DE2702501C2 - Oxime derivatives of 7-aminothiazolylacetamidocephalosporanic acid (syn isomers), their preparation and pharmaceutical compositions - Google Patents

Description

CO ₂ H

CH ₂ -OC-CH ₃

with R 'as above, and optionally transferring the connection Ib in the usual way in a claim I corresponding salt.

6. Pharmaceutical compositions, characterized by at least one pharmaceutical Suitable compound according to claim ί or 2 as an active ingredient.

The invention relates to oxime derivatives of T-aminothiazolylacetamidocephalosporanic acid (syn isomers) of general formula I.

NH ₁

Aw

C O NH

N
OR '

/ -V

CO ₂ A

CH ₂ -OC-CH ₃

R '= Ci to C ₄ alkyl
and

A = H, an alkali metal atom or an equivalent of an alkaline earth metal or one of an organic Amine base derived ammonium group.

The "OR" group is in the syn position.

R 'accordingly denotes methyl, ethyl, propyl, isopropyl, η-butyl, sec-butyl and tert-butyl.

A denotes in particular lithium, sodium, potassium, calcium or magnesium; Organic bases suitable for A are trimethylamine, triethylamine, methylamine, propylamine, N.N'-dimethylethanolamine or tris-hydroxymethyl) aminomethane, arginine and lysine.

The invention relates in particular to compounds of the general formula 1 in which A is a hydrogen ode a sodium atom or a diethylammonium group

represents.

Compounds of general formula I are especially the compounds described in the examples, and particularly:

(a) 3-Acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamidoJ-ceph - ^ - em ^ -carboxylic acid (syn isomer) and its sodium salt, especially the crystalline sodium salt;

(b) 3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-ethoxyiminoacetamido] -ceph- (3) -em-4-carbon- acid (syn isomer) and

(c) 3-Acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-isopropyloxyiminoaGetamido] -Geph- (3) -em-4-carboxylic acid (syn isomer) and the salts of these compounds fertilize with alkali metals, alkaline earth metals and organic amine bases.

The compounds according to the invention can either have the structure described by formula I. or in a structure according to the formula Iz:

55

60

65

N-H

C — C — NH

NO
OR '

CH, -O-C-CH ₃

CO ₂ A

(Iz)

in which R 'and A have the above meaning.

The invention also relates to a process for the preparation of compounds of the general formula I, which is characterized by what is known per se

- Implementation of S-acetoxymethyl-Z-aminocephalosporanic acid of formula 25

HjN

NH — R,

30th

CH ₂ -OC-CH ₃

CO ₂ H

with an anhydride of an acid of the formula II (syn- 35 isomer)

NH-R ₁

= / CO ₂ H (Π)

Il

N
OR '

with Ri = one by acid hydrolysis or hydrogenolysis easily removable group and R '= Cr to Gt-alkyl or with a mixed anhydride of Acid II and a monoalkyl carbonic acid ester to form a compound of the formula Ia (syn isomer)

CO NH _n (Ia)

N
OR '

with Ri and R 'as above and formation of the formula Ia to a compound of

acid hydrolysis or hydrogenolysis of the compound formula Ib (syn isomer)

NH ₂

/ ll \

C O NH

Il

N
OR '

away)

CH ₂ -OC-CH ₃

CO ₂ H

with R 'as above,

- as well as transferring the connection, if applicable Ib in the usual way in a (claim 1) corresponding salt.

Groups Rj which can easily be split off by acid hydrolysis or hydrogenolysis are, for example, t-butoxy ^ carbonyl, trityl, benzyl, dibenzyl, Tf ichlöf ethyl, carbobenzyloxy, Formyl, trichloroethoxycarbonyl and 2-tetrahydropyranyl.

The anhydride of the acid of the formula II can in situ by the action of isobutyl chloroformate or of Dicyclohexylcarbodiimide can be generated on the acid; it can also be used other anhydrides, which are generated in situ by the action of other alkyl chloroformates, Dialkylcarbodiimides or other dicycloalkylcarbodiimides can be made. When the implementation of S-acetoxymethyl-T-aminocephalosporic acid with a rni * isnhutylehlorfnrmiat Anhydride of the acid of the general formula II is carried out, it is preferably carried out in the presence of one basic agent.

Alkali metal carbonates or tertiary organic bases, for example, can be used as the basic agent are, for example N-methylmorpholine, pyridine or trialkylamines such as triethylamine.

Agents for acid hydrolysis with which the compounds of the formula Ia are optionally reacted are formic acid, trifluoroacetic acid and acetic acid. These acids can be anhydrous or in aqueous solution can be used. The zinc / acetic acid system is particularly suitable as a hydrolysis agent.

For splitting off t-butoxycarbonyl or trityl groups an acidic hydrolysis agent such as anhydrous trifluoroacetic acid is preferably used, aqueous formic acid or acetic acid.

The zinc / acetic acid system is preferred for splitting off the trichloroethyl group, the catalytic one Hydrogenation used to split off benzyl, dibenzyl and carbobenzyloxy groups.

The compounds of the formula Ib can be converted into salts by customary processes. the Salt formation can be caused, for example, by the action of inorganic bases such as sodium or potassium hydroxide or sodium hydrogen carbonate or substituted or unsubstituted aliphatic carboxylic acids by salts such as diethyl acetic acid, ethylhexanoic acid and especially acetic acid on the appropriate acids.

The preferred terms of those listed above Acids are the sodium salts.

The salt formation can also be caused by the action of an organic base such as triethylamine can be achieved.

To prepare the salts, the solvates of the free acids can also be used instead of the free acids Starting material can be used, for example the solvates with water, formic acid or alcohol can be obtained.

The solvates with an alcohol, in particular ethanol, can also be obtained in this way, for example that the solvate formed with formic acid is treated with a mixture as alcohol and water and the solution is then concentrated.

This salt formation is preferably in one Solvent or a mixture of solvents, for example in water, ethyl ether, methanol, ethanol or acetone.

The salts will
Reaction conditions
Maintain shape.

The crystallized

Depending on the applied in amorphous or crystalline

Salts are preferably prepared by using the free acids or their formed for example with formic acid or ethanol ten solvates can react with a salt of the abovementioned aliphatic carboxylic acids, preferably with sodium acetate.

When preparing a sodium salt, the reaction is carried out in a suitable organic solvent carried out, for example in methanol, the solvent containing small amounts of water can.

It is also possible to convert amorphous salts into the crystalline form. An amorphous sodium salt, which is in the form of a solvate with, for example, 0.5, 1 or 1.5 mol of water, in a suitable one organic solvents, preferably dissolved in a lower alcohol such as methanol; the Crystallization can then be carried out immediately or after adding other solvents, for example of ethanol, isopropanol, n-butanol,

Acetone, ethers and generally organic solvents that are miscible with methanol.

When the starting material, the solvent, or both contain water, the crystallized salt in Form of the hydrate can be obtained. The crystallized sodium salt of 3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido] -ceph- (3) -em-4-car- bonsäure (syn-isomer) can, for example, with 0.5, 1 or 1.5 mol of water can be isolated.

The compounds of formula II can be prepared by adding thiourea to a Allow compound of formula III to act

Cl-CH ₂ -C —C —COjalk (DT)

Il Il ο ν

OR '

in which R 'and alk are an alkyl group with 1 to 4 carbon atoms represent, and after treatment with a base, a compound of the following formula IV is obtained

NH ₂

CO, alk

OV)

N
OR '

with R 'and alk as above, which is treated with a functional derivative of a group which are easily split off by acid hydrolysis or by hydrogenolysis Vann ^ whereby a compound of the formula V results

x ^N CO ₂ alk

OR '

in which Ri is a group which can be easily split off by acid hydrolysis or hydrogenolysis and R ^{1 is} C [- to Gt-alkyl; the compound of the formula V is finally treated with a base and then with an acid and thus converted into a compound of the formula II:

NH - R.

CO ₂ H

N
OR '

In a preferred embodiment of the process., Potassium acetate is used as the base for the preparation of the compound of the formula IV, but there are also Carbonates and bicarbonates of alkali metals, dilute sodium hydroxide or potassium hydroxide usable.

The functional derivative of the group which can be easily split off by acid hydrolysis or hydrogenolysis preferably trityl chloride, which in the presence of triethylamine or other tertiary nitrogen bases, like other trialkylamines, N-methylmorpholine or Pyridine is used.

Furthermore, other functional derivatives with groups that can easily be hydrolyzed by acidic can also be used or can be split off by hydrogenolysis. Such derivatives are z. B. t-butyl chloroformate, which is in situ or t-butyl azidoformate, trichloroethyl chloroformate or benzyl chloroformate, the mixed anhydride of formic acid and acetic acid, the was made in situ. Benzyl or dibenzyl chloride or other benzyl or dibenzyl halides, phthalic anhydride or about N-carbethoxyphthalimide.

Sodium hydroxide is preferably used to saponify the compound of the formula V, but are

other bases such as potassium hydroxide or barium oxide can also be used.

To isolate the acid of the formula II, preference is given to using dilute hydrochloric acid, however (V) 5 acetic acid or formic acid can also be used.

The compounds of general formula Ib or their salts can be prepared in that a Salt of a compound of the formula yes either if Ri represents a group which can easily be split off by acid hydrolysis, is hydrolyzed with an acid, whereupon the The resulting acid is converted into a corresponding salt, if necessary, or, if Ri is one by hydrogenolysis Easily cleavable group means is reacted with an agent for hydrogenolysis, wherein a corresponding salt is obtained.

The acid which is used to hydrolyze the salts of the compounds of the formula Ia is are preferably formic acid, but trifluoroacetic acid or acetic acid can also be used will. The acids can be used in anhydrous form or in an aqueous solution.

As a means for hydrogenolysis, for. B. used catalytic hydrogenation.
The compounds Ib can also be prepared by reacting S-acetoxymethyl-T-aminocephalosporanic acid of the formula

(Π)

H ₂ N

CH ₂ -O-C -CH ₃

O ₂ H

with an anhydride of an acid of the formula Ha
NH-COCH ₂ Cl

, ^N CO ₂ H

(Ha)

N
OR '

uiitR '= C ₁ - to Q-alkyl to a compound of
Formula IX

NH-COCH ₂ Cl

(DQ

O ₂ H

CH ₂ OC-CH ₃

with R 'as above and reaction of the compound of the formula IX with thiourea to give the corresponding Compound of formula Ib.

The implementation of S-acetoxymethyl ^ -aminocephalosporanic acid with the acid of the formula Ha is carried out under the same conditions as the reaction of 3-acetoxymethyl-7-aminocephalosporanic acid carried out with the acid of formula II.

