CS219254B2 - Method of preparation of the /6r,7r/-7-/2-/2-amino-4-thiazolyl/-2-/methoimino/acetamido/-3-/substituted/-thio/-methyl/-8-oxo-5-thia-1-azabicyclo/4,2,0/okt-2-en-2-carboxyl acid - Google Patents

Description

The present invention relates to a process for the preparation of (6R, 7RJ-7- [2- (2-amino-4-thlazolyl) -2- (methoxyimino Jacetamido) -3 - {[[substituted J-thio] methyl] -B- oxo-S-hia-1-aabicyclo [4.2.0] oct-2-ene-2-carboxylic acid of formula I

CH ^ ON = C-

CONH

COOR, ¹ ω in which

X represents one of the groups

R 10 is alkanoyl-oxyalkyl of up to 7 carbon atoms or a cation of a pharmaceutically acceptable base; and

R 1 is the same as R 10 and further may also represent hydrogen, and pharmaceutically acceptable salts of the compound and hydrates of the compound or salts.

Esters of compounds of formula I may be compounds of formula I whose carboxy group is in the form of an easily hydrolyzable ester group. ^Examples: such esters are alkanoyloxy with up to 7 carbon atoms, such as acetoxymethyl, pi.valoyloxymethyl-, 1-acetoxyethyl and 1-pivaloyloxyeehylester; alkoxycarbonyloxyalkyl esters, for example methoxycarbonyloxymethyl-, 1-ethoxycarboneloxyethel- and 1-isopoxy-oxycarbonyloxyethyl; a lactonyl ester such as a phthalidyl and thiophthalidyl ester; alkoxymethelesters such as methoxymethyl ester; and alkanoelaminomethyl esters such as acetamidomethyl ester.

Other esters, such as benzyl and cyanomethyl ester, are also useful.

The ethers of the compounds of formula (I) may be compounds of formula (I) wherein X is 2,5-dihydro-6-hydroxy-2-methyl-5-oxo-as-triazin-3-el, the enol OH-group of which is in the form of easily ether groups. Suitable ether groups are the same as above for easily hydrolyzable ester groups. Examples of such ethers are also, for example, alkanoyloxyalkelethers with up to 7 carbon atoms, for example acetoxymethyl, pivaloyloxymethyl, 1 * acetoxyethel and 1-pivaloyloxyethyl ether, alkoxycarbonyloxyalkelethers, for example methoxycarbonyloxymethyl, 1-ethoxycarbonyloxyoxyethyl and 1-isopropoxyethyl; lactonyl ethers, for example phthalidyl and thiophthalidyl ether; alkoxymethylethers such as methoxemethelethel; and alkane-methylmethylethers, for example acetamidomethyl ether.

For example: salts of the compounds of formula I are alkali metal salts such as sodium or potassium salts; ammonium salt; alkaline earth metal salts such as calcium salt; salts with organic bases such as salts with amines, for example salts with N-ethelpiperidine, procaine, dibenzylamine, N, N ^, -dibenzylethylethylenediamine, alkylamines or dialkylamines, salts with amino acids such as salts with arginine or lysine. The salts may be mono-salts or di-salts. Further salt formation can occur with compounds having the hydroxy-residue of 2,5-dihedro-6-hedroxe-2-methyl-5-oxo-C 18 -triazin-3-el group.

The compounds of formula I also form addition salts with organic or inorganic acids. Examples of such salts are hydrohalides such as hedrochloride, hydrobromides, hydroiodides and other mineral acid salts such as sulfates, nitrates, phosphates and the like, alkyl and mono-arylsulfonates such as ethanesulfonates, toluenesulfonates, benzenesulfonates and the like, as well as other organic acid salts. such as acetates, tartrates, maleates, citrates, benzoates, salicylans, ascorbates and the like.

The compounds of formula I, including their salts, may be hydrated. The hydration may be carried out in the preparation process or sequentially as a result of the hygroscopic properties of the initially anhydrous product.

The products obtained by the process of the invention may be in syn-isomeric form

or in an anti-isomeric form

W-t — C- CONH—

СИ ₃ О or - may be present as mixtures of both forms.

Preferred is the syn-isomeric form or the mixture in which the syn-isomeric form predominates.

Preferred products are 166.R, 7R) -7 - ['2- [12-amino-4-thiazolyl} -2- (2-methoxyiminoacetamido)] 7 - [(2,5-dihydro-5-hydroxy-Z)]. methyl-5-oxo-as-triazin-3-yl) thio] methyl [8-oxo- [5-thienyl-1-azabicyclo [4 / 2.0] oct-2-ene-2-carboxylic acid and its salts and - the respective hydrates.

The process for the preparation of the compounds of the formula I is characterized in that - in the carboxylic acid of the formula II

CONH

AND

COOH (I) in which

X * - represents the above -groups a ^ b - neώο - d,

R represents a known cleavable protecting group on cephalosporin and the carboxyl group can be in protected form, cleaves the protecting group R and the optionally present carboxyl protecting group, whereupon the resulting carboxylic acid of formula I is optionally reacted with an alkanoyloxyalkyl halide of at most R carbon atoms to form an alkanoyloxyalkyl group R 1 having at most R carbon atoms, and when X represents a tautomeric group a ^ b, a -C group is obtained where R 10 is an alkanoyloxyalkyl group having at most R carbon atoms, the carboxylic acid obtained in formula I optionally reacted with an appropriate base to form R 1 cations, wherein when X represents a tautomeric group a-b, a c-group is obtained with R 10 = base cation, whereupon the product obtained is optionally converted into a salt or a summer-salt hydrate.

