Note: Descriptions are shown in the official language in which they were submitted.
~ RAN 4410/118
The present invention relates to acyl derivati~es. More
particularly, the invention is concerned with acyl derivatives,
a process for the manufacture thereof and pharmaceutical
preparations containing same.
The acyl derivatives provided by the present invention
are compounds of the general for.mula
R H H
R lON=C--CONH~
o~N~CH2~S--X ( I )
COOH
, wherein R represents furyl, thienyl or
phenyl optionally substituted by halogen,
hydroxy, lower alkoxy or lower alkyl, Rl
represents ~lkyl or aminocarbonylmethyl and
X represents a group of the formula
R2 R4 o
N~ ~ ~S~N~I~O ~NH2
R3
(a) (b~ (c)
in which one of the two s~mbols R2 and R3
or ~ and R5 represents hydrogen and the
Mn/20.4.1978
. . .
'
other represents lower aLkyl, carboxymethyl
or sulphomethyl,
r~ac~c~ C~ ?Cc~
~JI as well as~salts of said compounds and hydrates of said salts.
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 metal salts such as the calcium salt, salts
with organic bases such as salts with amines (e.g. salts with
N-ethyl-piperidine, procaine, dibenzylamine, N,N'-dibenzylethyl-
ethylenediamine, alkylamines or diaLkylamines) as well as salts
with amino acids (e.g~ salts with arginine or lysine). The
salts can be mono-salts or di-salts. The second salt formation
occurs at the tautomeric enol form of the triazine group (b),
said form being acidic.
..
The compounds of formula I also form acid addition salts
with organic or inorganic acids. Examples of such salts are
hydrohalides (e.g. hydrochlorides, hydrobromides and hydro--
iodides) as well as other mineral acid salts (e.g. sulphates,
nitrates, phosphates and the like), alkylsulphonates and mono-
arylsulphonates (e.g. ethanesulphonates, toluenesulphonates,
benzenesulphonates and the like) and other organic acid salts
(e.g. acetates, tartrates, maleates, citrates, benzoates,
salicylates, ascorbates and the like).
The salts of the compounds of formula I can be hydrated.
The hydration can be effected in the course of the manufacturing
process or can occur gradually as a consequence of the hygro-
scopic properties of an initially anhydrous salt of a compound
of formula I.
-- 4
4~n~
The aforementioned lower alkyl groups are either straight-
-chain or branched-chain and can contain up to 7 carbon atoms
(e.g. methyl, ethyl, n-propyl, isopropyl, n-pentyl and n-heptyl).
The lower alkoxy groups have an anaLogous significance. The
halogen atom is rluorine, chlorine, bromine or iodine with
chlorine and bromine being preferred.
Preferred gxoups denoted by R are furyl, thienyl and
phenyl, especially furyl. Rl preferably represents methyl.
X preferably represents the group of ~ormula (c) or a group of
formula (a) or (b) in which one of the two symbols R2 and R3 or
R4 and R5 represents hydrogen and the other represents methyl.
Especially preferred groups denoted by X are the 1,2,5,6-
-tetrahydro-2-methyl-5,6-dioxo-as-triazin-3-yl group and the
1,4,5,6-tetrahydro-4-methyl-S,6-dioxo-as-triazin-3 yl group.
Preferred acyl derivatives provided by the present
invention are the compound (R)-7-~2-(2-furyl)~2-~methoxyimino)-
acetamido]-3- r [(1,2,5,6-tetrahydro-2-methyl-5,6-dioxo-as-
-tria~in-3-yl)thio]methyl_/-3-cephem-4-carboxylic acid and
salts thereof as well as the hydrates of said salts.
The compounds of formula I as well as their salts and
hydrates of said salts can exist in the syn-isomeric form
R C--C O N H--s
N
OR1
or in the anti-isomeric form
. ~ ' .
R--C~ C O N H - -;
N
R10
or as mixtures of these two forms. The syn-isomeric form is
preferred as are mixtures in which the syn-isomeric form
predominates.
