2H, s), 9.1S (lH~
d, J=8Hz)
(6) 7-[2-(2-aminoakhiazol-4~yl)-2-hexyloxyimino-
acetamido~-3-cephem-4-carboxylic acid (syn isomer).
I. R. v Na~l : 32S0, 1760, 1640, 1600 cm 1
N.M.R.~(DMSO-d6, ppm) : 1.88 (3H, m), 1.1 -
1.9 (8H, m), 3.60 (2H, m), 4.06 (2H, t,
J=6Hz), 5.10 (lH, t, J=5Hz), 5.82 (lH,
dd, J=SHz, 8Hz~, 6.46 (lH7 m), 6.70 (lH,
s), 7.25 (2H, m), 9.56 tlH, d, J=8Hz)
~7~ 7-~2-(2-aminothiazol-4-yl~-2-allyloxyimino-
acetamid~-3-ceph~m-4-carboxylic acid (syn isomer).
I R v NUJol 3300, 17~0, 1660, 1630 cm
N.M.R. ~(DMSO-d6, ppm) : 3.67 (2H, d, J=4Hz),
.
.
' ~ ' ' ' ,
" .
4.67 (2H, m), 5.17,(lH~ d, J=5Hz~,
5.25 (lH, m), 5.50 ~lH, m)~ 5.90 ~lH,
dd, J-SHz, 8Hz), 6.G3 (lH, m)~ 6,55 (lH9
m), 6.80 (lH, s), 7.50 (2H, m), 9.68 (lH,
d, J=8Hz)
t8) 0 7-[2-(2-aminothiazol-4-yl)-2-propargyloxyimino-
acetamido]-3-cephem-4-carboxylic acid (syn isomer). :
I. R. v Nu~ol 3500, 3300, 1780, 1720, 1660,
1630 cm~l
N.M.R. ~(DMSO-d6, ppm) : 3.48 (lH, m)g 3.67 (2H,
m), 4.80 (2H, d, J=2Hz), 5.17 (lH9 d,
J=5Hz), 5.88 (lH, dd, J=5Hz, 8Hz), 6.55
~lH, m), 6.~5 (lH, s), 7.33 (2H, m),
9.73 (lH, d, J=8Hz)
~9) 7- ~2- (2-aminothiazol-4-yl~ - 2-cyclohexyloxyimino-
acetamido]-3-cephem-4-carboxylic acid (syn isomer)
I. R. ~ Na~l : 335-0, 1775~ 1665, 1620, 1540 cm 1
N.M.R. ~ (DMSO-d6, ppm) : 0.8-2.2 (lOH9 m), 3.60
;(2H, broad s~, 4.04 (lH, m), 5.09 (lH, .
d, J=SHz), 5.83 (lH, dd, J=5Hz, 9Hz),
6.45 (lH, t, J=4Hz), 6.67 (lH, s), 7.19
(2H, s), 9.48 (lH, d, J=9Hz) .
In this disclosure the expression 'such as' is
to be understood as meaning 'for example'.
.
.
, ,
,
. ~ ,
