Note: Descriptions are shown in the official language in which they were submitted.
1:~54~09
me present invention is concerned with novel oephalosporin deriva-
tives of the general formula
H H
CH30N= C--CONH ~S ~
N~ ~LN //LCH2--S--X
H2N S
COOH
in which X represents one of the groups
3 \N~ ~ ORl \ N--N
J~N O /~N ~\ COOH
(a) (b)
wherein Rl represents lower alkyl or (together with the oxygen) a
readily hydrolysable ether group,
as well as readily hydrolysable esters of the compounds of formula I wherein X
represents the group (b) or (a) in which Rl represents lower alkyl and salts of
these cc~pounds and hydrates of the compounds of formula I or of esters and
salts thereof.
. . ~ . .
~54~09
Rl in the group (a) can represent lower aIkyl, preferably a straight-
chain or branched alkyl group with 1-7 carbon atoms such as ethyl, n-propyl, iso-
propyl, n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl or, especially, methyl.
Rl can (together with the oxygen) also represent a readily hydrolysable ether
group. Thus, Rl can signify, for example: lower alkanoyloxyalkyl (e.g. acetoxy-
methyl, pivaloyloxymethyl, l-acetoxyethyl and l-pivaloyloxyethyl), lower alkoxy-
carbonyloxyalkyl (e.g. methoxycarbonyloxymethyl, l-ethoxycarbonyloxyethyl and
l-isopropoxycarbonyloxyethyl), lactonyl groups (e~g. phthalidyl and thio-
phthalidyl), lower aIkoxymethyl (e.g. methoxymethyl) and lower alkanoylam mo-
methyl (e.g. acetamidomethyl).
As readily hydrolysable esters of the compounds of formula I there areto be understood compcunds of formula I in which at least one carboxy group is
present in the form of a readily hydrolysable ester group. Examples of such
esters, which can be of the conventional type, are the lower aIkanoyloxyalkyl
esters (e.g. the acetoxymethyl, pivaloyloxymethyl, l-acetoxyethyl and
l-pivaloyloxyethyl ester), the lower aIkoxycarbonyloxyalkyl esters (e.g. the
methoxycarbonyloxymethyl, l-ethoxycarbonyloxyethyl and l-isopropoxycarbonyloxy-
ethyl ester), the lactonyl esters (e.g. the phthalidyl and thiophthalidyl ester)
the lower alkoxymethyl esters (e.g. the methoxymethyl ester) and the lower
aIkanoylaminomethyl esters (e.g. the aoe tamidomethyl ester). Other esters (e.g.
-
-b
,.
~ ~' "" , '
- '
3,154~0~
the benzyl and cyanomethyl esters) can also be used. The esters can be mono-
esters or diesters. me second ester formation can occur in the compound with
the carboxy group of the 3-carboxy-l-methyl-IH-1,2,4-triazol-5-yl group X [group
(b)].
Examples of salts of the compounds of formula I are aLkali metal salts
such as the sodium and potassium salt, the ammonium salt, alkaline earth m~etal
salts such as th~ calcium salt, salts with organic bases such as salts with
amines (e.g. salts with N-ethyl-piperidine, procaine, dibenzylamine, N,N'-
-dibenzylethylenediamine, alkylamines or dialkylamines) as well as salts with
an mo acids such as, for example, salts with arginine or lysine. The salts can
be mono-salts or also di-salts. me second salt formation can occur in com-
pounds with the carboxy group of the 3-carboxy-1-methyl-lH-1,2,4-triazol-5-yl
group X [group (b)].
me compounds of formula I also form addition salts with organic or
inorganic acids. Exa~ples of such salts are hydrohalides (e.g. hydrochlorides,
hydrobromides and hydroiodides) as well as other mineral acid salts such as sul-
phates, nitrates, phosphates and the like, alkylsulphonates and monoarylsulphon-
ates such as ethanesulphonates, toluenesulphonates, benzenesulphonates and the
like and also other organic acid salts such as acetates, tartrates, maleates,
citrates, benzoates salicylates, ascorbates and the like.
-- 3 --
1154~ 9
The compounds of formula I as well as their salts and readily hydrolys-
able esters can ke hydrated. The hydration can be effected in the course of the
manufacturing process or can occur gradually rlS a result of the hygroscopic pro-
perties of an initially anhydrous product.
The products in accordance with the invention can be present in the
syn-isomeric form
N C - CONH
/1 ~ N
H2N ~ S ~ \OCH3
or in the anti-isomeric form
1 ~ C - CONH
H2N S
CH30
or as a mixture of these tw~ forms~ The syn-isomeric fonm or mixtures in which
the syn-isomeric form predominates is/are preferred.
-- 4 --
'
:1154009
Preferred products are
- (6R,7R)-7-/2-(2-amino-4-thiazolyl)-2-[(Z)-methoxyimino]acetamido/-3-
-/[(2,5-dihydro-6-methoxy-2-methyl-5-oxo-as-triazin-3-yl)thio]methyl/-
-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-carboxylic acid,
- (6R,7R)-7-/2-(2-amino-4-thiazolyl)-2-[(Z)-methoxyimino]acetamido/-3-/
[(3-carboxy-1-methyl-lH-1,2,4-triazol-5-yl)thio]methy V-8-oxo-5-thia-
-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid,
- (6R,7R)-7-[2-(2-amino-4-thiazolyl)-2-((Z)-methoxylmm o)acetamido]-3-
-[[[2,5-dihydro-2-methyl-5-oxo-6-[(pivaloyloxy)methoxy]-as-triazin-3-
-yl]thio]methyl]-5-oxo-4-thia-1-azabicyclo[4.2.0]oct-2-ene-2-
-carboxylic acid
and their salts as well as the corresponding hydrates.
me above cephalosporin derivati~es can be m~nufactured in accordanoe
with the invention by
(a) cleaving off the protecting group R, and, if desired, carboxy protect-
ing groups which may be present, in a compound of the general formula
H H
CH3oN~= C - COMH ~ S ~ 1 II
~ ~ o \~CH2--S--X
~~\S
COO~
`- llS4V09
-- 6 --
in which X has the same significance
as X in formula I, except that the carboxy
group of group (b) can be protected,
R represents a cleavable protecting
group and the carboxy group in the 4-
-position of the cephalosporin moiety
can be present in protected form,
or
(b) reacting a halide of the general formula
H H
C H3 ON--C--CONH~/ ~ 1
I ~ N ~4 ~ -~H2 - S - X III
-` .. C H 2 Y
i.n which Xl has the significance
. given above and Y represents a
halogen atom and the carboxy group
in the 4-positlon of the cephalosporin
moiety can be present in protected form,
with thiourea~
G..'
(c~ .tor the manufacture of a readily hydrolysable ester
of a compound of formula I, subjecting a carboxylic acid of
formula I to a corresponding esterification,
or
. . .
1154~09
(d) for the manufacture of salts or hydrates of a compound
of formula I or hydrates of these salts, converting a compound
of formula I into a salt or hydrate or into a hydrate of
said salt.
If desired, carboxy groups present in the starting
materials of formulae II and III can be protected; for
example, by esterification to form a readily cleavable ester
such as a silyl ester (e.g. the trimethylsilyl ester)~ The
readily hydrolysable esters mentioned above also come into
consideration. The carboxy groups can also be protected
by salt formation with an inorganic or tertiary organic
base such as ~riethylamine. Possible R-protecting groups
arel for example, protecting groups which are:cleavable by
acid hydrolysis such as, for example, t-butoxycarbonyl or
trityl/ or protecting groups which are cleavable by hasic
hydrolysis such as, for example, trifluoroacetylO Preferred
R-protecting groups are chloroacetyl, bromoacetyl and
iodoacetyl, especially chloroacetyl. The last-mentioned
protecting groups can be cleaved off by treatment with
thiourea.
