CS209879B2 - Method of making the new alkyoxine derivatives of the 7-/2-/2-amino-4-thiazolyl /acetamido/cephalosporane acid - Google Patents

Description

The present invention relates to a process for the preparation of novel alkyloxime derivatives of 7- [2-amino-4-thlazolyl] acetamido] cephalosporanic acid having the general formula I

in which

R represents an acyl group having 2 to 4 carbon atoms or 2-methyl-1,3,4-thiadiazolyl or 1-methyltetrazolyl,

R 1 represents a hydrogen atom or a trityl radical,

R2 represents a saturated or unsaturated alkyl radical having 1 to 4 carbon atoms,

A represents a hydrogen atom or an equivalent of an alkali metal, an alkaline earth metal, magnesium or a pharmaceutically acceptable aminated organic base, a wavy line means that the OR 2 group may be in one of two possible syn or anti positions.

Acyl, which may be R, is acetyl, propionyl, n-butyryl or isobutyryl.

C 1 -C 4 alkyl is generally methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, vinyl, propenyl, ethynyl, or propargyl.

Possible meanings of the general substituent A include, in particular, the equivalent of sodium, potassium, lithium, calcium or magnesium.

pharmaceutically acceptable aminated organic bases which may be represented by the general substituent A include trimethylamine, triethylamine, ipethylamine, propylamine, Ν, Ν-dimethylethanolamine, tri (hydroxymethyl jaminoethane, arginine or lysine).

The compounds obtained by the process of the invention, having the form of the syn isomer, may be represented by the following general formula

Compounds obtained by the process of the invention having the anti-isomer shape may be represented by the following general formula

In particular, the present invention provides a process for the preparation of compounds having the general formula I - in which the group easily eliminated by hydrolysis by acidic or hydrogenolysis, which may be represented by R 1, is a trityl group, - wherein R represents either an acyl group having 2 to 4 carbon atoms; 2-methyl-1,3,4-thiodiazolyl or 1-methyltetrazolyl, - in which R1 represents a hydrogen atom or a trityl group, R2 represents a methyl residue, - in which R represents an acetyl residue of 2-methyl-1,3,4 -tetrazolyl and A is hydrogen, - wherein R 1 is hydrogen, R 2 is methyl, R is acetyl, 2-methyl-1,3,4-thiadiazolyl or 1-methyltetrazolyl and A is hydrogen or equivalent an alkali metal, an alkaline earth metal, magnesium or a pharmaceutically acceptable aminated organic base, - in which R 1 represents a hydrogen atom, represents a methyl radical, R represents an acyl radical having 2 to 4 carbon atoms and A represents a hydrogen atom or

L 'equivalent of alkali metal.

Compounds of formula I include the compounds listed below. examples and in particular:

7 - {[2- (2-amino-4-thlazolyl) -2-methoxyiminoacetyl] amino} -3- [2-methyl-1,3,4-thiadiazol-5-yl] thiomethyl] -3-cephem- 4-carboxylic acid 7 - {[2- [2-amino-4-thiazolyl) -2-methoxyiminoacetyl] amino} -3 - [(2-methyl-1,3,4-thiadiazol-5-yl] thiomethyl] -3-cefem-χ,

4-carboxylic acid, 3-acetylthiomethyl-7 - {[2- (2-amino-4-thiazolyl) -2-methoxyiminoacetyl] amino}

3-Cephem-4-carboxylic acid 7 - {[2- (2-amino-4-thiazolyl) -2-methoxyiminoacetyl] amino} -3 - [(1-methyltetrazol-5-yl) thiomethyl] -3- cefem-4-carboxylic acid, isomer syn, 7 - {[2- (2-amino-4-thiazolyl) -2-methoxyiminoacetyl] amino} -3 - [(2-methyl-1,3,4-thiadiazole-5-) yl] thiomethyl] -3-cephem-4-carboxylic acid, isomer syn, 3-acetylthiomethyl-7 - {[2- (2-amino-4-thiazolyl) -2-methoximinoacetyl] amino] -3-cephem-4- carboxylic acid isomer syn, 7-β- [2- (2-amino-4-thiazolyl] -2-methoxyiminoacetyl] ammonium-3 - [(2-methyl-1,3,4-thiadiazol-5-yl) acid, sodium salt thiomethyl-3-cephem-4-carboxylic acid, isomer syn, 7 - {[2- (2-amino-4-thiazolyl] -2-methoxyiminoacetyl] amino] -3- [1-methyltetrazol-5-yl) sodium salt thiomethyl] -3-cephem-4-carboxylic acid, isomer syn, 3-acetylthiomethyl-7 - {[2- (2-amino-4-thiazolyl) -2-methoxyiminoacetyl] amino] -3-cephem-4- sodium salt carboxylic acid, syn isomer, as well as crystalline sodium 3-acetylthiomethyl-7 - {[2 -, (2-amino-4-thiazole) acid] -2-methoxyiminoacetyl] amino-3-cephem-4-carboxylic acid isomer syn.

