DD143911A5 - PROCESS FOR THE PREPARATION OF NEW ACYL DERIVATIVES - Google Patents

Abstract

The invention relates to a process for the preparation of novel acyl derivatives, namely cephalosporin derivatives with antimicrobial activity of general formula I, in which X is 1,2,15,5-S, 6-tetrahydro-2-methyl-, 5,6-dioxo-as-triazine 3-yl group, their corresponding tautomeric form, the 2,5-dihydro-6-hydroxy-2-methyl-5-oxo-as-triazin-3-yl group, or the 1,4,5,6-tetrahydro-4-one methyl-5,6-dioxo-as-triazin-3-yl group, as well as slightly hydrolyzing esters, ethers and salts of these compounds and their hydrates »- formula I.

Description

Berlin, 14 ·· 9, 1979 55 ^ 87 11

Process for the preparation of new Äcylderivate

In the field of application of the invention

The present invention relates to a process for the preparation of novel acyl derivatives, namely oephalosporin derivatives having antimicrobial activity of the general formula

H H

OH ₃ OK ^ = 0-0QNH-- τ

I ₂ N

y / ' CH' S " ¹ X

COOH

in the X, the 1,2,5,6-tetrahydro-2-methyl-5,6-dioxo-astriazin-3-yl group, their corresponding tautomeric form, the 2,5-dihydro-6-hydroxy-2-methyl 5-oxo-astriazin-3-yl group, or the 1,4,5,6-tetrahydro-4-methyl-5,6-dioxo-as-triazin-3-yl group, as well as easily hydrolysable esters, easily hydrolyzable ethers and Salts of these compounds and hydrates of the compounds of the formula I or of their esters, ethers and salts.

^As readily hydrolyzable esters of the compounds of the formula I compounds of the formula I are to be understood, whose carboxy group is present in the form of a readily hydrolyzable Estergruppe. Examples of such esters which may be conventional are the lower alkanoyloxyalkyl esters, for example the acetoxymethyl, pivaloyloxymethyl, 1-acetoxyethyl and 1-pivaloyloxyethyl esters; the lower alkoxycarbonyloxyalkyl esters, for example the methoxycarbonyloxymethyl, 1-ethoxycarbonyloxyethyl and 1-isopropoxycarbonyloxyethyl esters; the lactonyl esters, eg the phthalidyl and thiophthalidyl esters; the lower alkoxymethyl esters, for example the methoxymethyl ester; and the lower alkanoylaminomethyl esters, eg the acetamidomethyl ester. Other esters, for example the benzyl and cyanomethyl esters, may be useful.

Suitable readily hydrolyzable ethers of the compounds of the formula I are compounds of the formula I in which X denotes the 2S-dihydro-e-hydroxy-methyl-S-oxo-as-triazine-S-yl group, the enolic OH of which Group is present in the form of a readily hydrolyzable ether group. Suitable ether groups are the same groups as mentioned above for the readily hydrolyzable ester groups. Representatives of such ethers are thus, for example, the lower alkanoyloxyalkyl ethers , for example the acetoxymethyl, pivaloyloxymethyl, 1-acetoxyethyl and 1-pivaloyloxyethylhydrogens.

 35 ether; the lower alkycarbonyloxyalkyl ethers, e.g. the

21 3 1

55 487 11

- 3 -

Methoxyoarbonyloxymethyl, 1-ethoxycarbonyloxyethyl and 1-isopropoxycarbonyloxyethyl ether; the lactonyl ethers, e.g. phthalidyl and thiophthalidyl ether; the lower alkoxymethyl ethers, e.g. the methoxymethyl ether; and the lower alkanoylaminomethyl ethers, e.g. the acetamidomethyl ether.

Examples of salts of the compounds of formula I are alkali metal salts, such as the sodium and potassium salts; the ammonium salt; Alkaline earth metal salt, such as the calcium salt; Salts with organic bases, such as salts with amines, ZcB. with N-ethyl-piperidine, procaine, dibenzylamine, N, iP-dibenzylethylethylenediamine, alkylamines or bialkylamines, as well as salts with amino acids, such as salts with arginine or lysine. The salts may be monosalts or else disalts. The second salt formation can occur in ^e compounds with the hydroxy group of 2,5-dihydro-6-hydrozy-2-methyl-5-oxo-as-triazin-3-YL ^ ^u PP.

The compounds of formula I also form addition salts with organic or inorganic acids. Examples of such salts are hydrohalides, for example hydrochlorides, hydrobromides, hydroiodides, and other mineral acid salts, such as sulfates, titrates, phosphates and the like, alkyl and monoarylsulfonates, such as ethanesulfonates, toluenesulfonates, benzenesulfonates and the like, and also other organic acid salts, such as acetates , Tartrates, maleates, citrates, benzoates, salicylates, ascorbates, and the like. "

55 487 11

The compounds of formula I (including their salts, readily hydrolyzable esters and ethers) may be hydrated. The hydration may take place in the course of the production process or may occur gradually as a consequence of hygroscopic properties of a product which is initially free of water.

The products according to the invention can be produced in the synisomeric form

or in the anti-isomeric form

N 0

(J)

0-CONH

Ho

¥ / e are present as mixtures of these two forms. The syn isomeric form or mixtures in which the syn-isomeric formin predominates is preferred.