According to the process provided by the present invention,
the acyl derivatives aforesaid (i.e. the compounds of formula I
~ I ,oh~r~ c ~ ~ * c ~ c c ~f~ ~/G
~31 as well as their~salts and hydrates of said salts) are
manufactured by
(a) reacting a compound of the general formula
H H
H2N~/~
10~ N ~ CH2 - S ~ X (II)
COOH
, wherein X has the significance given
earlier and the carboxy group can be
present in protected form,
with an acid of the general formula
15RlON = C - COOH (III)
, wherein R and Rl have the significance
given earlier,
or with a reactive functional deriv?tLve of this acid and, where
required, cleaving off the protecting group,
or
(b) reacting a compound of the general formula
R ON C CONH H H S ( IV)
~CH2 Y
COOH
, wherQin R and Rl have the significance given earlier and Y re-
presents a leaving group, with a thiol of the general formula
HS - X (V)
, wherein X has the significance given earlier, in the presence of
water and
(c) if desired, converting the reaction product into a salt or a
hydrate of such a salt.
The carboxy groups present in the starting material
of formula II can be protected if desired; for example, by ester-
ification to ~orm a readily cleavable ester such as a silyl
ester (e.g. the trimethylsilyl ester). The carboxy group can
also be protected by salt formation with an inorganic base or a
tertiary organic base such as triethylamine.
~ ~ -6-
~ 7 ~ '~
Examples of reactive functional derivatives of acids of
formula III include halides (i.e. chlorides, bromides and
fluorides), azides, anhydrides, especially mixed anhydrides
with strong acids, reactive esters, (e.g. N-hydroxysuccinimide
esters) and amides (e.g. imidazolides).
Examples of leaving groups denoted by Y in compounds of
formula IV include halogen atoms (e.g. chlorine, bromine or
iodine)~ acyloxy sroups (e.g. lower alkanoyl groups such as
acetoxy), lower alkylsulphonyloxy or arylsulphonyloxy groups
(e.g. mesyloxy or tosyloxy) and the azido group.
The reaction of a compound of formula II with an acid of
formula III or a reactive derivative thereof can be caxried out
in a manner known per se. Thus, for example, a free acid of
formula III can be condensed with one of the arorementioned
esters of a compound of formula II in the presence of a carbodi-
imide (e.g. dicyclohexylcarbodiimide) in an inert solvent (e.g.
ethyl acetate, acetonitrile, dioxan, chloroform, methylene
chloride, benzene or dimethylformamide~ and the ester group can
subsequently be cleaved off. Oxazolium salts (e.g. N-ethyl-S-
-phenyl-isoxazolium-3l-sulphonate) can be used in the place of
carbodiimidesO
According to another embodiment, a salt of an acid of
formula II (e.g. a trialkylammonium salt) is reacted with a
reactive functional derivative of an acid of formula III as
mentioned earlier in an inert solvent (e.gO one of the solvents
specified earlier).
According to a further embodiment, an acid halide,
preferably the acid chloride, of an acid of formula III is reacted
with an amine of formula II. The reaction is preferably carried
out in the presence of an acid-binding agent; for example, in the
presence of aqueous alkali, preferably sodium hydroxide, or in
the presence of an alkali metal carbonate such as potassium car-
bonate, or in the presence of a lower-alkylated amine such as
trimethylamine. Water is preferably used as the solvent, although
the reaction can also be carried out in an aprotic organic solvent
such as, for example, dimethylformamide, dimethyl sulphoxide or
hexamethylphosphoric acid triamide.
The reaction of a compound of formula II with a compound
of formula III or a reactive functional derivative thereof can
be carried out conveniently at a temperature between about -40C
and room temperature, for example at about 0-10C.
The reaction of a compound of formula IV with a thiol
of formula V can be carried out in a mannerknown per se; for
example, at a temperature between about 40C and 80C. conveniently
at about 60C, in water or in a buffer solution having a pH of about
6 to 7, preferably 6.5.
After completion of the reaction of a compound of formula
II with an acid of formula III or a reactive derivative thereof
: ~ -8-
any protecting group present is eleaved off. Where the protecting
group is a silyl group (silyl ester), this group ean be eleaved
ofE espeeially readily by treating the reaction produet with water.
