CS207575B2 - Method of preparation of new oxime derivates of the 7-aminothiazolylacetamidocephalsporan acid - Google Patents

Abstract

New cephalosporins are of formula (I) In the formula R and R1 are both H or both hydrolysable or hydrogenolysable gps. or R1 can also be opt. substd. 1-4C alkyl, A is H or an equiv. of an alkali(ne earth) metal or an amine. (I) have antibacterial activity against Gram-positive and -negative bacteria, esp. penicillin-resistant Staphylococci, E.coli, Klebsiella, Salmonella and Proteus spp. A typical cpd. is 3-acetoxymethyl-7-(2-(2-tritylamino-4-thiazolyl)-2-tritylhydroxyi- mino-acetamido)-3-cephem-4-carboxylic acid. In an example, this is prepd. starting from ethyl gamma-chloro-alpha-oximinoacetyl-acetate.

Description

The invention relates to a process for the preparation of novel oxime derivatives. 7-aminothiazolylacetaimidocephalosporanic acids having significant antibiotic activity on Gram positive bacteria such as staphylococci and streptococci, especially penicillin-resistant staphylococci, and Gram negative bacteria such as coliforms, Kleionellate Proteus, Klebsellate .

SUMMARY OF THE INVENTION The present invention provides a process for the preparation of novel oxime derivatives of 7-aminothiazolylacetamido-phthalosporanic acid of general formula Г

in which

Rc represents a hydrogen atom, a trityl group or a chloroacetyl group,

R 'c represents a hydrogen atom, a trityl group or an unsaturated alkyl group having 1 to 4 carbon atoms,

A represents a hydrogen atom or an equivalent of an alkali metal, an alkaline earth metal, magnesium or an amine organic base, and a wavy line means that the group OR'c can be in one or the other of the two possible syn or anti positions, where R ' c is trityl, then Rc is trityl, and when R'c is hydrogen, then Rc is hydrogen, which comprises reacting 7-aminocephalosporanic acid of the formula

CH 3 O-C-CH 3

With an acid of formula IIc

(II cl or with a functional derivative of this acid, wherein in said general formula IIc) in which:

Rd is trityl or chloroacetyl;

R 'd represents a trityl group or a saturated or' unsaturated alkyl group having 1 to 4 carbon atoms' to give the product of formula Ic nh-r

(lei in which

Rd and R’d are as defined above, which product corresponds to the product of formula Γ in which

Rc is trityl or chloroacetyl; and

R 'c represents a trityl group or an unsaturated or saturated alkyl group having 1 to 4 carbon atoms, whereupon the above product of formula Ic is hydrolysed in an acidic medium, or · hydrogenolysed or is converted to a. according to the meanings of the general substituents Rd and, R'd, .kk, to obtain the product of the formula. [0057] Ib

in which

R 't represents a hydrogen atom or a saturated or unsaturated alkyl group having a total number of carbon atoms of 1 to 4, said product corresponding to the product of formula Γ in which:

Rc and A are each hydrogen and

R 'c represents a hydrogen atom or a saturated or unsaturated alkyl group having 1 to 4 carbon atoms, whereupon the products of the formulas Ic or Ib are optionally converted into salts.

The present invention also relates to a process for preparing the products of formula (I)

(II in which

R is hydrogen or trityl,

R 'represents a hydrogen atom, a trityl group or a saturated or unsaturated alkyl group having 1 to 4 carbon atoms,

A represents a hydrogen atom or an equivalent of an alkali metal, an alkaline earth metal, magnesium or an amine organic base, and the wavy line indicates that the OR 'group may be present. in one or other of the possible positions syn or anti, wherein when R 'is trityl, then R is trityl, and when R' is hydrogen, then R is atom · Hydrogen, which comprises reacting 7-aminocalphalosporanic acid of the formula

with an acid of formula II

S ^ N

ORI (II) where

R _t represents a trityl group a

Rf represents a trityl group or a saturated or unsaturated alkyl group having 1 to 4 carbon atoms, or a functional derivative thereof, to give the product of formula (Ia)

_R 'and R _f' are as defined above, said product corresponds to a product of formula I wherein

R is trityl and

R 'represents a trityl group or a saturated or unsaturated alkyl group having 1 to 4 carbon atoms, whereupon the product of formula (Ia) is optionally hydrolysed in an acidic medium or * is hydrogenolyzed to give the product of formula (Ib)

(1Ы in which

Rj 'represents a hydrogen atom or a saturated or unsaturated alkyl group having a total number of carbon atoms of 1 to 4, said product corresponding to the product of formula (I) in which:

R and A are each hydrogen and

R 'represents a hydrogen atom or a saturated or unsaturated alkyl group having 1 to 4 carbon atoms, whereupon the products of formulas Ia and Ib are optionally converted into salts.

The present invention also provides a process for the preparation of the products of formula (Ib)

in which

Rf 'represents a hydrogen atom or a saturated or unsaturated alkyl radical having 1 to 4 carbon atoms, which product corresponds to the product of the general formula Г in which:

Rc and A are each hydrogen and

Rc 'means a hydrogen atom or a saturated or unsaturated alkyl group having 1 to 4 carbon atoms, characterized in that the acid is treated with a salt of the formula Ic in which:

Rd represents a trityl group and

R d represents a trityl group or a saturated or unsaturated alkyl group having 1 to 4 carbon atoms, to form the product of formula Ib, or treatment with a salt of the product of formula Ic in which

Rd represents a trityl group and

R d denotes a trityl group or a saturated or unsaturated alkyl group having a total number of carbon atoms of 1 to 4, to form a salt of the product of formula Ib, or treating with a thiourea the salt of the product of formula Ic in which

Rd represents a chloroacetyl group and a saturated or unsaturated alkyl group having 1 to 4 carbon atoms, to form a salt of the product of formula Ib.

The present invention also provides a process for the preparation of the products of formula (Ib)

(Ib / in which

Rf 'means a hydrogen atom or a saturated or unsaturated alkyl group having a total number of carbon atoms of 1 to 4, which product corresponds to the product of formula (I) in which:

R and A are each hydrogen and

R 'represents a hydrogen atom or a saturated or unsaturated alkyl group having a total number of carbon atoms of 1 to 4, characterized in that the acid is treated with a salt of the product of formula (Ia) in which:

R ₁₁ represents a trityl group and

Rf represents a trityl group or a saturated or unsaturated alkyl group having a total number of carbon atoms of 1 to 4, to form a product of formula Ib, or is treated with a hydrogenolyzing agent to form a salt of the product of formula Ia in which:

R 1 represents a trityl group and

R ₁ 'represents a trityl group or a saturated or unsaturated alkyl group having 1 to 4 carbon atoms, to give the product of formula (Ib).

Examples of the general substituents R c 'and R' denoting a saturated or unsaturated alkyl group having 1 to 4 carbon atoms include methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, vinyl, propenyl, butenyl, ethynyl and propargyl.

Examples of particular substituents of general substituent A include, but are not limited to, sodium, potassium, lithium, calcium, or magnesium; Examples of organic bases which may be represented by the general substituent A include trimethylamine, triethylamine, methylamine, propylamine, N, N-dimethylethanolamine, tris (hydroxyinethylamino) ethane, arginine or lysine.

Examples of products of formula (I) and products of formula (I) are those described in the Examples below, and in particular:

3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -4-methoxyiminoacetamido] -3-cephem-4-carboxylic acid, 3-acetoxymethyl-7- [2- (2) 3-acetoxymethyl-7- [2- (2-amino-4-thiazoilyl) -2-methoxyimino] -amino-4-thiazolyl) -2-methoxyethoxyacetamido] -3-cephem-4-carboxylic acid; oacetamido -] - 3-cephem-4-carboxylic acid, isomer · syn, - sodium 3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido] -3-cephemate 4-carboxylic acid, isomer syn, - 3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoactactamido] -3-cephem-4-carboxylic acid, - sodium 3-acetoxymethyl acid -7- [2- (2-Amino-4-thiazolyl) -2-methoxy-aminoacetamido] -3-cephem-4-carboxylic acid obtained as described in the example, - crystalline sodium salt of 3-acetoxyimethyl -7- [2- (2-31-410-4--10820-4) -2-methoxyiminoacetamido] -3-cephem-4-carboxylic acid isomer · 3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2- ethoxyiminoacetamido] -3-cephem-4-carboxylic acid isomer syn and its salts with alkali metals, alkaline earth metals, magnesium or amine organic bases - 3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2- ((2-propenyl) oxyimino) -4-tertamido] -3-oephem-4-carboxylic acid, isomer syn and its alkali metal salts, alkaline earth anchors, magnesium or amine organic bases and 3-acetoxymethyl-7 - [2- (2-Amino-4-th-azolyl) -2- [(1-methylethoxy- [eta] < 6 > -acetamide]] -3-cephem-4-carboxylic acid isomer syn and its alkali metal, metal salts alkaline earths, magnesium · or amine · organic bases.

It goes without saying that the products of formula I may exist:

- either in the form given by formula (I) or - in the form of products of formula (Iz)

in which

R, R ', and A are as defined above.

It goes without saying that the products of the general formula rovněž may also exist in the two forms indicated for the products of the general formula I.

In a preferred embodiment of the process for preparing the products of formula (I) or formula (I) according to the invention, the 7-aminocephalosporanoic acid is treated with a functional derivative of the acid of formula (IIc) or II, such as an anhydride or chloride thereof. in situ by treating the acid with isobutyl chloroformate or dicyclohexylcarbodiimide.

Other halides or other anhydrides formed in situ by the action of other alkyl chloroformates, dialkylcarbodiimides or other dicycloalkylcarbodimides may also be used. It is also possible to use other acid derivatives, such as, for example, acid azide, activated acid amide or activated acid ester formed, for example, with hydroxysuccinimide, p-nitrophenol or 2,4-dinitrophenol. In case the reaction of 7-amtoocefalosporanové haloigenidem performed with an acid of formula ¹ or II IIc · · or anhydride formed with a chloroformate isobutylnatým then * is preferably conducted in the presence of a basic · · agent.

As the basic agent, for example, an alkali metal carbonate or a tertiary organic base such as N-methylmorpholine, pyridine or trlalkylamine such as triethylamine can be used.

Examples of the acid hydrolyzing agent which optionally treats the products of formula Ic or Ia include formic acid, trifluoroacetic acid. or acetic acid. These acids can be used in anhydrous media or aqueous solutions. In particular, the zinc / acetic acid system is a hydrolyzing agent.

Preferably, anhydrous trifluoroacetic acid or an aqueous solution of formic acid or acetic acid is used to cleave the trityl groups.

Treatment of the product of formula Ic with thiourea in which

Rd represents a chloroacetyl group and

R 6 represents a chloroacetyl group or a saturated or unsaturated alkyl group having a total number of carbon atoms of 1 to 4, preferably in a neutral or acidic environment. This type of reaction has been described by Masaki [J. A. C. S. 90,4508 (1968)].

The products of formula (Ic), (Ia) or (Ib) may be converted to the corresponding salts by methods known per se. Conversion of said acids to the corresponding salts can be effected, for example, by the action of a mineral base such as sodium or potassium hydroxide or sodium bicarbonate, or a salt of a substituted or unsubstituted aliphatic carboxylic acid such as diethylacetic acid, ethylhexanoic acid, and in particular acetic acid.

Preferred salts of the above acids are sodium salts.

Conversion of acids to the corresponding salts can also be accomplished by treatment with an organic base such as triethylamine.

Solvates of the free acids can also be used as starting materials for the preparation of salts. Examples of such solvates are, for example, those obtained with water, formic acid or alcohol.

Solvates with alcohol, especially ethanoium, can also be obtained, for example, by treating the solvate formed with formic acid with a mixture of alcohol and water, and then concentrating the resulting solution.

Said conversion of the acids to the corresponding salts is preferably carried out in a solvent or a mixture of solvents such as water, ethyl ether, methanol, ethanol or acetone.

The salts are obtained in amorphous or crystalline form according to the reaction conditions used.

Crystalline salts are preferably prepared by reacting the free acids or solvates thereof formed, for example, with formic acid or ethanol, and one of the above-mentioned aliphatic carboxylic acid salts, preferably sodium acetate.

To prepare the sodium salt, the reaction is carried out in a suitable organic solvent such as methanol, and the solvent may contain a small amount of water.

In addition, amorphous salts can be converted to crystalline salts. To this end, the amorphous sodium salt, which may be in the form of solvates with, for example, 0.5, 1 or 1.5 moles of water, is dissolved in a suitable organic solvent, preferably in a low molecular weight alcohol such as methanol whereupon the crystallization is carried out either by direct addition or by addition of other solvents such as ethanol, isopropanol, n-butanol, acetone, ethers and, in general, water-miscible organic solvents.

If the starting product, the solvent, or both contain water, then a crystalline salt can be obtained in the form of a hydrate; for example, the crystalline N, N -cetoxymethyl-7- [2- (2-amine-4-thiazolyl ⁾ oil] -2-methoxyiminoacetamido] -3-cephem-4-carboxylic acid, found in in the isomeric form syn, may be isolated with, for example, 0.5, 1 or 1.5 moles of water.

The products of formula (IIc) in which the general substituents are as defined above may be prepared by reacting the product of formula (IV)

The products of formula (II) in which the general substituents are as defined above may be prepared by reacting the product of the general formula; of formula IV

OR (IV) wherein

R 'represents a hydrogen atom, a trityl group or a saturated or unsaturated alkyl group having 1 to 4 carbon atoms, and

Alk represents an alkyl group having 1 to 4 carbon atoms, acting either as a functional derivative of the trityl group or, in the case where 'R' represents a hydrogen atom or a saturated or unsaturated alkyl group having a total number of 'carbon atoms 1 to 4, a chloroacetyl group derivative to give the product of formula (Vc)

CO ^ Alk (ivl in which in which

R 'means hydrogen atom, trityl. or a saturated or unsaturated alkyl group having 1 to 4 carbon atoms; and

Alk represents an alkyl group having 1 to 4 carbon atoms, acting as a functional derivative of the trityl group to give the product of formula V

AND"

NH-Rd

, OR d (Vc)

Rd is trityl or chloroacetyl and

R 6 represents a trityl group or a saturated or unsaturated alkyl group having 1 to 4 carbon atoms, where R 6 represents a chloroacetyl group. then. Rd represents a chloroacetyl group, whereupon. the product of formula Vc · · is treated with a base and a. Then with an acid to give the product of formula T1e

NH-Rd with N 1 = 7 every ₂ h

D ORd (HO in which '

Rd and Rd are as defined above.