Preferred products are

(6R, 7R) -7 - ^ * "2- (2-amino-4-t] iiazolyl) -2- (Z-metho-3-cyimino) -

21 3 lit

^{Ä 8} ** 55 487

t r iaz in-3-yl) thio-7-methyl / -8-oxo-5-thia-1-azabicyclo- [2.07oct-2-ene-2-carboxylic acid and its salts, and the corresponding hydrates.

Characteristic of the known technical solutions

Cephalosporin derivatives having the 3-side chain of the compounds prepared according to the invention have already been proposed. (EE-PS 830 455).

Also, cephalosporin derivatives with a 7 ^ ~ 2- (2-amino-4-thiazolyl) -2- (Z-methoxy-amino) -acetamidoZ group have already been proposed (BS-PS 853 545).

Both gephalosporin derivatives are antimicrobial varksanu aim of the invention

The aim of the invention is to produce an antimicrobially more effective derivative of cephalosporin than those already known.

Explanation of the invention of the invention

The invention has for its object to produce new antimicrobial more effective cephalosporin derivatives »

The above acyl derivatives are prepared according to the invention by reacting

a) in a compound of the general formula

- 4b -

213 1

5548? 11

-Comb-f

CH _O --S Σ II

COOH

in which X has the meaning given above, R represents a cleavable protective group and the carboxy group may be present in slotted form,

the protective group R, optionally also a possibly present carboxy protective group, splits off or that one

b) for the preparation of a readily hydrolyzable ester or ether of a compound of the formula I, a carboxylic acid or an enol of the formula I of a corresponding.

Veresterung or etherification subjects or that one

c) for the preparation of salts or hydrates of a compound of the formula I or hydrates of these salts, a compound of the formula I is converted into a salt or hydrate or a hydrate of this salt.

The carboxy group present in the starting compound of formula II may optionally be protected, e.g. by esterification to an easily cleavable ester, such as the silyl ester, e.g. the trimethylsilyl ester. It is also the above-mentioned, easily hydrolyzable esters into consideration. The carboxy group can also be protected by salt formation with an inorganic or tertiary organic base such as triethylamine. Possible R-protecting groups are, for example, acid-hydrolytically removable protecting groups, such as e.g. t-butoxycarbonyl or "trityl" or also basic hydrolytically removable protecting groups such as trifluoroacetyl Preferred R-protecting groups are chloro, bromo and iodoacetyl, especially chloroacetyl The latter protecting groups can be cleaved off by treatment with thiourea.

The starting compounds of formula II can e.g.

be prepared by N-acylation of the corresponding 7-amino compound, by adding a compound of general formula.

21 3 189

H H

Ill -CH ₂ -S-X

COOH

in which X has the meaning given above and the carboxy group and / or the amino group may be present in protected form, with an acid of the general formula

CH ₃ ON = C-COOH

N / A

IV

Λ I

in which R has the meaning given above,

or with a reactive functional derivative: reacting this acid and optionally splitting off any carboxy protecting group present.

The carboxy group present in the 7-amino compound of the formula III can optionally be protected, in the manner described above for the starting compound of the formula II to be prepared. The amino group of the compound of formula III may e.g. be protected by a silyl protecting group such as trimethylsilyl.

As reactive functional derivatives of acids of formula IV are e.g. Halides, i. Chlorides, bromides and fluorides; azides; Anhydrides, in particular mixed anhydrides with stronger acids; reactive esters, e.g. N-hydroxysuccinimide ester, and amides, e.g. Imidazolides, into consideration.

213 1

-3 -

The reaction of the 7-amino compound of the formula III with the acid of the formula IV or a reactive functional derivative thereof can be carried out in a manner known per se. So you can, for example a free acid of formula IV with one of the mentioned esters according to forms]. III by means of a carbodiimide, such as dicyclohexylcarbodiimide, in an inert solvent such as ethyl acetate, acetonitrile, dioxane, chloroform, methylene chloride, benzene or dimethylformamide and then cleave the ester group. Instead of carbodiimides, oxazolium salts, e.g. N-ethyl-5-phenylisoxazolium-3'-sulfonate.

In another embodiment, a salt of an acid of formula III, e.g. a trialkylammonium salt, such as the triethylammonium salt, with a reactive functional derivative of an acid of formula IV as mentioned above in an inert solvent, e.g. one of the above, um.

According to a further embodiment, an acid halide, preferably the chloride of an acid of the formula IV, is reacted with the amine of the formula III. The reaction is preferably carried out in the presence of an acid binding agent, e.g. in the presence of aqueous alkali, preferably sodium hydroxide, or else in the presence of an alkali metal carbonate, such as potassium carbonate, or in the presence of a lower alkylated amine, such as triethylamine. The solvent used is preferably water, if appropriate in admixture with an inert organic solvent, such as tetrahydrofuran or dioxane. It may also be in an aprotic organic solvent, e.g. Dimethylformamide, dimethyl sulfoxide or hexamethylphosphoric triamide, to be worked. When using silylated starting compounds of formula III is carried out in anhydrous medium.

21 3 1

- 40 -

The reaction of the 7-amino compound of formula III with the acid ^ ^{he Forine} l ^IV or a reactive functional derivative thereof can be advantageously carried out at temperatures between about -40 ⁰ C and room temperature, beispiels-

'o

at about 0-10 C, take place.