Where the earboxyl group is protected by salt formation (e.g. with
triethylamine), then the cleavage of this salt-forming protecting
group can be earried out by treatment with acid. In this case
there can be used as the aeid, for example, hydroehloric acid,
sulphuric acid, phosphoric acid or citric aeid.
The starting materials of formula II hereinbefore can be
prepared by reaeting a eompound of the general Eormula
~H H
H2N ~ - ~ ~ (VI)
/~ N CH2 Y
O
COOH
,wherein Y has the signifieanee given earlier, with a thiol of
formula V hereinbefore. The reaetion ean be earried out under the
same eonditions as deseribed earlier in connection with the reaetion
of a eompound of formula IV with a thiol of formula V.
-- 10 --
A syn/anti mixture of a compound of formula I which may be
obtained can be separated into the corresponding syn- and anti-
-forms in the usual manner; for example, by recrystallisation
or by chromatographic methods using a suitable solvent or
5 solvent mixture.
The compounds of formula I, their salts and the h~ydrates
of said salts have antlbiotic, especially bactericidal, activlty.
They have a wide spectrum of activity against gram-positive and
gram-negative microorganisms, including ~lactamase forming
Staphylococci and various ~-lactamase forming gram-negatlve
bacteria such as, for example, Haemophilus influenzae,
Escherichia coli, Proteus species and Klebsiella species.
The compounds of formula I as well as their pharmaceutically
acceptable salts and hydrates of said salts can be used for the
treatment and prophylaxis of infectious diseases. In the case
of adults a daily dosage of about l g to about 4 g may be
administered. Administration by the paxenteral route is
especially preferred.
In order to demonstrate the antimicrobial activity of the
compounds provided by the present invention, the following
representative compounds were tested:
Compound A: (7R)-7-[2~(2-furyl)-2-(methoxyimino)acetamido]-3-
-~ [(l,2,5,6-tetrahydro-2-methyl-5,6-dioxo-as-
-1:riazin-3~yl)thio]methyl /-3-cephem-4-carboxylic
acid.
,:
Compound B: (7R)-7-[2-(2-~uryl)-2-(methoxyimino)acetamidol-3-
-/ [(l-amino-1,2 dihydro-2-oxo~4-pyr~midinyl)thio]-
methyl /-3-cephem-4-carboxylic acid.
Compound C: (7R)-7-[2-(phenyl)--2 (methoxyimino)acetamido]-3-
-/ [(1,2,5,6-tetrahydro-2-methyl-5,6-dioxo~as-
-triazin-3-yl)thio:lmethyl 7-3-cephem-4-carboxylic
acid.
Compound D: (7R)-7-[2-(2-furyl)-2-(methoxyimino)acetamido]-3-
-
-/ [(1,4,5,6-tetrahydro-4-methyl-5,6-dioxo-as-
-triazin-3-yl)thio]methyl 7~3-cephem-4-carboxylic
acid.
Compound E: (7R)-7-[2-(methoxyimino)~2-(2-thienyl)acetamido]-
-3-/ [(1,2,5,6-tetrahydro-2-methyl-5,6-dioxo-as-
-triazin-3-yl)thio]methyl /-3-cephem-4-carboxyllc
acid.
Compound F: (7R)-3-/ [(1-ethyl 1,4,5,6-tetrahydro-5,6-dioxo-
-as-triazin-3-yl)thio]methyl_7-7-~2-(2-furyl)-2-
-(methoxyimino)acetamido]-3-cephem-4-carboxylic
acid.
Compound G: (7R)-7-/ 2-[(carbamoylmethoxy)imino]-2-(2-furyl)-
acetamido_/-3- r [(1,4,5,6-tetrahydro-4-methyl-5,6-
-dioxo as triazin-3-yl)thio]methyl /-3-cephem~-4-
-carboxylic acid.
..
: ' , ' ~ '~: .
. ~ , .