It is understood that the compounds of formula I above may exist - either in the form indicated by the above formula I - or in the form of compounds of formula I ₂

Ν

STEED,

in which

R, R1, R2 and A are as defined above. The present invention provides a process for the preparation of a compound of formula (I), characterized in that the compound of formula (II) is produced

R 1 and R 2 are as defined above with a compound of formula III

R = SH (III) wherein

R is as defined above so as to obtain compounds of the general formula Γ

R 1, R 2 and R are as defined above, optionally converted into a salt to give compounds of formula I wherein A is an alkali metal, alkaline earth metal, magnesium or pharmaceutically acceptable aminated organic base equivalent.

The action of the compound of formula (III) on the compound of formula (II) is preferably carried out in a water-acetone mixture, but other aqueous solvents such as water-dioxane, water-tethahydrofuran or water-ethyl alcohol may be used equally.

The reaction is preferably carried out in the presence of a buffer to maintain a slightly acidic pH, such as a mixture of sodium phosphate and sodium bicarbonate, but the reaction can also be carried out without a buffer.

The salt formation of the compounds of the formula (Γ) can be carried out by the method described in the Examples section.

In particular, the invention relates to a process for the preparation of compounds of the formula I in which the group OR2 is in the syn position, characterized in that compounds of the formula II in which the group OR2 is in the syn position are used as starting materials.

The compounds of the formula I, in particular those in the form of sons, have very good antibiotic activity, on the one hand on Gram-positive bacteria, such as staphylococci and especially penicillin-resistant staphylococci, on the other on Gram-negative bacteria, especially coliforms, Klebsiella , Salmonella and Proteus.

These properties allow these products, pharmaceutically acceptable, to be used as medicaments in the treatment of diseases caused by germs and in particular in the treatment of diseases caused by staphylococci such as staphylococcal septicemia, malignant staphylococia of the face and skin, pyodermitis, infected or festering wounds, anthrax phlegmona , erysipelas, simple and influenza acute staphylococia, bronchopneumonia and pneumonia.

The aforementioned pharmaceutically acceptable substances can also be used in the treatment of colibacillosis and related infections, infections caused by Proteus, Klebsiella and Salmonella and other diseases caused by Gram-negative bacteria.

The pharmaceutically acceptable compounds of formula (I) may also be used for the preparation of pharmaceutical preparations containing as an active ingredient at least one of the aforementioned products, and in particular those having the structure syn.

Among these products, mention may be made in particular of those which contain, as an active ingredient, at least one of the pharmaceutically acceptable substances of the formula I - in which R represents an acyl group having 2 to 4 carbon atoms or the 2-methyl-1,3,4- thiadiazolyl or 1-methyltetrazolyl, wherein R1 is hydrogen or trityl, R2 is mythyl and R is acyl, 2-methyl-1,3,4-thiadiazolyl or 1-methyltetrazolyl and A is hydrogen or an equivalent of an alkali metal, an alkaline earth metal, magnesium or a pharmaceutically acceptable aminated organic base.

Among the latter, mention may be made in particular of pharmaceutically acceptable substances which contain, as an active ingredient, at least one compound of the formula I in which R1 represents a hydrogen atom, R2 represents a methyl group and R represents an acetyl group having 2 to 4 carbon atoms, represents a hydrogen atom or an alkali metal equivalent; of these, in particular the compounds in which the OR2 group is in the syn position.

In particular, pharmaceutical compositions comprising at least one of the following compounds as active ingredient:

7 - {[2- (2-Amino-4-thiazolyl) -2-methoxyiminoacetyl] amino} -3 - [(2-methyl-1,3,4-thiadiazol-5-yl) thiomethyl] -3- cefem-4-carboxylic acid, 3-acetylthiomethyl-7 - {[2- (2-amino-4-thiazolyl) -2-methoxyiminoacetyl] dicin, such as the simple tabacid 7 - {[2- (2-amino) 4-thiazolyl-2-methoxyiminoacetyl] amino} -3 - [(1-methylterazil-5-yl) thiomethyl] -3-cephem-4-carboxylic acid, isomer syn, 7 - {[2- (2-amino- 4-thiazolyl] -2-methoxyiminoacetyl] amino-3 - [(2-methyl-1,3,4-thiadiazol-5-yl) thiomethyl] -3-cephem-4-carboxylic acid, isomer syn, 3-acetylthiomethyl acid -7 - {[2- (2-amino-4-thiazolyl) -2-methoxyiminoacetyl] amino; -3-cephem-4-carboxylic acid, Isomer syn, sodium 7 - {[2- (2-, amino- 4-Thiazolyl-2-methoxyiminoacetyl-amino-3 - [[2-methyl-1,3,4-thiadiazol-5-yl] thiomethyl] -3-cephem-4-carboxylic acid, Isomer syn, 7-i [2- (2-Amino-4-thiazolyl-2-methoxyiminoacetyl) amino} -3 - [(2-methyl-1,3,4-thiadiazol-5-yl) thiomethyl] -3-cephem- 4-carboxylic acid isomer syn, 7 - {[2- (2-amino-4-thiazolyl] -2-methoxyiminoacetyl] amino] -3- [(1-methyltetrahydrazol-5-yl) thiomethyl] -3-cefem, sodium salt 4-carboxylic acid, isomer syn, 3-acetylthiomethyl-7 - {[2- (2-amino-4-thiazolyl] -2-methoxyiminoacetyl] amino} -3-cephem-4-carboxylic acid, isomer syn and crystalline 3-acetylthiomethyl-7 - {[24 2-amino-4-thiazolyl) -2-methoxyiminoacetyl] amino} -3-cephem-4-carboxylic acid sodium salt, isomer syn.