The starting materials of formula II can be preparedf
for example, by N-acylating the corresponding 7-amino
compound, namely by reacting a compound of the general
formula
- -' 11540(~
H H
IV
~ N ~ CH2--S--X
COOH
in which ~ has the significance given above and the carboxy group and/
or the amino group can be present in protected form,
with an acid of the general formula
CH30N--C ~--COOH
N--¦
RHN--~S ~ V
in which R has the significan oe given above,
or wqth a reactive functio.nal derivative of this acid and, if desired, cleaving
off carboxy protecting groups which may be present.
If desired, the carboxy groups present in the 7-am mo compound of
form~la IV can be protected in the manner mentio.ned above for the starting mate-
~S4~)9
rial of formula II to be prepared. me amLno group of the compound of formulaIV can be protected, for example, by a silyl protecting group such as trimethyl-
silyl.
As reactive functional derivatives of acids of formula V there come
into consideration, for example, halides, (i.e. chlorides, bromides and fluor-
ides), azides, anhydrides, especially mixed anhydrides with strong acids, reac-
tive esters (e.g. N-hydroxysuccinimide esters) and amides (e.g. imidazolides).
me reaction of the 7-amino o~mpound of formula IV with an acid of
formula V or a reactive functional derivative thereof can ke carried out in an
manner known per se. Thus, for example, a free acid of formula V can be con-
densed with one of the aforementioned esters corresponding to formula IV by
means of a carbodiimide such as dicyclohexylcarbcdiimide in an inert solvent
such as ethyl acetate, acetonitrile, dioxan, chloroform, methylene chloride,
benzene or dimethylformamide and subsequently the ester group can be cleaved off.
~xazolium salts (e.g. N-ethyl-5-phenyl-isoxazolium-3'-sulphonate) can also be
used as the condensation agent in place of carbcdiimides.
According to another emkodiment, a salt of an acid of formula IV (e.g.
a trialkylammonium salt such as the triethylammonium salt) is reacted with a
...
.
- ~ ~ ~
~154U09
reactive functional derivative of an acid of formula V as mentioned above in an
inert solvent (e.g. one of the solvents named above).
According to a further embodiment, an acid halide, preferably the
chloride, of an acid of formula V is reacted with the amine of formula IV. me
reaction is preferably carried out in the presence of an acid-binding agent, for
example, in the presen oe of aqueous alkali, preferably sodium hydroxide, or in
the presenoe of an alkali metal carbonate such as potassium carbonate or in the
presence of a lower alkylated amune such as triethylamine. As the sol~ent there
is preferably used water, optionally in admixture with an inert organic solvent
such as tetrahydrofuran or dioxan. m e reaction can also be carried out in an
aprotic or~anic solvent such as, for example, dimethylformamide, dimethyl sul-
phoxide or hexamethylphosphoric acid triamide. When silylated starting mate-
rials of formula IV are used, the reaction is carried out in an anhydrous medium.
The reaction of the 7-amino co~pound of formula IV with the acid of
formula V or a reactive functional derivative thereof can conveniently be
carried out at temperatures between about -40& and room temperature, for
example at about o-lo&.
-- 10 --
i' 'i
~ .
'
1~54~09
Starting materials of formula II wherein R is not monohalogenated (as
in bromo-, chloro and iodoaoe tyl) can also be prepared by thiolation, namely by
reacting a comFound of the general formula
H H
~a~2-Z VI
COOH
in which RO iS as R but cannot be monohalogenated, Z represents a leav-
ing group and the carboxy group can be protected by salt formation
with an inorganic or tertiary organic base,
with a thiol of the general formula
HS--Xl VII
h~ which Xl has the significance given above,
and, if desired, cleaving off a carboxy protecting group which may be present.
As the leaving group Z in a compound of formula Vl there come into con-
sideration, for example, halogens (e.g. chlorine, bromine or iodine), acyloxy
groups (e.g. lower aIkanoyloxy groups such as acetoxy), lower alkylsulphonyloxy
or arylsulphonyloxy groups such as mesyloxy or tosyloxy, or the azido group.
-- 11 --
:~:154VU9
me reaction of a oompound of formLla Vl with a thiol of formula VII
can be carried out in a manner kncwn per se; for example, at a temperature be-
tween about 40C and 80C, conveniently at about 60C, preferably in water or in
a buffer solution with a pH of about 6 to 7, preferably 6.5.
me carbcxy group of the resulting compounds of formula II can sub-
sequently be protected if desired (e.g. by salt formation or esterification).
me thiols of formula VII are in tautameric equilibrlum with the
corresponding thiones as shown in the follcwm g formulae
3 \ N ~ ~ ORl 3 \ N
5 1 N ~ O EIS /bN ~ O
VIIal VIIa2
- 12 -
.
.
. 1 540()9
wherein R has the above significance,
or
3 \ 3 \
S 1H COOH HS N COOH
VlIbl VIIb
wherein the carboxy group can be protected.
The preparation of these compounds is described in Examples 1,2 and 3.
For the preparation of thiols (thiones) correspanding to group (a) which are
etherified in the 6-position, the Rl- group is generally introduced by reacting
a S-protected thiol (e.g. by benzhydryl) with the halide containing the Rl-
group, preferably the iodide, in an inert organic solvent in the presence of an
acid-binding agent (e.g. potassium carbonate), preferably at about 10-50 &, and
cleaving off the protecting group (benzhydryl can be cleaved off with anisole
and trifluoroacetic acid at room temperature).
In accordance with variant (a~ of the process in accordance with the
invention, the amino protecting group R in a starting material of formula II is
- 13 -
, . ,
,:
l:~S~09
cleaved off. Protecting groups which are cleavable by acid hydrolysis are pre-
ferably removed with the aid of a lower alkanecarboxylic acid which may be
halogenated. In particular, formic acid or trifluoroacetic acid is used. This
cleavage is generally carried out at rcom temFerature, although it can be
carried out at a slightly higher or slightly lower temperature, for example, a
temperature in the range of about 0C to +40C. Protecting groups which are
cleavable under alkaline conditions are generally hydrolysed with dilute aqueous
caustic alkali at 0C to 30&. m e chloroacetyl, bromoacetyl and iodoacetyl
protecting groups can be cleaved off by means of thiourea in acidic, neutral or
aIkaline medium at about 0-30&. Hydrogenolytic cleavage (e.g. cleavage of
benzyl) is unsuitable in this case, since the oxime function would be reduced to
the amino group during the hydrogenolysis.
After carrying out process variant (a), carboxy protecting groups
which may be present in the reaction product can be cleaved off if desired.
~hen the protecting group is a silyl group (silyl ester), this group can be
cleaved off especially readily by treating the reaction product with water.
Lower aIkanoyloxyalkyl, alkoxycarbonyloxyaLkyl, lactonyl, alkoxymethyl and
alkanoylaminomethyl esters are preferably cleaved enzymatically with the aid of
a suitable esterase (at about 20-40C). When a carboxy group is protected by
- 14 -
i~54(~
salt formation !e.g. with triethylamine), then the cleavage of this salt-forming
protecting group can be carried out by treatment with acid. me acid which can
be used for this purpose can be, for example, hydrochloric acid, sulphuric acid,
phosphoric acid or citric acid.
me carboxy protecting groups can be cleaved off in the same manner as
just described also prior to the cleavage of the protecting group R.
The halide of formwla III used in accordance with the invention can be
prepared, for ex3mple, by reacting a co~pcund of formula IV with a halogenated
carboxylic acid of the general formula
CH30H = C - COOH
CO VIII
~H2Y
in which Y represents a halogen atom,
or with a reactive derivative of this compound. The halogenated carboxylic acid
of formLla VIII is used either in free form in the presenoe of a condensation
agent (e.g. a N,N'-disubstituted carbodiimide such as N,N'-dicyclohexylcarbodi-
imide or an azolide compc~nd such as N,N'-carbonyldiimidazole or N,N'-thionyl-
- 15 -
,. ..