The carboxyl group present in the - starting compound of formula II may be protected, for example by esterification to an easily cleavable ester, as a silyl ester, - for example, a trimethylsyl ester. The above-mentioned easily hydrolyzable esters are also contemplated. The carboxyl group can also be protected by salt formation with an inorganic - or tertiary organic base, such as triethylamine. Protecting groups R are, for example, acid-withdrawable protecting groups, such as, for example, tert-butoxycarbonyl or trityl, or else basic hydrolysis of a leaving-protecting group, such as trifluoroacetyl. Preferred protecting groups R are chloro, bromo and iodoacetyl, especially chloroacetyl. The latter protecting groups can be cleaved off with thiourea.

For example, the starting compounds of the formula II can be prepared by N-acylation - of the corresponding 7-amino compound - by reacting the compound of the general formula - III

H H

C00H ( _{f / f} ) in which

X is as defined above and the carboxyl group and / or the amino group may be in protected form with an acid of formula IV

CH 2 ON-C 2 C 4 C 4 OH

RHN

(IV) in which it has

R is as defined above, or with a reactive functional derivative of the acid, and optionally the carboxyl protecting group present is optionally cleaved.

The carboxyl group present in the - amino compound of formula III can be protected, in the manner explained above for the starting compound of formula II. For example, the amino group of the compound of formula III may be protected by a silyl protecting group such as trimethylsilyl.

Suitable reactive functional derivatives of the acids of the formula IV are, for example, halides such as chlorides, bromides and fluorides; azides; - anhydrides, - especially mixed anhydrides - with - strong - acids; reactive esters, for example N-hydroxysuccinimide esters, and amides, for example imidazolides.

The reaction of the 7-amino compound of formula III with an acid of formula IV or a reactive functional derivative thereof may be carried out in a manner known per se. For example, the free acid of formula IV can be condensed with said ester of formula III with a carbodiimide, such as dicyclohexylcarbodiimide, in an inert solvent such as acetate, acetonitrile, dtoxane, chloroform, methylene chloride, benzene or dimethylformamide and then cleaved. Instead of carbodiimide, oxazolium salts such as N-ethyl-5-phenyl-isoxazole-3‘-sulfate may also be used as a condensing agent.

In another embodiment, a salt of an acid of formula III, for example a trialkylammonium salt, such as a triethylammonium salt, is reacted with a reactive functional derivative of an acid of formula IV as described above in an inert solvent, for example one of the above.

According to another embodiment, the acid halide, preferably the acid chloride of the formula IV, is reacted with an amine of the formula III. The reaction is preferably carried out in the presence of an acid binding agent, for example in the presence of an aqueous alkali solution, preferably sodium hydroxide, or also in the presence of an alkali metal carbonate such as potassium carbonate or in the presence of a low amine such as triethylamine. The solvent is preferably water, optionally in admixture with an inert organic solvent such as tetrahydrofuran or dioxane. It is also possible to work in an aprotic organic solvent such as dimethylformamide, dimethylsulfoxide or hexamethylphosphoric triamide.

The silylated starting compound of formula III is used in an anhydrous medium.

The reaction of the 7-amino compound of formula III with an acid of formula IV or a reactive functional derivative thereof may advantageously be carried out at a temperature between -40 ° C and room temperature, for example at about 0 to 10 ° C.

The starting compounds of the formula II can also be prepared by reaction with a thiol, namely by reacting a compound of the formula V

CH _Ó OW == C—

CONH

COOH (V) in which

R is as defined above,

Y represents a terminal group and the carboxyl group may be in protected form with the thiol of formula VI

HS - X (VIJ, in which it has

X as defined above, and any carboxyl protecting group that may be present, are optionally cleaved.

Suitable end groups for the compounds of formula V are halides, for example chlorine, bromine or iodine, acyloxy radicals, for example alkanoyloxy radicals, such as acetoxy, low alkyl or arylsulfonyloxy radicals, such as mesyloxy or tosyloxy, or an azide radical. The compound of formula V can be protected at the carboxyl group in the manner illustrated by the starting compound of formula II.

Reaction of a compound of formula V with a thiol of formula VI may be effected in known manner, for example in a temperature range 4I0 ^1-80 ° C., preferably at 6'0 ° C in a polar solvent such as an alcohol, eg a lower alkanol in example , such as ethanol, propanol and the like, in dimethylformamide or dimethylsulfoxide, preferably in water or a buffer solution at a pH of about 6 to 7, preferably 6.5.

According to the process of the invention, the amino protecting group R in the starting compound of formula II is cleaved. The protecting groups cleavable by acid hydrolysis are preferably removed by means of a low alkanecarboxylic acid, which may optionally be halogenated. Especially formic acid or trifluoroacetic acid is used. Generally, the process is carried out at room temperature, although a slightly elevated or slightly lowered temperature can be used, for example in the range of 0 ° C to + 40 ° C. The alkali-cleavable protecting groups are generally hydrolyzed with dilute aqueous hydroxide solution at 0 to 30 ° C. The chloroacetyl, bromoacetyl and iodoacetyl protecting groups can be cleaved with thiourea in an acid, neutral or alkaline medium at 0 to 30 ° C. Hydrogenolytic cleavage, for example the cleavage of benzyl, is unsuitable here since the oxime function is reduced to the amino group during hydrogenolysis.

After carrying out the process, the carboxyl protecting group possibly present in the reaction product can optionally be cleaved off. If the protecting group is a silyl group (silyl ester), this may in particular be readily cleaved by the action of water on the reaction product. The alkanoyloxyalkyl-, alkoxycarbonyloxyalkyl-lactonyl-, alkoxymethyl- and alkanoylaminomethyl esters of up to 7 carbon atoms can preferably be cleaved off enzymatically by means of a suitable esterase (at about 20-40 ° C). If the carboxyl group is protected by salt formation, for example triethylamine, the cleavage of this salt-forming protecting group can be carried out by treatment with an acid. As the acid, for example, hydrochloric acid, sulfuric acid, phosphoric acid or citric acid can be used.