The starting compounds of the formula II can also be prepared by thiolation, namely a compound of the general formula

H H

CH ₃ ON = C - CONH

RHN

CH ₂ -Y

COOH

in which R has the meaning given above, y represents an leaving group and the carboxy group may be in protected form, with a thiol of the general formula

HS-X

where X has the meaning given above,

converts and optionally splits off any existing carboxy protective group.

As the leaving group Y of a compound of formula V, for example, halogens, e.g. Chlorine, bromine or iodine, acyloxy radicals, e.g. lower alkanovoloxy radicals, such as acetoxy, lower alkyl or arylsulfonyloxy radicals, such as mesyloxy or .tosyloxy, or the azido radical. The connection V

may be protected at the carboxy group in the manner explained for the starting compound of formula II.

The reaction of the compound of the formula V with the thiol of the formula VI can be described in

in a manner known per se, e.g. at a temperature between about 40 and 80 ° C, conveniently at about 60 ° C, in a polar solvent, for example in an alcohol, e.g. in a lower alkanol such as ethanol, propanol and the like, in dimethylformamide or dimethylsulfoxide, preferably in water or in a buffer solution having a pH of about 6 to 7, preferably 6.5.

In accordance with process variant a) of the process according to the invention, the amino-protecting group R is split off from a starting compound of the formula II. Protective groups cleavable by acid hydrolysis are preferably removed with the aid of a lower alkanecarboxylic acid, which may optionally be halogenated. In particular, formic acid or trifluoroacetic acid are used. The temperature is usually room temperature, although slightly elevated or slightly lower temperature can be used, for example in the range of about 0 ⁰ C to + 40 ⁰ C. Alkaline cleavable protecting groups are generally with dilute aqueous liquor at ^{0 0} C to Hydrolyzed 30 ⁰ C. The chloroacetyl, bromoacetyl and iodoacetyl protecting groups can be split off in about 0-30 ⁰ C by means of thiourea in acidic, neutral or alkaline medium, v / ground. Hydrogenolytic cleavage (eg cleavage of benzyl) is unsuitable here, since in the hydrogenolysis the oxime function is reduced to the amino group.

After carrying out process variant a), it is possible, if desired, to cleave off a carboxy protective group which may be present in the reaction product. When the protecting group is a silyl group (silyl ester), this group is particularly easy to treat

21 3 1

of the reaction product are split off with water. Lower alkanoyloxyalkyl, alkoxycarbonyloxyalkyl, lactonyl, alkoxymethyl and alkanoylaminomethyl esters are preferably cleaved enzymatically with the aid of a suitable esterase (at about 20-40 ° C). When the carboxyl group is formed by salt formation (e.g.

ethylamine), the removal of this salt-forming protective group can be effected by treatment with acid. As acid in this case, e.g. Hydrochloric acid, sulfuric acid, phosphoric acid or citric acid can be used.

The carboxy-protecting group can be split off in the same way as just described before the deprotection of R.

For the preparation of the readily hydrolyzable esters of the carboxylic acids of the formula I according to variant b), the carboxylic acid is preferably reacted with the corresponding halide containing the ester group, preferably with the iodide. The reaction can be carried out with the aid of a base, e.g. an alkali metal hydroxide or carbonate or an organic amine such as triethylamine are accelerated. In the presence of the 2,5-dihydro-6-hydroxy-2-methyl-5-oxoas-triazin-3-ylgruppe X with their enolic function, this is etherified to form a corresponding readily hydrolyzable ether. Preferably, an excess of the corresponding halide is used. The esterification / etherification reaction is preferably carried out in an inert organic solvent, such as dimethylacetamide, hexamethylphosphoric triaiuide, dimethyl sulfoxide or, preferably, dimethylformamide. The temperature is preferably in the range of about 0-40 C.

The preparation of the salts and hydrates of the compounds of the formula I or of the hydrates of these salts can be carried out in a manner known per se, e.g. by reacting the carboxylic acid of the formula I with an equivalent amount of the desired base, suitably in a solvent, such as

- 43 .

Water or in an organic solvent such as ethanol, methanol, acetone and others. When a second equivalent of base is used, salt formation also takes place on a possibly existing tautomeric enol form (2,5-dihydro-6-hydroxy-2-methyl-5-oxo-as-triazin-3-yl group X), a disalt being formed. The temperature of the salt formation is not critical, it is generally at room temperature, but may also be slightly above or below, for example in the range of O ⁰ C to +50 ⁰ C.

The hydrates are usually produced automatically in the course of the production process or as a consequence of hygroscopic properties of an initially anhydrous product. For the targeted production of a hydrate, a wholly or partially anhydrous product (carboxylic acid of the formula I or ester, ether or salt thereof) of a moist atmosphere, for example at about +10 ⁰ C to +40 ⁰ C, are exposed.

The 7-amino compounds of the formula used above

III can be prepared starting from a compound of formula

-CH ₂ -Y 20

COOH

in which Y represents an leaving group and the carboxy group may be in protected form,

be prepared with a thiol of the formula VT. The reaction can be carried out under the same conditions as those described for the reaction of the starting compounds V with VI. On the other hand, the compounds ' * of the formula ν can be prepared starting from a compound of the formula VII and an acid of the formula IV or a reactive functional derivative thereof under the same conditions as described for the reaction of the compounds of the formulas III and IV ,

A possibly obtained syn / anti mixture of a compound of the formula I can be separated into the corresponding syn and anti forms in the customary manner, for example by recrystallization or by chromatographic methods using a suitable solvent or solvent mixture.