~1~4~
diimidazole) or in the form of an acid halide such as the acid chloride or acid
bromide, in the form of an acid anhydride such as an acid anhydride with a
carbonic acid monoester (e.g. with monomethyl OE bonate or monoisopropyl carbon-
ate) or in the form of an activated ester such as the p-nitrophenyl ester, 2,4-
-dinitrophenyl ester, N-hydroxysuccinimide ester or N-hydroxyphthalimide ester.
The reaction is generally carried out in an inert organic solvent, for example
in a halogenated hydrocarbon such as chloroform, dichloromethane or carbon tetra-
chloride, in an ether (e.g. tetrahydrofuran or dioxan), in dimethylformamide,
dimethylacetamide, water or mixtures thereof. m e reaction temperature mainly
lies in the range of about -50C to +40 C, preferably at about -lo& to +lo&.
The reaction in accordance with the invention of the halide of formula
III with thiourea, variant (b) of the process in accordanoe with the invention,
is preferably carried out in an inert solvent such as, for example, in a lower
alkanol (e.g. ethanol), in a lcwer ketone such as aoe tone, in an ether as tetra-
hydrofuran or dioxan, in dimethylformamide, dimethylaoe tamide, in water or in
mixtures thereof. The reaction temperature generally lies in the range of about
0C to 60&, preferably at room temFerature. m e chloride, bromide, fluoride or
iodide can be used as the halide of formula III, the chloride or the bromide is
preferably used. me free acid of formula III or, if desired, also a salt there-
of can be used, whereby the same salts as the salts of the oompounds of formlla
I mentioned above come into considbration.
- 16 -
~S~ 9
In order to manufacture the readily hydrolysable esters of the
carboxylic acids of formula I in aocordance with variant (c), the carboxylic
acid is preferably reacted with the corresponding halide, preferably with the
iodide, containing the ester group. The reaction can be accelerated with the
aid of a base, for example an aIkali metal hydroxide or carbonate or an inorganic
amine such as triethylamine. When group (b) is present the carboxy function
thereof is also esterified, a readily hydrolysable diester being obtained. In
this case an excess of the corresponding halide is preferably used. The esteri-
fication is preferably carried out in an inert organic solvent such as dimethyl-
acetamide, hexamethylphosphoric acid triamide, dimethyl sulphoxide or, preferably,dimethylformamide. The temperature preferably lies in the range of about 0-40~.
me manufacture of the salts and hydrates of the co~pounds of formula
I or the hydrates of these salts can be carried out in a manner kncwn per se;
fo~ example, by reacting the carboxylic acid of formula I with an equivalent
amount of the desired base, conveniently in a solvent such as water or in an
organic solvent such as ethanol, methanol, acetone etc. When a second equivalent
of base is used salt formation is also effected on the carboxy function of a
- 17 -
-'``"` 115~00~
group (b) which may be present, a di-salt resulting. The te~perature at which
the salt formation is carried out is not critical. It is generally carried out
at room temperature but it can also be carried out at a temperature slightly
above or belcw room temperature (e.g. in the range of 0 C to +50&).
The manufacture of the hydrates usually takes place automatically in
the course of the manufacturing process or as a result of the hygroscopic proper-
ties of an initially anhydrous product. For the controlled manufacture of a
hydrate, a completely or partially anhydrous product (carboxylic acid of formLla
I or ester, ether or salt thereof) can be exposed to a moist atmosphere (e.g. at
about +10C to +40C).
The 7-amino compcunds of formula IV used above can be prepared start-
ing from a compound of the formula
H H
H2N ~ ~ IX
~ N ~ 2
O
CO~H
- 18 -
115400g
-- 19 --
in which Z represents a leaving group
and the carboxy group can be protected
by salt formation with an inorganic or
tertiary organic base,
with a thiol of formula VII. The reaction can be carried
out under the same conditions as those which have been
described for the reaction of the starting materials of
formulae VI and VII. On the other hand, the compounds of
formula VI can be prepared starting from a compound of
formula IX and an acid sf formula V or a reactive.functiona.l.
derivative thereof under the same conditions as have been
described for the reaction of the compounds of formulae IV
and V.
The carboxy group and/or the amino group of the
resulting compound of formula IV can s.ubsequently be
protected if desired, for example:by esterifying the
carboxy group or forming a salt thereon or by silylating
the amino group.
A syn/anti mixture of a ~ompound of formula I which
may be obtained can be separated into the corresponding
s~n~ and anti-forms in the customary manner, for example
by recrystallisation or by chromatographical methods using
a suitable solvent or solvent mixture.
.. .:....
- ~ ,, ~,
~Ll 54009
- 20 ~
The compounds of formulae I and II as well as the
corresponding readily hydrolysable esters and salts or
. the hydrates of these products have antibiotic, especially
bactericidal, activity. They possess a broad spectrum
of activity against gram-positive bacteria (e.g. Staphylccocci)
and against gram-negative bacteria such as, for example,
Haemophilus influenzae and Neisseria.gonorrhoeae, as well as
against various ~-lactamase-forming gram-negative organisms
such as Escherichia coli, Serratia marcescens, Enterobacter
cloacae, Pseudomonas aeruginosa, Proteus mirabilis and
Proteus vulgaris.
The compounds of formula I and II as well as the
corresponding readily hydrolysable esters and salts or
: the hydrates of these products can be used for the treat-
ment and prophylaxis of infectious diseases. A daily
dosage of about 0.1 g to about 2 g comes into consideration
for adults. The parenteral administration of the compounds
in accordance with the invention is especially preferred.
In order to demonstrate the antimicrobial activity
of the aforementioned products, the following representative
members were tested
.
1~540(~9
- 21
Product A: (6R,7R)-7-/2-(2-amino-4-thiazolyl)-2-[(Z)-
-methoxyimino]acetamido/-3-/[(2,5-dihydro-6-
-methoxy-2-methyl-5-oxo-as-triazin-3-yl)thio]-
methyl/-8-oxo-S-thia-l-azabicyclo[4.2.0]oct-
-2-ene-carboxylic acid sodium salt.
Product B: (6R,7R)-7-/2-(2-amino-4-thiazolyl)-2-[(Z)-
-methoxyimino]acetamido/-3-/[(3-carboxy-1-
-methyl-lH-1,2,4-triazol-5-yl)thio]methyl/-
-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-
-2-carboxylic acid disodium salt.
Product C: (6R,7R)--7-[2-(2-amino-4-thiazolyl)-2-((Z)~
-methoxyimino)acetami~o]~3-[[[2,5-dihydro~2=
-methyl-5-oxo-6-[(pivaloyloxy)methoxy]-as-
-triazin-3-yl]thio]methyl]-5-oxo-4-thia-1-
-azabicyclo[4~2oo]oct-2-ene-2~carboxylic acid
sodium salt.
Activity in vitro: Minimum inhibitory concentration
(~y/ml)
1~5400~
- 22 -
.__
Pathogenic organism A B C
._ _ . . __
Escherichia coli *) 0.04 0.08 0.10
Serratia marcescens *) 0.16 0.16 3.1
Enterobacter cloacae *) 1.2 10 25
Proteus mirabilis *) 0.04 0.01 0.4
Proteus vulgaris *) 0.02 0.01 0.2
Pseudomonas aeruginosa
strain 1 *) 40 40 50
strain 2 *) 80 40 50
Haemophilus influenzae 0.01 0.01 0.04
Neisseria gonorrhoeae o.01 0.0025 0.006
Staphylococcus aureus 2.5 10 1.6
*) ~-lactamase-forming strain
Compounds of formula I in which X represents group (b)
15 as well as the corresponding esters, salts and hydrates are
distinguished by a long half-life, i.e. they remain in the
organism for a long time and can accordingly more effectively
exert their gH~micidal functionO Thus, for example, the
disodium salt of (6R,7R)-7-/-2-(2-amino-4-thiazolyl)-2-[(Z)-
20 -methoxyimino]acetamido/-3-/[(3-carboxy-1-methyl-lH-1,2,4-
-triazol-5-yl)thio]methyl/-8-oxo-5-thia-1-azabicyclo[4.2.0]-
oct-2-ene-2-carboxylic acid has a half life ih rats of 22
minutes after the administration of 20 mg of subs~ance per
5~()09
- 23 -
kg body weight.
