DK161082B - ANALOGY PROCEDURE FOR PREPARING SYN-ISOMER OXIMER DERIVATIVES OF 7-AMINOTHIAZOLYLACETAMIDOCEPHALOSPORANIC ACID - Google Patents

Description

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DK 161082 B

This invention relates to an analogous process for the preparation of novel syn-isomeric oxime derivatives of 7-aminothiazolylacetamidocephalosporanoic acid of the general formula I as set forth in claim 1.

Among the values of R 'are mentioned methyl, ethyl, propyl, isopropyl, butyl, sec.-butyl, tert-butyl, vinyl, propenyl, butenyl, ethynyl and propargyl.

In particular, the values of A include an equivalent of sodium, potassium, lithium, calcium or magnesium.

Among the organic bases which may be represented by A, mention is made of trimethylamine, triethylamine, methylamine, propylamine, Ν, Ν-dimethylethanolamine, tris- (hydroxymethyl) aminomethane, arginine and lysine.

In particular, the invention relates to a process for preparing the compounds of formula I wherein A represents a hydrogen atom, a sodium atom or an equivalent diethylamine.

Among the compounds of formula I, in particular, are mentioned the compounds described below in the Examples, and in particular: - 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido) -ceph-3-em-4-carboxylic acid, syn-isomer, - the sodium salt of 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) 2-methoxyiminoacetamido) -ceph-3-em-4 -carboxylic acid, syn isomer, 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido) -ceph-3-em-4-carboxylic acid, as obtained by the method of Example 4, - The sodium salt of 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido) -ceph-3-em-4-carboxylic acid, as obtained by the procedure of Example 7 - the crystallized sodium salt of 3-acetoxy-methyl-7- (2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido) -ceph-3-em-4-carboxylic acid, syn-isomer, 35-3 acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2-ethoxyiminoacetamido) -ceph-3-em-4-carboxylic acid, syn isomer, and its

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- 2 - salts with alkali metals, alkaline earth metals, magnesium and organic aminated bases, - 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2 - ((2-propenyl) -oxyimino) -acetamido ) ceph-3-em-4-carboxylic acid, syn isomer, and its salts with alkali metals, alkaline earth metals, magnesium and organic aminated bases and - 3-acetoxymethyl-7- (2-amino-4-thiazolyl) -2 - (((1-methyletho-xy) -imino) -acetamido) -ceph-3-em-4-carboxylic acid, syn isomer, and its salts with alkali metals, alkaline earth metals, magnesium and organic aminated bases.

It will be understood that the above compounds of formula I may exist: - either in the form specified by formula I - or in the form of the products of formula I

Li 15 tf i x 20 (½) Ίτνί 25 cd2a where R 'and A have the same meaning as above.

It will also be understood that the products of formula 1 'may exist in the same two forms as indicated above for the products of formula I.

The process for preparing the products of formula 1 'is characterized by the characterizing part of claim 1.

The process for preparing the products of formula I is peculiar to the characterizing part of claim 1.

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In a preferred embodiment of the process for preparing the products of formula I, 7-aminocephalosporanoic acid is treated with a functional derivative of the acid of formula II such as an acid anhydride or acid chloride, the anhydride being formed in situ by the action of isobutyl chloroformate or dicyclohexylcodhexyl acid. Other halides or other anhydrides formed in situ may also be employed by the action of other alkyl chloroformates, a dialkylcarbodiimide or another dicycloalkylcarbodiimide. Other acid derivatives such as the acid azide, the activated acid azide or an activated acid ester formed with e.g.

hydroxysuccinimide, p-nitrophenol or 2,3-dinitrophenol.

In the case where the reaction of 7-aminocephalosporanic acid occurs with a halide of the acid of the general formula Ile or II or with anhydride formed with an isobutyl chloroformate, it is preferably employed in the presence of a basic agent.

As a basic agent, for example, choose an alkali metal carbonate or a tertiary organic base such as N-methylmorpholine, pyridine or a trialkylamine such as triethylamine.

As acidic hydrolyzing agent with which to treat the products of formula Ia optionally, mention must be made of formic acid, trifluoroacetic acid or acetic acid. These acids can be used anhydrous or in aqueous solution. As a hydrolysing agent, the system consisting of zinc and acetic acid must be mentioned in particular.

An acidic hydrolyzing agent such as anhydrous trifluoroacetic acid or formic acid or acetic acid, both in aqueous state, is preferably used to eliminate the tert-butoxycarbonyl or trityl groups.

The system consisting of zinc and acetic acid is preferably used to eliminate the trichloroethyl group 35 and a catalytic halogenation to eliminate the benzyl, dibenzyl and carbobenzyloxy groups.

The products of formula Ib can be converted into salts - 4 -

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according to conventional methods. The salt formation can e.g. is obtained by acting on these acids by an inorganic base such as e.g. sodium or potassium hydroxide or sodium bicarbonate or a salt of a substituted or unsubstituted aliphatic carboxylic acid such as diethylacetic acid, ethyl hexanoic acid or especially acetic acid.

The preferred salts of the above acids are the sodium salts.

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

For the preparation of the salts, the solvates of the free acids can also be used as starting products instead of the free acids. One can, for example. mention the solvates obtained with water, formic acid or an alcohol.

The solvates with an alcohol, especially ethanol, can also be obtained, e.g. treating with a mixture of alcohol and water the solvate formed with formic acid, this treatment being followed by evaporation of the solution.

This salt formation 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 depending on the reaction conditions used.

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The crystalline salts are preferably prepared by reacting the free acids or their solvates formed with e.g. formic acid or ethanol, with one of the salts of the aliphatic carboxylic acids mentioned above, preferably sodium acetate.

In the preparation of a sodium salt, the reaction is carried out in a suitable organic solvent such as e.g. methanol, which solvent may contain small amounts of water.

Furthermore, it is possible to convert the amorphous salts 35 to their crystalline form. For this purpose, an amorphous sodium salt which may be in the form of a solvate with e.g. 0.5, 1 or 1.5 moles of water are dissolved in a suitable -

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organic solvent, preferably a low molecular weight alcohol such as methanol, and the crystallization can then be carried out directly or by the addition of other solvents, e.g. ethanol, isopropanol, n-butanol, acetone, 5 ethers and generally methanol miscible organic solvents.

If the starting product, the solvent or the two components contains water, the crystalline salt can be obtained in hydrated form, e.g. it has been possible to isolate the crystalline sodium salt of 3-acetoxymethyl-7 ~ (2-amino-4-thiazolyl) -2-methoxyiminoacetamido) -ceph-3-em-4-carboxylic acid, syn isomer, with e.g. . 0.5, 1 or 1.5 moles of water.

The acid used for hydrolysis is the salts of the products of formula Ia, which represents an easily eliminable group by acid X, and R represents a group which is readily eliminable by acid hydrolysis or a saturated or unsaturated alkyl group having 1-4 carbon atoms, preferably formic acid. However, trifluoroacetic acid or acetic acid may be used. These acids can be used anhydrous or in aqueous solution.

As the hydrogenolysing agent by which the salts of the products of formula Ia are treated, wherein R ^ represents an X «by hydrogenolysis easily eliminable group and R represents a hydrogenolysis easily eliminable group or a saturated or unsaturated alkyl group of 1-4 carbon atoms can be mentioned. catalytic hydrogenation.

The products of formula II can be prepared by treating a compound of formula IV

30 NH2

/ NN

S N

\ = t / __ ^^ - C02alc (u)

N

OR 'wherein R' represents a hydrogen atom, one by acid hydrolysis

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or hydrogenolysis easily eliminable group or a saturated or unsaturated alkyl group of 1-4 carbon atoms, and alc represents an alkyl group of 1-4 carbon atoms, with a functional derivative of an acidic hydrolysis or hydrogenolysis 5 group, to obtain a product of formula V

NH-R * ·

A

10 A / ^ - COjalo (V) 11

N

i 1 '

OR

Wherein R 2 represents an acidic hydrolysis or hydrogenolysis 1

easily eliminable group and R represents an easily eliminable group by acid hydrolysis or hydrogenolysis or a saturated or unsaturated alkyl group of 1-4 carbon atoms, which product of formula V is treated with a base and then 20 with an acid to give a product of formula II

NH-R ^

A

S N

\ == / co2H

25 il

N

· * I i 'where R and R have the same meaning as above.

In a preferred embodiment of preparation 30 of the products of formula Ile and the products of formula II, the functional derivative of an acid-hydrolysis or hydrogenolysis group is easily eliminated preferably trityl chloride used in the presence of triethylamine or other tertiary aminated bases such as other trialkylamines, methylmorpholine or pyridine.