The agents may be administered orally, rectally, parenterally, intramuscularly or locally by topical application to the skin and wet areas.

These may be solid or liquid and may be administered in pharmaceutical forms commonly used in human medicine, such as simple capsule tablets, jellies, granules, suppositories, injectables, pomades, creams, gels. They are prepared by conventional methods. Conventional excipients such as talc, acacia, lactose, starch, magnesium stearate, cocoa butter, aqueous and non-aqueous carriers, fats of animal or vegetable origin, paraffinic derivatives, glycols, various wetting agents, dispersants or emulsifiers may also be used and preservatives.

The dose administered varies depending on the disease being treated, the patient, the mode of administration and the agent being considered. For example, it may be between 0.250 g and 4 g per day, by the oral route in humans for the substance described in Example 2, or between 0.5 g to 1 g three times a day intramuscularly or intravenously.

Compounds of formula II are prepared by treatment with an acid of formula IV

A / W- /? _{y with} A _n

H

N

T or _l (IV) in which

R 1 and R 2 are as defined above, to 7-aminocephalosporanic acid, which reaction may be followed by acid hydrolysis or hydrogenolysis.

An example of such a procedure is given in the following! example parts.

Similarly, compounds of formula (II) in which the group OR2 is in the syn position are obtained starting from compounds of formula (IV) in which the group OR2 is in the syn position and reacted with 7-aminocephalosporanic acid.

The process according to the invention is explained in more detail in the following examples, which are illustrative only and do not limit the scope of the invention.

Example 1

7-j [2- (2-Tritylamino-4-thiazolyl) -2-methoxyiminoacetyl] amino] -3- [(1-methyltetrazol-5-ylthiomethyl) -3-cephem-4-carboxylic acid

To 2.7 g of 3-acetoxymethyl-7 - {[[2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetyl] amino] -3-cephem-4-carboxylic acid is added 0.624 g of primary sodium phosphate; 51 g of sodium bicarbonate, 1.32 g of 1-methyl-5-mercaptotetrazole, 40 ml of water and 20 ml of acetone. The mixture was left at 48 ° C overnight and at 60 ° C for one hour. After purification, the desired product is obtained in the form of the syn isomer. A sample was chromatographed on thin layer silica gel using the following chromatographic systems were determined, these values R _f:

For acetone with 5% water and

0.5 for ethyl acetate / ethanol / water: 70-20-20

Infrared (chloroform) \

C = O β-lactam: 1785 cm ^-1 /

NMR Spectrum:

(CD 3) δ ppm ppm of the trityl group = 7.3, the proton at the 5-position of the thiazole-6.7.

Example 1 a d 2

7- ((2- (2-Amino-4-thiazolyl) -2-methoxyiminoacetyl) amino) -3 - [(1-methyltetrazol-5-yl) thiomethyl] -3-cephem-4-carboxylic acid

Starting from 1,7 g of 3-acetoxymethyl-7 - {[2- (2-amino-4-thiazolyl) -2-methoxyiminoacetyl] amino] -3-cephem-4-carboxylic acid, 0,624 g of primary sodium phosphate, 51 g of sodium bicarbonate, 1.32 g of 1-methyl-5-mercaptotetrazole, 40 ml of water and 20 ml of acetone; the mixture was left overnight at 48 ° C and one hour at 60 ° C. In this way the desired product is obtained in the form of the syn isomer. 3-Acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl-2-methoxyiminoacetyl jamino) -3-cephem-4-carboxylic acid, isomer syn used in Example 1 and 3-acetoxymethyl-7-) The (2- (2-amino-4-thiazolyl) -2-methoxyiminoacetyl] amino) -3-cephem-4-carboxylic acid isomer used in Example 2 was obtained as follows:

a) 3-Acetoxymethyl-7-β- [2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetyl] amino} -3-cephem-4-carboxylic acid.

3.06 g. Of 2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetic acid sodium salt is placed in 65 ml of methylene chloride and 6.5 ml of 2 N hydrochloric acid, washed in water, dried and concentrated to give free acid.

The free acid obtained is dissolved in 30 ml of methylene chloride. 0.78 grams of dicyclohexylcarbodiimide was added and stirred for one hour at ambient temperature. The resulting dicyclohexylurea was removed, cooled to -10 ° C, and a solution of 1.01 g of 7-aminocephalosporanic acid in 13 ml of methylene chloride and 0.9 ml of triethylamine was added. Allow to warm to ambient temperature, add 1 ml of acetic acid, dehydrate, wash in water with hydrochloric acid, dry, concentrate, take up with 10 ml of dioxane, add 1 ml of water and 3 ml of saturated sodium bicarbonate solution. Stir, dehydrate, wash and concentrate, then take up in methylene chloride, wash in 10 ml of water and 5 ml of normal hydrochloric acid, decant, wash with water, dry, partition in ether to obtain 1.747 g of crude material which is purified by dissolution in ethyl acetate and subsequent precipitation in ether. 1.255 g of pure material is obtained.