Toxicity
.__
Test substance A B
LDloo mg/kg i.v. , 500 > 500
s.c. >4000 >4000
- >5000 >5000 >5000
The products in accordance with the invention can
be used as medicaments, for example in the form o~ pharma-
ceutical preparations which contain them in admixture with
a pharmaceutical, organic or inorganic inert carrier material
which is suitable for enteral or parenteral administration
such as, for example, water, gelatin, gum arabic, lactose,
starch, magnesium stearate, talc, vegetable oils, polyalkylene-
glycols, Vaseline etc. The pharmaceutical preparations can
be made up in solid form (e.g. as tablets, dragées,
suppositories or capsules) or in liquid form (e~g. as solutions,
suspensions or emulsions). If necessary, they can be
sterilised and/or can contain adjuvants such as preserving,
stabilising, wetting or emu:lsifying agents, salts for varying
the osmotic pressure, anaesthetics or buffers. They can also
contain still other therapeutically valuable substances.
``= 11~400~
- 24 -
The compounds of formula I and their salts or hydrates are
preferably administered parenterally and for this purpose
are preferably prepared as lyophilisates or dry powders for
dilution with customary agents such as water or isotonic sodium
chloride solution. The readily hydrolysable esters of the
compounds of formula I and their salts or hydrates also come
into consideration for enteral administration.
'
54009
- 25 -
The following Examples illustrate the present
invention:
Example 1
Preparation of the sodium salt of (6R,7R)-7-/2-(2-
-amino-4-thiazolyl)-2-[(Z)-methoxyimino]acetamido/-3-/[(2,5-
-dihydro-6-methoxy-2-methyl-5-oxo-as-triazin-3-yl)thio]methyl!-
-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid.
34.5 g of the sodium salt of (6R,7R)-7-/2-L2-(2-
-chloroacetamido)-4-thiazolyl]-2-[(Z)-methoxyimino]-acetamido/-
-3-/[(2,5-dihydro-6-methoxy-2-methyl-5-oxo as-triazin-3-yl)-
thio]methyl/-8-oxo- 5-thia-1 azabicyclo[4.2 O]oct-2-ene-2~
-carboxylic acid are dissolved in 700 ml o~ water together
with 20 g of thiouxea and stirred overnight at 25C while
gassing w.th nitrogen, the pH-value of the reaction solution
being held at 7 by adding lN sodium hydroxide. The pH-value
is re-adjusted to 4 by adding lN hydrochlo~ric acid while
stirring vigorously. The material which thereby precipitates
is filtered o~f under suction and discarded. The yellow m~ther
li~uor is adjusted to pH 3 with lN hydrochloric ~cid. The
precipitated product is filtered o~f under suction, washed
with water, ethanol and low-boiling petroleum ether and dried
at gO~C in vacuo. There is obtained beige coloured (5R,7R)-
-7-/2-(2-amino-4-thiazolyI)-2-[(Z)-methoxyimino]acetamido/~-
-3-/[(2,5-dihydro-6-methoxy-2-methyl-5-oxo-as-triazin 3-
-yl)-thio]methyl/-8-oxo-5-thia-1-azabicyclo[4 r 2 ~ O] OcT~ 2-erne.
', .
. . . .
;. ~
- . ' ` ~ ' ' ~ ' '
.
-
~ 1~54009
- 26 -
-2-carboxylic acid which, for conversion into the sodium
salt, is suspended in 350 ml of methanol and treated with
20 ml of a 2N solution of sodium 2-ethylcaproate in ethyl
acetate A solution has resulted after ca 15 minutes and
this solution is diluted with 300 ml of ethanol. Thereby,
there precipitates a small amount of amorphous material
which is filtered off under suction and discarded. The
filtrate is concentrated strongly at 40C in vacuo. The
material which thereby precipitates is filtered off under
suction, washed with ethanol and low-boiling petroleum ether,
and dried overnight at 40C in a high vacuum. There is
obtained pure sodium salt of (6R,7R)-7-/2-(2-amino-4-
-thiazolyl)-2-[(Z)-methoxyimino]acetamido/-3-/[(2,5-dihydro-
-6-methoxy-2-methyl-5-oxo-as-triazin-3-yl)thio]methyl/-8-
lS oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carbo~ylic acid
as a yellowish powder with [a]25 = -140.9 (c = l in water).
The compound used as the starting material in the
above process can be prepared as follows:
~a) Preparation of tetrahydro-~-meth~1-5,6-dioxo-3-thioxo-
-as-tria~ine-1(2H~-carboxylic acid benzyl esteru
39~79 g of 1,2,5,6--tetrahydro-5 D 6-dio~o-3-mercapto-
-2-methyl-as-triazine are ta~en up in 500 ml of dimethyl-
formamide and treated with 35 ml of triethylamine. 39 ml
of benzyloxycarbonyl chloride are added dropwise during
-- 27
15 minutes, whereby the temperature of the reaction mixture
rises to 45C, triethylamine hydrochloride precipitates
and the suspension becomes yellow in colour. The mixture
is stirred at 25C for 2 1/2 hours and subsequently
5 evaporated at 70C in vacuo. The oily evaporation residue
is stirred with 500 ml of water for 1 hour, whereby it
becomes solid. The solid is filtered off under suction and
washed with 50 ml of water. The yellow filter material i5
stirred well with 250 ml of ethanol, filtered off under
10 suction, washed with ethanol and dried at 40C in vacuo.
There is obtained a colourless crude crystallisate which
is recrystallised from methanol and yields colourless
tetrahydro-2-methyl-5,6-dioxo-3-thioxo-as-triazine-1(2H)-
-carboxylic acid benzyl ester of melting point 150-153C.
,
15 (b) Preparation of 3- [(diphenylmethyl)thio]-5,6-dihydro-
-2-methyl-5,6-dioxo-as-triazine-1(2H)-carboxylic
acid benzyl ester.
13.2 g of tetrahydro-2-methyl-5,6-dioxo-3-thioxo-as-
~triazlne-1(2H)-carboxylic acid benzyl est.er are dissolved
~0 ir! 500 ml of ethyl acetate and treated with a solution of
diphe),yl~iazometl~ane :in 90 ml of low-boi:Ling petroleum
ether~ The initial:Ly ~riolet reaction solution is left to
stand at 25C for 40 hours, the colour becoming pink.
3 ml of glacial acetic acid are added and, after 1 hour,
`
"` 1154009
- 28 -
the mixture is evapor~ted at 40C in vacuo. A yellow
oil is obtained as the evaporation residue. This is
separated by means of column chromatography on silica gel
with the elution agents benzene, benzene/ethyl acetate 95:5
and benzene/ethyl acetate 90:10. The fractions containing
the desired substance are combined and evaporated at 40C
in vacuo. After recrystallisation from ethanol, there is
obtained colourless 3-[(diphenylmethyl)thio]-5,6-dihydro-2-
-methyl-5,6-dioxo-as-triazine-1(2H)-carboxylic acid benzyl
ester of melting point 90-92C.
. .
(c) Preparation of 3-[(diphenylmethyl)thio]-1,2-dihydro-
-2-methyl-as-triazine-5,6-dione.
; 6.9 g of 3-[(diphenylmethyl)thio]-5,6-dihydro~2-
-methyl-5,6-dioxo-as-triazine-1~2H)-carboxylic acid benzyl
ester are dissolved in 100 ml of ethyl acetate and stirred
well with 30 ml of an aqueous, 7~ ammonia solution at 25C
for lS minutes. The e.thyl acetate phase is separated and
discarded. The aqueous phase is treated with 7 ml of
concentrated hydrochloric acid and stirred in an ice-bath
~o t`or 30 minutes. The substance which thereby precipitates
is filtered off under suction, washed with water and
,mmediately recrystallised from ethanol. There is obtained
colourless 3-[(diphenylmethyl)thio]-1,2-dihydro~2-methyl-
-as-triazine-5,6-dione of melting point 180-182C.