R 1 represents a trityl group and

Rf represents a trityl group or a saturated or unsaturated alkyl group having 1 to 4 carbon atoms, after which the product of formula V is treated with a base and then. with an acid to give the product of formula II

NířR ₁

CO.H

ORj (II) in which

R1 and Rf are as defined above.

In the process for preparing the products of formula IIc or products of formula II according to the invention, trityl chloride is preferably used as a functional derivative of the trityl group in the presence of triethylamine or other tertiary amine bases such as trialkylamines, methylmorpholine or pyridine.

The functional derivative of the chloroacetyl group is preferably chloroacetic anhydride or a halide such as monochloroacetyl chloride.

When the reaction to obtain the product of formula (Vc) is carried out with chloroacetyl halide, the reaction is preferably carried out in the presence of a basic agent which is identical to the basic agents already cited above.

The base used to saponify the product of formula V or formula Vc is preferably sodium hydroxide; in addition, other bases such as potassium carbonate or barium hydroxide may be used.

Dilute hydrochloric acid is preferably used as the acid to be used for the isolation of the acid of formula (II) or formula (IIc); in addition, acetic acid or formic acid may also be used.

The products of formula (IV), in which the general substituents are as defined above, may be prepared by treating the product of formula (III) with thiourea.

СГС / ^ - С-С-СС ^^ Ц 'о и

OR (III) wherein

R 'represents a hydrogen atom, a trityl group or a saturated or unsaturated alkyl group having a total number of carbon atoms of 1 to 4, to form after the base treatment of the product of formula IV

in which

R 'and Alk are as defined above.

In a preferred embodiment, potassium acetate is used as the base to be used to obtain the product of formula (IV). It is also possible to use alkali metal carbonates and bicarbonates such as sodium carbonate or potassium carbonate in a dilute aqueous solution.

It has been found that the configuration of the syn or anti products of formula Г and the products of formula I is guided by the configuration of the products of formula IV, since the configuration of products obtained from products of formula IV can be maintained during synthesis.

The same is true, of course, of the products obtained from products of formula (IVa) and products of formula (IV), since these products are included in formula (IV).

The configuration of the products of formula IV depends on maintaining a certain number of conditions during the preparation of the products of formula IV.

It has also been found, for example, that if thiourea is applied to the products of formula (III):

(a) either in an aqueous solvent such as an aqueous acetone solution or an aqueous ethanol solution;

b) or at ambient temperature and using a substantially stoichiometric amount of thiourea at a relatively short reaction time of about 1 to 3 hours

(c) or, subject to all of the above conditions (a) and (b), the syn configuration isomer is obtained.

Thus, the products of formula (IV) prepared in Examples 3, 8, 10, 12 and 21 are syn.

The products of formula (I), in particular products of formula (I) in which the substituents are as defined above, in particular those of the syn configuration formulas, have very good antibiotic activity on Gram positive bacteria such as staphylococci and streptococci, especially penicillin resistant. staphylococci, and on the other hand Gram-negative bacteria, such as coiiform bacteria, Klebsiella, Salmonella and Proteus.

In view of the above characteristics, the products may be of use as medicaments in the treatment of diseases caused by germs, in particular in the treatment of diseases caused by staphylococcal infestations, such as staphylococcal blood poisoning, pyodermitis, septic and septic wounds, anthrax, phlegmones, erisipelia, influenza, bronchopneumonia and pneumonia.

Said pharmaceutically acceptable products may also be used as medicaments in the treatment of collibacilos and associated infections, in the treatment of infections caused by Proteus, Klebsiella and Salmonella microorganisms and in the treatment of other diseases caused by gram-negative bacteria.

The pharmaceutically acceptable products of formula (I) may be used in the preparation of pharmaceutical compositions containing, as an active ingredient, at least one of said products and in particular one of the syn configuration products described in the examples.

These preparations can be administered orally, rectally, parenterally, or topically topically by application to the skin or muscles.

Said preparations may be in solid or liquid form, as is commonly used in medicine, such as in the form of normal tablets or dragees, granules, suppositories, injectables, ointments, creams or gels; these preparations can be prepared by methods known per se.

The active compound (s) can be incorporated into the pharmaceutical preparations in the customary excipients used, such as talc, acacia, lactose, starch, magnesium stearate, cocoa butter, aqueous or non-aqueous vehicles, vegetable fats. or of animal origin, paraffin derivatives, glycols, various wetting agents, dispersing or emulsifying agents and preservatives.

The dose of the active ingredient depends on the type of disease being treated, the age and condition of the subject being treated, the mode of administration of the active ingredient and the active ingredient itself.

For example, the dosage may be 0.25 g to 4 g per day for oral administration of the product described in Embodiment 4 or 7 to a human, or 0.5 to 1 g for intramuscular administration three times daily.

In the following, the invention will be explained in more detail by way of examples, without being limited thereto.

Example 1

3-Acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl) -2-tritylhydroxyiminoacetamido] -3-cephem-4-carboxylic acid

Grade A

Ethyl [2- (2-amino-4-thiazolyl) -2-hydroxyimino] acetate

2 g of ethyl (χ-chloro-α-oximinoacetyl) acetate are added to 5 cm ^{3 of} ethanol and 0.76 g of thiourea and the mixture thus obtained is stirred at ambient temperature for a total of 16 hours, whereupon the hydrochloride crystallizes. ^The solid was washed with ethanol-ether (1: 1) and then with ether to give 1.55 g of the hydrochloride.

1.55 g of the hydrochloride obtained above are dissolved in a temperature of 40 to 50 DEG C. in 8 cm @ 3. The solution was neutralized to pH 5-6 by addition of sodium acetate. The free amine crystallizes. The reaction mixture was cooled in an ice bath, centrifuged, washed with water and dried

1.22 g of anti isomer.

Mp 154 ° C.

The mother liquors and washings were combined and the fraction so combined was concentrated and precipitated with water; After washing with ether, sodium bicarbonate is added and the mixture is centrifuged. The solid was then washed with water to give 1.9 g of product which was separated into two spots on thin layer chromatography. The product was purified by chromatography on silica eluting with ether. The pure syn isomer fractions were combined and concentrated. The residue was triturated with ether and centrifuged; the solid is then dried to give 50 mg of the isomer.

Grade В

Ethyl [2- (2-tritylamino-4-thiazolyl) -2-tritylhydroxyimino] acetate

To a solution of 5.4 g of the product prepared in Step A in 54 cm ^{3 of} chloroform and 7.5 cm ^{3 of} triethylamine, a solution of 15 g of trityl chloride in 30 cm ^{3 of} chloroform is added at 10 degrees Celsius. After standing for one hour at rest, the reaction mixture is washed with 40 cm ^{3 of} water and 20 cm ^{3 of} water containing 4 cm? hydrochloric acid, decanted, dried and concentrated to dryness.

The residue is extracted with 10 cm ^{3 of} ether; after addition of 50 cm ^{3 of} methanol, the mixture is stirred, centrifuged and the solid is washed with methanol to obtain 14.2 g of the desired product in two portions.

Grade C

2- (2-Tritylamino-4-thiazolyl) -2-tritylhydroxyiminoacetic acid

10.5 g of the ester obtained in step B are suspended at a temperature close to the reflux temperature in 55 cm @ ^{3 of} dioxane. The resulting supernatant is then slowly added with 17 cm @ ^{3 of} 2 N sodium hydroxide solution. The reaction mixture is then heated to moderate reflux, the mixture is cooled and the precipitated salt is centrifuged. The salt obtained is extracted with 60 cm @ ^{3 of} methylene chloride, 20 cm @ ^{3 of} water and 2 cm @ ³ of acetic acid. The resulting acid is centrifuged and washed with water to obtain a first crop of 7 g of acid.

Dioxane is evaporated from the mother liquor and 20 cm ^{3 of} methylene chloride, 10 cm ^{3 of} water and 1 cm ³ of acetic acid are added. A second crop of 1.5 g of the same product is then isolated. A total of 8.5 g is thus obtained.

20757S

Analysis for С _й НззОзМз8.0,5 H ₂ O:

calculated:

75.85% С, 5.03 θ / ο Η, 6.17% Ν, 4.7% S; found:

75.8 ° / ο C, 4.9% Η, 5.9% Ν, 4.6% S.

Grade D

3-Acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl) -2-tritylhydroxyinpnoacetamiido] -3-cephem-4-carboxylic acid

To a stirred suspension of 8.5 g of the acid prepared in Step C in 50 ml of methanol was added 5 ml of N-methylmorpholine. The mixture thus obtained is stirred for 10 minutes at 30 DEG C. and 30 ml of methylene chloride are then added thereto; The mixture is then concentrated, 100 ml of ether are added, the layers are separated, centrifuged, washed with ether and dried to give a first portion of 7.2 g of salt. The potion is concentrated to dryness, extracted with ether; a second crop of the same product is obtained.

4.24 g of the above morpholine salt are suspended in 60 ml of methylene chloride with stirring. The resulting mixture was stirred for five minutes, then cooled to -5 ° C and 6 ml of a molar solution of isobutyl chloroformate in methylene chloride was added to the cooled mixture. The mixture was then stirred for 15 minutes at-5 ° C, then cooled to -20 ° C; a solution of 1.36 g of 7-aminocephalosporanic acid in 25 ml of methylene chloride and 1.4 ml of triethylamine is then added to the cooled mixture.

The mixture obtained above was allowed to stand for one hour at room temperature and then washed with 50 ml of water containing 10 ml of 1N hydrochloric acid; Centrifuge, decant, wash with water and concentrate to dryness. The residue was triturated with ether, centrifuged and washed with ether to give 4.5 g of crude product.

The crude product is stirred at 10 DEG C. for 1 hour in 10 ml of methylene chloride. The insoluble matter is centrifuged and washed with methylene chloride. 50 ml of ether are then added to the filtrate and the resulting mixture is stirred. Then, the precipitate formed is centrifuged and washed with ether to give 2.29 g of the desired product.

A second crop (0.856 g) of product is obtained. Thus, a total of 3.146 g of the desired product is obtained.

Example 2

3-Acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-hydroxyiminoacetamido] -3-cephem-4-carboxylic acid

2.29 g of the product obtained in the example

1774 cm ⁴ (beta-lactam)

1740 cm ¹

1676 cm ¹

1630 cm ¹

1520 cm < -1 & gt ^{; of} Embodiment 1 is suspended in 18.4 ml of a 50% aqueous formic acid solution. The resulting mixture was then stirred vigorously at 55 ° C for 15 minutes.

After cooling, 10 ml of water are added, followed by centrifugation, washing with water, concentration in vacuo, addition of acetone, centrifugation, addition of 30 ml of ethanol, mixing of centrifugation and washing with ether to give 0.665 g of product. 0.123 g of product in crystalline form are also obtained; a total of 0.788 g of product is thus obtained.

0.735 g of product were dissolved in 7.5 ml of ethanol а к rezultujícímu ¹ solution was added 7.5 ml of acetone. 70 mg of activated carbon are then added, followed by centrifugation, removal of the solvents, addition and separation of ethanol, washing in ethanol; a first crop of 0.450 g of product is obtained; the second crop is 0.105 g.

Infrared spectrum:

\ C = O /

c = cnh ₂ NH

Example 3

3-Acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetamido] -3-cephem-4-carboixylic acid

Grade A

Ethyl [2- (2-amino-4-thlazolyl) -2-methoxyiminioacetate

To 3 ml of absolute ethanol was added 1 g of ethyl (χ-chloro-α-methoxyiminoacetyl acetate), followed by the addition of 0.42 g of triturated thiourea, and the resulting mixture was stirred at ambient temperature for two hours. Dilute with 60 ml of ether to crystallize the resulting hydrochloride, stir the mixture, centrifuge, wash and dry to obtain 685 mg of hydrochloride.

The hydrochloride obtained is dissolved in 4 ml of 50 ° C water, and potassium acetate is added to the resulting solution to reach a pH of solution 6, whereupon the amine liberated crystallizes. Cooling, centrifugation, washing with water and drying are then carried out to obtain 270 mg of the desired product. Mp 161 ° C. The obtained product has a syn configuration. NMR _(CDCl3 60 MHz) ppm:

(N — OCH ₃ )

6.7 (thiazole ring proton).

Grade В

Ethyl 2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetate

4.6 g of the product obtained in the preceding step are dissolved at 30 DEG C. in 92 ml of methylene chloride. The resulting solution was cooled to -10 ° C and triethylamine (2.9 ml) was added to the cooled solution; The mixture is then cooled to -35 ° C and 6.1 g of trityl chloride are added over 15 minutes. The reaction mixture was then allowed to stand at ambient temperature for two and a half hours. The reaction mixture was washed with water, then with 0.5 N hydrochloric acid and finally with aqueous sodium acetate solution. Drying, concentration, ether extraction, re-concentration, dissolution in methanol, addition of water and ether, crystallization, centrifugation, and washing with ether to give 6.15 g of the desired product. The melting point of the product is 120 ° C.

The obtained product has a syn configuration.

Grade C

2- (2-Tritylamino-4-thiazolyl) -2-methoxyiminoacetic acid

The ester (7.01 g) obtained in Step B is dissolved in 35 ml of dioxane. The solution thus obtained is heated to 110 DEG C. in an oil bath and 9 ml of 2 N sodium hydroxide solution are added to the solution over five minutes; the reaction mixture is then heated to reflux for 30 minutes. The sodium salt crystallizes. The reaction mixture is cooled, centrifuged; washing with dioxane and then ether gave a first salt of 5.677 g.

The mother liquor is then concentrated to give a second crop of 1.017 g; a total of 6.784 g of salt is thus obtained.

3.06 g of said salt are added to 65 ml of methylene chloride and 6.5 ml of 2 N hydrochloric acid; after washing with water, drying and concentration to dryness, the free acid is obtained in quantitative yield.

The obtained product has a syn configuration.