The obtained substance has a syn configuration.

bj 3-Acetoxymethyl-7 - {[2- (2-amino-4-thiazolyl) -2-methoxyiminoacetyl] amino] -3-cephem-4-carboxylic acid.

0.975 g of the substance obtained in part a) is stirred for 10 minutes at 55 ° C in 4 ml of 50% formic acid. Add 4 ml of water, centrifuge and concentrate to dryness under vacuum. It is partitioned in 2 ml of ethanol, centrifuged, washed with ethanol and then with ether to give 0.428 g of pure substance.

The obtained substance has a syn configuration.

NMR (DMSO 60 MHz)?

2.03 - (C — CH3) doublet 9.58, J = 8 Hz,

O

6.76 (thiazole cyclone proton.

Example 1 a d 3

3-Acetylthiomethyl-7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacetyl] amino) -3-cephem-4-carboxylic acid, isomer syn.

A solution of 4.55 g of 3-acetoxymethyl-7 - {[2- (2-amino-4-thiazolyl) -2-methoxyiminoacetyl] amino) -3-cephem-4-carboxylic acid isomer syn, 1.7 g of thioacetate is formed. potassium phosphate, 2 g of anhydrous primary sodium phosphate and 20 ml of distilled water. Maintain at 70 ° C for one and a half hours. It is acidified with 2 ml of formic acid and extracted four times with 60 ml of methyl acetate (the insoluble component is filtered).

The organic solution is washed with water, dried and concentrated to dryness. The residue is dissolved in 20 ml of acetone with 10% water. 0.5 g of activated carbon are added. Stir 5 minutes, freed from coal, washed with acetone with 10% water, and the filtrate concentrated. The residue is worked up in neat ethanol, dried, washed with alcohol and then with ether. 1.9 g of the expected product are obtained in pure form.

NMR Spectrum:

(DMSO 60MHz)

3.85 (N-OCH 3),

6.75 (thiazole cycle proton)

Example 4

7 - {[2- (2-Amino-4-thiazolyl) -2-methoxyiminoacetyl] amino] -3- [(2-methyl-1,3,4-thiadiazol-5-yl) thiomethyl] -3- sodium salt cef em-4-carboxylic acid isomer syn

To a solution of 2.3 g of 7 - {[2- (2-amino-4-thiazolyl) -2-methoxyiminoacetyl] amino-3 - [(2-methyl-1,3,4-thiadiazolyl-5-yl) thiomethyl] 3-Cephem-4-carboxylic acid (isomer syn) in 5 mL of acetone was added aqueous sodium bicarbonate to pH 7. 0.4 grams of charcoal was added to the resulting mixture, and the mixture was stirred for 5 minutes. The solid was centrifuged and the residue was washed with aqueous acetone (1: 1).

Ethanol (50 ml) was added to the bleached filtrate and the mixture was concentrated to dryness under reduced pressure at 30 ° C. The residue is taken up in 5 ml of ethanol, the crystals are triturated, centrifuged and washed with ethanol and then with ether. 1.3 g of the expected product are obtained. Analysis:

Calculated: 4.18% Na

Found: 4.70 ’% Na

Ultraviolet spectrum:

(ethanol)

Max. 235 nm Ει! = 360

265 nm Eif = 322

Inflection 280 nm Ed = 295 ε = 19,800 · ε = 17,700 (ethanol, N / 10 hydrochloric acid)

Inflection 218nm El ¹ = 244

Λ max 266-267 nm λ = 406

Inflection 280 nm λ = 363 ε = 22 300

Example 5

7 - {[2- (2-Amino-4-thiazolyl) -2-methoxyiminoacetyl] amino] -3- [(1-methyltetrazol-5-yl) thiomethyl] -3-cephem-4-carboxylic acid, sodium salt isomer syn

3.35 g of 7 - {[2- (2-amino-4-thiazolyl) -2-methoxyiminoacetyl] anilino] -3 - [(1-methyltetrazol-5-yl) thiomethyl] -3-cephem The 4-carboxylic acid (isomer syn) was mixed with 7 ml of methanol and 7 ml of a 1 molar aqueous solution of sodium bicarbonate. The resulting mixture was stirred for 5 minutes at ambient temperature, the insoluble matter was centrifuged and washed twice with 1: 1 methanol / water.

While stirring, 85 ml of ethanol are first added, the sodium salt crystallizes and 170 ml of ether are added; The mixture was stirred for 10 minutes and washed with ethanol / ether (1: 1) followed by ether and drying to give 3.25 g of the desired product.