: ~ '
. ~ "
~ 12 ~
Activity in vitro: Minimum Inhlbitory Concentration (~g/ml)
:
- E C D E F G
Haemophilus
influenzae strain 1 1.2 2.5 2.5 2.5 2.5 ~.5 5.0
strain 2 0.16 0.63 0.16 0.31 0.16 0.31 0.63
strain 3 O.L6 0.63 0.16 0.31 0~16 1.2 0.31
strain 4 0.16 0.:31 0.16 0.31 0.16 0.16 0.63
strain 5 0.08 0.31 0.08 0.16 0.08 0.08 0.31
strain 6 0.16 0.31 0.08 0.16 0.08 0.08 0.63
strain 7 0.08 0.31 0~08 0.16 0~08 0.16 0.63
. , _ _ _ _ __
Klebsiella pneumoniae 10 20 20 5 40 40 2.5
Escherichia coli
strain 1 0.63 2.5 1.2 0.16 2.5 1.2 0.31
~ strain 2 40 20 10 5 20 80 _
15 ~ Proteus mirabilis
strain 1 2.5 10 10 10 10 20 5
strain 2 5 40 20 20 20 40 10
Proteus vulgaris 5 40 10 1.2 10 20 1.2
._ _ , ,_ . . ~
Proteus rettgeri 0.63 2.5 0.63 0.63 2.5 10 0.63
Staphylococcus aureus
ATCC 6538 2.5 0.16 2.5 0.63 2.5 5 1.2
Penicillin resistant
strain ¦ 5 1.2 2.5 0.63 2.5 5 1.2
Activity _n vivo
Groups of 10 mice are infected intraperitoneally with an
aqueous suspension of Proteus mirabilis. One hour after the
infection the test compound is administered subcutaneously.
The number o~ surviving mice is determined on the 4th day.
Various dosages of test compound are administered and the dosage
~ 13 ~ 3~r~
at which 50% of the mice survive (CD50, mg/kg) is determined by
interpolation.
. _ . .,
Test compound A B C D E F G
CD50, mg/kg 0.09 3.0 0 60 0~70 1.15 0.850.80
Toxicity (micP, 24 hour values)
Test compound A B
.. .._ .
LD50, mg/kg
i.v. 500- 1000- 1000-500- 2000- 1000-~4000
1000 2000 20001000 40~0 2000
s.c. ~4000 ~4000
p40~ . ~=
Pharmaceutical preparations, preferably dry ampoules, can
contain the compounds of formula I, their pharmaceutically
acceptable salts ox hydrates of said salts, optionally in
admixture with another therapeutically valuable substance.
Such compounds, salts or hydrates are conveniently mixed with
pharmaceutical inorganic or organic inert carrier material,
especially one which is suitable for parenteral administration,
such as, fox example, water or gum arabic. The pharmaceutical
preparations are preferably made up in liqu~d form (e.g. as
solutions, suspensions or emulsions). ~he pharmaceutical
preparations may be sterilised and/or may contain adjuvants
such as preserving agents, stabilising agents, wetting agents,
emulsifying agents, salts for varying the osmotic pressure or
buffers.
- 14 ~
The foLlowing Exarnples illustrate the process provided by
the present invention:
.
Example 1
Pre~aration of the sodium salt o ~
S _ ~ tet=~bydro-2-methyl-5,6-
-dioxo-as-triazin-3-yl)thio]methyl 7-3-cephem-4-carboxylic acid
5.06 g of 2-meth~xyimino-2-furyl-acetic acid (syn/anti
mixture 80:20) are dissolved in 150 mL of benzene and treated
at 5-10C while gassing with nitrogen with 4.2 ml of triethyl-
amine, 2.6 ml of oxalyl chloride and 6 drops o~ dlmethylformamide.The mixture is stirred while gassing with nitrogen at 5-10C
for 1 hour and at 25C for 0.5 hour and then evaporated at 40C
in vacuo. The residue is suspended in 150 ml of acetone and
treated at 0C with a solution of 11.2 g of (7R)-7-amino-3-
-desacetoxy-3 ~ ,5,6-tetrahydro-2-methyl-5,6-dioxo-as-
-triazin-3-yl)thio]cephalosporanic acid in 15~ ml of water,
which has previously been adjusted to pH 7.5 using 2-N aqueous
sodium hydroxide. The mixture is stirred at 0-10C for 2.5
hours under nitrogen, the pH being held between 7.5 and 8.0 by
the addition of 2-N aqueous sodium hydroxide. 500 ml of ethyl
acetate are then added and the pH is adjusted to 1.5 with 2-N
aqueous hydrochloric acid. After separating the organic phase,
the aqueous ph~se is extracted once with ethyl acetate. The
combined organic phases are washed twice wlth saturated aqueous
sodium chloride solution, dried over sodi~m sulphate and
concentrated t:o a volume of ca lO0 ml. Insolubles are filtered
off and the orange coloured filtrate obtained is diluted with
1000 ml of e-ther. 'rhe amorphous (7R)-7-[2-(2-furyl)-2-
-(methoxyimino)acetamido]-3- ~[(1,2,5,6-tetrahydro-2-methyl-
5,6-dioxo-as-triazin-3-yl)thio]methyl7 -3-cephem-4-carboxylic
acid which thereby precipitates is filtered off under suction
and washed with ether and with low-boiling petroleum ether.