NMR Spectrum (DMSO, 60 MHz) p.

3.68 [N-OCH 3]

6.6 (thiazole ring proton).

Grade D

3-Acetoxymethyl 1-7 [2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetamido] -3-cephem-4-carboxylic acid

The dry acid obtained in Step C was dissolved in 30 ml of anhydrous methylene chloride. 0.78 Dicyclohexylcarbodiimide was added to the resulting solution, and the mixture was stirred at ambient temperature for one hour. The dicyclohexylurea formed is centrifuged; After cooling to -10 ° C, a solution of 1.01 g of 7-aminocephalosporanic acid in 13 ml of methylene chloride and 0.9 ml of triethylamine is added. The reaction mixture is then allowed to warm to ambient temperature and 1 ml of acetic acid is added; After centrifugation, washing with aqueous hydrochloric acid and water, drying, concentration to dryness and extraction with 10 ml of dioxane, 1 ml of water and 3 ml of saturated sodium bicarbonate solution are added. After stirring, centrifugation, washing, concentration to dryness, extraction with methylene chloride, washing with 10 ml of water and 5 ml of normal hydrochloric acid, decanting, washing with water, drying, addition and separation of ether, 1.477 g of crude product are obtained. and subsequent elimination after addition of ether. 1.255 g of pure product are thus obtained.

The obtained product has a syn configuration.

Ethyl (χ-chloro-α-methoxyiminoacetyl) acetate used as starting material in Example 3 was prepared as follows:

22.5 g of ethyl (γ-chloro-α-oximinoacetyl) acetate are added to 100 ml of methylene chloride. The resulting mixture is placed in an ice bath and a fresh solution of diazomethane (21.6 g / l) in a total amount of 275 ml is slowly added with stirring.

The reaction mixture was allowed to react and the excess diazomethane was decomposed with a small amount of aluminum. The mixture was then concentrated and purified by elution on silica using methylene chloride as eluent. 11.93 g of the expected product are obtained.

Ethyl [2- (2-amino-4-thiazolyl) -2-methoxyimino] acetate used in Example 3 was also prepared as follows:

Stage a

Ethyl (2-acetyl-2-methoxyimino) acetate

180 g of ethyl (2-acetyl-2-hydroxyimino) acetate in crude form are added to 900 ml of pure acetone. 234 g of potassium carbonate are added to the resulting mixture under nitrogen at 10 ° C; then 103 ml of dimethyl sulfate are added. The reaction mixture is stirred for three hours at ambient temperature, ice is added and the mixture is poured into 4 liters of water; After extraction with methylene chloride, washing with water and drying, the solvents are distilled off. 185 g of the expected product are obtained.

Step b

Ethyl (4-bromo-2-methoxyiminoacetyl) acetate

197 g of the product obtained in the previous step a are added to 1 liter of methylene

2075 75 chloride, 200 mg of p-toluenesulfonic acid are added to the mixture. One-tenth of a total solution of 191 g of pure bromine in 200 ml of methylene chloride is then added at 20 ° C. When the reaction was started, the remaining amount of bromine solution was added at 20 ° C for 1 hour. The temperature was then allowed to rise to 25 ° C. After washing with ice water, re-extraction with methylene chloride and drying, the solvent is distilled off. 268 g of the expected product are obtained.

Stage c

Ethyl [2- (2-amino-4-thiazolyl) -2-ethoxyiminojacetate g of thiourea is added to 270 ml of ethanol and 540 ml of water, and 268 g of the product obtained under nitrogen are added over a period of half an hour. in step b, in 270 ml of ethanol The reaction simulation was stirred at 20 ° C for one hour, then the mixture was cooled to 15 ° C and the cooled mixture was added in small portions of potassium bicarbonate to reach pH 5. After centrifugation, washing and drying, 133.8 g of the desired product are obtained, identical to the product obtained in Step A.

Example 4

3-Acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido] -3-cephem-4-carboxylic acid

0.975 g of the product obtained in Example 3 is stirred for 10 minutes at 55 ° C in 4 ml of a 50% aqueous formic acid solution. 4 ml of water are then added to the reaction mixture and the mixture is centrifuged and concentrated to dryness in vacuo. After partitioning in 2 ml of ethanol, centrifugation, washing with ethanol and then ether, 0.428 g of pure product is obtained.

Analysis calculated for C ^ ProHiyOyhHh:

% C, 42.19;% H, 3.76;% N, 15.37; found:

H, 4.1%; N, 15.2%; S, 13.0%.

The obtained product has a syn configuration.

NMR Spectrum (DMSO 60 MHZ) p / m:

2.03 (-CCH _3): doublet

II o

9.58 J = 8Hz (CONH)

6.76 (proton of the thiazole ring).

Example 5

3-Acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl) -2- (methoxyimino) acetamido] -3-cephem-4-carboxylic acid diethylamine salt

3-Acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl) -2- (methoxyimino) acetamido] -3-cephem-4-carboxylic acid, crude, obtained as described in Example 3 by acid condensation 2- (2-Tritylamino-4-thiazolyl) -2-methoxyiminoacetic anhydride, prepared with dicyclohexylcarbodiimide and 40.8 g of cephalosporanic acid, is dissolved in 350 ml of dioxane. 350 ml of sulfuric acid saturated ether and then 33 ml of diethylamine are then slowly added with stirring. The reaction mixture was then stirred for 20 minutes, after which the crystallized 2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetic acid diethylamine salt was centrifuged. The salt was washed twice with 30 ml of dioxane-ether to give 62.6 g of the salt.

The filtrate is concentrated to a syrupy consistency and two and a half liters of ether saturated with sulfuric acid are added to the concentrated filtrate. The reaction mixture was stirred and then centrifuged. 110.3 g of diethylamine salt are obtained. The obtained product has a syn configuration.

Example6

3-Acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2- (methoxyimino) acetamido] -3-cephem-4-carboxylic acid g of the product obtained in Example 5 was added to 180 ml of 50% of an aqueous solution of formic acid at 50 ° C. The reaction mixture is then stirred at 50 ° C for 20 minutes, after which the triphenylcarbinol formed is centrifuged. 180 ml of ethanol are added and concentrated to dryness under reduced pressure. The residue is extracted with a mixture of 100 ml of water and 20 ml of ethanol and concentrated again. The residue is leached with 100 ml of water and after stirring for 15 minutes at 15 DEG C., centrifugation and washing with water and then, with ether, 15.6 g of the desired product are obtained.

This product is identical to the product obtained in Example 4.

Example 1 a d 7

3-Acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2- (methoxyimino] acetamido] -3-cephem-4-carboxylic acid, sodium salt

45.55 g of 3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2- (methoxyimino) acetamido] -3-cephem-4-carboxylic acid obtained by the procedures described in Examples 4 or 6, is added to 100 ml of distilled water. 8 g of sodium bicarbonate and about 20 ml of ethanol are added successively to the mixture thus formed.

Ethanol (80 ml) and 4.5 g of activated carbon were added again. The resulting mixture was stirred for 5 minutes. Filtering, rinsing with ethanol and concentration to dryness under vacuum are then carried out. The residue is extracted with 100 ml of ethanol and, after concentration to dryness, the residue is dissolved in 100 ml of methanol. Two liters of acetone are added to the solution thus obtained, and after intensive stirring, centrifugation, rinsing with acetone and ether and drying under vacuum,

43.7 g of a white product which hydrates in air to obtain a total weight of 45.2 grams.

[α] _D = -I-55 ° + 2 ° (0.8% water)

Analysis: calculated:

C 40.24, H 3.38, N 14.67, S 13.43,

4.81% Na;

found:

40.3% C, 3.8% H, 14.4% H, 13.3% S,

4.84% Na.

The obtained product has a syn configuration. NMR Spectrum: (60 MHz D ₂ O) pp in:

2.01 _(COCH3), doublet at 9.53

J = 8 Hz (NHCO)

6.75 (thiazole proton).

Example 8

3-Acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl) -2 - [(2-propenyl) oxyimino] acetamido] -3-cephem-4-carboxylic acid

Grade A

Ethyl [2- (2-amino-4-thiazolyl) -2 - [(2-propenyl Joxyimino)] acetate

(a) 9.7 g of ethyl (2-hydroxyimino-4-chloroacetyl) acetate, 30 ml of acetone and 9.15 ml of 3-iodopropene are cooled in an ice bath and 27.5 ml of 2 N are added to the cooled mixture. sodium hydroxide solution and the reaction mixture is then allowed to stand at ambient temperature for two and a half hours.

b) 3.8 g of thiourea are added to the reaction mixture, whereupon the reaction mixture is heated at 60 ° C for 15 minutes and then for 45 minutes at ambient temperature. Acetone was distilled off and imethylene chloride, water and potassium carbonate were added. After stirring, decanting, reextracting with methylene chloride, drying and concentrating to dryness, 9.75 g of a residue is obtained which is chromatographed on silica eluting with ether; it is isolated

2.7 g of product which is taken up in isopropyl ether. The separated crystals are centrifuged, rinsed and dried. 783 mg of the expected product is obtained.

The melting point of the product is 100 ° C.

Grade В

Ethyl [2- (2-tritylamino-4-thiazolyl) -2 - [(2-propenyl) oxyimino]] acetate

511 mg of the product obtained in Step A are mixed with 0.92 ml of dimethylformamide,

1.8 ml methylene chloride and 0.29 ml trlethylamine. The mixture was cooled to -15 ° C and 615 µg of trityl chloride was added. After standing at room temperature for one and a half hours, 1N hydrochloric acid (2 ml) was added followed by 5 ml of water. After decantation, drying and concentration to dryness, 1.28 g of crude product is obtained.

The obtained product has a syn configuration.

Grade C

2- (2-Tritylamino-4-thiazoilyl) -2- [(2-propenyl) oxyimino] acetic acid

A mixture of 1.28 g of the product obtained in Step B, 6.2 ml of dioxane and 3 ml of 2 N sodium hydroxide solution is heated to 120 ° C. The mixture was then heated to reflux for one hour. The sodium salt crystallized, which was centrifuged, washed with ether-dioxane and dried, yielding 805 mg of product.

The product is taken up in 10 ml of methylene chloride and 3 ml of 1 N hydrochloric acid and the mixture is stirred until dissolved. After decantation, drying, concentration to dryness, transfer to ether, centrifugation, 715 mg of the desired product is isolated.

Mp 170 ° C. The obtained product has a syn configuration.

Grade D

3-Acetoxymethyl 1-7- (2- (2-tritylamino-4-thiazolyl) -2- [(2-propenyl) oxyimino] acetamido] -3-cephem-4-carboxylic acid

470 mg of the product prepared in Step C, 5 ml of methylene chloride and 130 mg of dicyclohexylcarbodiimide are added and the mixture is poured over with a small amount of methylene chloride and then stirred for one hour at ambient temperature; After centrifugation of the formed cyclocyclohexylurea, the filtrate is cooled and 136 mg of 7-aminocephalosporanic acid in solution in 2.4 ml of imethylene chloride and 0.14 ml of triethylamine are added under an inert gas atmosphere. The reaction mixture is allowed to stand at ambient temperature for one and a half hours, then 1N hydrochloric acid (2 ml) and water are added; after stirring, decanting, washing with water, drying and concentration, 610 mg of crude product is obtained.

The obtained product has a syn configuration.

Example 9

3-Acetoxymethyl-7- [2- (2-amino-4-thiazioilyl) -2- [(2-propenyl) oxyimino / acetamido] -3-cephem-4-carboxylic acid

A mixture of 610 mg of the product obtained in Example 8 and 3 ml of a 50% aqueous formic acid solution was heated to 60 degrees Celsius for 15 minutes, after which 4 ml of the voids were added; after stirring, centrifugation of triphenylcarbinol, rinsing with water, concentration in vacuo, transfer to water, centrifugation and rinsing, 120 mg of the desired acid is obtained.

The melting point of the product is about 160 ° C.

Ultraviolet spectrum:

In ethanol: max 236 nm Ef ¹ = 375 ε = 18 000 inflection 252 nm E _t ¹ = 316 inflection 295 nm! E ^ = 138 ε = 6600

In a mixture of ethane-N / 10 hydrochloric acid:

max 263 nm E! ¹ = 380 ε = 18 300 inflection 280 nm Ej ¹ = 317

NMR Spectrum (DMSO 90 MHZ) p.m .:

2.02 (OAc)

6.68 (thiazole proton).

The product thus obtained has a syn configuration.

Example 10

3-Acetoxymethyl 1-7- / 2- (2-tritylamino-4-thrazolyl) -2- (ethoxyimino) acetamido] -3-cephem-4-carboxylic acid

Grade A

Ethyl [2- (2-amino-4-thiazolyl) -2-ethoxyimino] acetate

(a) 19,4 g of ethyl (χ-chloro-α-oxyiminoaceto) acetate are added to 60 ml of acetone and 14,3 ml of diethyl sulphate. The mixture is then cooled in an ice bath and 55 ml of 2 N sodium hydroxide solution are added over 30 minutes. The reaction mixture is then stirred for <40 minutes.

b) 7.6 g of thiourea are added to the reaction mixture and the mixture is heated to 55 ° C for 20 minutes. The acetone is stripped off, transferred to ethyl acetate, added 6.9 g of potassium carbonate, stirred, decanted, reextracted with ethyl acetate, dried and concentrated to dryness. 17.4 g of residue are isolated, which is chromatographed on silica eluting with ether. Transfer of the isolated product to isopropyl ether, centrifugation, washing and drying afforded 2.8 g of the desired product. Mp 129 ° C.

The obtained product has a syn configuration.

Grade В

Ethyl [2- (2-tritylamino-4-thiazolyl) -2-ethoxy imino] acetate

3.16 g of the product obtained in Step A, 6 ml of anhydrous dimethylformamide, 12 ml of methylene chloride and 1.89 ml of triethylamine are mixed under an inert atmosphere. The mixture thus obtained was cooled to -15 C ^and а к cooled mixture was then added 2.98 g of trityl chloride. The reaction mixture was allowed to stand for half an hour, raising the temperature to 10 ° C; then, the mixture is maintained at ambient temperature for three and a half hours. Add 13 ml of 1N hydrochloric acid and decant, wash with hydrochloric acid (1N) and water. Extraction with methylene chloride, drying and concentration to dryness yielded 7.89 g of crude product.