The resulting product is purified as follows:

The above salt is dissolved in 40 ml of water, and 0.6 ml of acetic acid is added to the solution thus obtained to a pH of 6.8 to 7; the solution is then diluted with ethanol, the solvents are distilled off under reduced pressure at a temperature below 35 ° C. The residue was taken up in ethanol to remove water and the mixture was evaporated to dryness.

The residue is taken up in 16 ml of methanol, the mixture is diluted with 160 ml of acetone and the salt is crystallized. After stirring for 5 minutes, the solid was centrifuged and washed with acetone and then ether.

2.3 g of the expected product are obtained. [α] _D ²⁰ = -13.5 ° + 1 (1% solution in water). Analysis:

Calculated: 4.31% Na

Found: 4.8 ° / o Na

Example 1 a d 6

3-Acetylthiomethyl-7 - {[2- (2-amino-4-thiazolyl) -2-methoxyiminoacetyl] amino (-3-cephen) -4-carboxylic acid, isomer syn

4.5 g of 3-acetylthiomethyl-7 - {[2- (2-amino-4-thiazolyl) -2-methoxyiminoacetyl] amino]-3209879 is mixed at room temperature.

1S

4-carboxylic acid (isomer syn) with 9 ml of methanol and 9 ml of a 1 molar solution of sodium bicarbonate in water. The mixture is stirred for 5 minutes at ambient temperature, the solid is centrifuged and rinsed with a 1: 1 mixture of methanol and water. 110 ml of ethanol are then added with stirring, the sodium salt crystallizing out. The mixture was diluted with 220 ml of ethyl ether, stirred, centrifuged and washed with ethanol / ether 1: 1 and then with ether.

The obtained sodium salt is purified as follows:

The product is dissolved in 40 ml of water and a few drops of acetic acid are added to the solution thus obtained to obtain a pH of 6.8 to 7. The solution is diluted with 100 ml of ethanol and the solvents are evaporated off under reduced pressure at a temperature below 35 °. The concentrate is taken up twice in 50 ml of ethanol and then evaporated to dryness a second time. The residue is dissolved in 15 ml of methanol, the solid is filtered off and the methanolic solution is diluted with 150 ml of acetone.

The sodium salt crystallized, stirred for 5 minutes, then removed, washed with acetone and then with ether. The resulting residue is dried under reduced pressure to give 1.8 g of the desired product.

[α] ο ²⁰ = - 31 ° + (0,6% solution in water)

Calculated: 4.65%

Found: 4.9%

Ultraviolet spectrum:

ethanol)

Max 235 nm-El ¹ = 419;

Inflection 260 nm Ei ¹ = 343

Inflection 300 nm Ei ^{1 =} = 122 (ethanol / N / 10 hydrochloric acid)

Inflection 230 nm λ = 280

Max. 263 nm Eit = 391 ¢ = 19,700

Example 1 a d 7

(Microcrystalline) sodium salt of 3-acetylthiomethyl-7 - ([2- (2-amino-4-thiazolyl) -2-methoxyiminoacetyl] amino) -3-cephem-4-carboxylic acid isomer syn

471.5 mg of 3-acetylthiomethyl-7 - {[2- (2-amino-4-thiazolyl) -2-methoxyiminoacetyl] amino] -3-cephem-4-carboxylic acid (isomer syn) is mixed with 1.5 ml of a 1 molar solution of anhydrous sodium acetate in anhydrous methanol. To the obtained mixture is added

0.5 ml of ethanol, the insoluble matter is centrifuged, 2 ml of ethanol are added, the sodium salt crystallizes. This salt is centrifuged, washed with methanol and dried in a vacuum oven at 45 ° C.

0.25 g of n-hygroscopic salt is obtained. Analysis:

Calculated: 4.66%

Found: 4.6%

Example 8

7 - ((2- (2-Amino-4-thiazolyl) -2-methoxyiminoacetyl) amino) -3 - [(1-methyltetrazol-5-yl) thiomethyl] -3-cephem-4-carboxylic acid isomer syn g 3-acetoxymethyl-7 - ((2- (2-amino-4-thiazolyl) -2-methoxyiminoacetyl) amino) -3-cephem-4-carboxylic acid (isomer syn) is dissolved in 100 ml of water. 3.5 g of secondary sodium phosphate and 3 g of 1-methyl-5-mercaptotetrazole are then added to the solution.

The resulting mixture was heated to 60 degrees Celsius for two hours and thirty minutes, with 2 g of secondary potassium phosphate added at the end of the first hour.

The mixture is acidified with about 20 ml of 2 N hydrochloric acid to pH 3-4, centrifuged, washed and dried to give 8 g of crude product. This was extracted three times with 100 ml of 5% aqueous acetone solution and the insoluble material was centrifuged off. The solution is distilled to dryness; the residue is taken up in 20 ml of ethanol, after separation of the ethanol, 5.8 g of semi-purified product is obtained which is chromatographed on silica gel with a 10% aqueous acetone solution.

The product is identical to that of Example 2. NMR Spectrum:

[(CD5) 2SO] ppm 3.83 (N-OCH 3),

6.73 (thiazole cyclone proton.