The beige coloured product obtained is dissolved in 250 ml of
ethyl acetate and insolubles are f'iltered off. The orange
coloured filtrate is treated with 10 ml of a 2-N solution of
sodium 2-ethylcaproate in ethyl acetate, whereby the sodium salt
of (7R)-7-[2-(2-furyl)-2-(methoxyimino)acetamido]-3- ~[(1,2,5,6-
tetrahydro-2-methyl-5,6-dioxo-as-triazin-3-yl)thiolmethy,17 -3-
cephem-4-carboxylic acid precipitates. This is filtered off
under suction, washed with ethyl acetate and low-boiling
petroleum ether and dried at 25C in vacuo for 2 days. The
product obtained is a beige coloured powder (syn/anti mixture
80:20); [~]D = -108.6 (c = 0.5 in water); Rf value = 0.10
[thin-layer chromatography on Kieselgel*-F254-finished plates in
butanol/glacial acetic acid/water (4:1:1), visualisation with
ultraviolet light].
Example 2
Preparation of the sodium salt of (7R)-7-[2-(2-Euryl)-2-
-(meth_xyimino?acetamido]-3- ~ (1-amino-1,2-dihydro-2-oxo-4-
-pyrimidinyl)thio]metllyl ~-3-cephem-4-carboxylic acid
This salt is prepared in a manner analogous to that
described in Example 1 from 3.4 g of 2-methoxyimino-2-furyl-
acetic acid and 7.11 g of (7R)-7-amino-3-desacetoxy-3-[(1-
amino-1,2-dihydro-2-oxo-4-pyrimidinyl)thio]cephalosporanic
*Kieselgel is a generic
term meaning silicagel. - l-S~ -
.
.:
.
- 16 ~ ~ 'rv"3
acid. The product is a beige powder (syn/anti mixture 70:30);
Rf value = 0.40 [thin-layer chromatography on Kieselgel F254-
-finished plates in butanol/glacial acetic acid/water (4:1:1),
visualisation with ultraviolet light].
Example 3
~ of ~7R?-7-~2-(phenyl)-2-(methoxy-
imino)acetamido~-3-/ [(1,2,5,6-tetrahydro-2-methyl-5,6-dioxo-
-as-triazin-3-yl)thio]methyl 7-3-cephem-4-carbo~lic acid
This salt is prepared in a manner analogous to that
described in Example 1 from 1.8 g of 2-methoxyimino-phenyl-
- --acetic acid (syn/anti mixture 90:10) and 3.75 g of (7R)-7-
-amino~3-desacetoxy-3-[(1,2,5,6-tetrahydro-2~methyl-5,6-dioxo-
-as-triazin-3-yl)thio]cephalosporanic acid. The product is a
beige powder (syn/anti mixture 90:10); [a]20 = -135 (c = 0.5
in water); Rf value - 0.17 [thin-layer chromatography on
Kieselgel-F254-finished plates in butanol/glacial acetic acid~
water (4:1:1), visualisation with ultraviolet light].