The carboxyl-protecting group may be removed in the same manner as just described, also before the R-protecting group is removed.

The resulting carboxylic acids of the formula I can optionally be further converted as described below.

To prepare a readily hydrolyzable carboxylic acid ester of formula I, the obtained carboxylic acid of formula I is reacted, preferably with an appropriate halogen-containing ester group, preferably iodide. The reaction may be accelerated with a base, for example an alkali metal hydroxide or an alkali metal carbonate, or an organic amine such as triethylamine. In the presence of 2,5-dihydro-6-hydroxy-2-methyl-5-oxo-as-triazin-3-yl group X with an enol function, this can be etherified to form the corresponding, readily hydrolyzable -ether. Preferably, an excess of the corresponding halide is used in the reaction. The esterification / etherification reaction is preferably carried out in an inert solvent such as dimethylacetamide, hexamethylphosphoric triamide, dimethylsulfoxide or preferably dimethylformamide. The temperature is preferably in the range of 0 to 6 ° C.

For the preparation of salts of the .alpha.-and-hydrates of the compounds of the formula I or of the hydrates of these salts, the carboxylic acid of the formula I can be reacted in a known manner, for example with an equivalent amount of a suitable base, preferably in a solvent such as water or an organic solvent such as ethanol; acetone and others. When a second equivalent of base is used, the salt also forms on the optionally present tautomeric enol form (2,5-dihydro-6-hydroxy-2-methyl-5-toxo-as-riazin-3-yl group X), whereby the di- salt. The salt formation temperature is not critical. Generally, the reaction is carried out at room temperature, but the temperature may be slightly raised or lowered, in the range of about 0 ° C to + 50 ° C.

The preparation of the hydrates takes place mostly as an automatic process or as a result of the hygroscopic properties of the initially anhydrous product. For the preparation of Hydrate desired -se may expose wholly or partially anhydrous product (carboxylic acid of formula I or an ester, ether or salt) effect a moist atmosphere, e.g., about + 10 · -to +40 ^o C.

The above-used 7-amino compound of formula III can be prepared from a compound of formula VII

H H o i

COOH (Vil) in which it is

The Y terminal and the carboxyl group may be in protected form with the thiol of formula VI. The reaction can be carried out under the same conditions as described for the reaction of the starting compounds of the formula V with the compounds of the formula VI. On the other hand, the compounds of formula V can be prepared from the compound of formula - III and the acid - of formula IV or - their reactive functional derivatives under the same conditions as described for the reaction of the compounds of formula III and IV.

The desired dream / anti-mixture of the compound of formula I can be cleaved in the usual manner into the corresponding syn- and anti-form, for example, by recrystallization or chromatography techniques using a suitable solvent or solvent mixture.

The compounds of the formulas I and II and the corresponding readily hydrolyzable esters, ethers and salts or hydrates of these products are antibiotic, in particular bacterially active. They have a broad spectrum of activity against Gram-positive and Gram-negative microorganisms, including β-lactamase-forming staphylococci and various Gram-negative / Mactamase-forming bacteria, such as Pseudo-monas aeruginosa, Haemophilus influenzae, Escherichia coli, Serratia-marsceus and Elodylaus.

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

To demonstrate the antimicrobial activity of the products, the following selected compounds were tested:

Product A:

(6R, 7R) -7- [2- (2-amino-4'-thiazophyll) -2- (2-methoxyimino) acetamido] - 219254 {['(2,5-dihydro-6-yl) dr oxy-2-methyl-5-oxo-a-triazin-3-ylthio] -methyl) -8o-xo-S-thia-1-azabicyclo [4.2.0] oct-2-enecarboxylic acid acetamide 3-Ц (2,5-dihydro-6-hydroxy-2-methyl-5-oxo-α-triazin-3-yl) thio-1'-piperidin-1-yl-1 '- 1'; [eta] < 5 > -15-alpha: --- [2- (2-ethoxy) -cyclo [4.1.1] oct-2-ene-2-carboxylic acid

Product B:

In vitro activity:

(6R, 7R) -7- [2- [2- (2-chloro-etamido) -4-thiazolyl] -2- (2-methoxyimino) lowest inhibitory concentration (mg / ml)

Haemophilus influenzae

Tribe 1 0.08 1,2 Tribe 2 0.005 0.3 Tribe 3 0.005 0.16 Strain 4 0.005 0.16 Tribe 5 0,0025 0.08 Tribe 6 0,0025 0.16 Tribe 7 0,0025 0.16 Klebslella pneumoniae 1,2 10 Escherichia coli Tribe 1 0.02 0.16 Tribe 2 0.6 5 Proteus mirabiliis Tribe 1 <01.01 0,0 « Tribe 2 <0.01 0.16 Proteus vulgaris <0.01 0.16 Proteus rettgeri ¹ <0.01 0.16 Staphylococcus aureus ATCC 6538 strain 2.5 2.5 Strain resistant against penicillin 2.5 5 Pseudomonas aeruginosa Tribe 1 0.3 1,2 Tribe 2 10 > 80 Tribe 3 2.5 40 Strain 4 5 € Ю Tribe 5 5 80 Tribe 6 1'0 80 Tribe 7 5 80 Serratia marcescens 0.08 2.5

A'ktívíta in. in vivo

Groups of 5 mice are infected intraperitoneally with an aqueous suspension of Escherichia coli. Three times, i.e. 1 hour, 21/2 hours and 4 hours after infection, the test substance is administered subcutaneously in saline. On day 4, the number of surviving animals is determined. Different dosages were applied and the survival dose was determined by interpolation: 50% of the test animals [CD 50, mg / kgj.