Example 9

7 - {[2- (2-Amino-4-thiazolyl) -2-methoxyiminoacetyl] amino} -3 - [(1-methylteitrazol-5-yl) thlomethyl] -3-cephem-4-carboxylic acid sodium salt in the form of the isomer syn g of the semi-purified product obtained in the previous example 8 is stirred for 15 minutes in 17 ml of methanol and 28 ml of a 1 molar solution of sodium acetate in methanol are added thereto.

9 ml of ethanol are then added to the mixture and the insoluble impurity is centrifuged. Methanol is distilled off from the residue to give a small volume of solution. 20 ml of ethanol are added, centrifuged and rinsed with ether to give 6 g of crude product.

This product is chromatographed to give about 3.5 g of pure product which is identical to the product of Example 5. [α] D ²⁰ = -13.5 ° + 1 (1% solution in water)

Calculated: 4.31% Na

Found: 4.8% Na

Example 10

7 - {[2- (2-Amino-4-thiazolyl) -2-methoxyiminoacetyl] amino] -3- [(2-methyl-1,3,4-thiadiazol-5-yl) thiomethyl] -3-cephem 4-carboxylic acid isomer syn g 3-acetoxymethyl-7 - {[2- (2-amino-4-thiazolyl) -2-methoxyiminoacetyl] amino] -3-cephem-4-carboxylic acid sodium salt (isomer syn The mixture is dissolved in 120 ml of water, and 3.2 g of 1-methyl-5-mercapto-1,3,4-thiadiazazole and 3.5 g of secondary potassium phosphate are added to the solution thus obtained. The mixture was stirred for two hours at 60 ° C, at the end of the first hour 1 g of secondary potassium phosphate was added. The mixture was acidified with about 20 milliliters of 2N hydrochloric acid to reach a pH of 2-3 after being cooled to 35 ° C. The product formed is centrifuged, washed with water and dried to give 9.2 g of crude product.

This product is extracted three times with 100 ml of 5% aqueous acetone, each time insoluble impurities are centrifuged off.

The residue is concentrated to dryness, washed with 20 ml of alcohol to give 7 g of semi-purified product.

This product was chromatographed on silica gel using 10% acetone in water.

NMR Spectrum:

t (CD3) SO] ppm:

3.85 [= N - O - CH 3);

6.76 [thiazole cycle proton)

Example 11

7 - {[2- (2-Amino-4-thiazolyl) -2-methoxyiminoacetyl] amino [-3 - [[2-methyl-1,3,4-thiadiazolyl-5-yl) thiomethyl] sodium salt The 4-carboxylic acid isomer of syn g of the semi-purified product obtained in Example 10 above was mixed with 14 ml of methanol, and then 19 ml of sodium acetate in the form of a one molar solution in methanol was added. Then 6 ml of ethanol are added. The insoluble matter formed by impurities is centrifuged. The residue was concentrated and washed with ethanol to give 4.2 g of the sodium salt.

This product is chromatographed to give 2.5 g of product identical to that of Example 4.

Analysis:

Calculated: 4.18% Na

Found: 4.70% Na

Ultraviolet spectrum:

(ethanol)

Max. 235 nm E1 ^: = 360

265 nm? = 322

Inflection 280 nm E ^1: = 295 ε = 19 800 ε = 17 700 (ethanol / N / 10 hydrochloric acid)

Inflection 218 nm Ei ^x = 244

Max. 266-267 nm Ei * = 406

Inflection 280 nm λ = 363 ε = 22 300

Example 12

An injectable pharmaceutical preparation of the following composition is prepared:

- 7 - {[2- [2-amino-4-thiazolyl) -2-methoxyiminoacetyl] amino} -3 - [(1-methyltetrazol-5-yl) thiomethyl] -3-cephem-4-carboxylic acid 500 mg - sterile aqueous vehicle 5 ml

Example 13

An injectable pharmaceutical preparation of the following composition is prepared:

3-acetylthiomethyl-7 - {[2- [2-amino-4-thiazolyl) -2-methoxyiminoacetyl] amino] -3-cephem-4-carboxylic acid 500 mg - sterile aqueous vehicle 5 ml

Example 14

Gelatin tablets having the following composition are produced:

- 7 - {[2- (2-amino-4-thiazolyl) -2-methoxyiminoacetyl] amino] -3 - [(1-methyltetrazol-5-yl) thiomethyl] -3-cephem-4-carboxylic acid 500 mg - vehicle, supplemented to 400 mg

Example 15

Gelatin tablets of the following composition were produced:

- 3-acetylthiomethyl-7 - {[2- (2-amino-4-thiazolyl) -2-methoxyiminoacetyl] amino] -3-cephem-4-carboxylic acid 250 mg - vehicle, made up to 400 mg

Pharmacological tests with products according to the invention

A) In vitro potency

Dilution method in liquid medium

Pour the same amount of sterile nutrient medium into a series of tubes. Thereafter, increasing amounts of the test product were added to each nutrient tube and each tube was inoculated with the bacterial strain listed in the following tables.