Other functional derivatives of acid hydrolysis or hydrogenolysis may also be readily used.

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minerable groups. These derivatives include tert-butyl chloroformate prepared in situ or tert-butyl azidoformate, trichloroethyl or benzyl chloroformate, mixed formic acetic anhydride made in situ, benzyl or dibenzyl chloride or another halide, phthalic anhydride carbethoxyphthalimide.

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

The base used for hydrolysis of the product of formula V is preferably sodium hydroxide, but other bases such as potassium hydroxide or barium hydroxide may also be used.

The acid used to isolate the acid of formula II is preferably dilute hydrochloric acid, but acetic acid or formic acid can also be used.

The products of formula IV can be prepared by reacting thiourea with a product of formula III

C1-C2-C-C-CO2alc 20 "" 0 N (III) OR 'where R' represents a hydrogen atom, one by acid hydrolysis

Or by hydrogenolysis easily eliminable group or a saturated or unsaturated alkyl group of 1-4 carbon atoms, and alc represents an alkyl group of 1-4 carbon atoms, to obtain - after treatment with a base - a product of formula IV

30 ^ η2

Ah W, co23Ic <it> '11

N

"I

OR 'wherein R1 and alc have the same meaning as above.

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In a preferred embodiment of this process, the base used to obtain the product of formula IV is potassium acetate. However, the carbonates and bicarbonates of the alkali metals 5 as well as dilute sodium or potassium hydroxide can be used.

A variant of the process for preparing the products IV and of the formula Vb X1

10 S x N

C02alc Y (Vb> OR "b 15 where R4 represents an easily eliminable group by acidic hydrolysis or hydrogenolysis, R" b represents a saturated or unsaturated alkyl group of 1-4 carbon atoms, and alc represents an alkyl group of 1- And corresponding to a product of formula V defined as above, wherein R 1 represents an easily eliminable group by acid hydrolysis or hydrogenolysis, and R represents a saturated or unsaturated alkyl group of 1-4 carbon atoms. treating a product of formula IV "NH,

25 / W

S N

_jCOpalc II CIV ") ¥

OH

30

with an equivalent of a functional derivative of an easily eliminable group by acid hydrolysis or hydrogenolysis to obtain a product of formula X

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NHR S N

2 ° C / 2 ° C (X)

II

s N

5 i

OH

which is treated with an alkylating agent to obtain the expected product of formula Vb. In one embodiment of this process, the functional derivative of a group by acid hydrolysis or by hydrogenolysis is easily eliminated preferably trityl chloride. One then works in the presence of a base, preferably triethylamine. Other bases such as other trialkylamines, methylmorpholine or pyridine can also be used.

Other functional derivatives of acid-hydrolysis or hydrogenolysis easily removable groups such as tert-butyl chloroformate or azidoformate, trichloroethyl or dibenzyl chloroformate, in situ prepared anhydride of formic acid and acetic acid, benzyl or dibenzyl chloride or another halide can also be used. , phthalic anhydride or N-carbethoxyphthalimide.

The alkylating agent used to prepare the products of formula Vb is preferably an alkyl halide such as alkyl iodide or an alkyl sulfate.

A variant of the process for preparing the products IV and of formula IVa nh2

30 A

S N

> COo3lc (IVa) OR'a 35 - 10 -

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wherein alc represents an alkyl group of 1-4 carbon atoms and R'a represents a saturated alkyl group of 1-4 carbon atoms and corresponding to a product of formula IV wherein R 1 represents a saturated alkyl group of 1-4 carbon atoms consists of: processes a product of formula VI

CH3-C-C-C02alc

fl II

0 N (VI)

10H

wherein alc has the same meaning as above, with an alkylating agent to give a product of formula VII

CH3-C-C-C02alc

II II

15. 0 N (VII) OR'a which product is treated with a brominating agent to give a product of formula VIII 20 BrCH2-C-C-CO2alc

II II

0 (VIII) OR1a 25 on which product is allowed to act thiourea and then a base to obtain a product of formula IVa.

In one embodiment of this process, the alkylating agent used to convert the products of formula VI to products of formula VII is preferably an alkyl halide such as the alkyl chloride, bromide or iodide or an alkyl sulfate of 1-4 carbon atoms. .

The brominating agent used to convert the products of formula VII into products of formula VIII is preferably bromides.

The base that one uses after the impact of - 11 -

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the product of formula VIII on the thiourea is preferably an alkali metal carbonate or bicarbonate. However, dilute sodium or potassium hydroxide or potassium acetate may be used.

It has been found that the visual or anti-configuration of the products of formula I is bound to the configuration of the products of formula IV, because the configuration of the products obtained from the products of formula IV can be maintained through the course of the synthesis.

10 The same applies, of course, to the products obtained from the products of formulas IVa and IV ", since the latter are covered by formula IV.

The configuration of the products of the formula IV depends on a certain number of parameters which are involved in the manufacture of these products.

Thus, it has been found that when the effect of thiourea on the products of formula III is carried out either in an aqueous solvent such as aqueous acetone or aqueous ethanol or at room temperature, leaving a practically stoichiometric amount of thiourea to act for a fairly short period of the order of magnitude 1. -3 hours, or finally, when all the above conditions are combined, the syn isomer is obtained.

A process for preparing the products of formula IV above in syn form consists of reacting thiourea and then a base with a product of formula III 'X-CH 2 -C-C-CO 2 alk

II II

30 O N (III ') OR'b wherein X represents a chlorine or bromine atom and R'b represents a hydrogen atom, an acid by hydrolysis or hydrogenolysis easily eliminable group or a saturated or unsaturated alkyl group having 1-4 carbon atoms when X represents a chlorine - 12 -

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or R'b represents a hydrogen atom or a saturated alkyl group of 1-4 carbon atoms when X represents a bromine atom and, with respect to the reaction of the thiourea, either works in an aqueous solvent or works at room temperature in the presence of a convenient taken stoichiometric amount of thiourea and during the reaction for a limited period of time, or operating under the combination of all the above conditions.

A process for preparing the products of formula IV in syn form, reacting thiourea with a product of formula III as defined above, consists in either working in an aqueous solvent or working at room temperature in the presence of a practically stoichiometric amount of thiourea and during the reaction for a period of time limited to a few hours, or working under all conditions specified above.

A process for obtaining the products of Formula IVa as defined above wherein reacting thiourea with a product of Formula VIII consists in either working in an aqueous solvent or working at room temperature in the presence of a practically stoichiometric amount of thiourea and during the course of the reaction for a limited period or several hours or working under the conditions mentioned above.

Thus, the products of Formula IV obtained in Examples 3, 8, 10, 12 and 15 below have visual configuration.

The products of the general formula I ', especially the products of the general formula I, and especially those in the form of vision, have a very good antibiotic effect partly on gram-positive bacteria such as staphylococci and streptococci, especially penicillin-resistant staphylococci, and partly on gram-negative bacteria. , especially coliform bacteria, Klebsiella, Salmonella and Proteus.

These properties make the said pharmaceutical pro- 13

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drugs acceptable for use as medicaments in the treatment of susceptible germ disorders and in particular in the treatment of facial or skin staphylococci, pyodermitis, septic or suppurative ulcers, anthrax, phlegmone, erysipel, primitive or post-influenza acute staphylococci, and bronchopneumonia pulmonary suppurations.

These pharmaceutically acceptable products can also be used as medicaments in the treatment of coli bacilliosis and associated infections, in infections with Proteus, Klebsiella and Salmonella and other disorders caused by gram-negative bacteria.

The pharmaceutically acceptable products of formula I can be used to prepare pharmaceutical products containing as active ingredient at least one of said products and in particular one having a visual structure according to the examples.

In particular, among these products are the pharmaceutical products which contain as active ingredient at least one of the products of general formula I wherein R represents a hydrogen atom, R 'represents a hydrogen atom or a saturated or unsaturated alkyl group having 1 4 carbon atoms, and A represents a hydrogen or sodium atom.