The reaction of the compound IX with thiourea is preferably carried out in neutral or acidic Medium. This type of reaction is described in the literature (cf. Masaki, JACS 90 [1968] 4508).

The compounds of the formula Ha
NH-COCH ₂ Cl

when the reaction of the thiourea with the compounds of formula III in an aqueous Solvent such as aqueous acetone or aqueous ethanol or carried out at room temperature being, using an approximately stoichiometric amount of thiourea for a very short time of approximately Allow 1 to 3 h to react, or if both of the above measures are applied at the same time.

The compounds of the formula IV obtained in Examples 1, 4, 5 and 7 have corresponding properties syn configuration.

In that filed before the priority dates of the present invention and published only afterwards DEOS 25 56 736 are cephem compounds of the general formula

S N _

(Sa)

N
OR '

with R '= Ci- to Q-alkyl are produced by that a functional chloroacetic acid derivative with a compound of formula IVb

NH ₂

CH ₂ R ⁴ OOH

in which:

C O, alk

(IVb)

R 'NH ₂ or OH (free or protected),

R ² NH ₂ or OH or one in NH ₂ or OH

convertible group,
R ³ H, methoxy or a group which can be converted into methoxy,

R ⁴ H or a nucleophilic group and
R ⁸ H or halogen,

OR '

with R! as above and alk? = Ci- to d-alkyl to one Compound of the formula Va

NH-COCH ₂ Cl

(Va)

₌ / ^N CO ₂ alk

OR '

with R 'and alk is reacted as above, which is then reacted with a base and then with an acid is treated.

The syn configuration of the compounds of formula I is based on the configuration of the compounds of Formula IV back as the configuration of compounds prepared from compounds of formula IV is retained in the synthesis

The configuration of the compounds of formula IV depends on a number of parameters in the Making these connections.

It was found that the syn isomers obtained intermediates for these compounds and a process for their preparation is given.
For formula I it is defined that R ² also includes groups which can be converted into an amino group or a hydroxyl group, but the preferred compounds mentioned on page 22, line 15 ff., To page 23, line 4, as well as the Compounds of claims 2-7 and 10-13 excluding cc — NH ₂ or «—OH compounds, but no oxyimino compounds. Examples 2-5, 10, 12, 14, 41, 42, 47, 50, 55 and 71 also relate to corresponding end products with a — NH ₂ or a — OH groups.

The compounds of the formula VII listed on page 2, which have an oxyimino group or a may have corresponding protected group (see. Also p. 5, formulas XII, XVI and XV), serve as Starting compounds or intermediates.

The syn-anti isomerism of the corresponding oxyimino compounds consequently, no importance was attached to it, since this isomerism is responsible for the reduction of the Oxyimino group to the amino group and its subsequent diazotization is irrelevant This is on p. 19, 2nd paragraph, DE-OS 25 56 736 also expressly emphasized: There it is stated that the to the A-oxy-imino-2-substituted belonging to intermediates XIV and XVI (XIV is to be corrected there in XVI) ThiazoI-4-yl-acetic acid derivatives theoretically as syn- and anti-isomers to the oxyimino group may exist, but both isomers are equally good for this Reduction are suitable.

DE-OS 25 56 736 is therefore not based on the object of certain isomers (syn or anti) of

To provide connections I, also is no stereosis * selective synthesis for syn ^ isomers given. this will also supported by the fact that the anniversaries of DE-OS 25 56 736 later made another application (DE-OS 27 15 385) has submitted to the syn-isonation of certain Oxyiminocephalosporansäurederivate DE-OS 25 56 736 is directed.

It follows that between the present invention and the subject matter of DE-GS 25 56 736 none Identity exists.

From the JA-OS 42 885/76 of April 14th, 1976 goes also show that the anti-isomers of certain cephalosporanic acid derivatives have a higher antibacterial effectiveness than the corresponding syn compounds.

In contrast, the invention is based on the surprising finding that the isomerism on Oxyimino nitrogen plays a functionally crucial role, since the syn isomers according to the invention as Surprisingly proven pharmacologically more effective than the corresponding anti-isomers.

The compounds of general formula I have a very high antibiotic activity against both gram-positive bacteria such as staphylococci, streptococci, especially penicillin-resistant staphylococci, as well as gram-negative bacteria, especially coli-like bacteria, Klebsieila, Salmonella and Proteus.

As a result of these properties, pharmaceutically usable compounds according to the invention are suitable as Active ingredients for the treatment of diseases caused by sensitive pathogens, in particular of staphylococcal infections such as staphylococcal septicemia, malignant staphylococcal infections des Facial or skin, pyoderma, septic or purulent wounds, anthrax, phlegmon, erysipelas, simple acute or post-flu staph infections, bronchopneumonia and pneumonia.

These pharmaceutically suitable compounds according to the invention can also be used as active ingredients for Treatment of colibacillosis and concomitant infections caused by Proteus, Klebsiella and Salmonella Infections and other diseases caused by Gram-negative bacteria.

The superior pharmacological effectiveness of the compounds according to the invention is evident from the following Comparative tests.

The particularly favorable pharmacological effectiveness of the syn isomer of the sodium salt of 3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacet- amido] -ceph- (3) -em-4-carboxylic acid (short name HR 756, non-proprietary name CEFOTAXIM) is also an object of the following publications:

Y. A. Chabbert, A. Dang Van, and R. Labia, Abstracts the 18th tnterscience Conference on Antimicrobial Agents and Chemotherapy 1978, Atlanta, 6a, abstract 76, H.C Neu, N. Aswapokee and K.P. Fu, op. Cit., Abstract No. 77, J. Sosna, P. Murray and G. Medoff, loc. cit., Abstract No. 78, H.W. Van Landuyt and M. Pyckavet, op. Cit., Abstract No. 79, E. Schrinner, N. Kiesel, M. Limbert and A. Lutz, op. Cit., Abstract No. 80, W. Marget, B. Belohradsky and R. Roos, op. Abstract No. 296, K. H. Spitzy, W. Graniger, H. Pichler and J. Lackner, op. Cit., Abstract No. 297 and P. M. Shah, E. B. Helm and W. Stille, op. Cit., Abstract No. 298, J. M. T. Hamilton-Miller, W. Brumfitt and A.V. Reynolds, J. of Antimicrobial Chemotherapy 4 (1978) 437-414, R. Van Hoof, J.P. Butzler and E. Yourassowsky, The Lancet (1978) 209-210, Y. A. Chabbert and A. J. Lutz, Antimicrobial Agents and Chemotherapy 14 (1978) 749-754, B.C.

Stratford, The Lancet (1978) 528-529, H.C. Neu, N. Aswapokee, P. Aswapokee and K.P. F; i, Antimicrobial Agents and Chemotherapy 15 (1979) 273-281 and S. Masuyoshi, S. Arai, M. Miyamoto and S. Miisuhashi, Antimicrobial Agents and Chemotherapy 18 (1980) 1-8.

The invention accordingly also relates to pharmaceutical compositions which, as active ingredients, at least contain a pharmaceutically acceptable compound of the formula I (syn isomer), in particular those in the Examples of compounds described.

Examples are, in particular, pharmaceutical compositions that contain at least one active ingredient Compound of the general formula I contain, in which A represents a hydrogen atom or a sodium atom.

Such pharmaceutical compositions contain, for example, the following compounds as active ingredients:

3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamidoj-ceph-pj-em ^ -carboxylic acid (syn-isomer) (cf. Examples 1, 2, 6, 7) and its sodium salt (cf. Example 3) or the crystallized sodium salt of this acid (Examples 8, 9, 11, 12);
3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-ethoxyiminoacetamidoJ-ceph-fSl-em ^ -carboxylic acid (syn isomer) and

3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-isopro-

pyloxyiminoacetamido] -ceph- (3) -em-4-carboxylic acid

(syn isomer)

and the pharmaceutically suitable salts of these compounds with alkali metals, alkaline earth metals and organic amines.

The compositions can be buccal, rectal, parenteral or topically applied locally administered to the skin or mucous membranes.

They can be solid or liquid and in customary Pharmaceutical forms used in human medicine are present, for example as simple or coated tablets, gelatine capsules, granules, suppositories, injectable compositions, pomades, Creams and gels that can be prepared using conventional methods. The active ingredients can can also be incorporated into excipients commonly used for pharmaceutical compositions be e.g. B. in talc, gum arabic, lactose, starch, magnesium stearate, cocoa butter, aqueous ode. non-aqueous carriers, fatty substances of animal or vegetable origin, iParaffin derivatives, glycols, various Wetting agents, dispersants or emulsifiers and preservatives.

The dose administered depends on the disease being treated, the person being treated, the mode of administration and the respective compound and is for example 0.250 to 4 g / day when administered orally to humans (compound of Example 1 and

3) or 0.5 to 1 g if administered intramuscularly three times a day Administration.

The compounds of the formula III, which are not yet known, can be prepared from γ-chloro-α-oxyiminoacetyl acetic acid ethyl ester (described in J. of Medicinal Chemistry 1973, Volume 16, No. 9) as is explained below

These compounds are grounded by reaction of γ-chloro-a-oxyiminoacetyl acetic acid ethyl ester with ent-

speaking alkyl halides or sulphites.

In addition to the compounds described in the examples, the following compounds are according to the invention accessible:

a-Acetoxymethyl-Zp ^ -amino- ^ thiazolylJ ^ -propyloxyiminoacetamidoi-ceph - ^ - em ^ -carboxylic acid
(syn isomer);

3-Acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-butyloxyiminoacetamidoJ-ceph - ^ - em- ^ carboxylic acid
(syn isomer);

3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-i-butyloxyiminoacetamidoJ-ceph-iSJ-em-'t-carboxylic acid
(syr isomer) and

3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-t -butyI-oxyiminoacetamidopceph-CT-em-i-carboxylic acid
(syn isomer)

as well as the salts of these compounds with alkali metals, Alkaline earth metals and organic amine bases.

The following examples illustrate the invention.

example 1

3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -

2-methoxyiminoacetamido] -ceph- (3) -em-

4-carboxylic acid (syn isomer)

A.

3-acetoxymethyl-7- [2- (2-trityiamino-

T-thaazolyl) -2-methoxyiminoacetamido] -ceph-

(3) -em-4-carboxylic acid (syn isomer)

Level A:

2- (2-Amino-4-thiazolyI) -2-methoxyiminoacetic acid ethyl ester (syn isomer)

1 g of ethyl y-chloro-a-methoxyiminoacetoacetate, 3 ml of absolute ethanol and 0.42 g of powdered thiourea were mixed and at about 2 h Stirred at room temperature After dilution with 60 ml of ether, the hydrochloride crystallized out: after stirring, Centrifugation, washing and drying gave 685 mg of hydrochloride.