Test substance

CD50, mg / kg

Toxicity

Test substance

LD-0 mg / kg

i.v.

s.c.

after.

<0,005

And '250 to 5:00> 4000> 5000

0.16

(B)

250 'to 500 2000 to 4000> 5000

14

The products according to the invention can be used as medicaments, for example in the form of pharmaceutical preparations containing these products or their salts in admixture with a pharmaceutical, organic or inorganic inert carrier suitable for enteral or parenteral administration, such as water, gelatin, arabic gum, milk starch, magnesium stearate, talc, vegetable oils, polyalkylene glycols, petrolatum, etc. The pharmaceutical preparations may be in solid form, such as tablets, dragees, suppositories, capsules; or in liquid form, for example as solutions, suspensions or emulsions. Optionally, they are sterilized and / or contain excipients such as preservatives, stabilizers, wetting agents or emulsifiers, salts for varying the osmotic pressure, anesthetics or buffers. They may also contain other therapeutically valuable substances. The compounds of the formula I and their salts or hydrates are preferably suitable for parenteral administration and, for this purpose, they are preferably processed as lyophilisates or dry powder for dilution with conventional agents such as water or isotonic saline. Easily hydrolyzable esters or ethers of the compounds of the formula I and their salts or hydrates are also suitable for enteral administration.

Example 1

Preparation of (6R, 7R) -7- [2- (2-Amino-4-thiazolyl) -2- (2-methoxyimino) acetamido] trisodium salt ( ³ ( ^1, 2, 5-dihydro-6-hydroxy-2-methyl) ^; oxo-ast-triazin-5-yl) thio] methyl} -8-oxo-5-thia-1-azabicyclo [4.2.0] octane-2-carboxylic acid.

10.3 g · (6R, 7R) -7- {2- [2- (2-cycloacetamido) -4-thiazolyl] -2- (2-methoxyimino) acetamido-3- {| j [2,5 6-Hydroxy-6-hydroxy-2-methyl-5-oxo-as-triazol-3-yl] -thio] methyl | -8-oxo-5-thla-1-azabicyclo [4.2.0] oct-2-tert-carbonyl xylic acid (Fraction (See below) is suspended together with 5 g of thiourea in 150 ml of water, with good nitrogen gasification and stirring, the pH is adjusted to 5.8-7.0 with saturated sodium bicarbonate solution to give an orange-colored solution. The pH of the reaction solution is kept constant at 6.8 by addition of sodium bicarbonate solution for 6 hours, then 2.5 g of thiourea are added and the solution is stirred for a further 3 hours, while maintaining the pH by addition of saturated sodium bicarbonate solution. The red solution is then stored overnight in the refrigerator and darkened. 100% formic acid was added to 2.0-2.5 to remove the material, which was suction filtered and washed with 100 ml of 10% formic acid. The mother liquor is discarded. The brownish substance is suspended in 200 ml of water and the pH is adjusted to 7 with triethylamine to give a brown solution. This solution was stirred for 30 minutes with 2 grams of charcoal, the charcoal was filtered off, and the still brown filtrate was adjusted to pH 3.5 with good stirring with 10% formic acid. The precipitate is suction filtered, washed with 50 ml of 10% formic acid and discarded. The dark yellow filtrate was adjusted to pH 2 to 2.5 with 100% formic acid to precipitate. This was suction filtered, washed with ice water and dried. The obtained cephalosporic acid is suspended in a mixture of 40 ml of acetone and 40 ml of water for conversion to the disodium salt, and 20 ml of a 2N solution of sodium 2-ethylcaproic acid in acetate are added. 50 ml of acetone are added to the resulting orange-colored solution, which precipitates a brown resin which is filtered off. The yellow filtrate was stirred for 30 minutes, where the disodium salt crystallized. 50 ml of acetone are added in portions and stored in a refrigerator overnight. The crystal alisate is suction filtered and washed successively with acetone-water (80:10), pure acetone and low boiling petroleum ether and dried overnight at 40 ° C under vacuum. The title compound is obtained as beige crystals. [.Alpha.] D @ 20 = -444 DEG (c = 0.5 in water). The magnetic resonance spectrum and the microanalysis correspond to the above structure.

(6,7,7K) -7- (2--2--2 -halo-carbamido) -4-thiazolyl] -2- (2-methoxyethoxy) acetamido) -3 - {[[2 ' 5-Dihydrot-6-hydroxy-2-methyl- (oxo-tas-thiazol-3-yl) -thio] methyl] -B-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene The -2-carb'O'-xylic acid used as starting material can be prepared as follows:

22.24 g of 2- (2-chloroacetamido-4-thiazol-4-yl) -2- (2-methoxyimino) acetic acid are suspended in 240 ml of methylene chloride. To this suspension was added 13.3 (9 mL of triethylanine) to give a light brown solution. This solution was cooled to 0-5 ° C and 16.72 g of phosphorus pentachloride was added, stirred for 0 minutes at 0-5 ° C. The solution was evaporated in vacuo at 35 DEG C. The residue was shaken with n-heptane (2.times.) and decanted, 250 ml of tetrahydrofuran were added to the resin residue, and the undissolved triethylanine hydrochloride was filtered off, and the yellow filtrate contained the acid chloride.

g (7R) -thiamino-3-desacetoxy-3- [1- (2,5'-dihydrothihydroxy-2-methyl-5-oxo-as-triazine-3 ', 4') thio] cephalosporanic acid is suspended in a mixture of 300 ml. water and 150 ml of tetrahydrofuran. The slurry was added dropwise with good nitrogen gasification using an automatic 2 N sodium hydroxide titrator until a brown-red solution of pH 8 was formed. This solution was cooled to 0 to 0 ° C and the above prepared acid chloride solution in tetrahydrofuran was added dropwise over 15 minutes. . After stirring 2.0 hours at 25 ° C · ^Q