The obtained product has a syn configuration.

Grade C

2- (2-Tritylamino-4-thiazolyl) -2-ethoxyiminoacetic acid

A mixture of 7.89 g of the product obtained in Step B, 4.0 .mu.l of diolxane and 19.5 ml of 2N sodium hydroxide solution is heated at 110 DEG C. for 1 hour. Centrifuge, wash with ether-dioxane and then with potoim alone, and dry. 6.25 g of the sodium salt is obtained, which is taken up in 60 ml of methylene chloride and 20 ml of 1N hydrochloric acid; the two phases are mixed, 20 ml of methanol are added and decanted, washed again with water, re-extracted with methyl chloride-methanol, dried and concentrated; 5.85 g of 2- (2-tritylamino-4-thiazolyl) -2-ethoxyiminoacetic acid are isolated in pure form.

The obtained product has a syn configuration.

Grade D

3-Acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl) -2- (ethoxyimino) acetamido] -3-cephem-4-carboxylic acid

3.43 g of 2- (2-tritylamino-4-thiazolyl) -2-ethoxyxyiminoacetic acid prepared in step C are suspended in 34 ml of methylene chloride, and the resulting suspension is cooled and 970 mg of dicyclohexylcarbodiimide are added to the cooled suspension. . After washing with methylene chloride and stirring at room temperature for 1 hour, the dicyclohexylurea is centrifuged.

The filtrate was cooled to -20 ° C, and a solution of 1.02 g of 7-aminocephalosporanic acid in 18 ml of methylene chloride and 1.06 ml of triethylamine, cooled to -20 degrees Celsius, was added in one portion to the cooled filtrate.

The reaction mixture was allowed to warm for one and a half hours and 1.8 ml of acetic acid was added; after adding 9. ml of 1 N hydrochloric acid, stirring, decanting, washing with water, extraction with methylene chloride, drying and concentration, 4.56 g of the desired product are obtained.

The obtained product has a syn configuration.

Example 11

3-Acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2- (ethoxyimino) acetamido] -3-cephem-4-carboxylic acid

4.56 g of the product obtained in Example 10 are added to 23 ml of a 50% aqueous formic acid solution and the mixture thus obtained is heated to a temperature of 55 ° C for 15 minutes. The reaction mixture was then diluted with water (30 mL) and centrifuged with triphenylcarbinol. After concentration of the filtrate to dryness, transfer to water, stirring, centrifugation, washing and drying, 116 mg of impure product is obtained. Concentration of the filtrate gave a second crop of crystalline product of 674 mg; a total of 790 mg of product is thus obtained.

The following cleaning is performed:

1.063 g of crude product is triturated with 5 ml of water, the mixture is heated at 70 DEG C. for 5 minutes, then cooled, stirred again for half an hour, then the solid is centrifuged and dried to dryness. recovery of 815 mg of purified product. The 815 mg of purified proidect is taken up in 2 ml of water and 3 ml of acetone, the mixture is gently warmed, the solid is centrifuged, water is added in 3 ml, the mixture is heated to 60 ° C, acetone is blown off with nitrogen. The beads are centrifuged, rinsed with water and then with ether to give 438 mg of the desired product.

Analysis: For calculated:

% C, 43.49;% H, 4.08;% N, 14.92;% S, 13.66; found:

44.5% CI, 4.4% H, 14.8% N, 13.3% S. The product obtained is syn.

NMR Spectrum (60MHz DMSO) p.

2.05 (OAc)

6.75 (thiazole ring proton).

Example 12

39.8 g of ethyl (2-acetyl-2-hydroxyimino) acetate is added to 200 ml of pure acetone. . The mixture thus obtained is cooled in an ice bath and 52 g of potassium carbonate are added to the cooled mixture and then. within half an hour of 25 ml of 2-iodopropane. The mixture is then stirred for two hours, 800 ml of water and 500 ml of methylene chloride are added. after decantation, re-extraction with methylene chloride, drying, centrifugation and concentration, 41.5 g of the desired product are isolated.

Grade B '

Ethyl [4-bromo-2- (1-methylethoxyimino) acetyl] acetate

41.5 g of the product obtained in the preceding step are added to 190 ml of methylene chloride. containing traces of paratoluenesulfonic acid. The mixture thus obtained is stirred and then added. solution of 11.9 ml of bromine in 50 ml of methylene chloride was added at room temperature for one hour. After stirring, addition of ice water, decantation, re-extraction with methylene chloride, rinsing with ice water, drying and concentration, 55 g of the desired product are isolated.

Grade C

Ethyl [2- (2-amino-4-thiazolyl) -2- (1-methylethoxyimino) acetate

14.9 g of thiourea are added to 55 ml of ethanol and 105 ml of water, and a solution of 55 g of the product prepared in Step B in 55 ml of ethanol is then added over 40 minutes. The mixture is stirred at ambient temperature for two hours and thirty minutes, then 220 ml of a 10% aqueous sodium bicarbonate solution are added; the mixture obtained is stirred and, after centrifugation, washing and drying, 42.15 g of crude product is isolated, which is chromatographed on silica using ether as the eluent; the fractions rich in the desired product are collected, concentrated, the crystals formed are leached with isopropyl ether, centrifuged and washed to give 10.75 g of the desired product.

The obtained product has a syn configuration.

Grade D

Ethyl (2- (2-tritylamino-4-thiazolyl) -2- (1-methylethoxyimino)) acetate

3-Acetoxymethoxy-7- [2- [2-tritylamino-4-thiazolyl] -2- (1-methylethoxyiminoacetamido) -3-cephem-4-carboxylic acid

Grade A

Ethyl [2-acetyl-2- (1-methylethoxyimino)] acetate g of the product obtained in Step C is mixed with 20 ml of anhydrous dimethylformamide, 40 ml of methylene chloride and 6.2. ml of triethylamine. The resulting mixture was cooled and trityl chloride (13.2 g) was slowly added; The mixture is stirred for two and a half hours and then 43 ml of normal hydrochloric acid are added thereto, after stirring, decanting, washing with 40 ml of water, extraction with methylene chloride, drying, centrifugation and concentration to dryness, 7 g of the desired product. The obtained product has a syn configuration.

Grade E

2- (2-Tritylamino-4-thiazolyl) -2- [1-methylethoxyimino] acetic acid

A mixture of 27.7 g of the product obtained in Step D, 150 ml of dioxane and 65 ml of 2 N sodium hydroxide solution is heated to reflux for two hours. The sodium salt crystallizes, which, after cooling, is centrifuged, washed with ether-dioxane (1: 1) and dried to give 16.85 g of crude sodium salt.

15.9 g of this sodium salt are dissolved in 15.9 g of dimethylformamide, 100 ml of water and about 500 ml of methanol. 30 ml of a 2 N hydrochloric acid solution are then added to the solution thus obtained, after which methanol is stripped off; after dilution with water, centrifugation, washing and drying, 9.8 g of the obtained product are obtained; The viscous product is taken up in 220 ml of a 50:50 mixture of methylene chloride and methanol and, after concentration to dryness, taken up in ether, separating the layers, centrifuging the crystals, washing and drying, 4.9 g of the desired acid are obtained.

Mp 170 ° C.

Preparation of the analytical sample:

300 mg of the crude product is dissolved in 2 ml of methylene chloride and 1 ml of methanol, then the solution is diluted with water and methylene chloride, after stirring, centrifuging the crystals, washing with methylene chloride and water. Doiu and drying isolated 230 mg of pure product for analysis.

Analysis:

calculated:

C 68.77, H 5.34, N 8.91, S 6.8; found:

68.6%. C, 5.5% H, 8.8% N, 6.8% S.

The obtained product has a syn configuration.

Grade F

3-Acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl) -2- (1-methylethoxyimino) acetamiidio] -3-cephem-4-carboxylic acid

4.89 g of the acid obtained in step E are added to 13.5 ml of dimethylformamide under an argon atmosphere. After dissolution, the resulting solution was cooled in an ice bath and 1.62 g of dicyclohexylcarbodiimide in 16 ml of methylene chloride was added thereto. Dicyclohexylurea crystallizes. The mixture is then stirred in an ice bath and after centrifugation, washed with methylene chloride and dried, 1.424 g of dicyclohexylurea is isolated. After cooling in a methanol-ice bath, a solution of 1.41 g of 7-aminocephalosporanic acid in 30 ml of methylene chloride and 1.45 ml of triethylamine is added.

After stirring for three hours at ambient temperature, 20 ml of normal hydrochloric acid are added and after stirring, decanting, re-extraction with methylene chloride, drying and centrifugation, 9.05 g of the desired product-starting product mixture is obtained.

After transfer to methylene chloride, seeding, crystallization with stirring, the resulting crystals are centrifuged, washed and dried to give 1.6 g of pure parent product; After concentration to dryness, the residue is taken up in isopropyl ether with vigorous stirring, and 4.91 g of the insoluble viscous product which is the desired product is isolated.

The obtained product has a configuration, syn.

Example 13

3-Acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2- (1-methylethoxyimino) acetamido] -3-cephem-4-carboxylic acid

4.91 g of the crude product obtained in Example 12 are added to 30 ml of a 50% aqueous formic acid solution. The resulting mixture was stirred in a 60 ° C water bath, then diluted with water, and after centrifugation of the triphenylcarbinol formed, washing with water and drying, 1.39 g of triphenylcarbinoil was isolated. After concentration to dryness, transfer to water, centrifugation, washing with water and drying, 800 img of the desired product is obtained.

A sample for analysis is obtained by dissolving 972 mg of the crude product in 4 ml of methanol, diluting the solution with 20 ml of ether, centrifuging the insoluble matter, washing and drying to obtain 404 mg of the desired pure acid.

The melting point of the product is 200 ° C.

Analysis: Calculated: C 44.71, H 4.38, N 14.48, S 13.26; found *

44.5% C, 4.5% H, 14.1% N, 13.2% S. The product obtained is syn.

NMR Spectrum (60 MHz DMSO) p / m:

2.01 (CH ₃ CO);

doublet at 9.46 J 8 Hz (CONH);

6.7 (thiazole ring proton).

Example 14

3-Acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl) -2-ethoxyiminoacetamido] -3-cephem-4-carboxylic acid

Grade А

Ethyl [2- (2-tritylamino-4-thiazolyl) -2-hydroxyimino] acetate, anti isomer

11.5 g of ethyl [2- (2-amino-4-thiazolyl) -2-hydroxyimylnoacetate (anti-isomer) prepared in Step A of Example 1 are added to 90 ml of anhydrous dimethylformamide and 24 ml are then added to this mixture. trlethylamine. 47.6 g of trityl chloride are then added in small portions over a half hour period and the mixture is left to stand for two and a half hours under self-heating. Next, 150 ml of 2 N hydrochloric acid and 600 ml of water are added. The mixture was stirred for 15 minutes, triturated three times with ether, transferred to a mixture of methanol, triethylamine and water and stirred. After centrifugation, washing and drying, 60.2 g of the desired product is isolated.

The crude product (3.4 g) was recrystallized from methylene chloride / methanol to give pure product (3 g).

Mp 260 ° C.

Grade В

Ethyl [2- (2-tritylamino-4-thiazolyl) -2-ethoxyiminoacetate, anti-isomer

11.5 g of the product obtained in the previous well A and 5.85 g of potassium carbonate are added to 25 ml of anhydrous dtoethylformamiid. The mixture was cooled to 15 ° C, 16.7 mL of ethyl sulfate was added and the mixture was allowed to stand for 4 hours at ambient temperature. 420 ml of water and 250 ml of ethyl acetate are then added to the reaction mixture, followed by stirring, centrifugation, leaching with ethanol, crystallization, washing, trituration and drying to give 6.6 g of the desired product.

Mp 165 ° C.

This product can be recrystallized by the following! as follows:

797 mg of the product is dissolved in methylene chloride / ethanol (50:50) and 596 µg of pure product is isolated after centrifugation, concentration to evaporate the methylene chloride by crystallization, centrifugation, washing and drying.

Grade C

2- (2-Tritylamino-4-thiazolyl) -2-ethoxyiminoacetic acid, anti isomer

7.29 g of the product of Step B are added to 45 ml of dioxane and 9 ml of 2N sodium hydroxide solution. The mixture thus obtained is heated in a water bath with stirring at 50 ° C for one hour and 50 minutes, then cooled and seeded to crystallize in ice water; then centrifuging, washing; thereby obtaining

4.2 g of sodium salt. The salt is leached with 50 ml of methylene chloride, 40 ml of water and 11 ml of normal hydrochloric acid and after stirring, decanting, reextracting, washing, drying, centrifuging and concentrating to dryness, the residue is taken up in 50 ml of ether and centrifuged after stirring. the separated crystals are washed with ether to obtain 3.27 g of the desired product.

The resulting product melts with decomposition at about 200 ° C.

NMR (60 MHz CDC1 ₃₎ ppm =

7.66 (thiazole ring proton),

7.36 (proton of the trityl group).

Grade D

3-Acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl) -2-ethoxyiminoacetamido] -3-cephem-4-carboxylic acid, anti-isomer

4.1 g of the acid obtained in the previous step C are added to 36 ml of tetrahydrofuran, 27 ml of methylene chloride and 0.99 ml of N-methylmorpholine under argon. The mixture was cooled to -20 degrees Celsius and 1.17 mL of isobutyl chloroformate was added dropwise. The reaction mixture was left at this temperature for three minutes and then cooled to -35 degrees Celsius; to this cooled mixture is added a solution of 2.45 g of 7-aminocephalosporamic acid in 4.5 ml of methylene chloride,

2.52 ml of triethylamine. The reaction mixture is then allowed to stand by heating the mixture for two and a half hours, after which time the solvents are stripped off.

After transferring the residue to a mixture of methylene chloride and water, normal hydrochloric acid is added to a pH of 1 to 10

2. Extraction with methylene chloride, washing, drying, centrifugation, concentration, transfer to ethyl acetate, dilution with isopropyl ether, stirring, centrifugation, washing and drying afforded 4.87 g of the desired product (isoimer anti).