Also mentioned are the pharmaceutical products which contain as active ingredient: - 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido) -ceph-3-em-4-carboxylic acid , syn-isomer; this product in particular described in Examples 4, 6, 20 or 22, or the sodium salt of 3-acetoxymethyl-7- (2- (2-amino-4-30-thiazolyl) -2-methoxyiminoacetamido) -ceph-3-em-4 -carboxylic acid, syn-isomer; this product is described in particular in Example 7, or the crystalline sodium salt of 3-acetoxymethyl-7- (2- (2-amino-4-thiazoly) -2-methoxyiminoacetamido) -ceph-3-em 35 -4-carboxylic acid, syn-isomer, or at least one of the following products: - 14 -

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3-Acetoxymethyl-7- (2-amino-4-thiazolyl) -2-ethoxyiminoacetamido) -ceph-3-em-4-carboxylic acid, syn isomer, and its salts with alkali metals, alkaline earth metals, magnesium or pharmaceutically acceptable aminated organic bases, 5-3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2 - ((2-propenyl) -oxyimino) -acetamido) -ceph-3-em-4 carboxylic acid, synisomer, and its salts with alkali metals, alkaline earth metals, magnesium or pharmaceutically acceptable organic bases, 10-3-acetoxymethyl-7- (2-amino-4-thiazolyl) -2 - ((1-methyletho) - xyl-imino-acetamido-ceph-3-em-4-carboxylic acid, syn isomer, and its salts with alkali metals, alkaline earth metals, magnesium or aminated organic bases which are pharmaceutically acceptable.

These products may be administered by oral, rectal or parenteral route or topically by application to the skin and mucous membranes.

They may be solid or liquid and may be present in the pharmaceutical forms 20 commonly used in human medicine such as e.g. unsweetened or unsweetened tablets, gelatine capsules, granules, suppositories, injection preparations, pomade, cream or jelly. They are manufactured according to the usual methods. The active ingredient (s) may be incorporated therein together with commonly used additives such as talc, gum arabic, lactose, starch, magnesium stearate, cocoa butter, aqueous or non-aqueous carriers, fats of animal or vegetable origin, paraffin derivatives. glycols, various wetting, dispersing or emulsifying agents and preservatives.

The dose administered varies according to the condition being treated, the patient concerned, the method of administration and the product. It can e.g. range from 0.250 g to 4 g per per day orally in humans with the product described in Example 4 or 7, or between 0.500 g and 1 g, 3 times a day by intramuscular route.

The products of formula III, which are unknown - 15 -

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tea, can be prepared from gamma-chloro-alpha-oximinoacetyl-acetic acid ethyl ester described in J. of Medicinal Chemistry 1973, vol. 16, No. 9, as set forth below.

The products where R 'is a group which is easily eliminated by acid hydrolysis or by hydrogenolysis are obtained by the classical reactions which use functional derivatives of these groups.

The products where R 'is a saturated or unsaturated alkyl group having 1-4 carbon atoms are obtained by acting on gamma-chloro-alpha-oximinoacetylacetic acid ethyl ester of the corresponding alkyl halides or sulfates.

The products of formula III ', wherein X represents a bromine atom and R'b represents a hydrogen atom, can be obtained by treatment with a brominating agent of products of formula 15 vi CH3-C-C-CO2alc

11 II

O N (VI)

20H

under the same conditions as described for the treatment of the products of formula VII.

In addition to the products described in the Examples, the following products are additional substances obtainable by the invention: - 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2-propyloxyiminoacetamido) -ceph-3- em-4-carboxylic acid, syn isomers, and its salts with alkali metals, alkaline earth metals, magnesium or aminated organic bases, 30-3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2-butyloxyiminoacetamido ) -ceph-3-em-4-carboxylic acid, syn-isomer, and its salts with alkali metals, alkaline earth metals, magnesium or aminated organic compounds, - 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) - 2- (2-methylpropyloxyimino) -aeetamido) -ceph-3-em-4-carboxylic acid, syn isomer, and its salts with alkali metals, alkaline earth metals, magnesium or aminated organic bases,

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3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2- (1,1-dimethylethoxy-imino) -acetamido) -ceph-3-em-4-carboxylic acid, syn isomer, and its salts with alkali metals, alkaline earth metals, magnesium or aminated organic bases, 5-3-acetoxymethyl-7- (2- (2-amino-4-t hi az o ly 1) -2 - (2-but enyloxyimino) - -acetamido) -ceph-3-em-4-carboxylic acid, syn isomer, and its salts with alkali metals, alkaline earth metals, magnesium or aminated organic bases, - 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) ) -2- (3-Butenyloxyimino) -LO-acetamido) -ceph-3-em-4-carboxylic acid, syn isomer, and its salts with alkali metals, alkaline earth metals, magnesium or aminated organic bases, - 3-acetoxymethyl-7- (2- (2-Amino-4-thiazolyl) -2- (2-butynyloxyimino) acetamido) -ceph-3-em-4-carboxylic acid, syn-isomer, and its salts L5 with alkali metals, alkaline earth metals, magnesium or aminated organic bases, - 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2- (3-butynyloxyimino) -acetamido) -ceph-3-em-4-carboxylic acid, syn isomer . and its salts with alkali metals, alkaline earth metals, magnesium or aminated organic bases, - 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2- (vinyloxyimino) -acetamido) -ceph-3 em-4-carboxylic acid, syn-isomer, and its salts with alkali metals, alkaline earth metals, magnesium or aminated organic bases, 25-3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2- (1- propenyloxyimino) -acetamido) -ceph-3-em-4-carboxylic acid, syn isomer, and its salts with alkali metals, alkaline earth metals, magnesium or aminated organic bases, - 3-acetoxymethyl-7- (2- (2-amino) 4-thiazolyl) -2- (1-propynyloxyimino) -10-acetamido) -ceph-3-em-4-carboxylic acid, syn isomer, and its salts with alkali metals, alkaline earth metals, magnesium or aminated organic bases, - 3-acetoxymethyl -7- (2- (2-amino-4-thiazolyl) -2- (1-butenyloxyimino) -acetamido) -ceph-3-em-4-carboxylic acid, syn isomer, and its salts with alkali metals, alkaline earth metals , magnesium or aminated organic bases, - 3-acetoxymethyl-7- (2- (2-amino-4-thi) azolyl) -2- (1-ethynyloxyimino) acetamido) -ceph-3-em-4-carboxylic acid, syn-isomer, and its salts with alkali metals, alkaline earth metals, magnesium or aminated organic bases,

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- 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2- (2-propynyloxyimino) -acetamido) -ceph-3-em-4-carboxylic acid, syn-isomer, and its salts with alkali metals, alkaline earth metals, magnesium or aminated organic bases, r 5-3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2- (1-butynyloxyimino) -acetamido) -ceph-3-em. 4-carboxylic acid, syn isomer, and its salts with alkali metals, alkaline earth metals, magnesium or aminated organic bases,

The following examples illustrate the method of the invention.

15 20 25 30 35

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Example 1 3-Aethoxymethyl 1-7- (2- (2-tritylamino-4-thiazolyl) -2-tritylhydroxy-iminoaoetamido) -ceph-3-em-4-oreboxylic acid.

Step A; 2- (2-amino-4-thiazolyl ·) -2-hydroxyiminoeddikesyreethylester.

2 g of γ-chloro-α-oximinoacetylacetic acid ethyl ester are introduced into 5 ml of ethanol and 0.76 g of thiourea and stirred at room temperature for a total of 16 hours. The hydrochloride crystallizes out and is diluted with 5 ml of ether, suction, rinsed with a mixture of ethanol and ether in a ratio of slurry and then with ether to give 1.55 g of 10 hydrochloride.

The 1.55 g hydrochloride is dissolved at 40-50 g in 8 ml of water and then neutralized to a pH of 5-6 by the addition of sodium acetate. The free amine crystallizes out. One is cooled and then suctioned, washed with water, dried and 1.22 g of anti-isomer with m.p. 154 ° C.

The mother liquors and washings are combined from several experiments, evaporated, absorbed in water, washed with ether, sodium bicarbonate added, suctioned, washed with water and obtained 1.9 g of product with second stains by thin layer chromatography and purified by chromatography on silica gel 20 eluting with ether. The pure fractions of the syn isomer are combined, evaporated, ethered, suctioned, dried and 50 mg of this isomer obtained.

Step B: 2- (2-Tritylamino-4-thiazolyl) -2-tritylhydroxyimino acetic acid ethyl ester.

To a solution of 5.4 g of product A of step A in 54 ml of chloroform and 7 »5 ml of triethylamine is added at 10 ° C a solution of 15 g of trityl chloride in 30 ml of chloroform. After standing for 1 hour, wash with 40 ml of water and 20 ml of water containing 4 ml of 1 M hydrochloric acid, then decant, dry and evaporate to dryness.

30

The residue is taken up in 10 ml of ether and 50 ml of methanol are added, stirred, suctioned, washed with methanol and, in two yields, 14.2 g of the expected product are obtained.