The compounds of the formula I and II and the corresponding readily hydrolyzable esters, ethers and salts or the hydrates of these products are antibiotics, in particular

ΙΟ bactericidal effect. They have a broad spectrum of activity against Gram-positive and Gram-negative microorganisms, including 0-lactamase-producing staphylococci and various 0-lactamase-producing Gram-negative bacteria, e.g. Pseudomonas aeruginosa, Haemophilus influenzae,

] _5 Escherichia coli, Serratia marcescens, Proteus and Klebsiella species.

The compounds of the formula I and II and the corresponding readily hydrolyzable esters, ethers and salts or the hydrates of these products can be used for the treatment and prophylaxis of infectious diseases. For adults, a daily dose of about 0.1 g to about 2 g is considered. Parenteral administration of the compounds of the invention is particularly preferred.

To demonstrate the antimicrobial efficacy of the mentioned products, the following were representative

Representative tested:

Product A: (6R, 7R) -7- [2- (2-amino-4-thiazolyl) -2- (Z-methoxyimino) acetamido] -3 - / [(2,5-dihydro-6-hydroxy-2 -methyl-5-oxo-as-triazin-3-yl) thio] methyl / 8-oxo-5-thia-1-azabicyclo [4 .2.0] octane

2-ene-2-carboxylic acid

Product B: (6R, 7R) -7- / 2- [2- (2-chloroacetamido) -A-

thiazolyl] -2- (Z-methoxyimino) acetamido / -3 - / [^, S-dihydro-o-hydroxy - ^ - methyl-S-oxo-astriazin-3-yl) thio] methyl / -8-oxo- 5-thia ~ lazabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

Activity in vitro; Minimum inhibitory concentration (Mg / ml)

A B Haemophilus influenzae Starara 1 strain 2 strain 3 strain 4 strain 5 strain 6 strain 7 0.08 0.005 0.005 0.005 0.0025 0.0025 0.0025 1.2 0.3 0.16 0.16 0.08 0.16 0.16 Klebsiella pneumoniae 1.2 10 Escherichia coli strain 1 strain 2 0.02 0.6 0.16 5 Proteus mirabilis strain 1 strain 2 50.01 <0.01 0.08 0.16 Proteus vulgaris £ 0.01 0.16 Proteus rettgeri <-0,0l 0.16 Staphylococcus aureus strain ATCC 6538 Penicillin resistant strain 2.5 2.5 2.5 5 ί Pseudomonas aeruginosa strain 1 strain 2 strain 3 strain 4 strain 5 strain 6 strain 7 0.3 10 2.5 5 5 10 5 1,2> 80 40 80 80 80 80 y Serratia marcescens 0.08 2.5

- At? - activity in vivo

Groups of 5 mice are challenged with an aqueous suspension of Escherichia coli intraperitoneally. Three times, i. 1 hour, 2 1/2 hours and 4 hours after infection, the test substance is administered subcutaneously in physiological saline. The number of surviving animals is determined on the 4th day. Different dosages are applied and by interpolation the dose is determined in which 50% of the experimental animals survive mg / kg).

test substance V.C.O. A B CD ₅₀ , mg / kg toxicity ^ 0.005 0.16 test substance A B ^LD 50 ' ^m 9 / ^k 9 i · p. 250-500> 4000> 5000 250-500 2000-4000> 5000

The products according to the invention can be used as remedies, e.g. in the form of pharmaceutical preparations which use them or their salts in admixture with a pharmaceutical, organic or inorganic inert carrier suitable for enteral or parenteral administration, e.g. Water, gelatin, gum arabic, lactose, starch, magnesium stearate, talc, vegetable oils, polyalkylene glycols, vaseline, etc. The pharmaceutical preparations may be in solid form, e.g. as tablets, dragees, suppositories, capsules; or in liquid form, e.g. as solutions, suspensions or emulsions. Optionally, they are sterilized and / or contain adjuvants such as preservatives, stabilizers, wetting or emulsifying agents, salts for altering the osirtotic pressure, anesthetics or buffers. They may also contain other therapeutically valuable substances. The Ver-

21 3 ISf

- Λ t -

Compounds of the formula I and their salts or hydrates are preferably suitable for parenteral administration and are preferably prepared for this purpose as lyophilisates or dry powders for dilution with conventional agents such as water or isotonic saline. The readily hydrolyzable esters or ethers of the compounds of the formula I and their salts or hydrates are also suitable for enteral administration.