The material was dissolved in 4 ml of water at 50 ° C and up to a pH of 6 with potassium acetate added, whereupon the liberated amine crystallized out. After cooling, suctioning off, washing with water and drying, 270 mg of the expected product were obtained obtain; 161 ° C.

The product obtained had a syn configuration.

NMR (60 MHz, CDCI ₃ ). ppm: 4.0 N-OCH ₃ , 6.7 proton of the thiazole ring.

Level B:

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

4.6 g of the product from Step A were dissolved in 92 ml of methylene chloride at 30 ° C. After cooling to -1O ⁰ C 2.9 ml of triethylamine were added; after further cooling to -35 ° C., 6.1 g of trityl chloride were added over the course of 15 minutes, whereupon the mixture was allowed to warm to room temperature again in 2.5 hours drying, evaporation, resuming in ether, again evaporating and dissolving in methanol was added water and ether, and after the Auskrjstallisieren of the product was filtered off and washed with ether, and it was 6.15 g of the expected product are obtained F. 120 ⁰ C..

The product obtained had a syn configuration.

Level C:

2- (2-TrityIaniino-4-thiazoIyl) -2-methoxyiminoacetic acid (syn isomer)

7.01 g of the ester obtained in Step B were dissolved in 35 ml of dioxane The reaction Lm oil bath was heated to 110 ⁰ C and within 5 min with 9 ml of 2 η hydroxide solution was then ίο 30 min refluxed, wherein the sodium salt crystallized. After cooling and suction, washing was carried out with dioxane and then with ether and a first fraction of 5.767 g of salt was obtained. After evaporation of the mother liquor, a second fraction of 1.017 g was obtained, which corresponds to a total yield of 6.784 g of salt

3.06 g of the salt were added to 65 ml of methylene chloride and 6.5 ml of 2 η hydrochloric acid; after washing with The water and drying were evaporated to dryness whereupon the free acid was quantitatively obtained.

The product obtained had a syn configuration.

NMR (60MHz, DMSO) ppm: 3.68 N-OCH ₃ , 6.6 proton of the thiazole ring.

Level D:

3-AcetoxymethyI-7- [2- (2-trityIamino-4-thiazolyI) -

2-methoxyiminoacetamido] -ceph- (3) -ern-4-carboxylic acid

(syn isomer)

The dry acid from Step C was dissolved in 30 ml of dry methylene chloride with 0.78 g of dicyclohexylcarbodiimide are added and 1 h at room temperature gerührt- After centrifuging off the dicyclohexylurea and cooling formed on - 10 ⁰ C was added a solution of 1.01 g of 7-aminocephalosporanic acid added in 13 ml of methylene chloride and 0.9 ml of triethylamine. The mixture was allowed to warm to room temperature and 1 ml of acetic acid was added; After centrifugation, washing with hydrochloric acid solution and then with water, drying, evaporation to dryness and taking up again in 10 ml of dioxane, 1 ml of water and 3 ml of a saturated sodium hydrogen carbonate solution were added to the product. After stirring, suction. Washing and evaporation to dryness were taken up again in methylene chloride, washed with 10 ml of water and 5 ml in hydrochloric acid, decanted off, washed again with water, dried and digested in ether: 1.747 g of crude product were obtained, which was purified by dissolving in ethyl acetate and subsequent precipitation with ether became.

1.255 g of pure product were obtained.

The product obtained had a syn configuration.
The ethyl y-chloro - «- methoxyiminoacetoacetate used as starting material was prepared as follows:

22.5 g of ethyl y-chloro-oxyiminoacetoacetate were placed in 100 ml of methylene chloride. The batch was slowly stirred in an ice bath with a fresh solution of diazomethane (21.6 g / l) up to one Total volume of 275 ml added. After a reaction time of 5 minutes, the excess was Diazomethane decomposed with some aluminum oxide. It was then evaporated and chromatographed Purified on silica while eluting with methylene chloride. There were 11.93 g of the expected Product received.

The ethyl 2- (2-Amino'4 * thiazolyr) -2-methoxyiminoacetate used above was made as follows:

230 246/318

Level α:
Ethyl 2-acetyl-2-methoxyiminoacetate

180 g of crude ethyl 2-acetyl-2-hydroxyiminoacetate was introduced into 900 ml of pure acetone; 234 g of potassium carbonate were added at 10 ^{° C. under nitrogen;} 103 ml of dimethyl sulfate were then added. After stirring for 3 hours at room temperature and adding ice, the mixture was poured into 41% of water, extracted with methylene chloride, washed with water and dried; the solvents were then distilled off. 185 g of the expected product were obtained.

Level ß:
4-Bromo-2-methoxyiminoacetoacetic acid ethyl ester

197 g of the product obtained in stage ca were added to 1 l of methylene chloride and 200 mg of p-toluenesulfonic acid were added. Subsequently, at 20 ^° C., one tenth of a solution of 191 g of pure bromine in 200 ml of methylene chloride was added Bromine solution was added ^{at about 20 ° C. within 1 h.} The temperature was then allowed to rise to 25 ° C to complete the reaction. It was then washed with ice water, extracted with methylene chloride and dried, whereupon the solvent was distilled off.

268 g of the expected product were obtained.

Level y:

Ethyl 2- (2-amino-4-thiazolyl) -2-methoxyiminoacetate (syn isomer)

80 g of thiourea were placed in 270 ml of ethanol and 540 ml of water under nitrogen were the by stirring for 1 h at about 2O ⁰ C within 0.5 h 268 g in stage β obtained product in 270 ml of ethanol added. After cooling to about 15 ^° C., potassium bicarbonate was added in small portions up to a pH of 5. It was then filtered off with suction, washed with water and dried, 133.8 g of the expected product being obtained. The product obtained was identical to that obtained in step A.

B.

0.975 g of the product obtained above were 10 min stirred at 55 ° C in 4 ml of 50% aqueous formic acid. After adding 4 ml of water was aspirated and evaporated to dryness in vacuo. It was then digested in 2 ml of ethanol, centrifuged off and washed with ethanol and then with ether.

0.428 g of pure product were obtained.

Analysis:

% -M

% N

% S

Calculated:
Found:

42.19
42.3

3.76
4.1

15.37
15.2

14.08 13.8

The product obtained had a syn * configuration.
NMR (60 MHz, DMSO), ppm:

2.03-C-CH ₃ , doublet 9.58

Il ο

J = 8 Hz (COHH), 6.76 protons of the thiazole ring.
Example 2

3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -

2-methoxyimmoacetamido] -ceph- (3) -em-

4-carboxylic acid (syn isomer)

A.
Diethylammonium salt of 3-acetoxymethyl

7- [2- (2-trityiamino-4-thiazolyl) -

2-methoxyiminoacetamido] -ceph- (3) -em-

4-carboxylic acid (syn isomer)

Crude 3-acetoxymethyl-7- [2- (2-trityIamino-4-thiazolyl) -2-methoxyiminoacetamido] -ceph- (3) -em-4-carboxylic acid, which is obtained by condensation of with dicyclohexylcarbcdiiiTiid 2- ^ 2 -Trit "! Arriino-4-thiazol ' ^{l /} l ^" 2-methoxyiminoacetic anhydride with 40.8 g of 3-acetoxy-7-aminocephaIosporanäure was dissolved in 350 ml of dioxane

Thereafter, 350 ml of diethyl ether and then 33 ml of diethylamine were slowly added with stirring added

After stirring for 20 min, the crystallized diethylammonium salt of 2- (2-tritylamino-4-thiazo-Iyl) -2-methoxyiminoacetic acid became sucked off. Djeses salt

was twice with 30 ml of the above dioxane-ether mixture washed; it was obtained in an amount of 62.6 g.

The filtrate was made to a syrupy consistency

evaporated and treated with about 2.5 1 diethyl ether, stirred and centrifuged.

110.3 g of dietary ammonium salt were obtained. The product obtained had a syn configuration.

B.

36 g of the product obtained in A. aq formic acid were added to 180 ml of 50%, which was maintained at 5O ⁰ C.

After stirring at 50 ^° C. for 20 min, the triphenylcarbinol formed was centrifuged off. 180 ml of ethanol were then added, followed by evaporation to dryness under reduced pressure. The residue was taken up in a mixture of 100 ml of water and 20 ml of ethanol and evaporated again. It was then taken up again with 100 ml of water, stirred for 15 min at 15 ° C., filtered off with suction and washed with water and then with ether.

15.6 g of product were obtained.

The product was identical to that of Example 1.

Example 3

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

4-thiazolyl) -2-methoxyiminoacetamidol-ceph-

(3) -em-4-carboxylic acid (syn isomer)

60 To 45.55 g of pure 3-acetoxymethyl-7- [2- (2-amino-4-th "tazolyl) '2-methGXyiminoacetamido] -ceph- (3) -em-4 * carboxylic acid (obtainable according to Example 1 or 2) 100 ml of distilled water were added, then 8 g of sodium hydrogen carbonate and about 20 ml of ethanol were gradually added.

After adding another 80 ml of ethanol and 4.5 g Activated charcoal was stirred for 5 min, filtered, washed with ethanol and evaporated to dryness in vacuo.

It was then taken up with 100 ml of ethanol and evaporated to dryness; the residue was in 100 ml of methanol dissolved. After adding 21% acetone and stirring vigorously, the mixture was filtered off with suction and with acetone and then washed with ether.

After drying in vacuo, 43.7 g of a white product were obtained which redispersed in air hydrated and reached a final weight of 45.2 g.

[ec] f = + 55 ± 2 ° (0.8%, water).

Analysis:

10

% C

%H

% N

% S

%N / A

15th

Calculated
Found:

40.24
40.3

3.38
3.8

14.67
14.4

13.43
13.3

4.81 4.84

The product obtained had a syn configuration.

NMR (60 MHz, D ₂ O) ppm: 2.01 COCH ₃ , doublet at 9.53 J = 8 Hz NHCO, 6.75 'proton of the thiazole ring.

Example 4

3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -

2-ethoxyiminoacetamido] -ceph- (3) -em-

4-carboxylic acid (syn isomer)

A.