Step C: 2- (2-Tritylamino-4-thiazolyl) -2-tritylhydroxyiminoacetic acid 35.5.5 g of ester of Step B is suspended at near reflux temperature in 55 ml of dioxpn. 17 ml of 2 R sodium hydroxide solution are slowly added. A slight reflux is continued, it cools and sucks the salt. The salt is taken up in 60 ml of methylene chloride, 20 ml of water and 2 ml of acetic acid. The acid is extracted, washed with water and an initial yield of 7 g of acid is obtained.

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The dioxane is evaporated from the mother liquor and 20 ml of methylene chloride, 10 ml of water and 1 ml of acetic acid are added. A second yield of 1.5 g of the same product, ie a total of 8.5 g, is isolated.

Analysis. C ^^ HH ^^ 0O ^ 3J ^S, 0.5 HgO 5 calculated: C # 75.85 H # 5.03 6.17 S # 4.7 found: 75.8 4.9 5.9 4.6 lane D: 5-acetoxymethyl-7- (2- (2-tritylamino-4-thiazolyl) -2-trityl-hydroxyiminoacetamido) -ceph-3-em-4-carboxylic acid.

A suspension of 8.5 g of acid from step C is stirred in 50 ml of methanol and 5 ml of N-methylmorpholine is added. Stir for 10 minutes at 30 ° C, add 30 ml of methylene chloride, evaporate, add 100 ml of ether, comminute, suction, wash with ether, dry and get an initial yield of 7.2 g of salt. It evaporates to dryness, absorbs in ether and gives another yield of the same product.

While stirring under inert gas, 4.24 g of the above morpholine salt is suspended in 60 ml of methylene chloride.

Stir for 5 minutes, cool to -5 ° C and add 6 ml of 1 M solution of isobutyl chloroformate in methylene chloride. Allow to stand for 15 minutes with stirring at -5 ° C, cool to -20 ° C and 20 add a solution of 1.36 g of 7-aminocephalosporanoic acid in 25 ml of methylene chloride and 1.4 ml of triethylamine. Allow to stand for 1 hour at room temperature, wash with 50 ml of water containing 10 ml of 1S hydrochloric acid, suction, decant, wash with water and evaporate to dryness. Ether is purged, suctioned, washed with ether and 4.5 g of crude product are obtained.

The crude product is stirred for 1 hour at 10 ° C in 10 ml of methylene chloride. The insoluble material is suctioned off and rinsed with methylene chloride. 50 ml of ether are added to the filtrate, stirred, suction precipitated, washed with ether to give 2.29 g of the expected product.

A further yield of 0.856 g, i.e. 3.146 g of the expected product.

3-Aethoxymethyl 1-7- (2- (2-amino-4-thiazolyl) -2-hydroxyiminoacetamido) - ce ph-3-em-4-carboxylic acid.

2.29 g of the product obtained in Example 1 are suspended in 18.4 ml of 50 # aqueous formic acid. It is heated under vigorous stirring for 15 minutes to 55 ° C.

You cool, add 10 ml of water, suck off, wash with

-20- DK 161082 B

water, evaporates in vacuo, adds acetone, sucks, adds 30 ml of ether, stirs, sucks, washes with ether and obtains 0.665 g of product # An additional 0.123 g of crystallizing product is obtained, ie, 0.888 g. g of product in 7.5 ml of ethanol and 7.5 ml of ace tone. 70 mg of activated charcoal is added, suctioned off, the solvents evaporated, ethanol fines, washed with ethanol and 0.450 g in a first yield and then 0.105 g in a second yield.

I.R. Spectrum: 10) c = 0 1774 cm "1 (β-lactam) 1740 cm” 1 1676 cm ”1 C = C KH₂ 1630 cm” ”UH 1520 cm” 1

Example 3

5-acetoxymethyl-7- (2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetamido) -ceph-3-em-4-carboxylic acid.

Step A: 2- (2-Amino-4-thiazolyl) -2-methoxyiminoacetic acid ethyl ester. 20 g of γ-chloro-α-methoxyiminoacetylacetic acid ethyl ester are introduced into 3 ml of absolute ethanol and 0.42 g of powdered thiourea is added. 2 hours. The obtained crystalline hydrochloride is diluted with 60 ml of ether, stirred, suctioned, washed, dried and 685 mg of hydrochloride are obtained. It is dissolved in 4 ml of water at 50 ° C, potassium acetate is added until a pH of 6 and the released amine crystallizes out. You cool, suck, wash with water, dry and get 270 mg of the expected product. Mp. 161 ° 0th

The obtained product has syn configuration, H.M.B. spectrum 30 (GD01, 60 MHz) p.p.m.i 4 (H-OCH ^), 6.7 (proton in thiazole ring).

Step B: 2- (2-Tritylamino-4-thiazolyl) -2-methoxyiminoacetic acid ethyl ester.

4.6 g of the product obtained in the previous step is dissolved at 30 ° 0 in 92 ml of methylene chloride. Cool to -10 ° C, add 2.9 ml of triethylamine, cool further to -35 ° 0, add over 15 minutes 6.1 g of trityle chloride, allow to re-enter room temperature, all within 2 hours 30 minutes . Wash with water and then with 0.5 Ή hydrochloric acid and with sodium acetate in water. Dry, evaporate, take up ether, evaporate again, dissolve in methanol, add water and ether, crystallize, suction, wash with

-21- DK 1610 8 2 B

ether to obtain 6.15 g of the expected product. Mp. 120 ° C.

The product obtained has a vision configuration.

Step G; 2- (2-Tritylamino-4-thiazolyl) -2-methoxyiminoedetic acid.

Dissolve 7.01 g of ester B in 35 ml of dioxane. The oil bath is heated to 5 to 110 ° C and 9 ml of 2 IT sodium hydroxide solution are added over 5 minutes and allowed to stand for 30 minutes under reflux and with stirring. The sodium salt crystallizes out. It is cooled, suctioned, washed with dioxane and then with ether to give an initial yield of 5.767 g of salt. The mother liquor is evaporated and a second yield of 1.017 g, ie a total of 6.784 g of salt, is obtained.

3.06 g of salt are introduced into 65 ml of methylene chloride and 6.5 ml of 2 H hydrochloric acid, washed with water, dried and evaporated to dryness to give the free acid quantitatively.

The product obtained has syn configuration, N.M.E. spectrum 15 (DISO, 60 MHz) p.m .: 3.68 (N-OCH 2), 6.6 (proton in thiazole ring).

Step D: 3-Acetoxyme thyl-7- (2- (2-tritylamino-4-thiazolyl) -2-methoxyminoacetamide o) -ceph-3-em-4-carboxylic acid.

The dry acid from step C is dissolved in 30 ml of dry methylene-2Q chloride. 0.78 g of dicyclohexylcarbodiimide is added and stirred for 1 hour at room temperature. The resulting dicyclohexyl urea is filtered off, cooled to -10 ° C, a solution of 7,01 g of 7-aminoophalosporanoic acid is added in 13 ml of methylene chloride and 0.9 ml of triethylamine. It is allowed to re-enter room temperature, add 1 ml of acetic acid, suck __ off, wash with water added hydrochloric acid and with water, dry, evaporate to dryness, take up in 10 ml of dioxane and add 1 ml of water and 3 ml of saturated sodium bicarbonate solution. You stir, suck, wash and evaporate to dryness. It is taken up in methylene chloride, washed with 10 ml of water and 5 ml of 1 N hydrochloric acid, decanted, washed with water, dried 3Q, ether, and obtained 1.747 g of crude product which is purified by dissolving in ethyl acetate followed by precipitation in ether. 1,255 g of pure product is obtained.

The product obtained has a vision configuration. The γ-ehloro-oc-methoxyiminoacetylacetic acid ethyl ester used as the starting product in Example 3 is prepared as follows:

22.5 g of γ-chloro-α-oximinoacetylacetic acid ethyl ester are introduced into 100 ml of methylene chloride.

Place in an ice bath and slowly add with stirring a fresh solution of diazomethane (21.6 g per liter) or 275 ml.

You leave in contact for 5 minutes and destroy the surplus

-22- DK 161082 B

diazomethane with a little alumina. Evaporate and purify by elution on silica gel with methylene chloride. 11.93 g of the expected product is obtained.

2- (2-Amino-4-thiazolyl) -2-methoxyiminoacetic acid ethyl ester 5 used in Example 3 is prepared as follows:

Step a: 2-acetyl-2-methoxyiminoacetic acid ethyl ester.