"` 1~54~09
- 29 -
(d) Preparation of 3-[(diphenylmethyl)thio]-6-methoxy-
-2-methyl-as-triazin-5(2H)-one.
3.9 g of 3-[(diphenylmethyl)thio]-1,2-dihydro-2-
-methyl-as-triazine-5,6-dione are dissolved in 40 ml of
dimethylformamide and treated with 1.68 ml of triethylamine
as well as 4 ml of methyl iodide. After stirring for 2 hours
at 25C, 1.38 g of potassium carbonate are added to the
mixture in order to accelerate the reaction. After stirring
for a further 2 hours at 25C, the starting material has
reacted completely. The resulting red suspension is poured
into water and extracted three times with ethyl acetate.
The combined ethyl acetate extracts are washed with water,
dried over sodium sulphate and evaporated at 40C i~ vacuoq
The red evaporation residue is purified by means of colum~
chromatography on silica gel with ethyl acetate as the
elution agent. Recrystallisation from ethyl acetate/ether/
petroleum ether yields yellowish 3-[(diphenylmethyl)thio]~
-6-methoxy-2-methyl-as-triazin-5(2H)-one of melting point
135-138C.
2~ (e) Preparation of 3,4-dihydro~6-methoxy-2-methyl-3-
-thioxo--as~triazin-5(2H)-one.
2.6 g of 3-[(diphenylmethyl)thio]-6-methoxy-2-methyl-
-as-triazin-5(2H)-one-are dissolved in 26 ml of anisole and
~ . j,
~.~,
.. . . . . .
~l.S~{)()g
- 30 -
50 ml of trifluoroacetic acid and stirred at 25C for 3
hours. The reaction solution is evaporated at 40C in vacuo.
The evaporation residue is stirred with a small amount of
ethyl acetate and a large amount of low-boiling petroleum
ether, filtered off under suction and recrystallised from
methanol. There is obtained 3,4-dihydro-6 methoxy-2-methyl-
-3-thioxo-as-triazin-5(2H)-one as colourless, lustrous
crystals of melting point 215-220C.
(f) Preparation of (6R,7R)-7-amino-3-/[(2,5-dihydro-
-6-methoxy-2-methyl-5-oxo-as-triazin-3-yl)thio]-
-- methyl/-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-
-2-carboxylic acid.
13.6 g of 7-aminocephalosporanic acid and 9.53 g of
3,4-dihydro-6-methoxy-2-methyl-3-thioxo-as-triazin-5(2~)-
~one are suspended in 250 ml of water. The pH-value of
the suspension is adjusted to 6.5 at 25~ with lN sodium
hydroxide. The suspension is heated to 60C and stirred
at 60C for 4 hours while gassing with nitrogen, the
pH-value being held at 6.5 with lN sodium hydroxide. After
1 hour a solution results, after 2 hours some materia:!
begins to precipitate, and sodium hydroxide is no longer
consumed. The mixture is cooled to 25~ and separate(l
from a small amount of insoluble material by filtration.
The orange filtrate is adjusted to pH 3.5 at 25C"~ with
concentrated hydrochloric acid. The precipitated su~stance
.,
. . .
-; ~1540~)~
- 31 -
is filtered off under suction, washed successively with
100 ml of water, 300 ml of acetone and 300 ml of low-boiling
petroleum ether and dried at 40C overnight in vacuo. There
is obtained a beige coloured substance which, for purification,
is suspended in 150 ml of water/methanol 1:1 and treated
dropwise with 3.6 ml of triethylamine for 1 hour, whereby at
pH 8.5 there results a dark solution which still contains
a small amount of insoluble material. After the addition
of 3 g of decolourising charcoal, the mixture is stirred at
25C for 15 minutes, filtered off from the decolourising
charcoal over a hard filter and back-washed with 50 ml of
water/methanol 1:1. The orange filtrate is adjusted to
pH 3 with ca 2 ml of concentrated hydrochloric acid. The
substance which thereby precipitates is filtered off under
suction, washed successively with 50 ml of water/methanol
lol, 100 ml of methanol, 50 ml of ether and 50 ml of
low-boiling petroleum ether and dried at 40C overnight
in vacuo. There is obtained pure beige coloured (6R,7R)-
-7-amino-3-/[(2,5-dihydro-6-methoxy-2-methyl-5-oxo-as-
-triazin-3-yl)thio]methyl/-8-oxo-5-thla-1-azabicyclo(4.2.0]-
oct-2-ene-2-carboxylic acid.
Preparation of the sodium salt of (6R,7R)-7-/2-
~2-(2-chloroacetamido)-4-thiazolyl]-2-[(Z)-methoxy-
_ _ imino]acetamido/-3-/[(2,5-dihydro-6-methoxy-2-methyl-
-5-oxo-as-triazin-3-yl)thio]methyl/-8-oxo-5-thia-1-
-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid.
.,
~''' .
: , :
'
'
lS~9
- 32 -
25 g of phosporus pentachloride are added to 200
ml of methylene chloride dried over calcium chloride. The
suspension is cooled to -10C while stirring and 46 ml of
N,N-dimethylacetamide are added ~ropwise for ca 5 minutes,
the temperature rising to -5C. The suspension is cooled
to -15C and stirred for 15 minutes. After cooling to
-20C, 33.3 g of 2-(2-chloroacetamido-4-thiazolyl)-2-(Z)-
-methoxyimino-acetic acid are added and the mixture is
stirred at -15C for 45 minutes, a yellow-orange solution
resulting. This solution is cooled to -30C, treated with
50 g of ice and stirred at -50C for 10 minutes. The
~.
methylene chloride phase containing the acid chloride formed
is separated and stored initially at ca -15C. This acid
chloride solution is added ~dxo~wise durin~ 30 minutes while
stirring vigorously to a solution, cooled to 0-5C, which
has been prepared by treating (6R,7R)-7-amino-3-/[2,5-dihydro-
-6-methoxy-2-methyl-S-oxo-as-triazin-3-yl)thio]methyl/-8-oxo-
-S-thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid,
prepared under (f), with 3N sodium hydroxide to pH 8.2 in
500 ml of water, the pH-value being held at 8.0-8.2 with
3N sodium hydroxide with the aid of an autotitrator. The
mi~tuxe is stirred at 25C for 2 hours. Thereafter, 300 ml
of methylene chloride and 2 1 of n-butanol are added. The
pH is re-adjusted to 2 with 3N hydrochloric acid while
stirring ~iyorously. The organic phase is separated,
washed three times with 1 1 of water each time, stirred for
15 minutes with 20 g of decolourising charcoal and filtered.
. ~ . .
liS4009
- 33 -
The light yellow filtrate is concentrated strongly at 60C
in vacuo, the reaction product precipitating. The pre-
cipitated product is filtered off under suction and washed
successively with n-butanol, ether and low-boiling petroleum
ether and dried at 40C overnight in vacuo. There is
obtained crude, beige ~6R,7R)-7-/2-[2-(2-chloroacetamido)-4-
-thiazolyl]-2-[(Z)-methoxyimino]acetamido/-3-/[(2,5-dihydro-
-6-methoxy-2-methyl-5-oxo-as-triazin-3-yl)thio]methyl/-8-
-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid.
For conversion into the sodium salt and purification, the
acid is dissolved in a mixture of 300 ml of acetone and
50 ml of methanol and treated with 35 ml of a 2N solution of
sodium 2-ethylcaproate in ethyl acetate. The precipitated
sodium salt is filtered off under suction, washed successively
with acetone and low-boiling petroleum ether and dried at ~.5C
in vacuo. There is obtained the beige coloured sodium
salt of (6R,7R)-7-/2-~2-(2 chloroacetamido)-4-thiazolyl]-2-
-[~Z)-methoxyimino]acetamido/-3-/[(2,5-dihydro-6~methoxy-2
-methyl-5-oxo-as-triazin-3-yl)thio]methyl/-8-oxo-5-thia
, 20 -1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid with
! [a]D5 = -131.5 (c = 1 in water).