3-acetoxymethyl-7- [2- (2-trityIamino-4-thiazoIyl) -

2-ethoxyim> noacetamido] -ceph- (3) -em-

4-carboxylic acid (syn isomer)

Level A:

Ethyl 2- (2-amino-4-thiazolyl) -2-ethoxyiminoacetate (syn isomer)

25th

30th

35

a)

19.4 g of ethyl y-chloro-α-oxyiminoacetoacetate were introduced into 60 ml of acetone and 143 ml of diethyl sulfate; after the batch had been cooled in an ice bath for 10 min, 55 ml of 2η NaOH solution were added over the course of 30 min; the mixture was stirred for 40 min, b) 7.6 g of thiourea were added to the reaction mixture and the mixture was kept at 55 ° C. for 20 min; after the acetone had been driven off and taken up again with ethyl acetate, 6.9 g of potassium carbonate were added; it was then stirred, decanted off, extracted again with ethyl acetate, dried and evaporated to dryness.
17.4 g of residue were isolated, which was chromatographed on silica eluting with ether, to The desired product was collected, taken up again with isopropyl ether, filtered off with suction, washed and dried; 2.8 g of the expected product were obtained. 129 ° C.
The product obtained had a syn configuration.

Level B:

2- (2-Tritylamino-4-thiazolyl) -2-ethoxyiminoacetic acid ethyl ester (syn isomer)

3.16 g of the product obtained in stage A, 6 ml of dry dimethylformamide, 12 ml of methylene chloride and 1.89 ml of triethylamine were brought together under inert gas , Left to stand for 5 h, during which the temperature rose to + 10 ° C .; then the batch was kept at room temperature for 3.5 hours. After adding 13 ml of 1 η hydrochloric acid and stirring, the mixture was decanted and washed with 1 η hydrochloric acid and then with water. It was then extracted with methylene chloride, dried and evaporated to dryness. 7.89 g of crude product were obtained.
The product obtained had a syn configuration.

Level C:

2- (2-Tritylamino-4-thiazoIyI) -2-ethoxyiminoacetic acid (syn isomer)

At 110 ^° C., 7.89 g of the product obtained in stage B, 40 ml of dioxane and 19.5 ml of 2η NaOH solution were brought together within 1 h. The mixture was then filtered off with suction, with an ether-dioxane mixture and then with ether alone washed and dried

6.25 g of sodium salt were obtained, which with 60 ml Methylene chloride and 20 ml of 1 η hydrochloric acid was taken up again; the two phases were stirred, then 20 ml of methanol were added. This was followed by washing again with water, with a Extracted methylene chloride-methanol mixture, dried and evaporated

5.85 g of pure 2- (2-tritylamino-4-thiazoIyl) -2-ethoxyiminoacetic acid were obtained isolated

The product obtained had a syn configuration.

Level D:

3-acetoxymethyl-7- [2- (2-trityIamino-4-thiazolyl) -

2-ethoxyiminoacetamido] -ceph- (3) -em-

4-carboxylic acid (syn isomer)

3.43 g of 2- (2-tritylamino-4-thiazolyl) -2-ethoxyiminoacetic acid from stage C were in 34 ml of methylene chloride brought in; the suspension was cooled and 970 mg of dicyclohexylcarbodiimide were added. Then became washed with methylene chloride and stirred for 1 h at room temperature. Thereafter, the dicyclohexylurea formed was sucked off.

The filtrate was cooled to -20 ⁰ C and once with a cooled to -20 ° C solution of 1.02 g of 3-acetoxymethyl-7-aminocephalosporanic acid in 18 ml of methylene chloride and 1.06 ml of triethylamine.

The mixture was then allowed to warm up again for 13 hours 1.8 ml of acetic acid and 9 ml of 1 n hydrochloric acid to; after stirring it was decanted off, washed with water, with Extracted methylene chloride, dried and evaporated.

4.56 g of the expected product were obtained.

The product obtained had a syn configuration.

B.

The 4.56 g of the product obtained under A. were introduced into 23 ml of 50% aqueous formic acid, Heated to 55 ° C. for 15 min and diluted with 30 ml of water; then the triphenylcarbinol formed sucked off. The filtrate was evaporated to dryness; afterwards it was resumed with water, stirred, filtered off with suction, washed and dried; 116 mg of impure product were obtained.

Concentration of the filtrate gave a second fraction of 674 mg of crystallized product, so a total of 790 mg.

The cleaning was done as follows:

1.063 g of the crude product were stirred in 5 ml of water and heated to 70 ° C. for 5 minutes; after that became cooled again, stirred for 0.5 h, filtered off with suction, washed and dried, whereupon 815 mg of purified product were isolated. These 815 mg were taken up in 2 ml of water and 3 ml of acetone; after easy

The insoluble material was aspirated by heating; after addition of 3 ml of water was heated to 6O ⁰ C, after which the acetone was expelled by passing nitrogen; after suctioning off the formed grains, washing was carried out with water and then with ether; 438 mg of the product were isolated

Analysis: Q ₇ H ₁₉ O ₇ N ₅ S ₂

% C

%H

% N

Calculated:
Found:

43.49
44.5

4.08
4.4

14.92
14.8

The product had a syn configuration.
NMR (60 MHz, DMSO) ppm: 2.05 OAc, 6.75 protons of the thiazole ring.

Example 5

3-acetoxymethyl-7- [2- (2-amino-4-tKazoIyI) -

2-isopropyloxyiminoacetamido] -ceph- (3) -em-

4-carboxylic acid (syn isomer)

A.

3-acetoxymethyl-7- [2- (2-trityiamino-

4-thiazolyl) -2-isopropyloxyiminoacetamido] -

ceph- (3) -em-4-carboxylic acid (syn isomer)

Level A:

2-AcetyI-2-isopropyIoxyiminoacetic acid ethyl ester

39.8 g of ethyl 2-acetyl-2-hydroxyiminoacetate were dissolved in 200 ml of pure acetone. After cooling 52 g of potassium carbonate were added in an ice bath, followed by 25 ml of 2-iodopropane in 0.5 h. This was followed by 2 h stirred, mixed with 800 ml of water and 500 ml of methylene chloride, stirred, decanted off, again with methylene chloride extracted, dried, filtered off with suction and evaporated, whereupon 41.5 g of product were isolated.

Level B:

Ethyl 4-bromo-2-isopropyloxyiminoacetate

The 11.5 g of the one obtained in the previous stage Product were taken along with traces of p-toluenesulfonic acid introduced into 190 ml of methylene chloride, while stirring, a solution of 11.9 ml of bromine in 50 ml Methylene chloride in 1!: Added at room temperature. Ice water was added with stirring, decanted off, extracted again with methylene chloride and washed with ice water, dried and evaporated, whereupon 55 g of the derivative mentioned were isolated.

Level C:

2- (2-Amino-4-thiazolyl) -2-isopropyloxyiminoacetic acid ethyl ester (syn-isomer)

14.9 g of thiourea were added to 55 ml of ethanol and 105 ml of water; then a solution of 55 g of the product prepared in stage B in 55 ml of ethanol was added over the course of 40 minutes. After stirring for 2.5 hours at room temperature, 220 ml of a 10% strength aqueous sodium hydrogen carbonate solution were added; after stirring, suctioning off, washing and drying, 42.15 g of crude product were isolated, which was chromatographed on silica while eluting with ether; the enriched fractions of the expected product were collected and evaporated; after the crystals were taken up with isopropyl ether, suction filtered and washed, 10.75 g of the expected product were obtained.
The product obtained had a syn configuration.

Level D:

2- (2-TrityIamino-4-thiazolyl) -2-isopropyloxyiminoacetic acid ethyl ester (syn isomer)

11 g of the product obtained in Step C became 20 ml dry dimethylformamide, 40 ml of methylene chloride and 6.2 ml of triethylamine. Then it became Mixture cooled and slowly added with 13.2 g of trityl chloride offset; after stirring for 2.5 hours, 43 ml of 1 η hydrochloric acid were added

is admitted; then it was stirred, decanted off again washed with 40 ml of water, extracted with methylene chloride, dried, filtered off with suction and dried to dryness evaporated; 27.7 g of the expected product were obtained.

The product obtained had an svs configuration.

Level E:

2- (2-TrityIamino-4-thiazolyl) -2-isopropyl-oxyiminoacetic acid (syn isomer)

A mixture of 27.7 g of the product obtained in stage D, 150 ml of dioxane and 65 ml of 2η NaOH solution was refluxed for 2 hours. After the sodium salt had crystallized out, it was cooled, washed with an ether-dioxane mixture (1: 1) and dried, whereby 16.85 g of the crude sodium salt were obtained. 15.9 g of this sodium salt were dissolved in 15.9 g of dimethylformamide, 100 ml of water and about 500 ml of methanol. After adding 30 ml of 2N hydrochloric acid, the methanol was expelled, diluted with water, washed with suction and dried; the 9.8 g of the viscous product obtained were taken up in 220 ml of a methylene chloride-methanol mixture (50:50); then it was evaporated to dryness, taken up with ether and digested; were rocknen after suction filtration of the crystals, washing and ^T, 4.9 g of the desired acid obtained; F. ~ 170 ° C.

An analysis sample was carried out by lorries from 300 mg of crude product in 2 ml of methylene chloride and 1 ml of methanol, Dilute with water and methylene chloride, stir, filter off the crystals with suction, wash with methylene chloride and water and dry to obtain 230 mg of pure product for analysis.

50 Analysis: % c %H % N yo S 68.77
68.6 5.34
5.5 8.91
8, i » 6.8
.6.8 Calculated:
Found:

The product obtained had a syn configuration.

Level F:

3-acetoxymethyl-7- [2- (2-tritylamino-

4-thiazolyl) -2-isopropvloxyiminoaceta:, iido] -

ceph- (3) -em-4-carboxylic acid (syn isomer)

4.89 g of the acid obtained in stage E were added to 13.5 ml of dimethylformamide under argon. After this Dissolving was cooled in an ice bath and washed with 1.62 g of dicyclohexylcarbodiimide in 16 ml of methylene chloride added, whereupon dicyclohexylurea crystallized

sierlc. Then it was stirred in an ice bath, centrifuged off, washed with methylene chloride and dried, whereupon 1.424 g of dicyclohexylurea were isolated. It was then cooled in a methanol-ice bath and with a solution of 1.41 g / 'aminocephalosporanic acid added in 30 ml of methylene chloride and 1.45 ml of triethylamine. After stirring for 3 h at room temperature and adding 20ml In hydrochloric acid stirred, decanted off, extracted again with methylene chloride, dried and filtered off with suction, after which 9.05 g a mixture of the expected product with the starting product.

It was then taken up in methylene chloride, seeded and allowed to crystallize with stirring; after the crystals had been filtered off with suction, washed and dried, 1.6 g of pure starting material were separated off. It was then evaporated to dryness, after which the residue would be treated with isopropyl ether while stirring vigorously. 4.9 g of insoluble, viscous product were obtained, which corresponded to the expected compound.
The product obtained had a syn configuration.

B.