21 3 189 ~ ^AS "

example 1

Preparation of the disodium salt of (6R, 7R) -7- [2- (2-amino-4-thiazolyl) -2- (Z-methoxyimino) acetamido] -3 - / [(2,5-dihydro-6-hydroxy) 2-methyl-5-oxo-as-triazin-3-yl) thio] -methyl-S-oxo-S-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

15.3 g of (6R, 7R) -7- / 2- [2- (2-chloroacetamido) -4-thiazolyl] -2- (rZ- methoxyimino) acetamido / -3- [[2,5-dihydro-6 -hydroxy-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 (Fraction I, see below) are suspended together with 5 g of thiourea in 150 ml of water. Under good nitrogen fumigation and stirring, the pH is adjusted to 6.8-7.0 with saturated sodium bicarbonate solution to give an orange solution. By means of autotitrator with the addition of sodium bicarbonate solution, the pH of the reaction solution is kept constant at 6.8 for 6 hours. Thereafter, 2.5 g of thiourea are added and the solution is stirred for a further 3 hours, while maintaining the pH at 6.8 with the addition of saturated sodium bicarbonate solution. Thereafter, the red solution is stored in the refrigerator overnight, whereby it becomes darker. The pH of this solution is adjusted to 2.0-2.5 by addition of 100% formic acid, whereby the substance precipitates. This is filtered off with suction and washed with 100 ml of 10% formic acid. The mother liquor is discarded. The brownish filter cake is suspended in 200 ml of water and the pH is adjusted to 7 with triethylamine, whereby a brown solution is formed. This solution is stirred with 2 g of activated carbon for 30 minutes, filtered from the carbon and the still brown filtrate with 100% formic acid with good stirring to pH 3.5. The precipitated substance is filtered off with suction, washed with 50 ml of 10% formic acid and discarded. The dark yellow filtrate is adjusted to pH 2-2.5 with 100% formic acid, the. Substance fails. This is filtered off, washed with ice water and dried. The obtained

£. \ v ^ I Cl W

- 45 -

Cephalosporin acid is, for the purpose of conversion into the disodium salt, suspended in a mixture of 40 ml of acetone and 40 ml of water and treated with 20 ml of a 2-n solution of 2-Aethylcapronsäure sodium salt in ethyl acetate.

To the resulting orange solution, 50 ml of acetone are added, whereby a brown resin precipitates, which is separated by filtration. The yellow filtrate is stirred for 30 minutes to crystallize the disodium salt. The mixture is added portionwise with 50 ml of acetone and stored overnight in the refrigerator. The crystals are filtered off with suction, washed successively with an acetone-water mixture (85:15), pure acetone and low-boiling petroleum ether and dried overnight at 40 C in vacuo. The title substance is obtained as beige crystals, [cc] -, = -144 ° (c = 0.5 in water). The nuclear magnetic resonance spectrum and the microanalysis correspond to the given structure.

The starting (6R, 7R) -7- / 2- [2- (2-chloroacetamido) -4-thiazolyl] -2- (Z-methoxyimino) acetamido / 3 - / [(2,5-dihydroxy) 6-hydroxy-2-methyl-5-oxo-as-triazin-3-yl) thio] methyl / -8-oxo-5-thia-l-azabicyclo [4.2.0] oct-2-ene-2- carboxylic acid can be prepared as follows:

22.24 g of 2- (2-chloroacetamido-thiazol-4-yl) -2- (Z-methoxyimino) -acetic acid are suspended in 240 ml of methylene chloride. 13.39 ml of triethylamine are added to this suspension, resulting in a light brown solution. This solution is cooled to 0-5 C and treated with 16.72 g Phospnorpentachlorid, stirred for 5 minutes at 0-5 ⁰ C and 20 minutes without cooling. The yellow solution is evaporated in vacuo at 35.degree. The evaporation residue is shaken twice with n-heptane and the latter is decanted off. The resinous residue is treated with 240 ml of tetrahydrofuran and the undissolved triethylamine hydrochloride is filtered off. The yellow filtrate contains the acid chloride.

21 3 1

22 g of (7R) -7-amino-3-desacetoxy-3 - [(2,5-dihydro-6-hydroxy-2-methyl-5-oxo-as-triazin-3-yl) -thio] -cephalosporanic acid suspended in a mixture of 300 ml of water and 150 ml of tetrahydrofuran. 2-n sodium hydroxide solution is added dropwise to the suspension under good nitrogen fumigation with the aid of the autotitrator until a brown-red solution of pH 8 is formed. This is cooled to 0-5 ° C and treated dropwise over 15 minutes with the above prepared solution of the acid chloride in tetrahydrofuran. Thereafter, the mixture is stirred for 2 1/2 hours at 25 ° C. The pH of the acylation mixture is kept constant at 8 with addition of 2N sodium hydroxide solution. The practically black solution is freed of tetrahydrofuran at 40 ^° C. under reduced pressure. Now, 100 ml of 2N sulfuric acid are added. The precipitated substance is filtered off, washed with water and filtered with suction. The moist, brown filter cake is dissolved in 1.5 l of acetone. The dark solution is filtered off from a little dark undissolved material over Hyflo, mixed with charcoal, stirred for 30 minutes and filtered again over Hyflo. The orange-red filtrate is dried over sodium sulfate, concentrated in vacuo and evaporated with ethyl acetate. A black resin precipitates, which is filtered off and discarded. The 2-phase, still water-containing filtrate is azeotroped three times with benzene under vacuum at 40 ° C. The precipitated substance is filtered off with suction and dried in vacuo at 40 ° C. The latter is stirred twice with 1 l of acetone each, leaving a brown resin which is discarded. The combined orange-colored acetone extracts are reduced in vacuo at 40 C to approx ^. 150 ml, whereby a brown resin is filtered off and discarded. The filtrate is mixed with 1 liter of ethyl acetate and concentrated in vacuo at 40 C. The precipitated substance is filtered off with suction, washed with ethyl acetate and then with ether [(6R, 7R) -7- / 2- [2- (2-chloroaceta-

-2 -. (Z ^ ire

hydroxy-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, fraction 1: a beige, amorphous acid]. This fraction I can directly

213 1

be used for the production of the desired end product.