The pH of the acylation mixture was kept constant at 8 by the addition of 2N sodium hydroxide. The tetrahydrofuran was removed from the practically black solution under vacuum at 40 ° C. 1C0 ml of a 2 N sulfuric acid solution was then added. The deposited substance is suction filtered, washed with water and suctioned off well. The wet, brown product was dissolved in 1.5 L of acetone. From the dark solution, the less dark undissolved m-arterial was filtered through hyflo, charcoal was added, stirred for 30 minutes and again filtered through hyflo. The orange-red filtrate was dried over sodium sulfate, concentrated in vacuo and evaporated with acetone. A black resin precipitates and is filtered off and removed. The two-phase filtrate still containing water was azeotroped three times with benzene under vacuum at 40 ° C. The precipitate formed is suction filtered and dried under vacuum at 40 ° C. Stir 2X with one liter of acetone leaving a brown resin which is removed. The combined orange-colored acetone extracts were concentrated to about 150 ml in vacuo at 40 ° C while the brown resin was filtered off and removed. Add 1 liter of acetate to the filtrate and concentrate in vacuo at 4-1 ° C.

The precipitate is filtered off under suction, washed with acetate and then with [(6R, 7R) -7- [2- [2- [2-chloroacetamido) -4-thiazolyl] -2- (Z-methoxyl) -4- [eta] &lt; 6 &gt; (2,4-dihydro-6-hydroxy-2-methyl-5-oxo-as-triazin-3-yl) thio ] -methyl-8-oxo-5-hia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid, fraction I: beige, amorphous acid This fraction I can be directly used to prepare the desired end product.

The acetate mother liquor is vigorously concentrated in vacuo at 40 ° C, diluted with ether and suction filtered.

[(6R, 7R) -7- {2- [2- (2-Aminoacetyl) -4-thiazolyl] -2- (2-carboxylic acid) - acetamido] -3 - {['(2,5'-dihydro-6-hydroxy-2-methyl-5-ox-o-as-triazin-3-yl) thio] methyl] -8-oxo-5-thia-1- azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid, fraction II: light beige amorphous acid, somewhat purer than fraction I according to thin-layer chromatogram.

To prepare the disodium salt, 3.5 g of acid (fraction II) is dissolved in a mixture of 20 ml of acetone and 11 ml of water. 7 ml of a 2N solution of sodium 2-ethylcaproic acid in acetate are added to the solution, whereby the disodium salt crystallizes. 25 ml of acetone are added in portions and the mixture is left in the freezer for 2 hours. The crystallizate is then filtered off with suction, washed successively with 215 ml of an ice-cold mixture of acetone-water (80:20), neat with acetone and low-boiling petroleum ether, and dried overnight under high vacuum at 40 ° C. There was obtained (6R, 7R) -7- [2--2- (2-chloroacetamido) -4-thiazolyl] -2- (2-methoxy-4-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-en-2-carboxylic acids as pale yellow crystals. [α] D ²⁰ = -112.7 ° (c = 1 in water); Magnetic - resonance spectrum and microanalysis - corresponds to the given structure.

Example 2

Preparation of disodium salts (6R, 7R1-7- [2- (2-amino-4-thiazolyl) -): 2- (methoxyamino) acetamide - () - 8-ox (-3 - ([ (1,4,5,6-tetrallydolo-4-methyl-5,6-dioxo-as-triazin-3-yl) thio] ethyl} -5-thia-1-azabicyclo [4.2.0i] oct- 2-ene-2-carboxylic acids.

g (.6R, 7RJ-7- [2- [2--2-Chloroacetyl) -4-thiazolyl] -2- (aethoxyimino) acetamide-8-oxo-3 - {[(1,4,5 6-tetrahydro-4-methyl-5,6-dioxo-as-triazin-3-yl) thio] methyl] -5-thia-1-aza-cycl-o-4,2Ό] oct-2-en- 2-carboxylic acids are suspended together with 9.5 g of thiourea in 150 ml of water. The pH was adjusted to 6.8 with a 5% sodium bicarbonate solution under nitrogen and stirring to give a yellow-orange solution. By means of an automatic titrator, the pH of the reaction solution is kept constant at 6.8 to 7.0 by addition of sodium bicarbonate solution for 6 hours. To the orange-colored solution was added 10Cl% formic acid to pH 3.5. The precipitated material is suction filtered and washed with 100 ml of 10% formic acid. This substance is labeled ф

The filtrate was adjusted to pH 2.5 by addition of 10% formic acid, whereupon the material precipitated. The mixture is left on an Iel bath for 1 hour, then the precipitate is suction filtered and washed with a little ice water. This is called fraction I. The orange-brown substance uje is suspended - in two hundred and fifty ml of water. The suspension is adjusted to pH 7 with 2N sodium hydroxide solution to give an orange-brown solution. To this solution was again added 100% formic acid to pH 3.5. The precipitated substance is sucked off under suction and removed. The filtrate was adjusted to pH 2.5 with 100% formic acid to precipitate again. The mixture is left on an ice bath for 1 hour, then the precipitate is suction filtered and washed with a small amount of ice water. This substance is designated Fraction II. Fractions I. and II. are suspended together in 500 ml of ethanol and the water is removed in a rotary evaporator. After addition of ether, it is suction filtered and washed successively with ether and low boiling petroleum ether. The title compound is obtained as a yellowish substance, which is labeled A.

Mother liquors and washing water from fractions I. and II. It is concentrated from a volume of about 1.7 l to 250 ml, the pH is adjusted to 2.5 with 100% formic acid and the solution is stored in a refrigerator overnight, whereupon the substance crystallizes again. This was suction filtered and washed with a small amount of water. The product was azeotroped with ethanol. A solid, practically colorless solid is obtained, which is labeled B. According to the thin-layer chromatogram, compound B is cleaner than compound A.