4.91 g of the crude product obtained in stage F were added to 30 ml of 50% strength aqueous formic acid and stirred in a water bath at 60 ° C; after dilution with water, the triphenylcarbinol formed was centrifuged off, washed with water and dried; 139 g of triphenylcarbinol were then isolated evaporated to dryness, taken up with water, digested, filtered off with suction, washed with water and dried; 800 mg of the expected product were obtained.

An analytical sample was given by Loser of 972 mg of the crude product in 4 ml of methanol, diluting with 20 ml of ether, centrifuging off the insoluble, washing - ■ and drying produced, resulting in 404 mg of the expected pure acid. F.- 200 ° C.

Analysis:

% C

%H

% S

Calculated:
Found:

44.71
44.5

4.5

14.48
14.1

13.26 13.2

The product had a 3yn configuration.
NMR (60 MHz, DMSO) ppm: 2.01 CH ₃ CO, doublet at 9.46 J = 8 Hz CONH, 6.7 proton of the thiazole ring.

Example 6

3-acetoxymethyl-7- [2- (2-ajnino-4-thiazolyl) -

2-methoxyiminoacetamido] -ceph- (3) -em-

4-carboxylic acid (syn isomer)

Level A:

Ethyl 2- (2-chloroacetainido-4-thiazoIyl) -2-methoxyiminoacetate (syn isomer)

453 g of ethyl 2- (2-amino-4-thiazolyl) -2-methoxyiminoacetate (syn-isomer) prepared as in Step A of Example I were in 200 ml Methylene chloride was introduced to dryness

20 ml of this was distilled, then the mixture was cooled to 10 ° C. and 50 ml of pyridine were added. Thereafter, 41 g of monochloroacetic anhydride were added, with slight warming until dissolution. After standing for 6 hours at 20 ° C. under nitrogen, 5 ml of water were added, whereupon the mixture was stirred and 300 ml of ice-cold 2 η hydrochloric acid was poured in. After decanting off, extraction with methylene chloride, washing with water, sodium hydrogen carbonate and again with water and drying, the mixture was passed over activated charcoal, evaporated and 300 ml of isopropyl ether were added, whereupon the product crystallized out. It was evaporated to the consistency of a thick paste, then cooled with ice, filtered off with suction, washed with isopropyl ether and dried. 45.4 g of product were obtained; F. 113 ⁰ C.

An analytically pure sample was obtained by recrystallization in a mixture of methylene chloride and Isopropyl ether produced.

F. 118 ⁰ C.

NMR (60 MHz ₁ CDCl ₃ ) ppm:

O (d)
(f) HNC-CH ₂ Cl

- ν γ.

V = K ^ Jl (C) (a)

(e) HC 0 -CH _{2 -CH 3}

OCH ₃ (b)

(a) triplet centered at 1.38, J = 7 Hr;

(b) singlet 4.05;

(c) quadruplex centered at 4.44, J = 7 Hz;

(d) singlet 4.33;

(e) singlet 7.27;

(f) singlet 9.95.

Level B:

2- (2-chloroacetamido-4-thiazoIyl) -2-methoxyiminoacetic acid (syn isomer)

46 g of the product obtained in step A were placed in 230 ml of absolute ethanol; then 30 ml of pure caustic soda were added at 2O ⁰ C under nitrogen. The product dissolved and the sodium salt began to crystallize; and after that the product was eliminated; after 16 h it was filtered off with suction and washed with ethanol. The obtained salt was dissolved in water; while cooling with ice, 100 ml of 2 η hydrochloric acid were added; then it was saturated with sodium chloride

and extracted with ethyl acetate containing 10% ethanol. After drying, the mixture was passed over activated charcoal, distilled in vacuo and the water was removed by azeotropic distillation with benzene; ! was then taken up in methylene chloride, to dryness eui ^ edarnnft in Meth ^v enchlorid recurrent ^cr enQmnien sucked cooled with ice, washed with methylene chloride and dried were 34.5 g of the expected product are obtained; F. ~ 200 ° G

The product was purified by recrystallization from an acetone / isopropyl ether mixture.

Analysis: C ₈ H ₈ O ₄ N ₃ ClS = 277.68

% C % H% N

% S

Calculated; 34.60 2.90 15.13 Found: 34.8 2.8 14.8

NMR (60 MHz, DMSO) ppm:
If)

Il
(e) HNC-CH ₂ Cl

12.77 12.6

11.55 11.5

.C N

\ / · CO ₂ H

(d) H.

OCH ₃ (a)

(a) singlet 3.92;

(b) singlet 4.38;

(c) singlet around 5;

(d) singlet 7.58;
OO singlet 12.6.

Stage C;

3-acetoxymethyl-7 ^ [2- (2-chloroacetamido-

4-thiazolyl) -2-methoxyiminoacetämido] -ceph-

(3) -em-4-carboxylic acid (syn-isomer)

15.3 g of the product obtained in the previous stage B were introduced into 80 ml of methylene chloride; at 5 ^° C. 8 ml of triethylamine were added. 3.8 ml of thionyl chloride and 26 ml of methylene chloride were then allowed to flow in under nitrogen at ^{0 ° C.} After standing at ⁰ ° C. for 15 minutes, 7 ml of triethylamine were added. Then 13.6 g of 7-aminocephalosporanic acid in 100 ml of methylene chloride and 14 ml of triethylamine were added ^{at 0 ° C. under nitrogen.} After the temperature was allowed to rise again to 20 ^° C., the mixture was stirred for 1 h. The solution was evaporated to dryness in vacuo at 30 to 35 ° C. The residue was dissolved in 250 ml of water, poured over activated charcoal and diluted with 50 ml. 2 π hydrochloric acid added. The resulting precipitate was filtered off with suction and washed with water. The crude product obtained was suspended in 80 ml of ethanol and 7 ml of triethylamine were added at + 5 ° C. Then, with stirring at 5 ^° C., 15 ml of 4 η sulfuric acid were added all at once; the product crystallized out after 15 minutes. It was then filtered off with suction, stirred with ethanol and then with ether and dried in vacuo; 18.6 g of the expected product were obtained.

[a] ² _D ° = +20 ± 1 ° (1%, dimethylformamide). NMR (60 MHz, DMSO) ppm:

(e) HNC-CH ₂ Cl

(d) H.

(b)

(a) singlet 2.03;

(b) singlet 3.90;

(c) singlet 4.38;

(d) singlet 7.45.

CH ₂

CO ₂ H

(a)
-OC-CH ₃

Level D:

3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido] -ceph- (3) -em-

4-carboxylic acid (syn isomer)

532 g of the acid obtained in stage C were suspended in 10.6 ml of water with 912 mg of thiourea; at 20 ⁰ C ί g Kaliumhydrogencarbonai was excellent give. After dissolution, the mixture was stirred under nitrogen ^{at about 20 ° C. for 6 h.} The gummy precipitate began to precipitate after about 1.5 hours; 30 ml of water and 3 ml of formic acid were then added. After cooling to 5 ° C., the mixture was filtered off with suction and washed with water containing 10% formic acid. The residue was dissolved at about 5 ° C. in 30 ml of water containing triethylamine 3 ml of formic acid were added; the precipitate was filtered off with suction and washed by stirring with water containing formic acid. The dark brown gummy product was separated off

treated; a light yellow solution resulted which was saturated with ammonium sulfate. The educated Precipitate was filtered off with suction, stirred up with water, again filtered off with suction and washed with water (= Precipitation A).

The mother liquors were saturated with ammonium sulfate; Appeared precipitate was removed by centrifugation and moreover ³ !! washed with water (= lower

stroke B).

Precipitates A and B were combined. It was then taken up with ethanol, stirred for 1 h at 2O ⁰ C and 16 O hbüi stand close jassen ⁰ C. It was then filtered off with suction, washed with ethanol and with ether and dried in vacuo; 3.47 g of the expected product were obtained (syn isomer).

NMR (60 MHz, DMSO) ppm:

NH

(C)

CH ₂ -OC-CH ₃

(a) singlet 2.03;

(b) singlet 3.55;

(c) doublet 5.19, J = 5 Hz;

(d) singlet 6.8.

The product was identical to that obtained in Examples 1 and 2.

Example 7

3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido] -ccph- (3) -em-

4-carboxylic acid (syn isomer)

A.

Diethylammonium salt of 3-acetoxymethyl

7- [2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetamido] -ceph- (3) -em-4-carboxylic acid

(syn-isomer)

Level A:
Ethyl 2-acetyl-2-methoxyiminoacetate

4.69 kg of ethyl 2-acetyl-2-hydroxyiminoacetoacetate (corresponding to 4.21 kg of pure product) were in 21 1 Pure anhydrous acetone introduced. At 20-25 ° C, 6.1 kg of potassium carbonate were added. The suspension was stirred for 10 min; then 3.72 kg of dimethyl sulfate were added at 20 to 25 ° C., whereupon 3 h at 20 was stirred to 25 ° C. Thereafter, desalinated water was poured into 1261 and with 4 χ 51 and then extracted with 21 methylene chloride and washed with 101 deionized water. After this Drying and suction was washed with 21 methylene chloride and then distilled in vacuo 4.88 kg of the expected product were obtained.

Rf = 0.7 (thin-layer chromatography on S1O2; eluent Methylene chloride-ethyl acetate 9: 1).

Level B:
4-Bromo-2-r.ethoxyim! Noacetoacetic acid ethyester

3.53 kg of the product obtained in stage A were added to 18.61 of methylene chloride with 3.5 g of p-toluenesulfonic acid. A solution of 2.96 kg of bromine in 3.5 l of methylene chloride was added to this solution over the course of 30 minutes while maintaining a temperature of 22 ± 1 ° C .; after 15 min of flowing in, the evolution of hydrogen bromide began. After 45 min of stirring at 22 ^° C., the mixture was poured over and washed twice with 141 demineralized ice water. The wash waters were extracted twice with 3.5 liters of methylene chloride. After drying, filtering, washing with methylene chloride and distilling in vacuo, 4.73 kg of the expected product were obtained.

Level C:

2- (2-Amino-4-thiazolyl) -2-methoxyiminoacetic acid ethyl ester (syn-isomer)

1.43 kg of thiourea was dissolved in 3.551 ethanol and 7.11 deionized water. After stirring for 10 min at 20 ⁰ C of the product prepared in Step B at 20 to 25 ⁰ C was added 4.73 kg in 3.551 of ethanol, followed by 3 h stirring at 20 to 25 ⁰ C. Then ⁰ C was cooled to 15 to 20 and about 1.61 ammonia (22 ° Be) to a pH value of 7 neutralized

The mixture was then stirred for a further 15 minutes at 20 to 25 ° C. and then filtered off with suction and added five times 1.8 1 deionized water washed. After drying, 2.947 kg of the expected product were obtained; M.p. 162 ° C.