The product is purified as follows:

4.87 g of the above product are dissolved in 10 ml of ethyl acetate with heating, then the resulting solution is diluted with isopropyl ether, stirred and centrifuged and washed and dried to give 4.53 g of purified product.

Example 15

3-Acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-ethoxyiminoacetamido] -3-cephem-4-carboxylic acid, anti-isomer

4.27 g of the purified product obtained in Example 14 are added to 20 ml of an aqueous formic acid solution (50:50). The mixture thus obtained was heated in a water bath at 60 ° C for 20 minutes, then cooled and diluted with water. Stirring is continued for 10 minutes, the triphenylcarbinol is centrifuged and washed with water to obtain 1.44 g of product. Ethanol was added and the mixture was concentrated in vacuo. The residue is taken up in ethanol, concentrated again in vacuo and 30 ml of water are added to the residue; The mixture was stirred for 1 hour in an ice bath. After centrifugation, washing with water and drying, 2.06 g of the desired product is obtained.

The product is purified as follows:

2.06 g of the above product was dissolved in 5 ml of aqueous bicarbonate solution (10% solution) and 5 ml of water. After. concentrating the turbidity and washing with water, concentrated formic acid is added dropwise to a pH of 3-4; After centrifugation of the crystals obtained after 12 hours of rest at ambient temperature, washing with water and drying, 1.73 pure product is obtained.

This product melts with decomposition at about 200 ° C.

NMR Spectrum (60 MHz, DMSO)?

2.04 (CH ₃ CO);

7.5 (diazole ring proton).

Example 16

3-Acetoxymethyl-7- [2- (2-tritylamino-4-thiazolilyl) -2- (1-methylethoxyimino) acetamido] -3-cephem-4-carboxylic acid, anti-isomer

Grade A

Ethyl [2- (2-triethylamino-4-thiazolyl) -2- (1-methylethoxyimino) acetate, anti-isomer

Under argon, 6.86 ethyl (2- (2-tritylamino-4-thiazolyl) -2-hydroxyimino) acetate (anti-isomer), prepared according to step A of Example 14, was mixed with 3.51 g of potassium carbonate with 15 ml of dimethylformamide and 7, 7 ml of isopropyl iodide is then left to stand for four and a half hours, 250 ml of distilled water and 150 ml of ethyl acetate are added, followed by stirring, decanting, washing, reextracting with ethyl acetate, drying, centrifuging, Concentration to dryness, transfer to ethanol, seeding for crystallization, centrifugation to give 3.26 of the desired product.

The melting point of the resulting product is 182 ° C.

Grade В

2- (2-Tritylamino-4-thlazolyl) -2- (1-methylethoxyimino) acetic acid, anti isomer

6.8 g of the product obtained in Step A are mixed with 41 ml of dioxane and 8.15 ml of 2N sodium hydroxide solution. The mixture thus obtained is heated in a water bath at 55 ° C for 1 h. 9.5 ml of 2 N hydrochloric acid is added (the solution then has a pH of 2 to 3). After; removal of dioxane, crystallization, dilution with water, stirring, washing, trituration with ether and drying afforded 5.87 of the desired product.

The resulting product melts at 240 ° C with decomposition.

NMR Spectrum (CDCl _3, 60 MHz) ppm =

7.66 (thiazole ring proton);

7.31 (trityl group).

Grade C

3-Acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl) -2- (1-methylethoxyimino) acetamido] -3-cephem-4-carboxylic acid, anti-isomer

5.66 g of the acid obtained in the preceding stage B, are added to a mixture of 48 ml of tetrahydrofuran, ¹ tetrahydrofurane, 48 ml of methylene chloride and 1.32 ml of N-methylmorpholine. The mixture thus obtained is heated to dissolve the solid, then cooled to -20 DEG C. and 1.56 ml of isobutyl chloroformate are added to the mixture thus cooled. The reaction mixture was allowed to stand for 10 minutes at a temperature of between -20 ° C and -10 ° C, then cooled to -35 ° C and 3.26 g of 7-aminocaphalosporanic acid in 60 ml was added in one portion. methylene chloride and

3.36 ml of anhydrous triethylamine. The mixture was allowed to warm and then stirred for three hours and thirty minutes. After stripping off the solvents, eluting with ethyl acetate, the solution is stirred after complete dissolution and then diluted with isopropyl ether; After centrifugation, washing and drying 5.42 g of the desired product is obtained.

The product is purified by dissolving 5.82 g of the product obtained as described above in 20 ml of ethyl acetate, then diluting it with 200 ml of isopropyl ether and drying 4.82 g after drying. of the desired product.

Example 17

3-Acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2- (1-methylethoxyimino) acetamido] -3-cephem-4-carboxylic acid, anti-isomer

3.62 g of the product obtained in Example 16 are added to 16 ml of an aqueous formic acid solution (50:50). The mixture was heated at 60 ° C for 20 minutes, cooled and diluted with 16 mL of water. After stirring, centrifuging the formed triphenylcarbinol, washing with water, drying, 1.23 g of product is obtained.

Concentrate to dryness under vacuum,

3β conversion to · ethanol,. Then, 1.68 g of the desired crude product is isolated, which is again stripped off in vacuo, taken up in water, seeded for crystallization, centrifuged to obtain a crystalline gum, washed with water and dried.

A pure sample of this product is obtained as follows:

g of said product is dissolved in ml of 10% bicarbonate solution and 5 ml of water and the resulting solution is stirred for 10 minutes. Centrifuge, rinse with water, add · formic acid until pH = 3, stir in ice bath · spin for one hour, centrifuge, rinse with water, dissolve gummy product in 10 ml ethanol hot, cool in water ice water, centrifugation and washing with ethanol and then ether. 748 mg of purified product is obtained. In addition, crystal formation in the filtrate is observed, which is diluted with ether without precipitation, after which the crystals are formed. centrifuged and washed with ethanol-ether. In this manner . 177 mg of pure crystalline product (anti-isomer) is obtained.

The melting point of this product is 200 ° C (dec.).

NMR Spectrum · (DMSO 60 MHZ) pm = 7.46 (thiazole ring proton);

4.43 (protonated tertiary isopropyl groups).

Example 18

Acid 3-acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl) -2- ^{(2-propenyloxyimino)} acetamide) ^ 3-cephem-4-carboxylic acid, anti isomer

Grade A

Ethyl [2- (2-tritylamino-4-thiazolyl) -2- (2-propenyloxyimino)] acetate, anti isomer

6.86 g of ethyl 42- (2-trityamethylamino-4-thiazolyl) -2-hydroxyiminoacetate (isomer anti) prepared according to the procedure described in Example 14, Step A, 3.51 potassium carbonate, 15 ml are mixed under argon atmosphere. dimethylformamide and 7 ml of allyl iodide. The mixture thus obtained is stirred for five hours at a temperature. 250 ml of water and 150 ml of ethyl acetate are added. After stirring, decanting, washing, reextracting with ethyl acetate,. drying, centrifugation, concentration to dryness, ethanol extraction, seeding for crystallization, cooling in ice water, half-hour crystallization with stirring and centrifugation and washing of the separated crystals, 4.72 g of the desired product are obtained.

The product is purified as follows:

215 mg of the above product are dissolved in 2 ml of ethanol and 2 ml of methylene chloride. The filtrate was concentrated and then diluted with ethanol and crystallized in ice water. After centrifugation, washing and drying of the deposited crystals, 70 mg of purified product is obtained.

Melting point 90 ° C (pa consistency), above 160 ° C the product is liquid.

Grade B

2- (2-Trimylamino-4-thiazolyl) -2 (2-ptopenyloxyimino) acetic acid, anti isomer

3.71 g of the product obtained in the previous step A are mixed with 22 ml of dioxane and

4.5 ml of 2 N sodium hydroxide solution. The mixture thus obtained is heated to a water bath temperature. 55 ° C for 50 minutes, then cooled and 5.25 ml · 2 N hydrochloric acid are then added to the cooled mixture to pH 2. After stripping of dioxane, a gum is obtained which is diluted with water. Cool in ice water, centrifuge, rinse with water, then triturate with ether three times. 2.85 g of the expected product are obtained.

Mp 198 ° C (decomposes).

NMR spectrum, m (CDCl3 90 MHz) p

7.64 · (thiazole ring proton);

7.27 (trityl proton).

Grade C

3-Acetoxymethyl 1-7- [2- (2-tritylamino-thiazolyl) -2- (2-propenyloxyimino) acetamido) -3-cephem-4-carboxylic acid, anti-isomer

2.82 g of the acid obtained in the preceding step B are added to 24 ml of anhydrous tettahydrofuran and 24 ml of methylene chloride, after which they are added to the resulting solution of 0.66 ml of N-methylmorpholine. The mixture was. It is then cooled to -20 DEG C. and 0.78 ml of isobutyl chloroformate are added to the cooled mixture.

The mixture is stirred for 3 minutes at -20 degrees Celsius, then cooled to -35 ° C and a solution of 1.63 g &lt; 3 &gt;

of 7-aminocephalosporanic acid in 30 ml of methylene chloride and 1.68 ml of 2-amino-amine. The mixture was allowed to warm, then the solvents were stripped off, the residue was taken up in methylene chloride and normal hydrochloric acid. After stirring, decanting, washing, reextracting, drying, centrifuging, rinsing, concentration to dryness, ethyl acetate precipitation, dilution with isopropyl ether, stirring, centrifugation and washing, 3.08 g of the desired product is obtained.

The product number is as follows. as follows:

20757S

3.59 g of the product obtained above are dissolved in 15 ml of ethyl acetate and diluted with isopropyl ether, stirring, centrifuging the precipitate and rinsing in isopropyl ether isolated 3.33 g of purified product.

Example 19

3-Acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2- (2-propenyloxyimino) acetaimido] -3-cephem-4-carboxylic acid, anti-isomer

2.53 g of the product obtained in Example 18 are added to 11.5 ml of aqueous formic acid (50:50). The mixture was heated to 60 ° C for 20 minutes, diluted with water, cooled to ambient temperature and centrifuged. After drying the formed triphenylcarbinol, 863 mg of this product is obtained. The mother liquors are concentrated in vacuo, ethanol is added and stripped of ethanol, the gum is taken up in 15 ml of water, centrifuged, washed with water and dried to give 1.275 g of the desired product.

The product was purified by the following method ^1:

1.63 g of the crude product are suspended in 15 ml of ethanol, then the resulting suspension is heated to reflux and centrifuged; after washing with ethanol, 877 mg of insoluble product is isolated. Ether (20 ml) was added to the filtrate, the insoluble matter was centrifuged, and after washing with ethanol-ether and drying, 97 mg of a second crop were isolated. The filtrate was concentrated to half volume, whereupon the product crystallized. After centrifugation, washing and drying, 162 mg of pure product (isomer anti) is isolated.

The melting point of the product is 180 ° C (pasty consistency).

Nuclear Magnetic Resonance Spectrum (MSO 60 MHz) p

7.48 (thiazole ring proton);

2.04 (proton of the acetyl group).

Example 20

3-Acetoxymethyl 1-7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamide] -3-cephem-4-carboxylic acid, isomer syn

Grade A

Ethyl [2- (2-chloroacetamido-4-thiazolyl) -2-methoxyimino] acetate

45.8 g of ethyl [2- (2-amino-4-thiazolyl) -2-methoxyimino] acetate (isomer syn), prepared as described in Step A of Example 3, is added to 200 ml of methylene chloride. It is distilled · 20 ml to dry and 50 ml of pyridine are added to cool to 10 ° C. 41 g of oetoic anhydride are added and the mixture is gently heated until the solids dissolve. The mixture was left under nitrogen at 20 ° C for 6 hours, 5 ml of water was added, the mixture was stirred and then poured into 300 ml of 2 N hydrochloric acid (ice-cooled). After decantation, extraction with methylene chloride, washing with water and sodium bicarbonate solution, drying, purification with charcoal, concentration 300 ml of isopropyl ether is added. The product crystallizes. The mixture is concentrated until a thick paste is obtained and, after cooling with ice, centrifugation, washing and drying, 45.4 g of product are obtained,

The melting point of the product is 113 ° C.

A pure sample of the product is obtained by recrystallization in methylene chloride / isopropyl ether.

The melting point of the purified product is 118 ° C.

NMR (60 MHz CDC1 ₃₎ t 7 o \ J i = J Ic) (*) Й

OCH ₃ (a) triplet centered at 1.38 ppm J = 7 Hz;

(b) singlet 4.05 µm;

(c) a quadruplet centered at 4.44 µm. J-7 Hz;

[d) singlet 4.33 µm;

(e) a 7.27 p. m singlet; (f) a 9.95 p. m singlet.

Grade В

2- (2-Chloroacetamido-4-thiazolyl) -2-methoxy-amino-acetic acid, isomer syn g of the product obtained in Step A above, is added to 230 ml of absolute ethanol and added thereto at 20 ° C and below. 30 ml of pure sodium hydroxide are added under a nitrogen atmosphere · The product dissolves and the sodium salt starts to crystallize to thicken the reaction medium After 16 hours, the crystals are centrifuged and washed The salt obtained is dissolved in water, frozen, 100 ml of 2 N acid are added. hydrochloric acid, saturated with sodium chloride and extracted with ethyl acetate containing 10% ethanol.After drying, transfer over activated carbon, distillation under vacuum, removal of water with benzene, transfer to methylene chloride, freeze-drying, centrifugation, washing and drying, 34.5 g of the desired product are obtained. product.

The melting point of this product is about 200 ° C.

This product is purified by recrystallization in acetone / isopropyl ether.

Pra CsHfANaClS (277.68) calculated:

C, 34.60; H, 2.90; N, 15.13; Cl, 12.77.

11.55% S;

found:

34.81%. C, 2.8% H, N 14.8%, Cl 12.6%,

11,5% S.

NMR (60 MHz DMSO) (e) Y is ^{H, C1 ť_CH} 2 ^(N ^ cl ICU 0CH ₃ (&>

(a) singlet 3.92 µm;

(b) singlet 4.38 µm;

(c) a singlet of about 5 µm;

(d) singlet 7.58 µm;

(e) singlet 12.6 µm.