Example 2
Preparation of the disodium salt of (6R,7R)-7-/2-
-(2-amino-4-thiazolyl)-2-[(Z)-methoxyimino]acetamido/-3-
25 -/[(3-carboxy-1-methyl-lH~1,2,4-triazol-5-yl)thio]methyl/-
~, -8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid.
... . .
~S~ 3~
53 g of ~6R,7R)-3-/[(3-carboxy-1-methyl-lH-1,2,4-triazol-5-yl)thio]-
methyl/-7-/2-[2-(2-chloroaoetamido)-4-thiazolyl)-2-[(Z)-methoxyimino]aoetamido/-
-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2~ene-2-carboxylic acid are suspended in
1 1 of water together with 34 g of thiourea. The pH is adjusted to 7 by the
dropwise addition of lN sodium hydroxide, a dark brcwn solution resulting. This
solution is stirred at pH 7 overnight while gassing with nitrogen, the pH being
held constant with IN sodium hydroxide with the aid of an autotitrator. m e pH
of the reaction solution is re-adjusted to 3.5 with lN hydrochloric acid while
stirring. The precipitated substanoe (fraction I) is filtered off under suction,
washed with 250 ml of water and dried at 40& in vacuo. Fraction I is brcwn in
colour and very much impure; it is discarded. The orange mother liquor is re-
-adjusted to p~I 2.65 with lN hydrochloric acid while stirring. me precipitated
material is filtered off under suction and washed with 250 ml of water. The
beige coloured filter material is azeotropically distilled with ethanol under re-
duced pressure, filtered off under suction, washed with ethanol and low-boiling
petroleum ether and dried at 40 & in vacuo. There is thus obtained a beige-
-brcwn fraction II which, for purification and conversion mto the disodium salt,
is suspended in 2 1 of methanol, treated with 50 ml of a 2N solution of sodium
2-ethylcaproate in ethyl acetate and subsequently stirred with 200 ml of water
for 30 minutes. A small amount of insoluble, brown material is filtered off and
discarded. Ca 1 1 is removed fram the orange coloured filtrate by evaporation
at 40C in vacuo, then 1 1 of ethanol is added and the mixture is concentrated
in vacuo to a volume of ca 500 ml. The solution is decanted of fram the viscous
brown resin, a heavy mixture, which thereby separates. me decanted-off yellcw-
-orange solution is treated with 1 1 of ethanol and concentrated strongly at 40C
in vacuo, substance partially precipitating. After adding 2.5 1 of methanol,
there is concentrated strongly at 40& in vacuo. The thereby precipitated
- 34 -
llS4009
disodium salt is filtered off under suction, washed with methanol and lcw-boiling
petroleum ether and dried at 35& in a high vacuum. There is obtained the pure,
beige coloured disodium salt of (6R,7R)-7-/2-(2-amino-4-thiazolyl)-2-[(Z)-
-methoxyimino]aoetamido/-3-/[(3-carboxy-l~methyl-LH-1,2,4-triazol-5-yl)thio]-
methyl/-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid with [~]25 =
-38.7 (c = 1 in water).
The compound used as the starting material in the akove pro oe ss can be
prepared as follcws:
(a) Preparation of 4,5-dihydro-1-methyl-5-thioxo-lH-1,2,4-triazol-3-
-carkoxylic acid methyl ester.
- 35 -
~i54009
- 36 -
12.9 g of 1-methoxyoxalyl-2-methyl-thiosemicarbazide
are added to a solution of 1.6 g o~ sodium in 200 ml of
methanol. Tl~e mixture is boiled under reflux for 4 hours.
The substance which precipitates after 1/2 hour is
separated. The mother liquor is evaporated at 40C in vacuo.
The evaporation residue is dissolved in 100 ml of water and
acidified with concentrated hydrochloric acid. The
crystallisate which thexeby separates out yields, after
recrystallisation from water, pure, colourless 4,5-dihydro-
-1-methyl-5-thioxo-lH-1,2,4-thiazole-3-carboxylic acid methyl
ester of melting point 188-190C (decomposition).
(b) Preparation of 4,5-dihydro-1-methyl-5-thioxo-lH-
-1,2,4-trlazole-3-carboxylic acidO
3.46 g of 4/5-dihydro-l-methyl-5-thioxo-lH-l~2 t 4~
-triazole-3-carboxylic acid methyl estex are dissol~ed in
50 ml of lN sodium hydroxide. After stirring for 1/2 hour
at 25C, the reaction solution is made acid with 25 ml of
2N hydrochloric acid and concentrated at 40C in ~acuo,
the desired acid crystallising. This acid is ~iltered
~o off under suction, washed with ice/water and dried at 45C
in a high vacu~m~ There is obtained colourless 4,5-dihydro-
-l~methyl-5-thioxo-lH-1,2,4-triazole-3-carboxylic acid of
melting point 177-178C (decomposition).
.
.
~546)09
(c) Preparation of (6R,7R)-7-amino-3-/[(3-carboxy-1-methyl-1,2,4-triazol-
-5-yl)thio]methyl/-8-oxo-5-thia-1-az~bicyclo[4.2.0]oct-2-ene-2-
-carboxylic acid.
34 g of 7-aminocephalosporanic acid are susFended together with 25.5 g
of 4,5-lihydro-1-methyl-5-thioxo-lH-1,2,4-triazole-3-carboxylic acid in 500 ml
of water. ~he pH is adjusted to 6.5 with 3N sodium hydroxide. The mixture is
stirred at 55 & for 4 hours while gassing with nitrogen, the p~ keing held con-
stant at 6.5 with 3N sodium hydroxide with the aid of an autotitrator. The dark
solution is ccoled to 25C and adjusted to pH 3.5 with concentrated hydrochloric
acid. After 15 minutes, the precipitated material is filtered off under suction,
suspended in 500 ml of water and brought into solution at pH 7 with 3N sodium
hydroxide. The orange-red solution is adjusted to pH 3 with conoe ntrated hydro-
chloric acid. The precipitated substance is filtered off under suction and
washed successively with 250 ml of water, 500 ml of aoe tone, 500 ml of low-
boiling petroleum ether and dried at 40-45C overnight in vacuo. There is ob-
tained (6R,7R)-7-amino-3-/[(3-carboxy-1-methyl-1,2,4-triazol-5-yl~thio]methy V~
-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid as a beige coloured
powd~r.
" " 1154009
(d) Preparation of (6R,7R)-3-/[(3-carboxy-1-methyl-LH-1,2,4-triazol-5-yl)-
thio]methyl/-7-/2-[2-(2-chloroacetamido~-4-thiazolyl]-2-[(Z)-methoxy-
imino]aoetamido/-8-ox~-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-
carboxylic acid.
m is compound is prepa~ed in accordan oe with paragraph (g) of Example
1 starting fram 100 g of 2-(2-chloroacetamido-4-thiazolyl)-2-(Z)-methoxyimino-
-aoe tic acid and 111.5 g of (6R,7R)-7-amino-3-/[(3-carboxy-1-mlthyl-1,2,4-
-triazol-5-yl)thio]methyl/-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-
carboxylic acid. There is obtained beige coloured (6R,7R)-3-/[(3-carboxy-1-
-methyl-IH-1,2,4-triazol-5-yl)thio]methyl/-7-/2-(2-chloroacetamido)-4-thiazolyl]-
-2-[(Z)-methoxyimino]aoetamido/-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-
-carboxylic acid.
Example 3
Preparation of the sodium salt of (6R,7R)-7-[2-(2-amino-4-thiazolyl)-
-2-((Z)-methoxyimino)aoe tamido]-3-[[[2,5-dihydro-2-methyl-5-oxo-6-[(pivaloyloxy)-
methoxy]-as-triazin-3-yl]thio]methyl]-5-oxo-4-thia-1-æ abicyclo[4.2.0]oct-2-ene-
-2-carboxylic acid.