Level D:

2- (2-Tritylamino-4-thiazolyl) -2-methoxyiminoacetic acid ethyl ester (syn-isomer)

3.41 kg of the product obtained in Step C in 17 1 of methylene chloride with triethylamine dissolved 2.2751 After 15 min stirring period of 1 hour with stirring and under nitrogen at 20 to 25 ⁰ C. 4.55 kg of trityl chloride was added After 20 h at 20 to

Stirred 25 ° C under nitrogen, with triethylamine hydrochloride crystallized.

Danr; was poured over and washed with 10.21 ice-cold 0.5 ή hydrochloric acid and twice with 10.21 deionized ice water. The wash waters are extracted successively with 1.71 methylene chloride; it was then dried, filtered and washed with 1.7 l of methylene chloride, whereupon it was evaporated to dryness in vacuo at a temperature below 50 ° C.
8.425 kg of crude product were obtained,
This product was redissolved in 8.41 of methanol ^{at 20-25 ° G;} then 2.8 l of deionized water were added over the course of 1 h at 20 to 25 ° C. with stirring and triggering crystallization. The mixture was stirred for a further 1 h, thinly centrifuged, stirred twice with 1.7 l of methanol with 25% water and dried at 40 ° C .; 7.165 kg of the expected product were obtained.

Level E:

Sodium salt of 2- (2-triiy! Amino-4-ihiazoiyi) -2-methoxyiminoacetamido acid (svn isomer)

10

20th

25th

35

4.175 kg of the product obtained in step D was in Brought 20.9 1 of ethanol and refluxed with stirring and under nitrogen; from 55 ° C one resulted homogeneous solution.

While boiling under reflux and under nitrogen, 5.235 liters of about 2 η sodium hydroxide solution were added; crystallization occurred rapidly; it was stirred under reflux for 1 h under nitrogen. Then the The batch was cooled to 20 to 25 ° C. and kept at this temperature for 2 h, after which it was filtered off with suction four times

2.1 1 ethanol was washed and dried; it fell

4.02 kg of the expected product.

Level F:

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

500 g of the product obtained in stage E (corresponding to 440 g of dry product) were dissolved in 2.5 1 of methylene chloride given and within 2 min at 20 to 25 ° C with stirring and under nitrogen with 2 1 about 1 η Hydrochloric acid added. The mixture was then stirred at 20 to 25 ° C. under nitrogen for 2 h. The methylene chloride phase was tbdecanted and washed three times with 2 liters of deionized water; the washing waters were again with 11 Extracted methylene chloride. After drying, addition of 25 g of activated charcoal, suction, washing with methylene chloride and evaporation to dryness gave 481 g of crude product.

The crude product was taken up again with 2.1 l of isopropyl ether, filtered off with suction, twice with 420 ml Washed isopropyl ether and dried in vacuo to constant weight; there were 424.6 g of the expected product received.

Level G:
Diethylammonium salt of 3-acetoxymethyl

7- [2- (2-trityiamino-4-thiazolyl-

2-methoxyiminoacetamido] -ceph- (3) -om-

4-carboxylic acid (syn isomer)

200 g of 2- (2-TrityIamino-4-thiazoryl) -2-methoxyiminoacetic acid from Stage F were placed in 1200 ml of methylene chloride. The suspension was refluxed with stirring under argon, whereupon 600 ml of methylene chloride were distilled off at normal pressure. The batch was cooled to 18 to 20 ° C and then treated at this temperature with 54 g of dicyclohexylcarbodiimide in 54 ml of methylene chloride was then stirred for 1 hour at 18 to 2O ⁰ C under argon and then within 15 an immediately previously min at this temperature from 61.4 g of 7-aminocephalosporanic acid in 900 ml of methylene chloride and 63 ml of triethylamine freshly prepared solution were added. After stirring for 1.5 hours at 20 ° C. (pH 6.5 to 7), 5 microns of acetic acid were added; after 15 min stirring at 20 ⁰ C was suction filtered, the material used as the starting 7-aminocephalosporanic acid to be removed. It was washed four times with 200 ml of methylene chloride. The organic solution was washed three times with 400 ml of deionized water and then dried over magnesium sulfate. It was then filtered off with suction, washed twice with 200 ml of methylene chloride and evaporated to dryness under argon. The dry oily extract was dissolved in 700 ml of dioxane at 20 to 25 ° C. with stirring and under argon; then 300 ml of a mixture of dioxane and methylene chloride were distilled off in vacuo and under argon at a temperature below 30.degree. The batch was then brought to 20 ± 2 ° C. and 500 ml of ether were added, whereupon 52 ml of diethylamine were added. After about 10 minutes, the diethylammonium salt of 2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetic acid crystallized out; which was left to stand under argon at 20 ° C. for 1 h. It was then filtered off with suction and washed three times with 100 ml of a solution of dioxane and ether. The diethylammonium salt obtained was obtained in an amount of 113.6 g after drying. The organic solution was precipitated in 3.25 l of isopropyl ether with stirring over the course of 30 minutes. After stirring for 15 min, suction was carried out using a suction filter, washed twice with 400 ml of isopropyl ether and dried in vacuo.
182 g of product were obtained.

B.

182 g of the product obtained in A. were under stirring and Ar <? On at 28 to 3O ⁰ C in 347 ml of formic acid and 87 ml of deionized water were charged. It completely dissolved and triphenylcarbinol crystallized out. The mixture was stirred under argon for 2.5 h at 28 to 30 ° C. and then precipitated within 1 ^ min with stirring with 1740 ml of deionized water and 847 g of ammonium sulfate, whereupon the mixture was stirred for 30 min. After suction filtration, washing twice with 174 ml of deionized water and drying in vacuo at 25 to 30 ° C., 147 g of a mixture of the expected product with triphenylcarbinol were obtained.

The crude product was stirred in 735 ml of ether ^{at 18 to 20 ° C. for 1 h;} it was then filtered off with suction, washed twice with 147 ml of ether and dried at 25 to 30 ° C., after which 89 g of the expected product were obtained. This product was stirred in 445 ml of ethanol with stirring and under nitrogen. The suspension was heated to 45 to 50 ° C. with stirring and kept under these conditions for 1 h; then ⁰ C, 1 h at 18 to 20 stirred mixture was then suction filtered, washed twice with 45 ml ethanol and dried in vacuo at 20 ⁰ C with 76.85 g of the expected product were obtained.

230 ml of acetic acid were added to this product, the mixture was stirred under nitrogen for 15 min and then with 77 ml demineralized water added; then about 700 ml of water were added to this solution. After 1 h Stirring at 18 to 20 ° C., 269 g of ammonium sulfate were added over the course of about 10 minutes; one let Stand for 15 min and then added 3.85 g of activated charcoal.

Found: 39.9 3.5

14.5 13.1 4.8

ίο

15th

After 15 min of stirring, the mixture was filtered off with suction, washed with 77 ml of deionized water containing 25% acetic acid and, at 18 to 20 ^° C., while stirring, 154 ml of formic acid were added to 20 ⁰ C and then 2 h at 0 to + 5 ° C, then suction filtered and washed four times with 77 ml of deionized water with 5% formic acid and dried at 20 to 25 ° C in vacuo. 49.45 g of product were in the form of the format.

The formate was mixed with stirring for 1 h at 45 to 50 ° C in 250 ml of ethanol and then allowed to stand for 1 h at 18 to 20 ⁰ C. It was then filtered off with suction, washed twice with 50 ml of ethanol and ^{dried at 20 °} C. in vacuo and then at 35 to 40 ^° C. for 10 to 15 hours. 43.45 g of the expected product were obtained.

[ _Ä ] g> = + 64.5 ° (0.5%, water with 0.5% NaHCO ₃ ).

The product was identical to that of Examples 1, 2 and 6.

Example 8

Sodium salt of 3-acetoxymethyl-7- [2- (2-amino-4-thiazoIyl) -2-methoxyiminoacetamido] -ceph-

(3) -em-4-carboxylic acid (crystallized syn isomer)

19.8 g of 3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamidoj-ceph-pj-em-'t-carboxylic acid (syn isomer) (see. Examples 1, 2,6 or 7) were in 65 ml of a 1 M sodium acetate solution in methanol dissolved The mixture was left to stand for 35 minutes at room temperature for crystallization; then 40 ml Ethanol was added, whereupon the mixture was stirred for 2.5 h in an ice-water bath, filtered off with suction and twice with 10 ml of one Methanol-ethanol mixture (1: 1), twice with 10 ml Ethanol and then washed twice with 20 ml of ether. After drying (2 h at 45 ° C im Vacuum and 48 hours in vacuo over sulfuric acid) 16.191 g of crystallized product were obtained.

By working while avoiding any contact with humidity, a product was made with receive the following physical properties:

H ₂ O (Karl Fischer) 0.2%

Methanol £ 0.1% 1 gas chromatography

Ethanol 0.45% J determined graphically

Analysis: C ₁₆ H ₁₆ O ₇ N ₅ S ₂ Na = 4773

% C% H% N% S% Na

Calculated: 40.24 3.38 14.67 13.43 4.81

Example 9

Sodium salt of 3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido] -ceph-

(3) -em-4-carboxylic acid
(crystallized syn isomer)

Level A:

Solvate of 3-acetoxymethyl-7- [2- (2-amino-

4-thiazolyl) -2-methoxyiminoacetamido] -ceph-

(3) -em-4-carboxylic acid (syn isomer) with

Formic acid

In small portions and with stirring, 87.2 g of diethylammonium salt of 3-acetoxymethyl-7- [2- (2-tritylamino-'t-thiazolylj ^ -methoxyiminoacetamidoj-ceph- (3) -em-4-carboxylic acid, which according to Example 2, A, was added to a mixture of 220 ml of pure formic acid and 220 ml of water After 30 minutes of stirring at 50 ° C., the mixture was cooled and the triphenylcarbinol formed was filtered off in an amount of 30.1 g Poured in water: a slight precipitate was removed with activated charcoal, then ^{the mixture was evaporated in vacuo at 40 °} C. After adding 200 ml of anhydrous ethanol, the mixture was cooled with ice, filtered, washed with ethanol and with ether and dried in a vacuum

31.1 g of the expected product were obtained.