Example 4
Pre aration of the sodium salt of (7R)-7-[2-(2-furyl)-2-
P _ , ,
~
-5,6.-dioxo-as-triazin-3-yl)thio]methyl 7-3-cephem 4-carboxylic
acid
This salt is prepared in a manner analogous to that
described in Example 1 from 3.4 g of 2-methoxyimino-2-furyl-
-acetic acid (syn/an=i mlxture 80:20) and 7.46 g oi ~7R~-7-
,
-amino-3-desacetoxy-3-~(1,4,5,6-tetrahydro-4-methyl-5,6-dioxo-
as triazin-3-yl)thio]cephalosporanic acid. The product is a
beige powder (syn/anti mixture B0:20); Ca]20 3 -44 (c = 0.5 in
water); Rf value = 0.34 ~thin-layer chromato~raphy on Kieselgel-
-F254-finished plates in butanol/glacial acetic acid/water
(4:1:1), visualisation with ultraviolet light].
Example 5
Preparation of the _ odium salt of (7R) -?- ~2-(methoxyimino)-2-
-(2-thienyl)acetamido]-3-/ ~(1,2,5,6-tetrah~dro-2-met~1-5,6-
-dioso-as-triazin-3-yl)thio]methyl 7-3-cephem-4-carboxylic ac d
This salt is prepared in a manner analogous to that
described in Example 1 from 1.85 g of 2-methoxyimino-2-thienyl-
-acetic acid (syn/anti mixture ca 70:30) and 3.71 g of (7R)-7-
-amino 3-desacetoxy-3-~(1,2,5,6-tetrahydro-2-methyl-5,6-dioxo-
-as-triazin-3-yl)thio]-cephalosporanic acid. The product is
a beige powder (syn/anti mixture ca 70 30); [~20 = -128.2
(c = 0.5 in water); R~ value = 0.21 ~thin-layer chromatography
on Kieselgel-F254-finished plates in butanol/glacial acetic acid/
water (4:1:1), visualisation with ultraviolet light].
Exam~le 6
Preparation of the disodium salt of (7R)-7-[2-(2-furyl)-2-
.-(methoxyimino)acetamido]-3-/ ~(2~5-dihydro-2-methyl-5-oxo-6-
-hydroxy-as-triazin-3-yl)thio]methyl 7-3-cephem-~-carboxylic
acid
This salt is prepared in a manner analogous to that
described in Example 1 from 10.12 g of 2-methoxyimino-2-furyl-
- 18 ~
-acetic acid (syn isomer) and 18.7 g of (7R)-7~amino-3-
-desacetoxy-3-[(1,2,5,6 tetrahydro-2-methyl-5,6-dioxo-as-
-triazin-3-yl)thio]-cephalosporanic acid. ~or the salt-
-formation there are used 20 ml (2 equivalents) of a 2-N
S solution of sodium 2-ethylcaproate in ethyl acetate. The
product is an almost colourless powder (sy~ isomer); [a]20 =
-141.6 (c ~ 0.5 in water); Rf value = 0.14 [thin~layer
chromatography on Kieselgel-F254-finished plates in butanol/
glacial acetic acid/wa~er (4:~:1), visualisa~ion with ultraviolet
light].
Example 7
Preparation of the sodium salt of (7R)-3~ ethyl-1,4,5,6-
tetrahydro-5,6-dioxo-as-triazin-3-yl)thio]methyl 7-7-~2-(2-
-furyl)-2-(methoxyimino)acetamido]-3-cephem-4-carboxylic acid
... . __ . ~ _ . . . . _. ... . . _ . . .
This salt is prepared in a manner analogous to that
described in Example 1 from 1.69 g of 2-methoxyimino-2-furyl-
-acetic acid (syn isomer) and 3.85 5 o (7R)-7-amino-3-
-desacetoxy-3-[(1-ethyl-1,4,5,6-tetrahydro-5,6 dioxo-as-
-triazin-3-yl)thio]-cephalosporanic acid. The product is a
beige powder (syn/anti mixture ca. 70:30); [a]20 = -47.2
(c = 0.5 in water); Rf value = 0.47 [thin-layer chromato-
graphy on Kieselgel-F254-finished plates in butanol/
~lacial acetic acid/water (4:1:1), visualisation with
ultraviolet light].
-- 19 ~
Example 8
Pre~aration of the sodium salt of (7R)-7-/ 2-~(carbamoylmethoxy)-
imino]-2-(2-furyl)acetamido 7-3-/ [(1,4 ! 5,6-tetrahydro-4-methyl-
.