180 g of crude 2-acetyl-2-hydroxyiminoacetic acid ethyl ester are introduced into 900 ml of pure acetone. At 10 ° C and under nitrogen atmosphere, 234 g of potassium carbonate are added. 10 103 ml of dimethyl sulfate is then introduced. Stir for 3 hours at room temperature, add ice, pour into 4 liters of water, extract with methylene chloride, wash with water, dry and distill off the solvents. 185 g of the expected product is obtained.

Step β: 4-Bromo-2-methoxyiminoaoethyl acetic acid ethyl ester.

197 g of the product obtained in step α is placed in 1 liter of methylene chloride and 200 mg of p-toluenesulfonic acid is added. At 20 ° C, 1/10 of a solution of 191 g of pure bromine in 200 ml of methylene chloride is introduced. Once the reaction is started, the rest of the bromine-2Q solution is introduced over 1 hour at approx. 20 ° 0th The temperature is then allowed to rise to 25 ° 0 to complete the reaction. Wash with ice-cold water, re-extract with methylene chloride, dry and distill off the solvent. The expected product is 268 g.

Step y: 2- (2-Amino-4-thiazolyl) -2-methoxyiminoacetic acid ethyl ester.

80 g of thiourea are introduced into 270 ml of ethanol and 540 ml of water. Under nitrogen atmosphere, 268 g of the product obtained in step β in 270 ml of ethanol are introduced over 30 minutes. Stir for 1 hour at approx. 20 ° 0th Cool to approx. 15 ° C and in small portions introduce potassium bicarbonate up to a pH of 5. It is sucked off, washed with water, dried and 133.8 g of the expected product are obtained.

The product obtained is identical to that obtained in step A.

Example 4

5-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido) - ceph-3-em-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 50 $ aqueous formic acid. 4 ml of water are added, suctioned off, evaporated to dryness in vacuo and comminuted in 2 ml of ethanol, suctioned off, washed with ethanol and then with ether

-23- DK 161082 B

to obtain 0.428 g of pure product.

Analysis: ^ 16 ^ 17 ^ 7 ^ 5 ^ 2 Calculated: C # 42.19 3.76 15.37 14.08 Found: 42.3 4.1 15.2 13.8 5 The product obtained has a vision configuration, NME spectrum (DMSO, 60 MHz) ppm: 2.03 (-COCH 5), doubled 9.58, J = 8 Hz (COM) 6.76 (proton in thiazole ring).

Example 5

The piethylamine salt of 3-acetoxymethyl-7- (2- (2-tritylamino-4-thiazolyl) -2-methoxyimino) -acetamido) -ceph-3-em-4-carboxylic acid.

The crude 3-acetoxymethyl-7- (2- (2-tritylamino-4-thiazolyl) -2- (methoxyimino) acetamide o) -ceph-3-em-4-carboxylic acid prepared as described in Example 3 from the condensation of 2- (2-tritylamino-4-15 -thiazolyl) -2-methoxyiminoacetic acid in the form of anhydride prepared by dicyclohexylcarbodiimide and 40.8 g of 7-aminoeephalosporanic acid are dissolved in 350 ml of dioxane. 350 ml of ether and 33 ml of diethylamine are slowly added with stirring. Stir for 20 minutes and extract the diethylamine salt of 2- (2-tritylamino-4-thiazolyl) -20 -2-methoxyimino acetic acid, which is crystallized out. This salt is washed twice with 30 ml mixture of dioxane and ether above to give 62.6 g. The filtrate is evaporated to syrupy consistency and added approx. 2.5 liters of ether. You stir and suck. 110.3 g of det. Is obtained. expected diethylamine salt. The product obtained has 25 vision configuration.

Example 6

3-Acetoxymethyl-7- (2- (2-amino-4-thiazolyl) - 2- (methoxyimino) -acetamide o) -ceph-3-em-4-carboxylic acid.

36 g of the product of Example 5 are added to 180 ml of 50 $ aqueous formic acid, which is kept at 50 ° C. The mixture is stirred at 50 ° C for 20 minutes and the resulting triphenylcarbinol is extracted. 180 ml of ethanol is added, evaporated to dryness under reduced pressure and the residue is taken up in a mixture of 100 ml of water and 20 ml of ethanol and evaporated again. It is taken up in 100 ml of water, stirred for 15 minutes at 15 ° C, suctioned off, washed with water and then with ether to give 15.6 g of the expected product.

The product is identical to that obtained in Example 4.

- 24 - DK 161082 B

Example 7 »

Sodium any one of 3-ethoxymethyl-7- (2- (2-amino-4-thiazoly) -2- (methoxy-amino) -acetamide o) -ceph-3-em-4-carboxylic acid.

45.55 g of pure 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2-5 - (methoxyimino) -acetamino) -ceph-3-em-4-carboxylic acid obtained according to the procedures of Example 4 or 6 is added 100 ml of distilled water. Gradually, 8 g of sodium bicarbonate is added with addition of approx. 20 ml of ethanol. -

80 ml of ethanol, 4.5 g of activated charcoal are again added, stirred for 5 minutes, filtered, rinsed with ethanol and evaporated to dryness under vacuum. It is taken up in 100 ml of ethanol, evaporated to dryness and the residue is dissolved in 100 ml of methanol. Add 2 liters of acetone, stir vigorously, suction and rinse with acetone and then with ether. After drying under vacuum, 43.7 g of a white product is obtained which is rehydrated in the air to reach a final weight of 45.2 g = + 55 ° + 2 ° (0.8 ° water).

Analysis calculated: 0 # 40.24 H # 3.38 N # 14.67 S # 13.43 Na # 4.81 found: 40.3 3.8 14.4 13.3 4.84 The product obtained has syn configuration, NMR spectrum (60 MHz, D 2 O) ppm: 2.01 (COCH +), doubled at 9.53, J = 8 Hz (NHCO) 6.75 (thiazole proton).

Example 8.

3-ace toxyme thyl-7- (2- (2-tritylamino-4-thiazolyl) -2- ((2-propenyl) oxyimino) -acetamide o) -ceph-3-em-4-carboxylic acid .

Step A: 2- (2-Amino-4-thiazolyl) -2 - ((2-propenyl) oxyimino) -acetic acid ethyl ester.

a) 9.7 g of 2-hydroxyimino-4-chloroacetylacetic acid ethyl ester, 30 ml of acetone and 9.15 ml of 3-iodopropene are cooled in ice, and 27.5 ml of 2N sodium hydroxide solution are added and allowed to stand for 1 hour 30 minutes at room temperature.

b) Add 3.8 g of thiourea to the reaction environment and warm to 60 ° C for 15 minutes followed by 45 minutes at room temperature, then evaporate the acetone and add methylene chloride, water and potassium carbonate and stir, decant, re-extract with methylene chloride. dries, evaporates to dryness and yields 9.75 g of residue which is chromatographed on silica gel eluting with ether, then isolates 2.7 g of product which is taken up in isopropyl ether. The crystals are sucked off, rinsed and dried. There are available

-25- DK 161082 B

783 mg of the expected product. Mp. 100 ° 0th The product obtained has a vision configuration.

lane B: 2- (2-tritylamino-4-thiazolyl) -2- ((2-propenyl) oxyimino) acetic acid ethyl ester.

511 mg of the product prepared in Step A, 0.92 ml of dimethylformamide, 1.8 ml of methylene chloride and 0.29 ml of triethylamine are mixed. It is cooled to -15 ° C and 615 mg of trityl chloride is added. Allow to stand at room temperature for 1 hour 30 minutes, add 2 ml of 1N hydrochloric acid and then 5 ml of water, decant, dry, evaporate to dryness and give 1.28 g of crude product.

The product obtained has a vision configuration.

lane C: 2- (2-tritylamino-4-thiazolyl) -2- ((2-propenyl) oxyimino) -acetic acid.

To 120 ° C 1.28 g of product from step B, 6.2 ml of dioxane and 3 ml of 2 L sodium hydroxide solution are heated. Reflux for 1 hour. The sodium salt crystallizes and sucks, rinses with a mixture of ether and dioxane, dries and isolates 8Ο5 mg of product.

It is taken up in 10 ml of methylene chloride and 3 ml of 1N hydrochloric acid, stirred until dissolved, decanted, dried, evaporated to dryness, taken up in ether, suctioned and obtained 715 mg of the expected product. Mp. 170 ° C.

The product obtained has a vision configuration.

Saline D: 5-acetoxymethyl-7- (2- (2-tritylamino-4-thiazolyl) -2 - ((2-propenyl) oxyimino) oacetamide o) -ceph-3-em-4-carboxylic acid.