13.5 g of (6R,7R)-7-[4-bromo-1-[(Z)-methoxyimino]aoe toaoetamido]-3-
-[[[2,5-dihydro-2-methyl-5-oxo-6-[(pivaloyloxy)methoxy]-as-triazin-3-yl]thio]-
- 38 -
.
--` ` 115~009
methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-c æboxylic acid and 2.96 g
of thiourea are dissolved in 80 ml of ethanol. me solution is stirred at 25C
for 2 hours and then treated with 22 ml of a 2N solution of sodium 2-ethyl-
caproate in ethyl acetate, the sodium salt precipitating immediately. me mix-
ture is now diluted to 1000 ml with ethanol. 500 ml of methanol are added, a
yellow-brcwnish solution resulting. Ca~ 600 ml of solvent are distilled off at
40& in vacuo. me material which thereby precipitates is filtered off under
suction, washed with ethanol and low-boiling petroleum ether and dried at 40&
in a high vacuum. There is obtained a brcwnish fraction I which is discarded.
A further 250 ml of solvent are distilled off from the mother liquor
at 40 & in vacuo. The material which thereby precipitates is filtered off under
suction, washed with ethanol and lcw-boiling petroleum ether and dried at 40 C
overnight in a high vacuum. A beige fraction II is obtam sd, For purification,
this substance is dissolved in 100 ml of water and treated with 2 1 of ethanol.
This solution is strongly concentrated at 40C in vacuo. me precipitated mate-
rial is filtered off under suction, washed with ethanol and low-boiiing petroleum
ether and dried at 40& in a high va~uum. There is obtained a fraction III
which is discarded. The mDther liquor is, for the remaval of water, treated
- 39 -
,, .
:, :
l~S4009
twioe with 1 1 of ethanol each time and strongly concentrated at 40 & in vacuo.
Finally, acetone is added. me substance which thereby precipitates is filtered
off under suction, washed with acetone and lcw-boiling petroleum ether and dried
at 40& overnight in a high vacuum. There is thus obtained pure, non-crystalline
title substance with [~]25 = 124.6 (c = 1 in water). me nuclear resonance
spectrum and the microanalysis correspond to the given structure.
The (6R,7R)-7-[4-bromo-2-[(Z)-methoxyimino]acetoacetamido]-3-[[[2,5-
-dihydro-2-methyl-5-oxo-6-[(pivaloyloxy)metho~y]-as-triazin-3-yl]thio]methyl]-8-
-oxo-5-thia-1-azabicylo[4.2~0]oct-2-ene-2-carboxylic acid used as the starting
material in the above process can be prepared as follows:
(a) Prep æation of tetrahydro-2-methyl-5,6-dioxo-3-thioxo-as-triazine-
-1(2H)-carboxylic acid benzyl ester.
39.79 g of 1,2,5,6-tetrahydro-5,6-dioxo-3-mercapto-2-methyl-as-
-triazine æe taken-up in 500 ml of dimethylformamide and treated with 35 ml of
triethylamine. 39 ml of benzyloxycarbonyl chloride are added dropwqse during 15
minutes, whereby the temperature of the reaction mixture rises to 45C, triethyl-
amine hydrochloride precipitates and the suspension becomes yellow in colour.
- 40 -
1154009
me mixture is stirred at 25 C for 2 1/2 hours and subsequently evaporated at
70 C in vacuo. The oily evaporation residue is stirred with 500 ml of water for
1 hour, whereby it becomes solid. me solid obtained is filtered off under suc-
tion and washed with 50 ml of water. The yellow filter material is stirred well
with 250 ml of ethanol, filtered off under suctionr w~shed with ethanol and
dried at 40& in vacuo. m ere is obtained a colourless crude crystallisate
which is recrystallised frcm methanol and yields colourless pure substance of
m.p. 150-153 &.
(b) Preparation of 3-[(diphenylmethyl)thio]-5,6-dihydro,2-methyl-5,6-dioxo-
-as-triazine-1(2H)-carboxylic acid benzyl ester.
13.2 g of tetrahydro-2-methyl-5,6-dioxo-3-thioxo-as-triazine-1(2H)-
-carboxylic acid benzyl ester are dissolved in 500 ml of ethyl acetate and
treated with a solution of diphenyldiazomethane in 90 ml of low-boiling petro-
leum ether. m e mitially violet reaction solution is left to stand at 25 & for
40 hours, the colour beooming pink. 3 ml of glacial acetic acid are added and,
after 1 hour, the mixture is evaporated at 40& in vacuo. A yellow oil is oh-
tained as the evaporation residue. This is separated by means of column
chromatography on silica gel with the elution agents benzene, benzene/ethyl
aoetate 95:5 and benzene/ethyl aoe tate 90:10. The fractions containing the de-
- 41 -
`-`" 1:15'~QO~
sired substan oe are combined and evaporated at 40 C in vacuo. After recrystal-
lisation from ethanol, there is obtained colourless pure substan oe of m.p. 90-
92C.
(c) Preparation of 3-[(diphenylmethyl)thio]-1,2-dihydro-2-methyl-as-
-triazine-5,6-dione.
6.9 g of the substanoe prepared under (b) are dissolved in 100 ml of
ethyl acetate and stirred well with 30 ml of an aqueous, 7~ ammonia solution at
25& for 15 minutes. The ethyl aoe tate phase is separated and discarded. The
aqueous phase is treated with 7 ml of con oe ntrated hydrochloric acid and stirred
in an ice-bath for 30 m mutes. m e substanoe which thereby precipitates is
filtered off under suction, washed with water and immediately recrystallised
from ethanol. m ere is obtained colourless pure substan oe of m.p. 180-182&.
(d) Preparation of 3-[(diphenylmethyl)thio]-6-pivaloyloxymethoxy-2-methyl-
-as-triazin-5(2H)-one.
3.25 g of the ccmpound prepared under (c) are dissolved in 40 ml of
dlmethylformamide and treated with 1.52 g of potassium carbonate. 2.66 g of
pivaloyloxymethyl iodide are added to this mixture in one portion, a pale yellcw
solution resulting. m e reaction mixture is stirred at 25 & for 4 hours and
- 42 -
. . .~
ki7~
1154V09
on oe again treated with 2.66 g of pivaloyloxymethyl iodide. This mixture is
stirred at 25 & for 15 hours and subsequently evaporated at 35C in a high
vacuum. m e resulting evaporation residue is partitioned between 100 ml of
water and 100 ml of ethyl acetate. The aqueous phase is extracted twioe with 50
ml of ethyl acetate each time. me oombined ethyl acetate phases are dried over
sodium sulphate and evaporated at 35& in vacuo. me residual yellcw oil is
chromatographed on a silica gel column with ethyl aoetate as the elution agent.
m e fractions containing the desired product are oombined and evaporated at 35 &
in vacuo. me residual pale blue oil is dried at 25 & for 1 hour in a high
vacuum, the pure substance being obtained as a pale blue resin.
(e) Preparation of [(2,3,4,5-tetrahydro-2-methyl-5-oxo-3-thio~o-as-triazin-
-6-yl)oxy]methyl pivalate.
3.6 g of the compound prepared under (d) are stirred at 25C for 2 1/2
hours in 18 ml of anisole and 18 ml of trifluoroacetic acid. The solution is
then evaporated at 50C in vacuo. m e oily residue is stirred with 50 ml of lcw-
boiling petroleum ether, crystallisation occurring. m e crystallisate is
filtered off under suction and washed with low-boiling petroleum ether. mere
are obtained white crystals which, after recrystallisation from ether/low-boiling
- 43 -
. .0
`- 1154~9
petroleum ether, yield white, crystalline pure substance of m.p. 112-113&. The
nuelear resonan oe spectrum and the microanalysis correspond to the given strue-
ture.
(f) Preparation of (6R,7R)-7-amino-3-[[[2,5-dihydro-2-methyl-5-oxo-6-
-[(pivaloyloxy)methoxy]-as-triazin-3-yl]thio]methyl]-8-oxo-5-thia-1-
-azabieyclo[4.2.0]oct-2-ene-2-earboxylic acid.