After incubation for 24 and 48 hours in an oven at 37 ° C, the inhibition of bacterial strain growth is evaluated in transmitted light by determining the minimum inhibitory concentration (MIC), expressed in µg / ml.

The results are shown in the following tables.

Table 1

Bacterial strain The product of Example 10

MIK (tg / ml) hours 48 hours

Staphylococcus aureus ATCC 6538, non-resistant against penicillin 1 1 Staphylococcus aureus UC 1128, penicillin resistant 2 2 Staphylococcus aureus n ° 54 146 2 2 Streptococcus pyogenes A 561 š0.02 š0.02 Streptococcus faecalis 5 432 1 2 Streptococcus faecalis 98 F 74 2 5 Bacillus subtilis ATGC 6,633 0.6 1 Tetracycline-resistant Escherichia Coli, ATCC 9637 0.6 0.6 Escherichia Coli resistant to tetracycline, ATCC 11 303 0.1 0.2 Escherichia Coli Exp. ΤΟ & βΒβ 0.4 0.4 0.4 Gentamicin and Tobramycin resistant Escherichia Coli R 55 123 D 0.4 Klebsiella pneumoniae Exp · 52,145 0.05 0.1 Klebsiella pneumoniae 2,536 gentamycin resistant 0.6 1 Proteus mirabilis (indole—) A 235 0.2 0.2 Salmonella typhlmurium 420 | 0.6 0.6

Table 2

Bacterial strain

The product of Example 2 MIK ((Ug / ml) hours 48 hours)

Staphylococcus aureus ATCC 5,638 Stainless Steel to Penicillin Staphylococcus aureus UC 1128 resistant 1 and against penicillin 2 2 Staphylococcus aureus exp. n ° 54 146 2 2 StreptóCbccliS pyogénfeS A 561 š0.02 š0,2 streptococcus faecalis 5432 2 2 Streptococcus faecalis 98 F 74 2 10 Bacillus subtilis ATCC 6633 2 5 Escherichia Coli ATCC 9637 resistant to Tetracycline 0.4 0.4 Escherichia Coll tetracycline resistant ATCC 11,303 0.05 0.05 Escherichia Coll. Exp. TO26B6 0.2 0.2 Escherichia Coli resistant to gentamycin and tobramycin R 55 123 Ď 0.2 0.2 Klebsiella pneumoniae Exp. 52 145 0.02 0.02 Klebsiella pneumoniae 2536 gentamycin resistant 0.4 0.6 Proteus mirabilis (indole—) A 235 0.05 0.05 Proteus vulgaris (indole +) A 232 3 20 May Enterobacter cloacae 681 40 40 Providencia Du 48 2 5 Serratia Resistance to Gentamycin 2,532 1 1 Salmonella typhiihurium 420 0.2 0.2 Enterobacter cloacae 681 20 May 20 May Providencia Du 48 2 2 Pseudomonas 3,935 Exp. SG 20 May 40 Gentamycin-resistant Serratia 2532 1 1

208879

Table 3

Bacterial strain

The product of Example 3 MIC (pg / ml) hours 48 hours

Staphylococcus aureus

ATCC 6538 resistant to penicillin 2

Staphylococcus aureus

UC 1128 resistant to penicillin 5

Staphylococcus aureus exp. n ° 54 146 2

Streptococcus pyogenes

A 561 š0.02

Streptococcus faecalis> 5432 5

Streptococcus faecalis

5432 5

Streptococcus faecalis * 99 F 75 5

Bacillus subtilis

ATCC 6633 1

Escherichia Coli resistant to tetracycline

ATCC 9637 '0.8

Escherichia Coli resistant to tetracycline

ATCC 11303 0.1

Escherichia Coli

Exp. T26B6 0.4

Gentamycin and Tobamycin resistant Escherichia Coli R 55 123 D 0.6

Klebsiella pneumoniae

Exp. 52 145 0.1

Gentamycin 1 Klebsiella pneumoniae 2536

Proteus mlrabilis (indole—)

A 235 0.05

Proteus vulgaris (indole +)

A 232 5

Salmonella typhimurium

420 0,4

Enterobacter cloacae

J 682 10

Providencia

Du 48

Gentamycin-resistant Serratia 2,532 1 w0.02 ·

0.6

0.1

0.4

0.6

0.1

0.1

0.4

1S

Table 4

Bacterial strain

Example 6 product _: MIC (Mg / ml) hours 48 hours

Staphylococcus blanc pathogene 69,396 Staphylococcus blanc 21,976

Staphylococcus dore 68 666

Escherichia Coli 73

Escherichia Coli 72,806

Escherichia Coli 69,916

Escherichia Coli 69 885

Escherichia Coli 68 697

Escherichia Coli 68,956

Escherichia Coli 21,976

Escherichia Coli 19,976

2 2 3 5 3 5 0.2 0.2 1 1 0.4 0.4 0.6 1 0.4 0.4 0.4 0.4 0.6 0.6 0.6 0.6

B) Experimental infection of Escherichia Coli TO26B6

In this test, the effect of the product of Example 3 on experimental infection of Escherichia Coli in mice was studied. Groups of male mice of an average weight of 22 g were infected by intraperltoneal injection of 0.5 ml of a 24-hour culture of Escherichia Coli strain

TO26B6 (Institut Pasteur) on broth, diluted to 1 / 5.5 with distilled water.