Mix 470 mg of product from step C, 5 ml of methylene chloride and 130 mg of dicyclohexylcarbodiimide, rinse with a little methylene chloride and leave to stir for 1 hour at room temperature, extract the resulting dicyclohexylurea, cool the filtrate and add, under inert gas 136 mg 7-amine in 2.4 ml of methylene chloride and 0.14 ml of triethylamine. Allow to stand for 1 hour 30 minutes at room temperature, add 2 ml of 1N hydrochloric acid and water, stir, decant, wash with water, dry, evaporate and give 610 mg of crude product.

The product obtained has a vision configuration.

-26- DK 161082 B

EXAMPLE 9 3-Acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2- ((2-propenyl) -oxyimino) -acetamido) -oeph-3-em-4-carboxylic acid.

The 610 mg product of Example 8 and 3 ml of 50 $ 5 aqueous formic acid are heated to 60 ° C for 15 minutes, 4 ml of water is added, stirred, extracted with the triphenylcarbinol, rinsed with water, evaporated to dryness under vacuum, absorbed in water. , comminuted, sucks from rinses and gets 120 mg of the desired acid. Mp. ca. 160 ° C * U.V. Spectrum (ethanol) s 10 max 236 nm E ^ = 375 ε = 18,000 turning point 252 nm E ^ = 316 turning point 295 nm E ^ = 138 ε = 6,600 IT. ?. spectrum (ethanol and HCl, U / 10): max 263 nm = 380 ε = 18,300 turning point 280 nm E + = 317 HM11 spectrum (DMSO, 90 MHz) ppm,: 2.02 (OAc), 6.68 (thiazolpro-ton).

The product has a visual configuration.

Example 10.

5-acetoxymethyl-7- (2- (2-tritlamino-4-thiazolyl) -2- (ethoxyimino) -acetamide o) -ceph-3-em-4-carboxylic acid.

Step A: 2- (2-Amino-4-thiazolyl) -2-ethoxyiminoacetic acid ethyl ester.

a) 19.4 g of γ-chloro-α-oxyiminoacetoacetic acid ethyl ester are placed in 60 ml of acetone and 14.3 ml of diethyl sulfate. It is cooled for 10 minutes in an ice bath and 55 ml of 2N sodium hydroxide solution are added over 30 minutes and stirred for 40 minutes.

b) Add to the reaction environment 7.6 g of thiourea, heat to 55 ° C for 20 minutes, evaporate the acetone, absorb in ethyl acetate, add 6.9 g of potassium carbonate, stir, decant, re-extract with ethyl acetate, dry and evaporates to dryness. 17.4 g of residue is isolated which is chromatographed on silica gel eluting with ether.

The expected product is collected, taken up in isopropyl ether, sucked off, rinsed, dried and obtained 2.8 g of the expected product. Mp. 129 ° 0, The product obtained has visual configuration.

Step B: 2- (2-Tritylamino-4-thiazolyl) -2-ethoxyiminoacetic acid ethyl ester.

3.16 g of product from step A, 6 ml of dry dimethylformamide, 12 ml

- 27 - DK 161082 B

methylene chloride and 1.89 ml of triethylamine are placed under inert atmosphere. It is cooled to -15 ° C and slowly added 3.98 g of triethyl chloride. Allow to stand for 30 minutes, the temperature rises to 10 ° C, then stay for 3 hours 30 minutes at room temperature.

5 ml of 1N hydrochloric acid are added, stirred, decanted, washed with 1H hydrochloric acid and then with water. Extract with methylene chloride, dried, evaporated to dryness to give 7.89 g of crude residue.

The product obtained has a vision configuration.

Step C: 2- (2-Tritylamino-4-thiazolyl) -2-ethoxyiminoacetic acid.

To 110 ° C 87.9 g of product from step B, 40 ml of dioxane and 19.5 ml of 2N sodium hydroxide solution are heated for 1 hour. One sucks, rinses with a mixture of ether and dioxane and then with ether alone and dries. 6.25 g of sodium salt are obtained, which are taken up in 60 ml of methylene chloride and 20 ml of 1 L hydrochloric acid, stirring the two phases, adding 20 ml of methanol, decanting, washing again with water, re-extracting with a mixture of methylene chloride and methanol, dries, evaporates and isolates 5.85 g of pure 2- (2-tritylamino-4-thiazolyl) -2-ethoxyiminoacetic acid.

The product obtained has a vision configuration.

8-5-Acetoxymethyl-7- (2- (2-triethylamino-4-thiazolyl) -2- (ethoxyimino) -acetamide o) -ceph-3-em-4-carboxylic acid.

3.43 g of 2- (2-tritylamino-4-thiazolyl) -2-ethoxy-iminoacetic acid from step C is applied to 34 ml of methylene chloride, the suspension is cooled and 970 mg of dicyclohexylcarbodiimide is added, rinsed with methylene chloride and stirred for 1 hour. room temperature. The icyclohexylurea is extracted.

The filtrate is cooled to -20 ° C and a solution at -20 ° C of 1.02 g of 7-aminocephalosporanoic acid in 18 ml of methylene chloride and 1.06 ml of triethylamine is added at one time. Heat for 1 hour 30 minutes, add 1.8 ml of acetic acid, add 9 ml of 1N hydrochloric acid, stir, decant, wash with water, extract with methylene chloride, dry, evaporate and get 4.56 g of the expected product.

35 The product obtained has a visual configuration.

Example 11.

5-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2- (ethoxyimino) aceto-amido) ceph-3-em-4-carboxylic acid.

The 4.56 g product of Example 10 is placed in 23 ml

"28 ~ DK 161082 B

50 $ aqueous formic acid, heat for 15 minutes to 55 ° C, dilute with water (30 ml), extract the triphenylcarbinol, Evaporate the filtrate to dryness, absorb in water, stir, suction, rinse, dry and get 116 mg of impure product . A second yield of 674 mg of product is obtained, which crystallizes by evaporation of the filtrate, ie a total of 790 mg.

The following cleaning is performed:

1.063 g of crude product is poured into 5 ml of water, heated to 70 ° C for 5 minutes, cooled, stirred for 30 minutes, suctioned, rinsed, dried and isolated 815 mg of purified product. These 815 mg are taken up in 2 ml of water and 3 ml of acetone, heat slightly, extract insoluble material, add 3 ml of water, heat to 60 ° C and evaporate the acetone by bubbling nitrogen, extract the formed grains, rinse with water and then with ether and isolates 438 mg of the expected product.

Calcd: 43.49 W ° 4.08 W 14.92 S # 13.66 Found: 44.5 4.4 14.8 13.3

The product has vision configuration, N.M.R. spectrum (60 MHz, 20 DMSO) p.p.m,: 2.05 (OAc), 6.75 (proton in the thiazole ring).

Example 12.

3-aoetoxymethyl-7- (2- (2-tritylamino-4-thiazolyl) -2- (1-methylethoxy-imino) acetamido) ceph-3-em-4-carboxylic acid.

Step A: 2-Acetyl-2- (1-methylethoxyimino) -acetic acid ethyl ester.

39.8 g of 2-acetyl-2-hydroxyiminoacetic acid ethyl ester are applied in 200 ml of pure acetone. Cool in an ice bath and add 52 g of potassium carbonate and then in 30 minutes 25 ml of 2-iodopropane. Stir then for 2 hours, add 800 ml of water and 500 ml of methylene chloride, stir, decant, re-extract with methylene chloride, dry, suction, evaporate and isolate 41.5 g of the expected product.

Step B: 4-Bromo-2- (1-methylethoxyimino) -aoethyl acetic acid ethyl ester.

35 The 41.5 g of product from the previous step is placed in 190 ml of methylene chloride with traces of p-toluenesulfonic acid. A solution of 11.9 ml of bromine in 50 ml of methylene chloride is stirred and introduced over 1 hour at room temperature. Stir, add ice-cold water, decant, re-extract with ethylene chloride, wash again with ice-cold water, dry, evaporate and isolate 55 g of the expected derivative.

-29- DK 161082 B

Step C: 2- (2-Amino-4-thiazolyl) -2- (1-methylthoxymimino) -acetic acid - ethyl ester.

14 »9 g of thiourea are added in 55 ml of ethanol and 105 ml of water and a solution of 55 g of product prepared in step B in 55 ml of ethanol is added over 40 minutes. Stir for 2 hours 30 minutes at room temperature, add 220 ml of 10 2 aqueous sodium bicarbonate solution, stir, suction, rinse, dry and isolate 42.15 g of crude product which is chromatographed on silica gel eluting with ether. The fractions that are rich in the expected product are collected, evaporated, the crystals taken up in isopropyl ether, suctioned, rinsed and 10.75 g of the expected product obtained.