30th

35

40

45

Analysis: Ci ₆ Hi ₇ N ₅ O ₇ S ₂ • HCO ₂ H • H ₂ O = 541 ^ % N % S % H ₂ O % C %H 13.48
13.2 12.34
12.8 3.46
4.15 Calculated: 39.3
Found: 39.2 4.08
4.1

Level B:
Crystallized sodium salt

15 g of the solvate freshly prepared in stage A were dissolved in 75 ml of methanol; then the solution was treated with 43 g of sodium acetate and 3 g of activated charcoal. After filtering, 5 ml of isopropanol were added with stirring. After 16 h at 0 ° C the crystals were separated, washed with ethanol and with ether and dried for 2 h at 50 ⁰ C in vacuo.

755 g of product were obtained.

The product was in the air for a short time; it provided the following analysis results:

60

The product reabsorbed water on standing in the air.

The X-ray analysis (Debye-Scherfer) confirms this crystalline structure of the product obtained.

Instead of ethanol, isopropanol could also be used to achieve the crystallization of the desired Product to achieve.

C ₁₆ Hi ₆ N ₅ NaO ₇ S ₂ · H ₂ O KC = 4953 % N %N / A % S ■ 38.78 14.14 4.64 12.94 se 38.6 KH 13.8 4.6 13.2 Sä Calculated: 3.66 230 246/318 Found: 3.7 I. I.

Example 10

Sodium salt of 3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido] -ceph- (3) -em-4-carboxylic acid (amorphous syn isomer)

Level A:

Solvate of 3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacet3mido] -ceph- (3) -em-4-carboxylic acid (syn isomer) with ethanol

52 g of the solvate obtained in Step A of Example 9 with formic acid were dissolved in a mixture of 3 liters of 96% ethanol and 350 ml of water; then was concentrated in vacuo to a volume of about 300 ml. The solvate started on evaporation crystallize out. It was then cooled for 1 h in an ice bath, filtered, washed with a little ethanol and im Vacuum dried over concentrated sulfuric acid at room temperature

44 g of the expected product were obtained.

Analysis: C ₁₆ H ₁₇ N ₅ O ₇ S ₂ x 0.8 mol C ₂ H ₅ OH = 492.3

10

25th

% c

% K

% N

% S

Calculated:
Found:

42.94
43.0

4.46
4.4

14.23 14.1

13.02 12.9

30th

Level B:
Amorphous sodium salt

Ci ₆ H ₁₅ N ₅ NaO ₇ S ₂ 1.5 H ₂ O = 504.47 %H % N % C 3.8
3.9 13.88
13.6 Calculated:
Found: 38.09
38.2

Example 11

Sodium salt of 3'Acetoxymethyl-7- [2- (2 * amino-

4-lhiazolyl) -2'-methoxyiminoacetamido] -ceph- (3) -em-4 * carboxylic acid (crystallized

syn isomer)

To 4.95 g of 3'Acetoxymethyl-7- [2 '(2-amino'4'thiazolyl) -2'methoxyirninoacetamido] ⁱ ceph- (3Venv4-carboxylic acid (syfi'lsömer) (cf. Examples 1, 2 , 6 or 7)

60

65

5 ml of ethanol and then 10 ml of an aqueous 1 M sodium hydrogen carbonate solution in an ice water band were added while stirring admitted. After dissolution, 15 ml of ethanol were added, whereupon at 30 ° C im Evaporated in vacuo, taken up again with ethanol and dried to constant weight. A powder was obtained which was taken up with 15 ml of methanol. After inoculation for Crystallization was left in the refrigerator overnight. 3.407 g of crystallized product were obtained isolated.

15th

20th

Example 12

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

4-thiazolyl) -2-methoxyiminoacetamido] -ceph-

(3) -em-4-carboxylic acid (crystallized.

syn isomer)

0.5 g of the amorphous sodium salt obtained according to Example 10 was dissolved in 2 ml of methanol; 0.25 ml of n-butanol was then slowly added with stirring and the mixture was kept at about 6 ° C. in the refrigerator for 48 hours. The crystals were washed with a little cold methanol and ^{dried in vacuo at 40 °} C. over concentrated sulfuric acid for 3 h.

0.2 g of crystallized product was obtained.

The product was then in the air for a short time.

Analysis: C ₁₆ H, 6N ₅ NaO ₇ S ₂ · 1.5 H ₂ O = 504.47

35 % c

%H

% N

% S

3 g of the solvate with ethanol of Step A were added at 0 ⁰ C in 60 ml of water; then 0.504 g of sodium hydrogen carbonate dissolved in 6 ml of water were added with stirring. The neutral solution was filtered and immediately freeze-dried. The product was then in the air for a short time.

The following analysis results were obtained:

45 Calculated:
Found:

38.09
38.4

3.8 3.8

13.88
13.8

26.96 27.1

55 When working under analogous conditions, slightly different crystalline forms are obtained. B. OJ mol of water or 1 mol of water and 1 mol of methanol.

X-ray analysis (Debye-Scherrer) confirms the crystalline nature of the above products.

Pharmacological study of the
compounds of the invention

Effectiveness in vitro according to the method of dilution in a liquid medium:

In this process, a number of test tubes with equal amounts of sterile nutrient medium are added to the investigating compound introduced in increasing amounts! after that, each test tube is filled with a Bakieriensfamm inoculated.

After 24 or 48 hours of incubation at 37 ° C., the Inhibition of growth assessed by fluoroscopy, resulting in the determination of the minimal Hemrnkonzentra ^ (MIC) allowed.

The MIC values are given in μg / ml,

The following results were obtained;

35 36

tribe

Compound of Example 1, 2, 6 and 7

MIC (µg / ml) 24h 48h

Connect and from example 3

MIC fcg / ml) 24 h 48 h

Comparison connection *)

MIC ((ig / ml) 24h 48h

40

Staphylococcus aureus ATCC 5638 = UC 1061, penicillin sensitive Staphylococcus aureus UC 1128, penicillin resistant Staphylococcus aureus exp. No. 54146 Streptococcus pyogenes A 561 Streptococcus faecalis 5432 Streptococcus faecalis 99 F 74 Bacillus subtilis ATCC 6633 Escherichia coli ATCC 9637 = UC 1020, tetracycline sensitive Escherichia coli ^ sTCC 11 303 = UC 1261,

LCiTacycxinrcsiSwwui

Escherichia coli Exp. TO ₂₆ B ₆ Escherichia coli R 55 123 D, gentamycin and tobramycin resistant Klebsiella pneumoniae Exp. 52 Klebsiella pneumoniae 2536, gentamycin resistant Proteus mirabilis (indole) A Proteus vulgaris (indole +) A Sahnonella typhidencia 48 Pseudomonas 3935 Exp. SG Serratia 2532, gentamycin-resistant Enterobacter cloacae 681 Pseudomonas 8951 RGT Staphylococcus Smith Escherichia coli gall Escherichia coli E ₆ Escherichia coli T 25 575 Escharichia coli T 96 875

*) 3-Acetoxymethyl-7- [2- (2-amino-4-thiazolyl> 2-methoxyiminoacetamido] -ceph- (3) -em-4-carboxylic acid (anti isomer).

2 2 2 3 > 40 > 40 3 3 2 2 40 40 0.02 0.05 0.01 0.01 0.4 0.4 2 2 1 3 > 40 > 40 2 3 3 10 > 40 > 40 0.2 1 1 2 20th 20th 0.2 0.2 0.1 0.1 20th 40 0.02 0.02 <0.02 <0.02 2 2 0.1 0.1 0.05 0.05 5 5 0.6 0.6-
1 0.1 0.1 10 10 0.02 X
0.02 <0.02 <0.02 1 1 0.6 0.6 6.2 0.2 > 40 > 40 0.02 0.02 <0.02 <0.02 2 2 2 40 0.2 0.6 > 40 > 40 0.2 0.2 0.1 0.1 20th 20th 5 5 1 2 40 > 40 10 40 10 20th > 40 > 40 1 1 1 1 > 40 > 40 40 40 40 > 40 1 1 0.1 0.2 0.05 0.05 0.05 0.05 0.05 0.05

tribe

connection of 4th 48 h Comparison 48 h example MIC fcg / ml) 2 link*) 20th 24 hours 3 MIC (μ g / ml) 40 2 2 24 hours 40 2 0.02 20th 0.4 2 10 40 > 1OÖ 0.02 20th 40 > 100 2 0.4 0.4 10 2 1 > 100 > 100 0.4 0.2 > 100 10 1 0.4 10 40 0.2 > 100 0.4 10 20th

Staphylococcus aureus ATCC 5638, penicillin sensitive Staphylococcus aureus UC 1128, penicillin resistant Staphylococcus aureus exp. No. 54 Streptococcus pyogenes A 561 Streptococcus faecalis 5432 Streptococcus faecalis 99 F 74 Bacillus subtilis ATCC 6633 Escherichia coli ATCC 9637, tetracycline sensitive Escherichia coli ATCC 11 303, tetracycline resistant Escherichia coli Exp, TO ₂₆ B ₆

37

Fa rtsemi ng

Eschenchia coli R 55 123 D resistant to gentamycin and tobramycin Klebsiella pneumoniae Exp. 52 Klebsiella pneumoniae 2536, gentamycin-resistant Proteus mirabilis (indole) A Proteus vulgaris (indole +) A Salmonella typhimurium 420 Providencia Du 48 Pseudomonas 3935 Exp. SG Serratia 2532, gentamycin resistant

*) 3-acetoxymethyl-7- [2- (2-amino-4-thiazoIyl) -2-ethoxyiminoacetaniido] -ceph- (3) -em-4-carboxylic acid ι

connection of 4th 48 h Comparison 48 h example MIC (μ g / ml) 0.6 link*) 40 24 hours 0.05 MIC (ng / ml) 20th 0.6 2 24 hours > 100 0.05 0.05 40 5 2 2 20th > 100 0.05 1 > 100 100 1 5 5 > 100 1 40 > 100 > 100 5 2 100 > 100 20th 100 2 > 100 > 100

connection of
Example 5 5 Comparison
link*) 48 h MIC ((ig / ml)
24 h 48 h 5 MIC ^ g / ml)
24 hours 20th 2 5 20th 40 3 0.02 40 40 2 40 40 1 0.02 20th 1 > 100 5 2 > 100 > 100 5 2 > 100 > 100 0.5 0.5 20th > 100 2 1 > 100 20th 0.5 1 20th 100 1 0.2 100 100 1 10 100 40 0.2 0.5 40 > 100 10 1 > 100 20th 0.5 2 10 > 100 0.5 5 > 100 100 2 40 100 > 100 5 5 > 100 > 100 40 ' 40 > 100 > 100 2 > 100 100 40 100

Staphylococcus aureus ATCC 5638, penicillin sensitive Staphylococcus aureus UC 1128, penicillin resistant Staphylococcus aureus Exp. 54 Streptococcus pyogenes A Streptococcus faecalis 5432 Streptococcus faecalis 99 F Bacillus subtilis ATCC 6633 Eschenchia coli ATCC 9637, tetracycline-sensitive Escherirhia coli ATCC 11 303, tetracycline-resistant Eschenchia coli Exp. TO ₂ ^ Bb Escherichia coli Exp 52 Klebsiella pneumoniae 2536, gentamycin-resistant Proteuä mirabiiis (indole-) A Proteus vulgaris (indole +) A Salmonella typhimurium 420 Providencia Du 48 Pseudomonas 3935 Exp

*) 3-Acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-isopropyloxyiminoacetamido] -ceph- (3) -ern-4-carboxylic acid (anti-isomer).