One hour, five hours, and twenty-four hours after said infection, the test animals are given the amount of test product administered or administered os orally.

Mortality of the test animals is observed over the next eight days; the results obtained are shown in Table 5 below.

Table 5 (subcutaneous application)

Mortality after Survived mice after

21.5 li 24 h 24.30 h 25.30 h 28 li 47 h 54 h 70 h 5 days 6 days 8 days o

τ-Η

IR rH cd cd o

r-1 £

o r-l

O

F — t F — H

IH> — — I>--i> SI n§ cu, N o N + -> s / + - ¹ 3 co 3 ό, g 0 0 ti ¹ ti CU CO Oj

The CO honors cu cuI

-O «

oo ea ^χ 3 ^ 3Χ co 3 3 co, Ό O Ό,

I S x g 1 co Oj co CU co

9987 9

C] Experimental infection of Klebsiella pneumonlae

In this test, the effect of the product of Examples 2 and 3 on experimental infection of Klebsiella pneumoniae strain in mice was studied. Groups of ten male mice of 21 g average weight were infected by intraperitoneal injection of 0.5 ml culture of Klebsiel22 la pneumoniae 52145 on nutrient broth, diluted to 1/1000 with distilled water.

Half an hour, four hours, eight hours, twenty-four hours and three hours after infection, the animals were dosed subcutaneously with a given amount of test product.

Animal mortality was observed for the next ten days, and the results are shown in Table 6 below.

# 8873

Mortality after Survived mice after

28.5 h 32 h 46 h 48 h 49 h 49.5 h 5th day 10th day

Control animals 1 1 5 111 0/10

The product of Example 2

O ό O'· © © rd rd rd rd rd σ> O cb O rd rd ^! T ~ ť rd

Co what

T3 the one that the one. n SO N> - <

and

S ^N and 60 N

4Ti with Ti '60 S ^{N H} ^ 4 in RSJ io 3 rd 3 cm @ 3 * - »rf RFTI 3 O 3 3 rt tf uOyOyOyQy

NμΛμΛ · _Η Λ ^ Λ tnMnftinfcinfcw bp

D) Experimental infection with Klebsiella pneumoniae

In this test, the effect of the products of Examples 4 and 6 on experimental infection of the Klebsiella pneumoniae strain in mice was studied. Groups of ten male mice of 21.5 g average weight were infected by intraperitoneal injection of 0.5 ml of a 24-hour culture of Klebsiella pneumoniae 52 145 strain on nutrient broth, diluted to 1/1000 with distilled water.

One hour, five hours, and twenty-four hours after the infection, the test animals were injected subcutaneously with the determined amount of test product.

The mortality of the test animals was observed over the next 10 days and the results are shown in Table 7 below.

Table 7 o

> tn 3>. t gxj o d> N

O

PH pi a

Td d

d

Td

ÍO d

d

Td

ID

M

CZ)

About £ xj

S o .O t>.

what

LIMB

Q what

CM xfl 'O, S ca o

rH2O

O O O O O O O O O tH r-1 rH rH rH rH rH rH rH CD O O d CD CD O O O rH rH rH rH rH rH

d

Td ca d

«3 «and ι — I

WHAT WHAT WHAT WHAT d d d d d d Ό Td Ό Td Td Td I d ca ca ca ca ř — H with 3 -hd £! £! > C > I-J E > C and cu CU cu cx cu N N N N N N

co d

Ό ca

Claims (2)

SUBJECT OF THE INVENTION 1. A process for the preparation of the novel alkoxy derivatives of 7- [2- (2-amino-4-thiazolyl) -acetamido] -cephalosporanic acid of the formula I: NH-Ri O II C 'NH N OR in which R represents an acyl group having 2 to 4 carbon atoms or a 2-methyl-1,3,4 thiadiazolyl or a 1-methyltetrazolyl radical, R 1 represents a hydrogen atom or a trityl radical, R2 represents a saturated or unsaturated alkyl radical having 1 to 4 carbon atoms, A represents a hydrogen atom or an alkali metal, alkaline earth metal, magnesium or a pharmaceutically acceptable amine organic base equivalent, wherein the wavy line means that the OR2 group may be in one of two possible syn or anti positions, A compound of the formula (II) in which they are present R 1 and R 2 as defined above have the effect of R 3-SH (III) wherein: R is as defined above to give a compound of formula (Γ), optionally converting it into a salt, to give a compound of formula (I) wherein R, R 1 and R 2 are as defined above and A is an alkali metal, alkaline earth metal equivalent, magnesium or a pharmaceutically acceptable aminated organic base. 2. A process according to item 1, for the preparation of a compound of formula I, wherein R, R1, R2 and A are as defined in item 1 a. Wherein the group OR2 is in the syn position, characterized in that use a compound of formula (II) wherein R 1 and R 2 are as defined in point 1 and a group OR2 is in the syn position.