To obtain the pure desired substance, the acid B is suspended in 150 ml of methanol and, with stirring, Ю ml of a 2N solution of sodium 2-ethylcaproic acid in acetate is added. After about 10 minutes, a solution is formed to which 100 ml of ethanol are added. The mixture was vigorously concentrated in vacuo at 40 ° C. Addition of ethanol precipitates the sodium salt amorphously. This is suction filtered, washed successively with ethanol and low-boiling petroleum ether and dried for 24 hours at 40 ° C under high vacuum. The title compound is obtained as a practically colorless amorphous powder. [Α] D 20 - -42.9 ° (c = in water).

According to the magnetic resonance spectrum, the desired substance is present as a Z / E mixture (90 = 10). Microanalysis also coincides with the given structure.

(6R, 7R) -7-] 2- [2--2 - (; chloroacetamoyl-4-thiazolyl) -2- (methoxyimino) acetamido] -8-oxo-13 - {[(. tetrahydro-f-methyl-N-dioxo-as-triazin-3-yl) thiomethyl-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid used as the starting compound may be prepare as follows:

g (7R) -7-Amino-3-desacetoxy-3 - [(1,4, S, 6-tetrahydro-4-methyl-5,6-dioxo-as-triazin-3-yl) - thio] cephalosporic acid is suspended in a mixture of 600 ml of water and 300 ml of tetrahydrofuran. A 2 N sodium hydroxide solution was added dropwise to the slurry with good nitrogen gasification, until a brown solution with a pH of 7.8 was formed. This solution was cooled to 0-5 ° C and 2- (2-chloro-ω-3-amido-thiazol-4-yl) -2- (2-methoxyimino) chloride solution was added dropwise over 15 minutes. -acetic acid in tetrahydrofuran (prepared from 44.5 g of the corresponding acid according to Example 1. It is then stirred for 21/2 hours at pH 8 and 25 ° C. The pH of the acylation mixture is kept constant by addition of 2N sodium hydroxide using an automatic titrator. at 7.8 to 8. The dark solution is freed of tetrahydrofuran in vacuo at 40 DEG C. The solution is then diluted with water to a volume of 21 liters. The pH is adjusted to 2 with 2N sulfuric acid. 1 l of water and dried for 2 days in vacuo at 40 ° C. For purification, this substance is first dissolved in a mixture of 100 ml of water and 3010 ml of acetone, and the dark solution is diluted with acetone to a volume of 2 liters. The filtrate was treated with 1 L of acetate and evaporated in vacuo at 40 ° C. Then dilute the solution with 2 · 1 acetate and remove the beige-brownish substance. The filtrate was vigorously concentrated in vacuo at 40 ° C. The crystallized acid is suction filtered. For crystallization, the acid is first dissolved in 800 ml of methanol under reflux. The solution was cooled to 25 ° C and a small amount of orange colored material was filtered off. The yellow filtrate was stirred in an ice bath for 1.5 hours while the acid crystallized. This was suction filtered, washed successively with methanol and low boiling petroleum ether and dried under vacuum at 25 ° C.

The starting compound is obtained as beige crystals. According to the magnetic resonance spectrum, the starting compound is present as a Z / E-mixture (7: 5: 25) · [α] _D ²⁰ = -127.9 · ° (c =

1 in dimethylformamide).

Example 3

Preparation of methylene- (6R, 7R) -7- [2- [2-amino-4-hazolyl] -2- [2-methoxyimino] -cyclohexyl] -34 [&lt; - &gt; 2,5-dihydro-2-methyl-5-oxo-N - ((pivaloyloxy) methoxy) -as-triazin-3-yl) -thio] methyl} -8-oxo-5-thia-1-azabicyclo-.

[.4.2] oct-2-ene-carboxylate pivalate.

1.85 g of cephalosporin disodium salt prepared according to Example 1 are suspended in 50 ml of dimethylformamide and 1.35 g of pivaloyloxymethyl iodide are added at 0-5 ° C under nitrogen. The reaction mixture was stirred at 0-5 ° ^C for 30 minutes and then poured into 500 mL of acetate. The mixture was washed with 3X water, 2X 5% sodium bicarbonate solution and finally with water again. The solution was dried over sodium sulfate and strongly concentrated in vacuo at 35 ° C. Upon addition of the ether, the desired material precipitated amorphously. This was suction filtered, washed with ether and low boiling petroleum ether and dried under high vacuum overnight at 25 ° C. The title compound is obtained as a beige amorphous powder. The magnetic resonance spectrum and microanalysis are consistent with the given structure.

Example 4

Preparation of dry ampoules for intramuscular administration. Ly-olfilzate 1 g of (6R, 7R) -7- disodium salt is prepared in a conventional manner

- [2- (2-amino-4-thiazplyl) -2- (2-methoxyimino) aceamido-C - {[1 (2,5-dihydro-6-hydroxy-2-methyl-5-oxo-as) -] - Riazin-3-yl) thio] methyl} -8-oxo-E5thia- ^; l · azabicyclo [ ^! 4.0 ml of oct-2-ene-2-carboxylic acid and filled into ampoules. 2.5 ml of a 2% aqueous lidocaine hydrochloride aqueous solution was added to the lyophilizate prior to administration.