The product obtained has a vision configuration.

Step D: 2- (2-Tritylamino-4-thiazolyl) -2- (1-methylethoxyimino) -acetic acid phyl ester.

15 g of Step C product are applied in 20 ml of dry dimethylformamide, 40 ml of methylene chloride and 6.2 ml of triethylamine. The mixture is cooled and slowly added 13.2 g of trityl chloride, stirred for 2 hours 30 minutes, added 43 ml of 1N hydrochloric acid, stirred, decanted, washed again with 40 ml of water, extracted with methylene chloride, dried, evaporated to dryness and dried. receives 27.7 g of the expected product.

The product obtained has a vision configuration.

Step E: 2- (2-Tritylamino-4-thiazolyl) -2- (1-methylethoxyimino) -25-acetic acid.

A mixture of 27.7 g of product from step D, 150 ml of dioxane and 65 µl of 2 L sodium hydroxide solution is heated to reflux for 2 hours. The sodium salt crystallizes, it cools, sucks, rinses with a mixture of ether and dioxane (1: 1), dries and gives 16.85 g of crude sodium salt. 15.9 g of this sodium salt is dissolved in 15.9 g of dimethylformamide, 100 ml of water and ca. 500 ml of methanol. 30 ml of 2N hydrochloric acid are added, the methanol is evaporated, diluted with water, suction, rinsed, dried, the 9.8 g of viscous product is taken up in 220 ml of a mixture of methylene chloride and methanol (50:50), evaporated to dryness. , absorbs in ether, fines, sucks the crystals, rinses and dries. 4.9 g of the expected acid are obtained.

Mp. ca. 170 ° C.

An analytical sample is obtained by dissolving 300 mg of crude product in 2 ml of methylene chloride and 1 ml of methanol, diluting with water and with methylene chloride, stirring, extracting the crystals, rinsing

- 30 - DK 161082 B

with methylene chloride and with water, drying and isolating 230 mg of analytical product.

Analysis: calculated: C # 68.77 W ° 5.34 H # 8.91 6.8 Found: 68.6 5.5 8.8 6.8

The product obtained has a vision configuration.

grin ff: 3-acetoxymethyl-7- (2- (2-tritylamino-4-thiazoly) -2- (1-methyl-ethoxyimino) -acetamido) -oeph-3-em-4-carboxylic acid.

4.89 g of the acid obtained in step E is placed in 13.5 ml of dimethylformamide under argon atmosphere. After dissolving, cool in an ice bath and add 1.62 g of dicyclohexylcarbodiimide in 16 ml of methylene chloride. Dicyclohexylurea crystallizes. Stir in an ice bath, suction, rinse with methylene chloride, dry and isolate 1,424 g of dicyclohexylurea. One is cooled in a bath of methanol and ice and a solution of 1.41 g of 7-aminocephalosporanic acid in 30 ml of methylene chloride and 1.45 ml of triethylamine is added. Stir for 3 hours at room temperature, add 20 ml of 1 L hydrochloric acid, stir, decant, re-extract with methylene chloride, dry, suction and obtain 9.05 g of mixture of the expected product and the starting product.

20

It is taken up in methylene chloride, initiates crystallization, lets crystallize with stirring, sucks the crystals, rinses, dries, obtains 1.6 g of pure starting product, evaporates to dryness, absorbs the residue in isopropyl ether with vigorous stirring and isolates 4.91 g of viscous insoluble product. , which is the expected product. The product obtained has a visual configuration.

Example 13

3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2- (1-methylethoxyimino) -acetamido) -ceph-3-em-4-carboxylic acid.

4.91 g of crude product from Example 12 is placed in 30 ml of 50 $ aqueous formic acid. Stir in a water bath at 60 °, dilute with water, extract the resulting triphenylcarbinol, rinse with water, dry and isolate 1.39g of triphenylcarbinol. It evaporates to dryness, absorbs in water, fines, sucks, rinses with water, dries and 35 gets 800 mg of the expected product.

An assay is obtained by dissolving 972 mg of crude product in 4 ml of methanol, diluting with 20 ml of ether, extracting insoluble material, rinsing, drying and 404 mg of the expected pure acid. Mp. ca. 200 ° C.

'31 - DK 1610 8 2 B

Analysis: Calculated: C / ° 44.71 B # 4.38 N / o 14.48 S # 13.26 Found: 44.5 4.5 14.1 13.2

The product has syn configuration, H.M.R. spectrum (60 MHz, 5 DMSO) p.p.m,: 2.01 (CH 2 CO), doubled at 9.46, J = 8 Hz (COBH), 6.7 (proton in thiazole ring).

Example 14.

3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido) -10-ceph-3-em-4-carboxylic acid, syn is r.

Step A: 2- (2-Chloroacetamido-4-thiazolyl) -2-methoxyiminoacetic acid ethyl ester, syn isomer.

45.8 g of 2- (2-amino-4-thiazolyl) -2-methoxyimino acetic acid ethyl ester, syn isomer prepared according to step A of Example 3, is introduced into 200 ml of methylene chloride. 20 ml of it is distilled off to dry, cooled to 10 ° C and 50 ml of pyridine is added. 41 g of monochloroacetic anhydride are added and warmed slightly until dissolved. Allow to stand for 6 hours at .20 ° C under nitrogen, add 5 ml of water, stir and pour into 300 ml of ice-cold 2H hydrochloric acid. It is then decanted, extracted with methylene chloride, washed with water and with sodium bicarbonate, dried, treated with activated charcoal, evaporated and 300 ml of isopropyl ether are added. This product crystallizes. Evaporate until a thick paste is obtained, ice-cooled, suctioned, washed, dried and 45.4 g of product are obtained with m.p. 113 ° C, 25 A pure sample is obtained by recrystallizing a mixture of methylene chloride and isopropyl ether, m.p. 118 ° C.

H.M.R. spectrum (CDCl3, 60 MHz): Q

(f) H-M “i-CH₂Cl1 S ri o (a) <e) i 23 35 CH OCH₂ (b) (a) triplet centered around 1.38 p.p.m., J = 7 Hz (b) singlet 4.05 p.p.m.

(c) quadruplet centered around 4.44 p.p.m, J = 7 Hz (d) singlet 4.33 p.p.m.

(e) the singlet 7.27 p.p.m.

(f) the singlet 9.95 p.p.m.

- 32 - DK 161082 B

lane B: 2- (2-chloroethamido-4-thiazolyl) -2-methoxyiminoacetic acid. syn isomer.

46 g of the product obtained in step A above is placed in 230 ml of absolute ethanol. 5 ml of pure sodium hydroxide solution are added at 20 ° C under nitrogen. The product dissolves and the sodium salt begins to crystallize, after which the environment solidifies. After 16 hours, it sucks and dries. The resulting salt is dissolved in water, ice-cooled, 100 ml of 2 U hydrochloric acid is added, nights with sodium chloride and extracted with ethyl acetate containing 10 $ 10 ethanol. Dry, treat with activated charcoal, distill under vacuum, immerse the water with benzene, take up in methylene chloride, distill to dryness, take up in methylene chloride, ice cool, suction, wash, dry and get 34 »5 g of the expected product. Mp. about 200 ° C. The product is purified by recrystallization from a mixture of acetone and isopropyl ether.

Analysis: GgHgO2 U2 ClS = 277.68 calculated: 0 $ 34.60 H $ 2.90. 15.13 01 $ 12.77 S $ 11.55 found: 34.81 2.8 14.8 12.6 11.5 HMR spectrum (DMS0, 60 MHz): 20 Q (b) (e) HN-Å -CH 2 Cl

Sr \ \ = J C02H (c) 25 γ (d) N 'NSsOCH3 (a) (a) singlet 3.92 p.p.m.

(B) the singlet 4.38 p.p.m.

(c) the singlet approx. 5 p.p.m.

(d) the singlet 7.58 p.p.m.

(e) the singlet 12.6 p.p.m.

Step 0: 3-Acetoxymethyl-7- (2- (2-chloroacetamide o-4-thiazolyl) -2- methoxyiminoacetamido) -ceph-5-em-4-oarboxylic acid. syn isomer. 15.3 g of the product obtained in step B above is introduced into 80 ml of methylene chloride. At 5 ° C, 8 ml of triethylamine are added.

At 0 ° C, 3.8 ml of thionyl chloride and 26 ml of methylene chloride are introduced under a nitrogen atmosphere. Allow to stand for 15 minutes at 0 ° C and then 7 ml of triethylamine are added. It is introduced at 0 ° C under nitrogen.