Grade C

3-Acetoxymethyl-7- [2- (2-chloroacido-4-thiazolyl) -2-methoxyiminoacetamido] -3-cephem-4-carboxylic acid, isomer syn.

15.3 g of the product obtained in Step B above are added to 80 ml of methylene chloride. To the mixture thus obtained, cooled to 5 ° C, 8 ml of triethylamine are added. 3.8 ml of thionyl chloride and 26 ml of methylene chloride are then added at 0 ° C under a nitrogen atmosphere. The mixture was allowed to stand for 15 minutes at 0 ° C and 7 ml of triethylamine was added. 13.6 g of 7-aminocephalosporanic acid in 100 ml of methylene chloride and 14 ml of triethylamine are then added at a temperature of 0 DEG C. and under a nitrogen atmosphere. The temperature of the reaction mixture was allowed to rise to 20 ° C and the mixture was then stirred for one hour. The solution thus obtained is distilled to dryness under vacuum at 30 to 35 ° C. Dissolve the residue in 250 ml of water, pass the solution through activated carbon, and add 50 ml of 2 N hydrochloric acid to the treated solution. The precipitate is centrifuged and washed with water.

The crude product thus obtained is suspended in 80 ml of ethanol. 7 ml of triethylamine are added at 5 ° C. 15 ml of 4N sulfuric acid are then added in one portion at 5 [deg.] C. with stirring, after which the product crystallizes after 15 minutes. The crystals are centrifuged, washed with ethanol by trituration of the product with ethanol and then with ether and dried under vacuum to give 18.6 g of the desired product.

[axis] n = + 26 ° (± 1 °) At conc. 1% in dimethylformamide.

NMR Spectrum- (DMSO 60 MHz)

O (c)

H-N-C-CH 2 's- + n ° -I-Aa>

(dl ^ _OCH ^ ^ ^^^^, с о-с _{3 3} (ЫCOJJ O (a) singlet 2.03 ppm;

(b) singlet 3.90 µm;

(c) singlet 4.38 µm;

(d) singlet 7.45 p. m.

Grade D

3-Acetoxymethyl-7- [2- (2-amino-4-γg (Яу1) -2-dimethoxyiminoacetic acid)] - 3-cephem-4-carboxylic acid, isomer syn

5.32 g of the acid obtained in the preceding step are suspended in 10.6 ml of water and 912 mg of thiourea. 1 g of potassium bicarbonate is then added at 20 ° C. After dissolution, the solution was stirred for six hours at 20 ° C under a nitrogen atmosphere. After about one and a half hours, the elimination of the gum product begins. 30 ml of water and 3 ml of formic acid are added. The reaction mixture was then cooled to 0 ° C. After removal, the product is washed with water containing 10% formic acid. The residue is dissolved at about 5 ° C in 30 ml of water containing triethylamine. At 5 ° C, 3 ml of formic acid are added, after which the precipitate formed is centrifuged, washed by trituration with water containing formic acid.

The dark brown gum was collected. The aqueous phases were combined and passed through activated carbon. A light yellow solution is obtained which is saturated with ammonium sulfate. The precipitate formed is centrifuged, triturated with water, centrifuged and washed again with water to obtain precipitate A.

The mother liquors were also saturated with ammonium sulfate, resulting in the formation of a precipitate which was centrifuged and washed three times to obtain precipitate B.

The precipitate A and precipitate B are combined. The combined fraction was taken up in ethanol and the mixture was stirred at 20 ° C for 1 hour and then held at 0 ° C for 16 hours. After centrifugation, washing with ethanol and then. ether and drying under vacuum to give 3.47 g of the desired product (isomer syn).

NMR Spectrum · (DMSO 60 MHz):

(a) singlet 2.03 µm;

(b) 3.55 µm singlet;

(c) doublet 5.19 bp J = 5 Hz (d) singlet 6.8 bp.

Example 21

3-Acetoxymethyl-7- [2- (2-tritylamino-4-hiazolyl) -2-methoxyiminoacetamido] -3-cephem-4-carboxylic acid dimethylamine salt, isomer syn

Grade A

Ethyl (2-acetyl-2-methoxyimino) acetate

4.69 kg of ethyl (2-acellyl-2-hydroxyiminoacetyl acetate) (equivalent to 4.21 kg of pure product) is added to 21 liters of anhydrous pure acetone and 6.1 kg is added at 20-25 degrees Celsius. The resulting suspension was stirred for 10 minutes, then 3.72 kg of dimethylsulfate was added at 25 ° C and the reaction mixture was stirred for 3 hours at 20-25 ° C. 126 liters of demineralized water and extracted four times with 5 liters and then 2 liters of methylene chloride, then washed with 10 liters of demineralized water, dried, centrifuged, washed with 2 liters of methylene chloride ¹ and distilled under vacuum to obtain 4.88 kg of the desired product.

Rf = 0.7 (thin layer chromatography on silica gel; eluent: methylene chloride / ethyl acetate · 9: 1).

The product obtained is identical to the product obtained in step a of Example 3.

Grade B

Ethyl (4-bromo-2-nitroxylaminoethyl) acetate

3.53 kg of the product obtained in Step A are added to 18.6 liters of methylene chloride and 3.5 grams of paratoluenesulfonic acid. A solution of 2.96 kg of bromine in 3.5 liters of methylene chloride is added to the resulting solution over a period of 30 minutes while maintaining the temperature in the range of 22 ° C + 1 ° C. Release of hydrobromic acid was observed after 15 minutes of addition of said solution. The reaction mixture was stirred at 22 ° C for 45 min, poured from the vessel into the vessel, and then washed twice with 14 L of ice-cold demineralized water.

The washings are re-extracted twice with 3.5 liters of methylene chloride. After drying, filtering, washing and distilling under vacuum, 4.73 kg of the desired product is obtained.

This product is identical to the product obtained in Step B of Example 3.

Grade C

Ethyl [2- (2-amino-4-hiazolyl) -2-methoxyimino] acetate, isomer syn

1.43 kg of thiourea are mixed with 3.55 liters of ethanol and 8.1 liters of demineralized water. The mixture thus obtained is stirred for 10 minutes at 20 DEG C., and then 4.730 kg of the product prepared in Step B in 3.55 liters of ethanol are added at 20 DEG to 25 DEG. The mixture is stirred for three hours at 20-25 ° C, then cooled to 15-20 ° C and neutralized to pH = 71.6 liters of ammonia (184.6 g / l).

The mixture was stirred for 15 minutes at 20-25 ° C, then centrifuged and washed five times with 1.8 liters of demineralized water. 2,947 kg of the expected product is obtained.

under nitrogen and stirring. Already at 55 ° C complete dissolution is achieved.

To the refluxing mixture under a nitrogen atmosphere was added 5.235 liters of about 2 N sodium hydroxide solution. Rapid crystallization occurs. The reaction mixture is stirred for one hour at reflux and under nitrogen atmosphere. The temperature is then reduced to 20-25 ° C and maintained at this temperature for two hours. After centrifugation, washing four times with 2.1 liters of ethanol and drying, 4.02 kg of the desired product is obtained.

Mp 162 ° C. Grade F

The product obtained is identical to the product obtained in steps A and C of Example 3.

Grade D

Ethyl [2- (2-tritylamino-4-thiazolyl) -2-methoxyethoxyacetate, isomer syn

3.41 kg of the product obtained in the previous step C are mixed with 17 liters of methylene chloride and 2.275 liters of triethylamine. The mixture thus obtained is stirred for 15 minutes, and then 4.55 kg of trityl chloride are added thereto under stirring under nitrogen atmosphere at 20-25 ° C for one hour. The mixture is stirred for 20 hours at 20-25 ° C under a nitrogen atmosphere, whereby the mixture is stirred for 20 hours. triethylamine hydrochloride crystallizes.

The mixture is washed with 10.2 liters of 0.5 N hydrochloric acid (glacial) and twice 10.2 liters of demineralized water (also glacial). The washings were re-extracted in a series of 1.7 liters of methylene chloride. After drying, filtering, rinsing with 1.7 liters of methylene chloride, and distilling to dryness under vacuum at a temperature below 50 ° C, 8.425 kg of crude product is obtained.

This product is dissolved at 20-25 ° C in 8.4 liters of methanol, added over one hour and, with stirring, to promote crystallization of 2.8 liters of demineralized water. The mixture is then stirred for a further hour, then centrifuged, triturated twice with 1.7 liters of methanol containing 25% water, dried at 40 ° C and isolated. 7.165 kg of the desired product.

The product obtained is identical to the product obtained in Step B of Example 3.

Grade E

2- (2-Tritylamino-4-thiazolyl) -2-methyl-ethoxyiminoacetamidoacetic acid sodium salt, isomer syn

4.175 kg of the product obtained in step D are mixed with 20.9 liters of ethanol. The mixture is then heated to reflux

2- (2-Tritylamino-4-thiazolyl) -2-methoxyiminoacetic acid, syn isomer

500 g of the product obtained in the preceding step E (corresponding to 440 g of dry product) are mixed with 2.5 liters of methylene chloride. Thereafter, 2 liters of about 1N hydrochloric acid were added at 20-25 ° C with stirring under nitrogen. The mixture was stirred for two hours at 20-25 ° C under a nitrogen atmosphere. The chloromethylene fraction was separated by decantation and washed three times with 2 liters of demineralized water. After re-extraction in a series of one liter of methylene chloride, drying, addition of 25 g of activated carbon, centrifugation, rinsing and distillation to dryness, 481 g of crude product are isolated. This product was transferred to 2.1 liters of isopropyl ether. After centrifugation, washing with 420 ml of isopropyl ether (twice) and drying under vacuum to constant weight, 424.6 g of the desired product are obtained.

The product obtained is identical to the product obtained in Step C of Example 3.

Grade G

3-Acetoxymethyl-7- [2- (2-tritylamino-4-hiazolyl) -2-methoxyiminoacetamido] -3-cephem-4-carboxylic acid diethylamine salt, isomer syn

200 g of acid are mixed in a round-bottomed flask

2- (2-tritylamino-4-hiazolyl) -2-methoxyiminoacetic acid obtained in the previous step F with 1200 ml of methylene chloride. The mixture thus obtained is then heated to reflux under argon and 600 ml of methylene chloride are then distilled off under normal pressure. The temperature of the reaction mixture is thereafter ^. The mixture is cooled to 18-20 ° C and 54 g of dicyclohexylcarbodiimide in 54 ml of methylene chloride are added at this temperature. The mixture was stirred for one hour at 18-20 ° C under argon, then a solution prepared by dissolving 61.4 g of 7-aminocephalosporanic acid in 900 ml of methylene chloride and 63 ml of triethylamine was added over 15 minutes at that temperature. The mixture was stirred for one and a half hours at 20 ° C (pH = 6.5-7). 50 ml of acetic acid are then added to the mixture, the mixture is stirred for 15 minutes at 20 ° C and then centrifuged to remove the starting 7-aminocarboxylic acid.

Wash four times with 200 ml methylene chloride. The organic solution was washed three times with 400 ml of demineralized water and then dried over magnesium sulfate. After centrifugation, washing is carried out with 200 ml of methylene chloride (twice) and vacuum distillation under argon.

The dry oily extract was dissolved in 700 ml dioxane at 20-25 ° C under stirring and under argon. Then 300 ml of the dioxyx-methylene chloride mixture are distilled off under vacuum and under argon at a temperature below 30 ° C. The temperature of the mixture is lowered to 20 ° C + 2 ° C and 500 ml of sulfuric acid-saturated ether are added. Then 52 ml of diethylamine are added. After about 10 minutes, the diethylamine salt of 2- [2-tritylamino-4- (4-thia-2,4-dimethyl) -2-methoxyiminoacetic acid crystallized.

The mixture was left under argon at 20 ° C for one hour. After centrifugation, wash three times with 100 mol of sulfuric acid saturated dioxanether solution. After drying the isolated diethylamine salt, 113.6 g of this salt are obtained. The organic solution is poured into 3.25 liters of isopropyl ether with stirring over 30 minutes. The mixture was stirred for 15 minutes and centrifuged in vacuo. After washing with 400 ml of isopropyl ether (twice) and drying under vacuum, 182 g of product are obtained, which is identical to the product obtained in Example 5.

Example 22

S-acetoxymethyl-4- (4-amino-4-thiazolyl) -2-methoxyiminoacetamido] -3-cephem-4-carboxylic acid, isomer syn

182 g of the product prepared in the preceding example 21 are added to 347 ml of formic acid and 87 ml of demineralized water under stirring and under argon at 28-30 ° C. The triphenylcarbinol crystallizes completely. The mixture was stirred under argon at 28-30 ° C for two and a half hours and then poured into 1740 ml of demineralized water and 847 g of ammonium sulfate over 15 minutes with stirring. The mixture was then stirred for 30 minutes. After centrifugation, washing twice with 174 ml of demineralized water and drying under vacuum at 25-30 ° C, 147 g of a mixture of the desired product and triphenylcarbinol are obtained.

The crude product was triturated for 1 hour at 18-20 ° C with 735 ml of sulfuric acid-saturated ether. After centrifugation, washing twice with 147 ml of saturated sulfuric acid and drying at 25-30 ° C, 89 g of the desired product are obtained.

This product was suspended in 445 ml of ethanol under stirring and under a nitrogen atmosphere. The resulting suspension is heated to 45-50 ° C with stirring and maintained under these conditions for one hour. It is then stirred for one hour at 18 to 20 ° C. After centrifugation, washing twice with 45 ml of ethanol and drying under vacuum at 20 DEG C., 76.85 g of the desired product are obtained.

This product is mixed with 230 ml of acetic acid. The mixture is stirred for 15 minutes under a nitrogen atmosphere, then 77 ml of demineralized water are added. Then about 700 ml of water is added to this solution. The mixture was stirred at 18-20 ° C for one hour, after which about 269 g of ammonium sulfate was added over 10 minutes; The mixture was allowed to stand for 15 minutes, then 3.85 g of activated carbon was added. The mixture is then stirred for 15 minutes, centrifuged and washed with 77 ml of demineralized water containing 25% acetic acid. Thereafter, 154 ml of formic acid and a piece of the final product are added with stirring at 18 to 20 ° C, whereupon crystallization by scratching the bar against the vessel wall is promoted. The mixture is left under stirring at 18 to 20 ° C for two hours and then at 0 to 5 ° C for two hours. After centrifugation, wash (4 times) 77 ml of demineralized water containing 5% formic acid. After drying under vacuum at 20-25 ° C, 49.45 g of the formate product is obtained.