-5,6-dioxo-as-triazin-3-yl)thio]methyl 7-3-cephem-4-carboxylic
a _
9.76 g of r a ~(carbamoylmethoxy)imino]furfuryl 7cephalo-
sporin sodium salt (syn/anti mixture ca 70:30) are suspended
together with 4.77 g of 1,4,5,6-tetrahydro-4-methyl-5,6-dioxo-
-3-mercapto-as-triazine in 200 ml of phosphate buffer having a
pH of 6.4. The pH is adjusted to 6.4 using l-N sodium hydroxide
while gassing with nitrogen, whereby a dark solution is obtained.
This solution is stirred at pH 6.4-6.5 for 6 hours at 55-60C
while gassing with nitrogen, the p~ being held constant with the
~ aid of an autotitrator with the addition of l-N sodium hydroxide.
lS The solution is cooled to 0-5C and the pH is adjusted to 2
with 2-N hydrochloric acid, whereby the product separates out
as the acid. This is filtered off under suction, washed with
ice/water and dried at 40C overnight in vacuo. The product
is obtained in the form of the crude acid. For purification,
this crude acid is dissolved in 150 ml of methanol and the
solution is boiled with active carbon for 2 minutes. The
mixture is filtered through a ~luted filter and the orange
coloured filtrate is concentrated in vacuo. The resin which
thereby precipitates is separated and rejected. The
concentrated methanolic solution is poured into ether. The
acid which thereby precipitates is filtered off under suction
and washed with ether and with low-boiling petroleum ether.
The product is obtained in the form of the pure acid which, for
- 20 -
&
conversion into the sodium salt, is dis$;olved in 100 ml of
methanol and treated with S ml o~ a 2-N solution of sodium
2-ethylcaproate in ethyl acetate. A small amount o~ insolubles
is filtered off and the orange coloured filtrate is concentrated
at 40C in vacuo. This concentrated solution is added to
ethanoL, whereby the sodium salt precipitates. This salt is
filtered off under suction, washed with ethanol and low-boiling
petroleum ~ther and dried at 40C overnight in vacuo, There i5
obtained the sodium salt of (7R)-7-/ 2-~(carbamoylmethoxy)-
imino]-2-(2-furyl)acetamido 7-3- r [(1,4,5,6-tetrahydro-4-methyl-
-5,6-dioxo-as-triazin-3--yl)thio]methyl 7-3-cephem-4-carboxylic
acid in the fo~n of a beige powder (syn/anti mixture ca 70:30);
[a]20 = -30.1 (c = 1 in water)j Rf value = 0.29 [thin-layer
chromatography on Kieselgel-F254-finished plates in butanol/
glacial acetic acid/water (4:1:1), visualisation with ultra~iolet
lig,ht].
When the starting materials used in the preceding
paragraph are replaced by equivalent amounts f r a-[(methoxy)-
imino]furfuryl /cephalosporin and 1,2,5,6-tetrahydro-2-methyl-
-5,6-dioxo-3-mercapto-as-triazine, then there is obtained under
otherwise similar conditions the sodium salt of (7R)-7-[2-(2-
-furyl)-2-(methoxyimino)acetamido]-3-/ [(1,2,5,6-tetrahydro-2-
-methyl 5,6-dLoxo-as-triazin-3-yl)thio]methyl /-3-cephem-4
-carboxylic acid. This salt is identical with the salt
obtained according to Example 1.
The folLowing Example illustrates the preparation of a
pharmaceutical preparation provided by the present invention:
- 21 -
Preparation of dry ampoules for intramuscular admin.istration
A lyophiLisate of l g of the sodium salt of (7R)-7-[2-(2-
furyl)-2-methoxyimino)acetamido]-3-/ [(1,2,5,6-tetrahydro-2-
S -methyl-5,6-dioxo-as-triazin-3-yl)thlo]methyl 7-3-cephem-
-carboxylic acid is prepared in the usual manner and filled into
an ampoule. Prior to the administration, the latter is treated
with 2.5 ml of a 2% aqueous lidocaine hydrochloride solution.