The ethyl acetate mother liquor is strongly concentrated in vacuo at 40 ° C, diluted with ether and the precipitated substance is filtered off with suction [(6R, 7R) -7- / 2- [2- (2-chloroacetamido) -4-tMazoiyl] -2- (Z -me thoxyimino) acetamido / -3- / [(2,5-dihydro-6-hydroxy-2-methyl-5-oxo-as-triazin-3-yl) thio] methyl / 8-oxo-5 -thia-lazabicyclo [4.2.0] oct-2-ene-2-carboxylic acid, fraction II: a pale beige amorphous acid, thin-layer chromatography slightly purer than fraction I].

To prepare the disodium salt, 3.5 g of acid (fraction II) are dissolved in a mixture of 20 ml of acetone and 11 ml of water. The solution is treated with 7 ml of a 2-N solution of 2-Aethylcapronsäure sodium salt in ethyl acetate, whereby the disodium salt crystallized. Now 25 ml acetone are added in portions and the mixture kept for 2 hours in a freezer. Thereafter, the crystals are filtered off with suction, washed successively with 25 ml of an ice-cold acetone-water mixture (80: 2O) _{1 of} pure acetone and low-boiling petroleum ether and dried overnight at 40 C in a high vacuum. The disodium salt of (6R, 7R) -7- / 2- [2- (2-chloroacetamido) -4-thiazolyl] -2- (Z-metlioxyimino) acetamido / - 3 / [(2, S-dihydro -e-hydroxy- ^ -methyl-S-oxo-astriazin-3-yl) thio] -methyl / -8-oxo-5-thia-1-azabieyclo [4. 2.0] oct-2-ene-2-carboxylic acid as pale yellow crystals. [Cx]. _n =

-142.7 ° (c = 1 in water). The nuclear magnetic resonance spectrum and the microanalysis correspond to the given structure.

Example 2

c Preparation of the sodium salt of (6R, 7R) -7- [2- (2-amino-4-thiazolyl) -2- (me thoxy imino) acetamido] -8-OXO-3- / [(1,4,5 , 6-tetrahydro-4-methyl-5,6-dioxo-as-triazin-3-yl) -thio-methyl / -5-thia-1-azabicyclo [4. 2.0] oct-2-ene-2-carboxylic acid.

19 g (6R, 7R) -7- [2- [2- (2-chloroacetamido) -4-thiazolyl] -

21 3 1

2- (me thoxyimino) acetamido] -8-0x0-3- [[(1,4,5,6-tetrahydro-4-methyl-5,6-dioxo-as-triazin-3-yl) thio] methyl ] -5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid are suspended together with 9.5 g of thiourea in 150 ml of Viasser. Under nitrogen gassing and stirring, the pH is adjusted to 6.8 with 5% sodium bicarbonate solution to give a yellow-orange solution. By means of autotitrator with addition of sodium bicarbonate solution, the pH of the reaction solution is kept constant at 6.8-7.0 for 6 hours. To the orange-colored solution, 100% formic acid is added to pH 3.5. The precipitated material is filtered off with suction and washed with 100 ml of 10% formic acid. This filter is called (l) . The filtrate is adjusted to pH 2.5 by addition of 100% formic acid, again precipitating substance. The mixture is kept for 1 hour in an ice bath, then the precipitated substance is filtered off with suction and washed with a little ice water. This is called fraction I. The orange-brown filter Qj is suspended in 250 ml of water. The suspension is adjusted to pH 7 with 2N NaOH to give an orange-brown solution. 100% formic acid to pH 3.5 is again added to this solution. The failed material is sucked off and discarded. The filtrate is adjusted to pH 2.5 with 100% formic acid, again precipitating substance. The mixture is

Held for an hour in an ice bath, the precipitated substance is filtered off with suction and washed with a little ice water. This is called fraction II. The fractions I and

II are suspended together in 500 ml of ethanol and evaporated on a rotary evaporator to remove water.

After addition of ether, it is filtered off with suction and washed successively with ether and low-boiling petroleum ether. Thus, the title substance is obtained as a yellowish solid which is designated A.

The mother liquors and washings of fraction I and II are concentrated from a volume of about 1.7 1 to 250 ml, the pH adjusted to 2.5 with 100% formic acid

213 189

and the solution stored overnight in the refrigerator, again crystallized substance. This is filtered off with suction and washed with a little water. The filter cake is azeotroped with ethanol. This gives solid, practically colorless title substance, which is designated B. By thin-layer chromatography, B is more pure than A.

To obtain pure title substance, the acid B is suspended in 150 ml of methanol and, while stirring, 10 ml of a 2N solution of 2-ethyl-caproic acid sodium salt in ethyl acetate are added. After about 10 minutes, a solution is formed, which is mixed with 100 ml of ethanol. The mixture is concentrated strongly at 40 C under vacuum. After addition of ethanol, the sodium salt precipitates amorphous. This is filtered off with suction, washed successively with ethanol and low-boiling petroleum ether and dried at 40 C under high vacuum for 24 hours. The title substance is obtained as a virtually colorless, amorphous powder. [Cc] _n = -42.9 (c = 1 in water).