The formate was suspended with stirring during one hour at 45-50 ° ^C in 250 ml of ethanol and the suspension is left for one hour at 18-20 ° C. After centrifugation, washing (twice) with 50 ml of ethanol, and drying first at 20 DEG C. under vacuum and then for 10-15 hours at 35 DEG C., 45.45 g of the desired product are obtained.

[α] D = + 64.5 ° at conc. 0.5% in water containing 0.5% sodium bicarbonate *.

The product obtained is identical to the product obtained in Examples 4, 6 and 20.

Example 2 3

Sodium salt of 3-acetoxymethyl-7- [2- (2-amino-4-thiazclyl) -2-methoxyiminoacetamide-3 '] -3-cephem-4-carboxylic acid, isomer syn

19.8 g of 3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido] -3-tert-4-carboxylic acid (isomer syn), obtained according to Example 4, 6, 20 or 22 in ml of a one molar solution of sodium ootane in methanol. The mixture is left to crystallize for 35 minutes at ambient temperature and 40 ml of ethanol are then added in one hour. After two and a half hours. Stir in an ice bath, centrifuge, wash twice with 10 ml methanol-ethanol (1: 1j, wash twice with 10 ml ethanol, wash twice with 20 ml ether, dry (2 hours at 45 ° C under vacuum) and finally dry over acid. 16.191 g of the desired crystalline product is obtained in vacuo.

Maintaining the absence of air humidity, a product with the following physical properties is obtained:

H2O [Karl Fischer] = 0.2%

Determination of methanol and ethanol by gas chromatography:

Methanol 0,1%

Ethanol 0,45%

Analysis for C PjH CN ^N ^Na (477.5) calculated:

C 40.24, H 3.38, N 14.67, S 13.43,

4.81% Na;

found:

39.9% C, 3.5% H, 14.5% N, 13.1% S, 4.8% Na.

The product hydrates in air.

The X-ray spectrum (Debye Scherrer) makes it possible to confirm the crystalline nature of the product obtained.

It is also possible to use isopropanol instead of ethanol to crystallize the desired product.

Example 24

3-Acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido] -3-cephem-4-carboxylic acid sodium salt in crystalline form, isomer syn

Grade A

3-Acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido] -3-cephem-4-carboxylic acid solvate (syn isomer) and formic acid

87.2 g of 3-acetoxymethyl-4- [2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetamido] -3-cephem-4-carboxylic acid diethylamine salt prepared as described in Example 5 was added in small 220 ml of pure formic acid and 220 ml of water are added to the mixture in portions. The resulting mixture was stirred for 30 min at 50 ° C, cooled, and 30.1 g of triphenylcarbinol collected by filtration. The filtrate was poured into 450 ml of water, the coal was removed and the solution was concentrated at 40 ° C under vacuum until a precipitate formed. Add 200 ml of anhydrous ethanol and cool with ice, filter, wash with ethanol and ether and dry under vacuum.

31.1 g of the expected product are obtained.

For C16H17N5O7S2, HCO2H, H2O (541.5) calculated:

C, 39.3; H, 4.08; N, 13.48; S, 12.34; H, 3.46;

found:

39.2% C, 4.1% H, 13.2% N, 12.8% S,

4.15% H2O.

Grade B

Crystalline sodium salt g of the solvate, freshly prepared as described in Step A, is dissolved in 75 ml of methanol and then 3 g of activated carbon is added to the resulting solution. After filtration, 5 ml of isopropanol are added with stirring. ^R After 16 h at 0 ° C the resulting crystals collected. The crystals were washed with ethanol and then with ether and then dried under vacuum at 50 ° C for two hours.

7.95 g of the expected product is obtained.

Following brief exposure to air, the following analysis is obtained:

· C for ₅ NaO7S2 _ie H16N. 1 H2O (495.5) calculated:

38.78% C, 3.66% H, 14.14% N, 4.64% Na,

12.94% S;

found:

38.6% C, 33%, 14% N, 33.8% N, 4.6% Na,

13.2% S.

Example 25

Sodium salt of 3-acetoxymethyl-7t [2t - (2-amino-4-thiazolyl) -2-methoxy-aminoacetamido] -3-cephem-4-carboxylic acid in amorphous form (isomer syn)

Grade A

3-Acetoxymethyl-7- [2t (2-amino-4-thiazolyl) -2-methoxyiminoacetamido. Dissolve 3 liters of ethanol (96 why) and 350 ml of water in a mixture, concentrate in vacuo to a volume of 300 ml, crystallize during concentration, then cool in an ice bath for one hour and filter. with a small amount of ethanol and drying under vacuum at ambient temperature

44 g of the desired product are obtained.

For C ₁₆ H ₁₇ N ₅ O ₇ S ₂ 0.8 mol C ₂ H ₅ OH (492.3) calculated:

% C, 42.94;% H, 4.46;% N, 14.23; found:

% C, 43.0%;% H, 4.4%;

Grade В

Amorphous sodium salt g solvate. with ethanol obtained in step A is added at 0 DEG C. to 60 ml of water, and then 0.504 g of sodium bicarbonate dissolved in 6 ml of water are added to the mixture thus obtained with stirring. The neutral solution was filtered and the filtrate was immediately lyophilized. The resulting product is then exposed to brief air exposure.

Analysis calculated for: C ₁₆ H | ₀ N _with NaO7S · ,. 1.5 H ₂ O (504.47) calculated:

% C, 38.89;% N, 3.8%; found:

C 38.2%, H 3.9%, N 13.6%

Example 26

Sodium salt of 3-acetoxymethyl-7- [2-

- (2-Amino-4-thiazolyl) -2-methoxyiminoacetamide] -3-cephem-4-carboxylic acid in crystalline form (isomer syn)

To 4.95 g of 3-acetoxymethyl-7- [2-

- (2-Amino-4-thiazolyl) -2-methoxyiminoacetamido) -3-cephem-4-carboxylic acid (isomer syn) obtained according to Example 4, 6, 20 or 22 is added with 5 ml of ethanol. The mixture is then added with stirring in a water bath. 10 ml of a 1 molar aqueous sodium bicarbonate solution. After dissolution, 15 ml of ethanol are added, after which, after concentration under vacuum at 30 DEG C., leaching with ethanol and drying to constant weight, a powder is obtained which is taken up in 15 ml of methanol. The mixture was seeded for crystallization and then placed in the refrigerator overnight. 3.407 g of product are isolated in crystalline form.

Example 27

Sodium salt of 3-acetoxymethyl-7- [2-

- (2-amino-4-thiazolyl) -2-methoxyimino-acetamido] -3-cephem-4-carboxylate in crystalline form (isomer syn)

0.5 g of amorphous sodium salt, obtained as described in Example 25, is dissolved in 2 ml of miethianol, 0.25 ml of n-butanol is then added with little stirring and cooled to about 6 ° C in a refrigerator. . The crystals were washed with a small amount of cold methanol and dried under vacuum at 40 ° C for 3 hours in the presence of concentrated sulfuric acid.

0.2 g of crystalline product is obtained.

After a short delay of the resulting air product, the following analysis is obtained:

For C ₁₆ H ₁₆ N ₅ NaO ₇ S ₂ · 1.5 H ₂ O (504.47) calculated:

38.09 ° C, 3.8% H, 13.88% N, 29.96% O; found:

C 38.4%, H 3.8%, N 13.8%, O 27.1%.

When operating under analogous conditions, slightly different crystalline forms are obtained, containing, for example, 0.5 mol of water or 1 mol of water and 1 mol of methanol.

The X-ray spectra (Debye Scherrer) of the products obtained above confirm the crystalline nature of the products obtained.

Example 28

An injection preparation having the following composition is prepared:

3-Acetoxymethyl-7- [2- (2-amino-4-thiazoilyl-2-hydroxyiminoimino-acetamido) -3-cephem-4-carboxylic acid 500 mg

Sterile aqueous excipient q. p. 5 mil

Example 29

An injection preparation having the following composition is prepared:

3-Acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido] -3-cephem-4-carboxylic acid 500 mg

Sterile aqueous excipient q. p. 5 ml

Example 30

An injection preparation having the following composition is prepared:

3-Acetoxy-ethyl-7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido] -3-cephem-4-carboxylic acid sodium salt 500 mg Sterile aqueous excipient q. p. 5 ml

Example 31

An injection preparation having the following composition is prepared:

3-Acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2- (1-methylethoxyimino) 207575 acetarnido] -3-cephem-4-carboxylic acid 500 mg

Sterile aqueous excipient q. p. 5 ml

Example 3 2

The following gelatin capsules are prepared:

3-Acethoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido] -3-cephem-4-carbamic acid 250 mg

Excipient q. for one 400 mg capsule

Example 1 3

Gelatin capsules are prepared as follows:

3-Acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido] -3-cephem-4-carboxylic acid, sodium salt 250 mg

Excipient · q. for one 400 mg capsule

Example 34

The following gelatin capsules are prepared:

3-Acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2- (1-methyl-ethoxyimino) -acetamido] -3-cefeim-4-carboxylic acid 250 mg

Excipient q. for a capsule of 400 · mg

Pharmacological studies of the products prepared by the process of the invention.

In vitro potency.

Dilution method in liquid medium.

An equal amount of sterile nutrient medium is dispensed into a series of tubes. A certain amount of test product is then placed in the first test tube, and increasing amounts of the test product are placed in each subsequent test tube in series. All tubes are then inoculated with a particular bacterial strain.

After a 24 hour or forty-eight hour incubation in a thermostat at 37 ° C, the inhibition of bacterial strain growth in the backlight is evaluated in individual tubes to determine the minimum inhibitory concentrations (MIC, expressed in µg / cm 3).

The results obtained are shown in the following tables (product A) is 3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido] -3-cephem-4- carboxylic acid in the form of an isomer of antjj.

T a b u 1 k a '1

3-Acetoxymethyl-7- [2- (2-a-amino-4-thiazolyl) -2-hydroxyiminoacetamido] -3-cephem-4-carboxylic acid, anti-isomer

MIC (JU / ml) hours hours

Bacterial strain

Staphylococcus aureus' UC 1061 not resistant to penicillin 5 5 Staphylococcus aureus UC 1128 resistant to penicillin 10 10 Staphylococcus aureus Exp. No 54146 5 10 Streptococcus pyogenes A 561 0.5 0.5 Bacillus subtilis ATCC 6633 2 5 Escherichia Coli ST UC 1020 3 3 Escherichia Coli RT, US 1261 1 1 Escherichia Coli Exp. 1¾¾ 2 2 Escherichia Coli RG R55 123D 5 10 Klebsiella pneumoniae Exp. 52145 0.6 0.6 Klebsiella pneumoniae 2536 R 20 May 40 Proteus mirabilis (indole-) A 235 2 2 Salm-onella typhimurium 420 2 2 Enterobacter cloacae 681 40 40

Table 2 Bacterial strain The product of the examples The product of the example Product A . 4, 6, 20, 22 7 MIC ((ig / rnl) 24 h 48 h 24 h 48 h 24 h 48 h

Staphylococcus aureus ATCC 5638

Penicillin UC 1061 Staphylococcus aureus UC 1128, resistant to 1 1 1 2 40 40 penicillin Staphylococcus aureus exp. 2 2 2 3 > 40 > 40 No. 54 146 3 3 2 2 40 40 Streptococcus pyo enes A 561 0.02 0.05 0.01 0.01 0.4 0.4 Streptococcus faecalis 5432 2 2 1 3 > 40 > 40 Streptococcus faecalis 99 F 74 2 3 3 10 > 40 > 40 Bacillus subtilis ATCC 6633 Escherichia Coli resistant 0.2 1 1 2 20 May 20 May against tetracycline ATCC 9637 = UC1020 0.2 0.2 0.1 0.1 20 May 40 Escherichia Coli resistant to tetracycline ATCC 11 303 = UC 1261 0.02 0.02 <0.02 <0.02 2 2 Escherichia Coli Exp. TO26B ₀ Escherichia Coli resistant 0.1 o'l 0.05 0.05 5 5 against Gentamicin and Tobramycin R 55 123 D 0.6 0,6—1 0.1 0.1 10 10 Klebsiella pneumoniae Exp. 52 145 0.02 0.02 <0.02 <0.02 1 1 Gentamicin resistant Klebsiella pneumoniae 2,536 0.6 0.6 0.2 0.2 > 40 > 40 Proteus mirabilis (indole-) A 235 0.02 0.02 <0.02 <0.02 2 2 Proteus vul / aris (indole +) A 232 2 40 0.2 0.6 > 40 > 40 Salmonella typhimuriuin 420 0.2 0.2 0.1 0.1 20 May 20 May Providencia Du 48 5 5 1 2 40 > 40

'Bacterial strain The product of the examples 4, 6, 20, 22 The product of the example 7 MIK 24 h 48 h Product A 24 h 48 h 24 h 48 h Pseudomonas 3,935 Exp. SG. 10 40 10 20 > 40 > 40 Serratia resistant Gentamicin 2 532 1 1 1 1 > 40 > 40 Enterohacter cloacae 681 40 40 Pseudomonas 8951 RGT 40 > 40 Staphylococcus Smith 1 1 Escherichia Coli 0.1 0.2 Escherichia Coli Ec 0.05 0.05 Escherichia Coli T 25,575 0.05 0.05 Escherichia Coli T 96 875 0.05 0.05