Strain MIC-Werf (Vg / ml)

Connection of
Example 4

CEFUROXIM

Staphylococcus aureus SG 511 0.781 1.25 Micrococcus luteus ATCC 9341 0.039 1.25 Streptococcus durans D. 0.156 6.25 List, monocytogenes 10 50

tribe

continuation

tribe

Bacillus subtilis ATCC 6633 Escherichia coil 026 Escherichia coli 055 Escherichia coli 086 Escherichia coli 012ö Escherichia coli V6311 / 65 Escherichia coli TEM Escherichia coli 1507 E Citrobacter freundii ATCC 8090 Salmonella paratyphi A Salmonella typhi murium Salmonella Oranienburg Salmonella typhi

Klebsiella pneumoniae ATCC 10 Klebsiella aerogenes 1522 E Enterobacter aerogenes Hafniaalvei ATCC 11 Proteus mirabilis 112/3 Proteus mirabilis 174/3 Yersinia pseudotuberculosin Past, multocida 40

MIC value compound from example 4

1.563 0.031 0.039 0.031 0.039 0.007 0.195 0.078 1.25 0.015 0.019 0.078 0.039 0.007 0.039 0.078 0.31 j 0.156 0.007 0.007 0.003

CEFUROXIM

6.25

0.078

1.25

0.625

6.25 10

0.31

2.5

2.5

2.5

0.125

2.5

1.25

5 10

0.313

0.25

0.062

Compound of example 4

MIC ^ g / ml) 24 h 48 h

Compound of example 5

MIC ^ g / ml) 24 h 48 h

Comparison compound CEFUROXIM

KiHK fcg / ml) 24 h 48 h

Escherichia coli ATCC 9637, tetracycline sensitive Escherichia coli ATCC 11 303, tetracycline resistant Escherichia coli Exp. TO ₂₆ B ₆ Escherichia coli R 55 123 D, gentamycin resistant, tobramycin resistant

Klebsiella pneumoniae Exp. 52 145 Klebsiella pneumoniae 2536, gentamycin resistant Proteus mirabilis (indole) A 235 Proteus vulgaris (indole +) A 232 Salmonella typhimurium 420 Enterobacter cloacae 681

Providencia Du 48

Serratia 2532, gentamycin resistant

0.2 0.4 0.6

0.2 0.4 0.6

0.5 1 1

2 2

0.05 0.05 0.5

2 1 40 5 2

0.5 2 40 5 2

0.5 1 1

0.05 0.05 0.2 0.2

10

0.5

2 40

10

5 10

0.2 10 0.5> 40 10

> 40> 40> 40

> 40 20> 40> 40> 40

Experimental infection with Escherichia coli (T) O ^ Bs

A) The effectiveness of the compound of Examples 1, Z. and 7 was investigated in an experimental infection with Escherichia coli in mice. Male mice with a mean weight of 21.5 g were in groups of 10 mice each by intraperitoneal injection of 0, 5 ml of a 24-hour culture of the strain Escherichia ecu (T) QxB _{6 from} the Pasteur Institute in a nutrient medium which had been diluted to </ 6 with distilled water

41

42

h, 5 h and 24 h after the injection would be a certain amount of the compound to be investigated is injected subcutaneously or administered orally.

Then the mortality was within 8 days

determined.

The following results were obtained:

dose

Mortality after 21.5 h 28.5 h 32 h on the 8th day

survivors

Mice

Comparison group 9 0 1 0/10

0.05 mg subcutaneously 0 0 0 10/10

0.1 mg subcutaneously 0 0 0 10/10

0.25 mg subcutaneously 0 0 0 10/10

0.1 mg orally 0 10 9/10

0.25 mg orally 0 0 0 10/10

0.5 mg orally 0 0 0 10/10

B) it was geürueiiei under annual conditions, taking mice with a mean weight of 22.5 g by intraperitoneal injection with a 24-hour culture of the Escherichia coli strain (T) O26B6, which was diluted 1: 5.5, infected

Würucfi.

The following results were obtained with the compound of Examples 1, 2, 6 and 7:

dose Darrei mortality after 23 h 24 hours 25 h ? fih 31 H 37. h 46 h 70 h 4ci On the 8th day ching- 7h 9h 22 h 50 50 30th 30th survivors art 45 35 30th Mice

Comparison group
X 0.5 ml
water

0.015 mg
0.025 mg
0.05 mg
0.05 mg
0.1mg
0.25 mg

s.c.

sc
sc
sc
po
po
po

1 1

0
0
0
0
0
0

0
0
0
0
1
0

2
0
0
4th
0
0

2 0 0 0 0 0

0 0 0 1 1 0 0/10

4/10
6/10

10/10
2/10
5/10

10/10

Experimental infection with
Salmonella typhimurium

The effectiveness of the compound of Example 3 was demonstrated in an experimental infection with Salmonella typhimurium studied in mice

Mice with a mean weight of 20.5 g were administered intraperitoneally in groups of 10 mice Injection of 0.5 ml each of a 24-hour culture of the Salmonella typhimurium 5210 strain in one Culture medium diluted 1:75 with distilled water infected.

At hours, 4 hours, 8 hours, 24 hours and 32 hours after the injection, a certain amount of the compound to be tested was obtained administered by subcutaneous injection

The following results were obtained:

Mortality after

21h 30 25h 28h 29h 45h 56h 70h 4d

30 5d 6d 7d 8d 9d

On the 10th day

survivors

Mice

Comparative 1 1 1 1 1 1 4th 0 0 0 0 0 0 0/10 gmppe 0.1mg 0 0 0 0 0 0 0 0 0 3 3 1 0 3/10 0.25 mg 0 0 0 0 0 0 0 0 0 0 2 1 2 5/10 04 mg 0 0 0 0 0 0 0 0 1 1 1 0 0 7 / Ί0 lmg 0 0 0 0 0 0 0 0 0 0 0 1 0 9/10 2 mg 0 0 0 0 0 0 0 0 0 0 0 0 0 10/10

Experimental infection with Proteus mirabüis

The effectiveness of the compound of Example 4 was demonstrated upon experimental infection with Proteus lnirabilis investigated in mice.

Mice with a mean weight of 22 g were given intraperitoneally in groups of 10 mice each Injection of 0.5 ml each of a 24 ^ IvKuItUr of the strain Proteus mirabilis A 23 in a nutrient medium that was diluted 1: 4, infected.

At 1 hour, 5 hours and 24 hours after the injection, a certain amount of the compound to be tested was obtained administered by subcutaneous injection.

The following results were obtained:

Mortality after

6 h 55 21 h
30th

22 h
15th

24 h 20

28 h 31 h
45

46 h 53 h

70 h

On the 8th day

survivors

Mice

Comparison group

0.025 mg
0.05 mg
0.1 mg
0.25 mg
0.5 mg
1 mg

The acute toxicity of the compound of Example 3 was determined to be 19 to 21 g (Souris Swiss CDI, Specific Pathogen Free [Elevage CH. River-France]) in groups of 8 to 10 animals.

The animals were kept in a cage at a temperature of 21 ± rC, constant humidity and air over-

1 9 0 0 0 0 0 0 0 0/10 0 3 1 1 2 0 2 1 0 0/10 0 0 1 1 0 1 0 0 2 5/10 0 0 0 0 1 0 2 0 0 7/10 0 0 0 0 0 0 0 0 0 10/10 0 0 0 0 0 0 0 0 0 10/10 0 0 0 0 0 0 0 0 0 10/10 acute toxicity

pressure maintained with food and drinking water ad libitum. The compound of Example 3 became one Concentration of 100 mg / ml dissolved in distilled, sterile, apyrogenic water. The solution was in different volumes are injected into a caudal vein at a rate of 1 ml / min.

Results:

Dose (mg / kg)

Mortality according to (d) 12 3

Total
mortality

500 0 0 0 0 0/10 1000 0 0 0 0 0/10 2000 0 0 0 0 0/8

The lethal dose was over 2000 mg / kg.

Symptoms

No symptoms of intoxication were observed either during the injection or during the observation period.

Claims (5)

Patent claims: 1. Oxime derivatives of 7-aminothiazolyIacetamidocephaIosporanäure (syn isomers) of the general formula I NH ₂ κιι \ C O NH OR ' (D CH ₂ -OC-CH ₃ CO ₂ A with
R '=
and
A = Cr to Q-Aikyl H, an alkali metal atom, or an equivalent an alkaline earth metal or one derived from an organic amine base Ammonium group. 2. 3-Acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido] -ceph- (3) -em-4-carboxylic acid re (syn isomer) and its sodium salt. 3. 3-Acetoxymethyl-7- [2- (2-amino-4-thiazo! Yl) -2-sthoxyiminoacetamidoJ-cepn-iSJ-ern ^ -carboxylic acid (syn-isomer). 4. 3-AcetoxymethyI-7- [2- (2-amino-4-thiazolyI) -2-isopropyIoxyiminoacetamido] -ceph- (3) -em-4-car- boric acid (syn isomer). 5. Process for the preparation of the compounds according to claim 1. characterized by per se acquaintance - Implementation of S-acetoxymethyl ^ -aminoce- phalosporanic acid of the formula H ₂ N \ CH ₂ -OC-CH ₃ CO ₂ H 50 with an anhydride of an acid of the general formula II (syn isomer) NH-R ₁ (II) v_ / ^N CO ₂ H
C.
N
OR ' Ri = one by acid hydrolysis or Hydrogenolysis easily cleavable group
and R '= Ci-toQ-alkyl
or with a mixed anhydride of the acid II and a Monoalkylkohlensäureesler to a compound of the general formula Ia (syn isomer) NH-R ₁ C 0 NH Ii OR ' CH ₂ = OC-CH ₃ with R | and R 'as above and acid hydrolysis or hydrogenolysis of the connection of the forms! Yes to a connection of the general formula Ib (syn isomer) NH ₂ / n \ C O NH Il N
OR ' (Ib)