According to the nuclear magnetic resonance spectrum, the title substance is present as a Z / E mixture (90:10). The microanalysis also agrees with the indicated structure.

The starting compound (6R, 7R) -7- [2- [2- (2-chloroacetamido) -4-thiazolyl] -2- (methoxyimino) acetamido] -8-0x0-3 - [[(1,4, 5,6-tetrahydro-4-methyl-5,6-dioxoas-triazin-3-yl) thio] methyl] -5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid may be as be prepared follows:

44 g of (7R) -7-amino-3-desacetoxy-3 - [(1,4,5,6-tetrahydro-4-methyl-5,6-dioxo-as-triazin-3-yl) -thio] - cephalosporanic acid are suspended in a mixture of 600 ml of water and 300 ml of tetrahydrofuran. 2-n sodium hydroxide solution is added dropwise to the suspension under good nitrogen fumigation with the aid of the autotitrator until a brown solution of pH 7.8 is formed. This is cooled to 0-5 ° C and added dropwise over 15 minutes with a solution of 2- (2-Chloracetamido-

thiazol-4-yl) -2- (Z-methoxyimino) -acetic acid chloride in tetrahydrofuran (prepared from 44.5 g of the corresponding acid according to Example 1). Thereafter, it is stirred at pH 8 and 25 ° C for 2 1/2 hours. The pH of the acylation mixture is kept constant at 7.8-8 with the addition of 2N sodium hydroxide solution using the autotitrator. The dark solution is freed of tetrahydrofuran in vacuo at 40.degree. Thereafter, the solution is diluted with water to a volume of 2 1. The pH is adjusted to 2 with 2N sulfuric acid. The precipitated substance is filtered off, washed with 1 1 of water and dried for 2 days in a vacuum at 40 ° C. For purification, this is first dissolved in a mixture of 100 ml of water and 300 ml of acetone. The dark solution is diluted with acetone to a volume of 2. The thereby precipitated dark material is filtered off and discarded. The filtrate is treated with 1 1 ethyl acetate, and in vacuo at 40 ° C, 1 1 of solvent is evaporated. Now the solution is diluted with 2 1 ethyl acetate. The resulting beige-brown substance is discarded. The filtrate is concentrated in vacuo at 40 ° C. The crystallized acid is filtered off with suction. For ümkristaliisation the acid is first dissolved in 800 ml of methanol at reflux. The solution is cooled to 25 ° C and filtered off from a little orange-colored substance. The yellow filtrate is stirred for 1 1/2 hours in an ice bath, whereby the acid crystallizes. This is filtered off with suction, washed successively with methanol and .tief boiling petroleum ether and dried in vacuo at 25 C. Thus, the starting compound is obtained as beige crystals. According to the nuclear resonance spectrum, the starting compound is present as a Z / E mixture (75:25). [a]. ^ ⁰ = -127.9 ° (c = 1 in dimethylformamide). , '

Example 3

Preparation of methylene (6R, 7R) -7- [2- (2-amino-4-thiazolyl) -2- (Z-methoxyimino) acetamido] -3- [[[2,5,5-dihydro-2-methyl] 5-OXP-6 - [(pivaloyloxy) methoxy] -as-triazin ~ 3 ~ yl] -

^{1 m}

thio] methyl] -8-0x0-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylate pivalate.

1.85 g of the cephalosporin disodium salt prepared according to Example 1 are suspended in 50 ml of dimethylformamide and admixed with nitrogen at -5 ° C. with 1.35 g of pivaloyloxymethyl iodide. The reaction mixture is stirred for 30 minutes at 0-5 C and then poured onto 500 ml of ethyl acetate. The mixture is washed 3 times with water, twice with 5% sodium bicarbonate solution and finally again with water. The solution is dried over sodium sulfate and strongly concentrated in vacuo at 35.degree. After addition of ether, the title substance precipitates amorphous. This is filtered off with suction, washed with ether and low-boiling petroleum ether and dried overnight at 25 ° C. in a high vacuum. The title blank is obtained as a beige amorphous powder. The nuclear magnetic resonance spectrum and the microanalysis are consistent with the indicated structure.

Example 4

Preparation of dry ampoules for intramuscular administration:

A lyophilizate of 1 g of the disodium salt of (6R, 7R) -7- [2- (2-amino-4-thiazolyl) -2- (Z-methoxyimino) acetamido] -2- [[( 2,5-dihydro-6-hydroxy-2-methyl-5-oxo-as-triazin-3-yl) thio] methyl / 8-oxo-5-thia-1-azabicyclo [4.2.0joct-2-ene -2-carboxylic acid prepared and filled into an ampoule. Before administration, the lyophilizate is treated with 2.5 ml of a 2% aqueous lidocaine hydrochloride solution.