Example 1 a d 5

Gelatin hard capsules of the following composition are prepared in a conventional manner:

methylene (6R, 7R) -7- [2- (2- -amino-4-thiazolyl) -2- (2-methoxyimino) acetamido] -3 - {[1 (1), 2 '- (2') - (1 '- (2-oxo) -1' (1'-ethyl) - 6 - [(pivaloyloxy) methoxy] -as-triazin-3-ylthiomethyl) -8-oxo-5-hia-1-aza-bicyclo [4.2.1] oct-2-ene-2-carb <oxylate pivalate 500 ' mg luviskol (water-soluble vinylpyrrolidone poles) 20i mg mannit 20 May mg talc (15. mg magnesium stearate 2 mg

557 mg

219: 215 4

- Example - 8

When - in Example 1 - instead of - (6-R, 7R) -7- {24 2- [2-chloroacetamido] -44-thiazolyl] -2- (2-methoxyimino) -acetamide-1-B- - '(2,5-dihydro-6-hydroxy-ix> hydroxy-2-Mett -yl ^ - ^ 5 ^ XC ^ -a ^ s ^ ^-ti? _{thiazine-3-1-yl)} thio'] methyl-e-oxo ---- hia-1-azabicyclo [-4.2.01] -oct-3-er-2-carbonylbutyric acids - starting materials - one of the following compounds:

- (6R, 7R) -7f (2- [2- (2-Biphenyl-4-thiazolyl) -4-thiazolyl] -2t (2-meth) xylamino) acetyl amido-3 - 1 [(, 2,5-dihydro-6-hydroxy-2-methyl-5-oxo-as-trlazan-3-yl) -thio-methyl} -88E-α-5-thka-1 -azabicyclo; 1 '(8R, 7R-) -7--2- [2- (2-iodo-Getamido) -. -2- (1'-imethoxyinino) - ac-1-amino-N- (4-O-dihydro-O-hydroxy) -1 (m-1-oxo-aa) -thiazole (3a) ^ .l ') thio] mёthyl | A8 .0xo-5 "ahla- L-azabic.yklo? _[i i42 i (>} (- (kt2.ein2-lkarboxytová acid diester: yield se.za - JMA-k - tejných -.podmínek. ^(reaction with 'aqueous - sodium - - thiourea - (same product - ij. dvojsQdná sulphonyl-6RR RR) 77. [22 (| 2-thiazolyl sminoiá 1 ) 22- (Zimsthoxy imino) -

i.aDst-amino ΐ ^ ^ o ^] ^ _Э: Цl.22 misery ^ ^ ^^ d ^ IHY ro'8í ^ i ^ J ^ ljdгc ^ y ^^ 721-methyl-5-Oxx-as-triazin. 2'-yl] methyl-8-oxo-5-hia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid, which is each identical to the product obtained in Example 1.

Example 7.

When - in example -1 - instead of - (8R ', 7R ·) -7- | 21 ^ 24 (.2-chloroacetamide) - 44hiazcllyl] -acetamido-3- ([. ^; 2,5М1hydro * '. 6 ^hydroxy.n--; methyl-'S · roχ. (> ras .traazin.4 & al :)) etc. <óaI№ehyl} aш'Oithiaal-T88 - azabacykI lo [4.2 .0} oct-2-en-2-carboxylic acid-starting material one of the following compounds:

- '(6R, 7R') -7- {2- [2- (2-butoxycarbonylamino) -4-thiazolyl] -2- (Zm-ethoxyimino Jacetamido) -3 '- ((2', 5- dihydro [6 [2-h.ydroχ.y _{methylI5-oxo-as-tгia;} zin-3-yl -] - thio} with methyl-l ^ - ^ (^} ^ C ^ -t ^ ^ l ^ -TT 1'-azabicyclo [2.2.1.0 &lt; 2,7 &gt;] oct-2-ene-2-oxoic acid, - (OR4R4) -4- [2- (1,2-triethylamino) -4-thiazolyl] -2- (. Methoxyimino) acetamide: 33- [2,5,5-dihydroxy-6-hydroxy] -2-methyl-5-oxa-aza-thiazin-3-yl} -thio} [m-ethyl] - (-) - (8R, 7R (2- (2-formamamido) -44-thiazolyl) -2- (Z) -) 4-Methoxyimino) acetamido] -1 - [- (2 ₎ 5-dihydro-6-thiidopyloxy-2-methyl-5-oxo-phenyl] -furan-3-yl-1-yl} -mol | -8-oao-5-hia-1-aza-cyclo- [4.2.0-] oct [2-pent-2-carboxylic acid, obtained - under otherwise identical conditions) - but - interchanged - with - aqueous - -thiourea solution - -for reaction - - with - - & 0% aqueous solution - acid - - formic acid - disodium -salt (i6R, 7R) -7- [2- (2-Amino-4-thiazolyl) -2- (Z [methoxyimino] -acetoamino] _-1 Z - {[; (З.З ^ ЬуОro - 6-hyo-rox.y'r24eihth-51-S-'Oxo 'a'S + tT.iazin <2 •• * yl Jihlo] -ethyilaaeoxo + δ-thia ^; [4-.10] -oct-2-ene-2-carboxylic acid, which is identical in each view to the product obtained according to -example - 1.

Translation ^: -8

When - used - in Example 1 - instead of (8R, 7R) -7- (2- [2- (2-halomethylamido) -1,4-thiazolyl] -1- [2- [Z [methyl] aminoacetamide.0) -e-зЗf {[- ('2,5-dihxdro * 6-hydroxy-2'-methyl-5 * №O-triazin [3-yl]) thio] methyl | -8τoxot5-thiazol-abic 10 [4.2iO] .Oct-2tert-2-carboxylic acid - as a starting material - [η 6.R _/ 7.R] t7τ (2 ^ [2- (2 [trifluoroacetyl tamido]) 4-thi-acolyl-2-iodopentanediamine ([eta] &lt; 5 &gt; -acetamido [-3- {[; (1,2,5-dihydro-8-thydroxy-2-methyl-5-)] oxcoas-triazin-3-yl) thio] methyl [8-oxo-1-thia-1- [azabicyclo [4.2.0.0 &lt; 2,7 &gt;] oct-2-carboxylic acid is obtained under the same conditions, wherein: However, the reaction is reacted with an aqueous solution of thiourea to react with 0.1 N aqueous sodium hydroxide solution. - [2 - '(2-Thiamino-4-thiazolyl) -2- (2-methoxy-amino) acetamido}, 3] [[2,5-dihydroxy] [6-hydroxy] oxy] - .2-Methyl-5-oxo-as-tert-azin [2 [yl] thio] methyl (-8-oxo [-5-thia-1-azabicyclo] -o [4 _{; 2} .1.] oct-2-en-2-carbo-xylic acid which is - in each case identical - to - the product obtained according to - example - 1 .