3.0 g of the compound prepared under (e~ are suspended in 50 ml of
water together with 2.72 g of 7-aminocephalosporanic acid. ~hile gassing with
nitrogen the suspension is adjusted to pH 6.5 with lN sodium hydroxide and
stirred for 4 hours at pH 6.5-7 and at 55-60&. m e desired compound preeipit-
ates out and is filtered off under suction after cooling the mixture to 25 &.
After washing with water, aeetone and low-boiling petroleum ether and drying at
40 C overnight in a high vaeuum, there is obtained the pure substanee, the miero-
analysis and nuelear resonanoe spectrum of which correspond to the given struc-
ture.
(g) Silylation of (6R,7R)-7-amino-3-[[[2,5-dihydro-2-methyl-5-oxo-6-
-[(pivaloyloxy)methoxy~-as-triazin-3-yl]thio]methyl]-8-oxo-5-thia-1-
-azabicyelo[4.2.0]oet-2-ene-2-earboxylie aeid.
- 44 -
`` ~154U09
14.56 g of the compound prepared under (f) are suspended in 300 ml of
ethyl aoe tate and treated with 30 ml of N,O-bis-(trimethylsilyl~-acetamide. m e
mixture is stirred at 35-40C Eor 30 minutes, a dark solution resulting. m is
solution is oooled to -5& to -lo& and stirred with the exclusion of m~isture
until the acylation has taken place.
(h) Preparation of (Z)-2-hydroxyimino-3-oxo-butyric acid tert.-butyl ester.
592.1 g of acetoacetic acid tert.-butyl ester are dissolved in 560 ml
of glacial acetic acid. A solution of 290.6 g of sodium nitrite in 655 ml of
water is added dropwise to this solution at 5-10 & during 2 1/2 hours. me re-
sulting yellcw suspension is stirred at 20& for 30 minutes, treated with 940 ml
of water and stirred for a further 2 hours. The mixture is treated with 900 ml
of water and 900 g of ice and extracted in a stirrer vessel three times with 1 1
of ethyl a oe tate each time. m e combined ethyl acetate extracts are washed
three times with 1 1 of water each time, then ~reated with 5 1 of water and the
pH-value is adjusted to 6.8 with sodium hydrcgen carbonate. After separating
the aqueous phase, the organic phase is washed once with water. Thereafter, the
ethyl aoe tate solution is dried over sodium sulphate and evaporated at 40C in
vacuo. There is obtained (Z)-2-hydroxyimino-3-oxo-butyric acid tert.-butyl
ester as a yellow oil which is
- 45 -
11~4UV9
- 46 -
dried at 40C for a further 9 hours in a high vacuum.
(i) Preparation of (Z)-2-methoxyimino-3-oxo-butyric acid
tert.-butyl ester.
626.65 g of (Z)-2-hydroxyimino-3-oxo-butyric acid
tert.-butyl ester are dissolved in 2.86 1 of acetone. The
solution is cooled to 5C and treated portionwise with 703.5 g
of potassium carbonate. 322 ml of dimethyl sulphate are then
added dropwise to the yellow suspension without cooling during
1 hour, whereby the temperature of the mixture should not rise
above 25C. The light beige suspension is stirred at 20-25C
for ca. 4 hours until starting material is no longer detected
by thin-layer chromatographyO Thereafter, the mixture is
poured into 7 1 of water and extracted three times with 1 1
of ethyl acetate each time. l'he. combined ethyl acetate
extracts are washed three times with 1 1 of water each time,
dried over sodium sulphate and evaporated at 40C in vacuo.
The residual, yellow oil is dried at ~0~ for a further 6
hours in a high vacuum. There is obtalned (Z)-2-methoxy-
imino-3-oxo-butyric acid tert.-butyl ester
as a yellow oil with a boiling point o~ 57C at 0.02 mm Hg.
.: :
1~5~
(j) Preparation of (Z)-2-methoxyimino-3-oxo-butyric acid.
86 g of (Z) 2-methoxyimino-3-oxo-butyric acid tert.-butyl ester are
dissolved in 400 ml of trifluoroacetic acid. The solution is left to stand at
25& for 1 hour and thereafter evaporated at 35C in vacuo. The oily residu is
crystallised from ether/petroleum ether. There is obtained yellcwish, water-
soluble (Z)-2-methoxyimino-3-oxo-butyric acid of m.p. 80-85&.
(k) Preparation of (Z)-4~brcmo-2-methoxyimino-3-oxo-butyric acid.
145 g of (Z)-2-methoxyimino-3-oxo-butyric acid are dissolved in 1000 ml
of alcohol-free anhydrous dichlorcmethane. 10 ml of 30~ hydrobromic acid in
glacial acetic acid are added to this solution. Then, a solution of 37.5 ml of
bromine in 112.5 ml of dichloromethane is added dropwise during ca. 2 hours, the
temperature of the reaction mixture being held at 20-25 C by means of slight
cooling. Now, in order to remove hydrogen bromide frcm the reaction mixture,
nitrogen is vigoL~usly blown through. Subsequently, 250 g of ice, 250 ml of
water and 2 1 of ether are successively
- 47 -
:1~54(~9
- 48 -
added. The aqueous phase is separated and discarded. The
organic phase is washed with 250 ml of water and 250 ml
of saturated sodium chloride solution, dried with sodium
sulphate and evaporated in vacuo. There remain behind 170 g
of a brown oil which is recrystallised from carbon tetra-
chloride. There is o~tained almost colourless (Z)-4-bromo-
-2-methoxyimino-3-oxo-butyric acid. ~
(1) Preparation of the acid chloride of (Z)-4-bromo-
-2-methoxyimino-3-oxo-butyric acid.
6.72 g of (Z)-4-bromo-2-methoxyimino-3-oxo-butyric
acid are dissolved in 70 ml of anhydrous, alcohol-free
methylene chloride and treated at 0-5C with 6.24 g of
phosphorus pentachloride. The reaction solution is stirred
at 0-5C for 15 minutes and without cooling for 45 minutes.
,
(m) Preparation of (6R,7R)-7-[4-bromo-2-[(Z)-methoxyiminoJ-
acetoacetamido]-3-[[[2,5-dihydro-2-methyl-5-oxo-6-
-[(plvaloyloxy)methoxy]-as-triazin-3-yl]thio]methyl]-
-8~oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-caxboxylic
acid.
.~
' ' :
:
-
i llS4~09
- 49 -
The solution prepared according to (1) is added
dropwise at -10C to oC during ca. 30 minutes to the
solution prepared according to (g). The mixture is stirred
at -10C to 0C for 30 minutes and without cooling for 1
hour. 300 ml of ethyl acetate and 200 ml of water are then
added. The precipitated material is filtered off under
suction and discarded. The aqueous phase of the filtrate
is separated and discarded. The organic phase is washed
6 times with 250 ml of water each time, dried with sodium
sulphate, concentrated strongly at 40C in vacuo and finally
poured into 500 ml of ether. The precipitated substance is
filtered off under suction, washed with ether and low-
-boiling petroleum ether and dried at 40C overnight in a
high vacuum. There is obtained beige, amorphous pure substance.
The nuclear resonance spectrum and the infrared spectrum
correspond to the given structure.
Example 4
Production of dry ampoules for intramuscular
administration:
A lyophilisate of 1 g of the disodium salt of (6R,7R)-
7-j2-(2-amino-4-thiazolyl)-2-[(Z)-methoxyimino)acetamido/-3-
-/[(3-carboxy-1-methyl-lH-1,2,4-triazol-5-yl)thio]methyl/~8-
., : :'
' ' ' '
- I154~09
- 50 -
-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid
is prepared in the customary manner and filled into an
ampoule. Prior to the administration, the lyophilisate is
treated with 2.5 ml of a 2~ aqueous lidocaine hydrochloride
solution.
-
'' ' ''-
',
,