Table 3

Bacterial strain The product of Example 9 MIK (µg / ml) 24 hours 48 hours Staphylococcus aureus ATCC 5 638 not resistant to penicillin 1 2 Staphylococcus aureus UC 1128 resistant to penicillin 2 2 Staphylococcus aureus exp. No. 54 146 2 2 Staphylococcus aureus Co · 15, resistant against Cephialexin 10 20 May Streptoicoccus pyo / enes A 561 0.02 0.02 Streptoco'ccus faecalis 5,432 2 10 Bacillus subtilis' ATCC 6,633 1 1 Escherichia Coli - resistant against tetracycline 1 1 Escherichia .. Coli - resistant tetracycline ATCC 11 303 0.1 0.1 Escherichia Coli Exp. ΤΟ ₂ βΒβ 1 1 Escherichia Coli - resistant to Gentaimicin and Tobramycin R 55 123 D 1 1 Klebsiella pneumoniae Exp. 52 145 0.05 0.1 Klebsiella pneumoniae 2,536 - resistant against Gentamicin 2 5 Proteus mirabilis (indole—) A 235 0.2 0.2 Proteus vul / aris (indole +) A 232 2 10 Salmonella typhimurium 420 1 10 Proividencia Du 48 3 5 Pseudomonas 3,935 Exp. SG 20 May 20 May Serratia Resistant Gentamicine 2,532 0.4 1

AND

The product of Example 11 The product of Example 15

MIC 24 h

Table 4

Bacterial strain h

h

Staphylococcus aureus ATCC

638 resistant to

penicillin 2 2 20 May 20 May Staphylococcus aureus UC 1128 resistant to penicillin 2 3 40 40 Staphylococcus aureus exp. No. 54 146 2 2 ' 40 40 Streptococcus pyogenes A 561 0.02 0.02 0.4 0, Streptococcus faecalis 5 432 2 10 > 100 > 100 Streptococcus faecalis 99 F 74 2 20 May 100 ALIGN! 100 ALIGN! Bacillus subtilis ATCC 6,633 0.4 0.4 10 10 Escherichia Coli resistant against tetracycline ATCC 9 637 1 1 > 100 > 100 Escherichia Coli resistant against tetracycline ATCC 11 303 0.2 0.2 10 10 Escherichia Coli Exp. TO2eB _c 0.4 0.4 20 May 40 Escherichia Coli resistant against Gentamlcin and Tobra- miciniu R55 123D 0.6 0.6 40 40 Klebslella pneumoniae Exp. 52 145 0.05 0.05 20 May 20 May Klebslella pneumoniae 2,536 resistant to Gentamicin 2 2 > 100 > 100 Proteus mlrabilis [indole—] A 235 0.05 0.05 5 5 Proteus vulgaris (indole +) A 232 1 2 > 100 > 100 Salmonella typhimurium 420 1 1 100 ALIGN! 100 ALIGN! Providencia Du 48 5 5 100 ALIGN! > 100 Pseudoimonas 3,935 Exp. SG 20 May 40 > 100 > 100 Serratia resistant Gentamicin 2,532 2 2 > 100 > 100 Ta bulka 5 Bacterial strain The product of Example 13 Example product: MIK Leag / ml) 24 h 48 h 24 h 48 1 Staphylococcus aureu ATCC 5,638 penicillin 2 5 20 May 20 May Staphylococcus aureus UC 1128 resistant penicillin 3 5 40 40 Staphylococcus aureus exp. No. 54 146 2 5 40 40 Streptococcus pyogenes A 561 0.02 0.02 1 1 Streptococcus faecalis 5 432 5 40 100 ALIGN! 100 ALIGN! Streptococcus faecalis 99 F 74 5 20 May > 100 > 100 Bacillus subtilis ATCC 6,633 0.5 2 20 May 100 ALIGN! Escherichia Coli resistant against tetracycline ATCC 9 637 2 2 > 100 > 100 Escherichia Coli resistant against tetracycline ATCC 11 303 0.5 0.5 20 May 20 May Escherichia Coli Exp. To2gB ₆ 1 1 100 ALIGN! 100 ALIGN!

h

AND

7-575

The product of Example 13 The product of Example 17

MIC (g g / rnl) li 48 h 24 h 48 h

Bacterial strain

Gentamicin and Tobramy resistant Escherichia Coli-

R 55 123 D 1 1 100 ALIGN! 100 ALIGN! Klebsiella pneumoniae '2,536 resistant to Gentamicin 10 10 > 100 > 100 Proteus mirabilis (indole—) A 235 0.5 0.5 10 20 May Proteus vulgaris (indole +) A 232 0.5 1 > 100 > 100 Salmonella typhimurium 420 2 2 100 ALIGN! 100 ALIGN! Providencia Du 48 5 5 > 100 > 100 Pseudomonas 3,935 Exp. SG 40 40 > 100 > 100 Serratia resistant Gentamicin 2,532 2 5 > 100 > 100 Enterobacter. chloacae '681 40 40 100 ALIGN! 100 ALIGN!

Experimental infection with bacterial strain Escherichia Coli (T) O ₂ 6B ₆ .

A] In this test, the effect of the products of Examples 4, 6, 20 and 22 on experimental infection of a bacterial strain of Escherichia Coli (T) O26Be in mice is studied.

Groups of ten male mice with an average body weight of 21.5 kg were infected by intraperitoneal injection of 0.5 ml of a 24 hour (in culture medium) bacterial culture. strain Escherichia Coli (t) O ₂ 6Bg (Institut Pasteur), diluted to 1/6 with distilled water.

One hour, five hours and twenty-four hours after. said injection was administered to the mice by subcutaneous injection or oral administration of a specified amount of the test product. Mortality of the mice was then observed. over the next eight days. The results obtained are shown in Table 6 below.

Table 6 Application Mortality after 21.5 h 28.5 h 32 h Mice survived after> 8 days Control group of mice 9 1 0/10 0.05 mg subcutaneously 10/10 0.1 mg subcutaneously 10/10 0.25 mg subcutaneously 10/10 0.1 mg per os '- 1 9/10 0.25 mg. per. pers 10/10 0.5 mg per os 10/10

B) Proceed as above. of the test, wherein mice with an average body weight of 22.5 g are infected with the bacterial strain Escherichia Coli (T) Ο ₂ βΒβ, diluted to 1 / 5.5, with the bacterial strain E.

Using the products of Examples 4, 6, 20 and 22, the following results are shown in Table 7.

0 7 3 7 4

Method Mortality po Number of mice application 7 hr 9 hr 22 hr 23 hr 24 hr 25 hr 26 hr 31 hr 32 hr 46 hr 70 hr 4 days that survived 45 min 35 min 30 min 50 min, 50 min 30 min '30 min 8. day

5β

Experimental infection with bacterial strain Sallnonella typhimurium.

In this test, the effect of the product of Example 7 on experimental infection of Salmonella typhimurium in mice was studied.

Groups of ten mice with an average body weight of 20.5 g were infected by intraperitoneal injection of 0.5 ml of a twenty-four hour culture on a bovine broth of the bacterial strain Salmonella typhimurium 5210 diluted 1 / ha with distilled water.

One hour, four hours, 8 hours 24 hours, and 32 hours after the above injection, mice were injected subcutaneously with a specified amount of test product.

The results obtained are summarized in Table 8 below.

Table 8

Application Mortality after Number of surviving mice

10. day h 25 h 28 h 29 h 45 h 5611 70 h 5 days 6 days 7 days 8 days 9 days min 30 min

>>

Experimental infection with bacterial strain Proteus Mirabilis.

In this test, the effect of the products of Examples 11 and 15 on experimental infection of a bacterial strain in mice was studied.

Groups of ten mice with an average body weight of 22 g are infected by intraperitoneal injection of 0.5 ml of a 24-hour culture on a bovine broth of the bacterial strain Proteus Mirabilis A 23, diluted to 1/4.

One hour, five hours and twenty-four hours after the administration, the mice are injected subcutaneously with a determined amount of the test product.

The results obtained are summarized in Table 9 below.

Mortality po Number of mice, h 21 h 22 h 23 h 23 h 23 h 24 h 28 h 29 h 31 h 46 h 53 h 70 h 94 h survivors min 30 min 15 min 10 min 30 min 45 min 20 min 45 min 8 day

<35 1Г5 rH

Acute toxicity study in mice by intravenous administration.

The acute toxicity for the product of Example 7 was determined in female mice weighing between 19 and 21 g of the Swiss CDI Specific Pathogen Fre (CH. River-France); groups of 8 to 10 animals were used for each dose.

The animals are kept for four days in an isolated area (21 ° C + 1 ° C, stable humidity; food and water are provided ad libitum to the animals).

A solution of the product of Example 7 at a concentration of 100 mg / ml in distilled sterile and pyrogenic water was prepared. This solution is then injected into the tail vein at varying volumes and at a rate of 1 ml / min.

The results obtained are summarized in Table 10 below.

Table 10

Dose (mg / kg)

Mortality after

Day 1 Day 2 Day 3 Day 4

Total mortality

500 1000 2000

0/10

0/10

0/8

No symptoms of poisoning were observed in experimental animals during the aria injection during the four-day observation period.

Claims (4)

OBJECT OF THE INVENTION 1. A process for the preparation of novel oxirin derivatives of 7-aminothiazolylacetamidocyanosporic acid of the general formula I 'in which: Rc represents a hydrogen atom, a trityl group or a chloroacetyl group, Rc 'represents a hydrogen atom, a trityl group or an unsaturated or saturated alkyl group having 1 to 4 carbon atoms, A represents either a hydrogen atom or an equivalent of an alkali metal, an alkaline earth metal, magnesium or an amine organic base residue, and a wavy line means that the group OR'c can be in one or the other of the two possible syn or anti positions, that R'c is trityl, then Rc is trityl and, in the case where R'c is hydrogen, then Rc is hydrogen, characterized in that 7-aminocephalosporanic acid of formula ^{20 is} reacted; CH 2 ^-O- C-CWj coaH ⁰ with an acid of formula IIc (IId wherein Rd is trityl or chloroacetyl and R'd represents a trityl group or a saturated or unsaturated alkyl group having 1 to 4 carbon atoms, or even a functional derivative thereof, to give the product of formula Ic NH-Rd (Ic) in which Rd and R'd have the meanings given above, wherein the product corresponds to the product of formula Γ in which Rc is trityl or chloroacetyl; and R'c represents a trityl group or a saturated or unsaturated alkyl group having 1 to 4 carbon atoms, and the compound obtained is optionally converted into a salt as defined above, whereupon the above product Ic is optionally hydrolyzed in an acidic medium or hydrogenolysed or It is reacted with thiourea according to the meanings of the substituents Rd and R'd to form a product of the formula (Ib) or an unsaturated alkyl group having from 1 to 4 carbon atoms, whereupon the product of the formula (Ib) is optionally · importance. 2. The method according to FIG. 1 for the preparation of products of the general formula I (!) CH ₂ OCOCH ₃ (1b) wherein R _t 'represents a hydrogen atom or a saturated or unsaturated alkyl group having a total number of carbon atoms of 1 to 4, said product corresponding to the product of formula Γ in which: R c and A are each hydrogen and R c is hydrogen or saturated in which; R is a hydrogen atom or a trityl group, R 'is hydrogen, trityl not alkylovlou saturated or unsaturated group having a total carbon number of ¹ 1 to 4, A represents either a hydrogen atom or an equivalent of an alkali metal, an alkaline earth metal, magnesium or an amine organic base residue and the wavy line means that the OR 'group may be in one or the other of possible syn or anti positions; that R 'is a trityl group, then R is a trityl group, and when R' is a hydrogen atom, then R is a hydrogen atom, characterized in that the reaction of 7-aminocephalosporanic acid of the formula / NH ~ R ₁ 'I. (II! ..... where R 1 represents a trityl group and Rf represents a trityl group or a saturated or unsaturated alkyl group having from 1 to 4 carbon atoms, or a functional derivative thereof, to give the product of formula (Ia) in which: R 1 and R 1 'are as defined above, said product corresponding to the product of formula (I) in which: R is trityl and R 'represents a trityl group or an unsaturated or saturated alkyl group having a total number of carbon atoms of 1 to 4, and the compound obtained is optionally converted into a salt as defined above, whereupon the product of formula I is hydrolyzed in an acidic medium or hydrogenolysed to give the product of the general formula (Ib) (3-s- ₃ ) in which R 1 "represents a hydrogen atom or a saturated or bi-unsaturated alkyl group having 1 to 4 carbon atoms, said product corresponding to the product of formula (I) in which: R and A are each hydrogen and R 'represents a hydrogen atom or a saturated or unsaturated alkyl group having from 1 to 4 carbon atoms, whereupon the product of the general formula Ib is optionally converted into the salts as defined above. A process according to item 1 for the preparation of products of the general formula Ib (Ib) in which: Rf 'represents a hydrogen atom or a saturated or unsaturated alkyl radical having a total number of carbon atoms of 1 to 4 as well as their salts, which product corresponds to the product of the general formula kterém in which: Rc and A are each hydrogen and R 'c represents a hydrogen atom or a saturated or unsaturated alkyl group having 1 to 4 carbon atoms, characterized by treating the salt of the product of formula (Ic) with an acid, in which: Rd represents a trityl group and R 'd represents a trityl group or a saturated or unsaturated alkyl group having 1 to 4 carbon atoms to form a product of formula Ib, or a hydrogenolyzing agent is treated with a salt of the product of formula Ic in which: Rd represents a trityl group and R 'd represents a trityl group or a saturated or unsaturated alkyl group having 1 to 4 carbon atoms to form a salt of formula Ib, or a thiourea is treated with a salt of the product of formula Ic in which: Rd represents a chloroacetyl group and a saturated or unsaturated alkyl group having a total number of carbon atoms of 1 to 4, to form a salt of the product of formula Ib. 4. The process of item 2 for preparing a product of formula Ib C ^{h is} 0 -CW ₃ o (Ib) wherein R "represents a hydrogen atom. or a saturated or unsaturated alkyl group having a total number of carbon atoms of 1 to 4 as well as their salts, the product corresponding to the product of formula (I) in which: R and A. each hydrogen atom and R 'represents a hydrogen atom or an' unsaturated 'or saturated alkyl group having 1 to 4 carbon atoms, characterized by treating the salt of the product of formula (Ia) with an acid in which: R 1 represents a trityl group and R f represents a trityl group or a saturated or unsaturated group. an alkyl group having a total number of carbon atoms of 1 to 4, to give a product of the general formula (Ib), or treatment with a hydrogenolyzing agent. to a salt of the product of formula (Ia) in which: R 1 represents a trityl group and R f represents a trityl group or a saturated or unsaturated alkyl group having a total number of carbon atoms of 1 to 4, to form a salt of the product of formula Ib.