Claims (9)

SUBJECT A process for the preparation of (6R, 7R) -7 - [2- [2-amino-4-thiazolyl] -2- (methoxyimino) acetamido] -3 - {[(substituted] -thio] -methyl] -8-INVENTION oxo-5-] hia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid of formula I CHON - C —— GONH. And cwj-x COORt (I) in which X represents one of the groups H ₃ G H CH ₅ d 6 & R 10 is alkanoyloxyalkyl of up to 7 carbon atoms. or a cation of a pharmaceutically acceptable base; and R 1 is the same as R 10 and can also be hydrogen,. and ‘salts of the compound and hydrates of the compound or salts, characterized in that in the carboxylic acid of the formula II CHJDN = C CONH RhN ^ _s S i in which X 'is ab or d, R is a cleavable protecting group on cephalosporin and the carboxyl group can be in protected form, R is removed and any carboxyl protecting group present, whereupon the resulting carboxylic acid of formula I is optionally reacted with an alkanoyloxyalkyl halide of not more than 7 carbon atoms to form an alkanoyloxyalkyl group R 1 of not more than 7 carbon atoms, and when X represents a tautomeric group ab, a group -c is obtained, wherein R 10 is alkanoyloxyalkyl of not more than 7 carbon atoms; reacting with an appropriate base to form a cationite R 1, wherein when X represents a tautomeric group ab, a group, c, where R 10 is a cation of a pharmaceutically acceptable base, is obtained, whereupon the product is optionally converted into a hydrate. 2. The process of claim 1, wherein the starting compound of formula II wherein R is chloroacetyl-and X &apos; is as defined in point 1 is reacted with -thiourea. 3. A process according to item 1 or 2 for the preparation of a compound of formula I, wherein X is ab and R is as defined in item 1, and salts thereof and hydrates of said compounds or salts, characterized in that the starting compound is of formula II, wherein X 'is a and b and R are as defined in 1. 4. A process according to item 1 or 2 for the preparation of compounds of formula I, wherein X represents a group d, and R is as defined in item 1, and salts of these compounds and hydrates of these compounds. or a salt, characterized in that the starting compound of formula II is used, wherein X ‘represents a group d and R has the meaning given in point .1. 5. The method of item 1 or 2 for the preparation of [6R, 7R] -7- [2- (n-amino-α-thiazolyl) -2- (2-methoxyimino) acetamido] -3 - [(2; 5-Dihydro-6-hydroxy-2-methyl-5-oxo-as-triazin-3-ylthio-methyl-.alpha.-oxo-S-thia-1-azabicyclo [4.2.0.] Oct-2 -en-2-carboxylic acid and salts thereof, and hydrates of the compound or salts, characterized in that the starting compound of the formula II is used, wherein X 'is a and b and R are as defined in point 1. 6. A process according to claim 1 or 2 for the preparation of acyl derivatives of formula I, wherein R1 is alkanoyloxyalkyl of up to 7 carbon atoms, and salts of these esters and hydrates of these esters and salts, characterized in that the carboxylic acid of formula I is obtained. represents hydrogen and X is as defined in point 1, subjected to esterification by reaction with an alkanoyloxyalkyl halide of not more than 7 carbon atoms and, where appropriate, the product obtained is converted into a salt or a hydrate or hydrate thereof. 7. A process according to claim 1 or 2 for the preparation of easily hydrolyzable ethers of the acyl derivatives of the formula I and their salts and the hydrates of these ethers and salts, characterized in that the obtained enol of the formula I wherein X represents a and b and R has as defined in (1), etherifies by reacting with an alkanoyloxyalkyl halide of not more than 7 carbon atoms and the product obtained is optionally converted into a salt or a hydrate or a hydrate of the salt. 8. A process according to claim 6 for the preparation of pivaloyl-methoxymethyl esters of a compound of formula I and salts thereof, and hydrates of said esters and salts, characterized in that the resulting carboxylic acid of formula I wherein R is hydrogen and X is as defined under 1 is reacted with pivaloyloxymethyl halide, preferably iodide, and the product obtained is optionally converted into a salt or a hydrate or a hydrate thereof. 9. The process of items 6 and 7 for preparing methylene- (6R, 7R) -7- [2- (2-amino-4-thiazolyl) -2- (2-methoxy-imino) acetamido] -3 - {[( 2, 5-Dihydro-2-methyl-5-oxo-6- [(pivaloyloxymethyl) -as-triazin-3-yl] thio] methyl] -8-oxo-6-thia-1-azabicyclo [4.2.0] ] oct-2-ene-2-carboxylate pivalate and salts of this ester and hydrates of this ester or salts thereof, characterized in that (6R, 7R) -7- [2- (2-amino-4-thiazolyl) - Zy (Z-Methoxylminoacetamido) -3 - [(2,5-dihydro-6-hydroxy-2-methyl-5-oxo-as-triazin-3-yl] thio] metal} -8- oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid, preferably in the form of its disodium salt, is reacted with a pivaloyloxymethyl halide, preferably iodide, and obtained If appropriate, the product is converted into a salt or a hydrate or a hydrate of the salt.