213189

Example 5

A gelatin plug-in capsule of the following composition is prepared in the usual way:

Methylene (6R, 7R) -7- [2- (2-amino-4-thiazolyl) -2- (Z-methoxyimino) acetamido] -3- [[[2 _/ 2-dihydro-2-methyl-5-oxo -6-C (pivaloyloxy) methoxy] -as-triazin-3-yl] thio] methyl] -8-oxo-S-thia-1-azabicyclo [4. 2.0] oct-2-ene-2-carboxylate pivalate 500 mg

Luviskol (water-soluble 20 mg

polyvinylpyrrolidone)

Mannitol Talc 15 Magnesium Stearate

557 mg

20 mg 15 mg 2 mg

Claims (2)

2 ί 3 1 55 487 11 Claim for invention 1. A process for the preparation of acyl derivatives of the general formula H H   CONH- CH ₂ -SX I COOH in the X, the 1,2,5,6-tetrahydro-2-methyl-5,6-diis; oas-triaζin-3-yl group, the 2,5-dihydro-6-hydroxy-2-methyl-5-- o: xo-as-triazin-3-yl group or the
1 »4? 5,6-tetrahydro-4-methyl-5,6-dioxo-as-triazinbedeutet, as well as slightly hydrolysable esters, light
hydrolyzable ethers and salts of these compounds and of hydrates of the compounds of the formula I or of their esters, ethers and salts, characterized in that a) in a compound of the general formula - 26- 21 3 1 55 487 11 COMH COOH in which X has the abovementioned meaning, R represents a removable protective group and the carboxy group may be present in protected form, the protective group R, optionally also a possibly present carboxy protecting group, splits off or that one b) subjecting a carboxylic acid or an enol of the formula I of a corresponding esterification or etherification for the preparation of a readily hydrolyzable ester or * ether of a compound of formula I, or that you c) for the preparation of salts or hydrates of a compound of the formula I or hydrates of these salts, a compound of the formula I is converted into a salt or hydrate or a hydrate of this salt. Procedure according to point 1 ». characterized in that a starting compound of the formula II in which R is chloroacetyl is treated with thiourea » * - 27 21 3 55 487 11 3 · Process according to item 1 or 2, for the preparation of a compound of formula I ₁ in which X is the 1,2,5t6-tetrahydro-2'-methyl-5 »6-dioxo-as-triazin-3-yl * or 2,5-dihydro ^ 5-hydroxy-2-methyl-5-oxo-as-triazin-3-yl group, as well as salts of these compounds and hydrates of these compounds or salts, characterized in that a starting compound of formula II, wherein X has this meaning. 4 _# Process according to item 1 or 2 for the preparation of compounds of formula I, wherein X represents the 1,4,5,6-2etrahydro-4-methyl-5i6-dioxo-as-triazin-3-yl group, and salts thereof Compounds and hydrates of these compounds or salts, characterized in that one uses a starting compound of formula II, wherein X has this meaning · 5 - Process according to item 1 or 2 for the preparation of (6R, 7I) -7 - ^ "" 2- (2-amino-4-thiazolyl) -2- (Z-methoxyimino) acetamido7-3- / 2 "(2 , 5-dihydro-6-hydroxy-2-methyl-5-oxo-as-triazin-3-yl) thio7-methyl / -8-o2o-5-thia-1-aza-bicyclo ^ T-.2.o7oct -2-en-2 »-carboxylic acid and of salts of this compound and of hydrates of this compound or salts, characterized in that employing correspondingly substituted starting compounds 6 _e method according to item 1 or 2, for the preparation of readily hydrolyzable esters of the acyl derivatives of the formula I and salts of these esters and of hydrates μ 28 3 189 55 48? 11 these esters and salts, characterized in that a carboxylic acid of the formula I is subjected to a corresponding esterification and, if desired, the resulting product is converted into a salt or hydrate or a hydrate of this salt. Process according to item 1 or 2, for the preparation of readily hydrolyzable ethers of the acyl derivatives of the formula I and of salts of these ethers and hydrates of these ethers and salts, characterized in that an enol of the formula I is subjected to a corresponding etherification and, if desired, the product obtained in a salt or hydrate or, converted a hydrate of this salt 8 »Process according to item 6 for the preparation of pivalolyloxymethyl esters of a compound of the formula I and of salts of these esters and hydrates of these esters and salts, characterized in that reacting a Garbonsäure of the formula I with a Pivalolylosymethylhalogenid, preferably with the iodide, and if desired the product obtained is converted into a salt or hydrate or a hydrate of this salt. 9 · Process according to item 6 and 7 for the preparation of methyleneC 6H, 7R) -7- (2-amino-4-thiazolyl) -2- (Z-methoxy-imino) acetamido7 " ⁼ 3 ^ra / 1 ' ~ (2 "5-dihydro-2- 3-yl) thiq ^ -methyi / '- S-oxo-b-thia-1'-azabicyclo / 4.2 * O / oct - 29 - 55 4-8? 11 2-ene-2-carboxylate pivalate so? Ae of salts of this ester and hydrates of this ester bzyj. of these salts, characterized in that (6R, 7R) -7- ^ * "2- (2-amino-4-th.iazolyl) -2- (Z-methoxyimino) acetamido 7 ~ 3 ~ // ~ (2, 5-dihydro-6-hydrozy-2-methyl-5-oxo-as-triazine ~ ~ 3-yl) methyl-thio_7 / 8-oxo-5-thia-lazabicycl'o ^ .2.07-oct-2-en ~ 2 carboxylic acid, preferably in the form of the disodium salt, with a pivalolyloxymethyl halide, preferably with the iodide, and, if desired, converting the product obtained into a salt or hydrate with respect to a hydrate of this salt.