'33 ~ DK 161082 B

gene atmosphere 13.6 g of 7-aminocephalosporanoic acid in 100 ml of methylene chloride and 14 ml of triethylamine. The temperature is allowed to rise again to 20 ° C and then stirred for 1 hour. # This solution is distilled to dryness under vacuum at 30-35 ° C. The residue is dissolved in 250 ml of water, treated with activated charcoal and 50 ml of 2 H hydrochloric acid . The precipitate is extracted and washed with water. The crude product obtained is suspended in 80 ml of water. At 5 ° C, 7 ml of triethylamine is added. 15 ml of 4 N sulfuric acid is added at one time with stirring at 5 ° C and the product crystallizes after 15 minutes. Suction is washed, washed with 10 ethanol by rubbing and then with ether, dried under vacuum to give 18.6 g of the expected product, = + 26 ° + 1 ° (1 $, dimethylformamide).

H.M.R. spectrum (DMS0, 60 MHz): Q (=> 15 H-N-C-CH-C1 (e)

/ V O

8 N

^ = h3 «(b) Y CH2 - ° fCH3 4o2h δ (a) singlet 2.03 p.p.m.

(B) the singlet 3.90 p.p.m.

(c) the singlet 4.38 p.p.m.

(d) the singlet 7.45 p.p.m.

Step D: 3-Acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido) -ceph-3-em-4-carboxylic acid. syn isomer.

30

5.32 g of acid from step C above are suspended in 10.6 ml of water and 912 mg of thiourea. At 20 ° C, 1 g of potassium bicarbonate is added. After dissolving, stir for 6 hours at ca. 20 ° 0 under nitrogen atmosphere. The rubbery precipitate begins after approx. 1 hour 30 minutes. 30 ml of water and 3 ml of formic acid are then added. It is cooled to 5 ° C, suctioned and washed with water containing 10 $ formic acid. You dissolve the residue at approx. 5 ° C in 30 ml of water containing triethylamine. At 5 ° C, 3 ml of formic acid is added, the precipitate is extracted, washed by rubbing with water containing formic acid and eliminated the dark brown rubber. The aqueous phases are combined and treated with activated charcoal. A pale yellow solution is obtained which is saturated with ammonium

- 34 - DK 161082 B

sulfate. You. sucks the precipitate out, cleans with water, sucks off, washes with water and gets a precipitate a.

The mother liquor one is saturated with ammonium sulphate, which gives a precipitate from which you suck and wash three times, giving 5 precipitate B.

The precipitates A and B are merged. It is taken up in ethanol, stirred for 1 hour at 20 ° C and left for 16 hours at 0 ° C. It is extracted, washed with ethanol and ether, dried under vacuum to give 3.47 g of the expected product in the form of the syn isomer.

10 N.M.R. spectrum (DMSO, 60 MHz): 15! <Cl h in Nm \ ksxfH (b)

(a> N I I

(a) (b) o T ch2-oc-ch3

20 COH

(a) singlet 2.03 p.p.m, (b) singlet 3.55 p.p.m, (o) doubled 5.19 p.p.m. J = 5 Hz (d) singlet 6.8 p.p.m.

This product is identical to that of Examples 4 and 6.

Example 15

Diethylamine salt of 3-acetoxymethyl-7- (2- (2-tritylamino-4-thiazolyl) -30-2-methoxyiminoaoetamido) -ceph-3-em-4-oreboxylic acid. syn isomer.

Step A: 2-Ethyl-2-methoxyiminoacetic acid ethyl ester,

4.69 kg of 2-acetyl-2-hydroxyiminoacetylacetic acid ethyl ester corresponding to 4.21 kg of pure product is introduced into 21 liters of pure anhydrous acetone. 6.1 kg of potassium carbonate is added at 20-25 ° C. The suspension is stirred for 10 minutes and then 3.72 kg of dimethyl sulfate is added at 20-25 ° C. Stir for 3 hours at 20-25 ° C. It is then poured into 126 liters of demineralised water and extracted with 4 liters of 5 liters and then 2 liters of methylene chloride. Wash with 10 liters of demineralized water. Dry, suction and rinse with 2 liters of methylene chloride. It is distilled under vacuum to obtain 4.88 kg

- 35 - DK 161082 B

of the expected product.

Rf = 0.7 by silica gel thin layer chromatography, eluent: methylene chloride and ethyl acetate (9ί1).

The product is identical to that prepared in Step α of Example 3.

Step B: 4-Bromo-2-methoxyiminoacetylacetic acid ethyl ester.

3.53 kg of product from step Z above is placed in 18.6 liters of methylene chloride and 3.5 g of p-toluenesulfonic acid. a solution of 2.96 kg of bromine in 3.5 liters of methylene chloride. Hydrogen bromide release is observed after 15 minutes of feeding. The mixture is stirred for 45 minutes at 22 ° C and then poured and washed with 2 times 14 liters of ice-cold demineralized water. The washings are re-extracted with twice 3.5 liters of methylene chloride. Dry, filter, rinse and distill under vacuum. 4.73 kg of the expected product is available.

This product is identical to that prepared in step β of Example 3.

Step 2- (2-Amino-4-thiazolyl) -2-methoxyimino®acetic acid ethyl ester, syn isomer.

1.43 kg of thiourea is introduced into 3.55 liters of ethanol and 7.1 liters of demineralized water. The mixture is stirred for 10 minutes at 20 ° C and then at 20-25 ° C 4,730 kg of the product prepared in step B above 25 is added in 3.55 liters of ethanol. Stir for 3 hours at 20-25 ° C, cool to 15-20 ° C and neutralize to a pH of 7 with approx. 1.6 liters of 22 ° B ammonia water.

Stir another 15 minutes at 20-25 ° C, suction and wash with 5 times 1.8 liters of demineralized water. 2,947 3Q kg of the expected product is available. Mp. 162 ° C.

The product is identical to that prepared in steps A and γ of Example 3.

Step D: 2- (2-Tritylamino-4-thiazolyl) -2-methoxyiminoacetic acid ethyl ester. syn isomer.

3.41 kg of the product obtained in step C above is placed in 17 liters of methylene chloride and 2,275 liters of triethylamine. The mixture is stirred for 15 minutes and 4.55 kg of trityl chloride are added under stirring and under a nitrogen atmosphere at 20-25 ° C for 1 hour. The mixture is stirred for 20 hours at 20 DEG-25 DEG C. under a nitrogen atmosphere and triethylamine hydrochloride crystallizes.

-36- DK 161082 B

It is poured and washed with 10.2 liters of 0.5 N ice holding hydrochloric acid and twice 10.2 liters of ice-cold demineralized water. The washing liquids are re-extracted in series with 1.2 liters of methylene chloride.

Dry, filter and rinse with 1.7 liters of methylene chloride. Distill to dryness under vacuum at a temperature below 50 ° C. . in

8,425 kg of crude product is obtained.

island

This product is redissolved at 20-25 G in 8.4 liters of methanol and over 1 hour at 20-25 ° C, with stirring, under the initiation of crystallization, 2.8 liters of demineralized water are added. Stir for an additional hour, suck off, spray twice with 1.7 liters of methanol with 25 $ water, dry at 40 ° C and obtain 7 * 165 kg of the expected product.

The product is identical to that of Step B of Example 3.

Step E; Sodium any one of 2- (2-tritylamino-4-thiazolyl) -2-methoxy-iminoacetamidoic acid syn isomer.

4,175 kg of product from step D above is introduced into 20.9 liters of ethanol. It is heated to reflux under stirring and under a nitrogen atmosphere. A complete solution from 55 ° C is obtained.

During reflux and under nitrogen atmosphere 5.235 liters approx. 2 N sodium hydroxide solution. Rapid crystallization occurs. The mixture is stirred for 1 hour under reflux under a nitrogen atmosphere. Bring to 20-25 C and keep at this temperature for 2 hours. It sucks, washes with 4x 2.1 liters of ethanol, dries and gets 4.02 kg of the expected product.

Step ff: 2- (2-Tritylamino-4-thiazolyl) -2-methoxyiminoacetic acid, syn isomer.

500 g of the product obtained in step E above is placed corresponding to 440 g of dry product in 2.5 liters of methylene chloride. During stirring and under nitrogen atmosphere, 2 liters are added over 2 minutes at approx. 1 N hydrochloric acid. Stir for 2 hours at 20-25 ° C under a nitrogen atmosphere. The methylene chloride phase is decanted off and washed with 3 times 2 liters of demineralised water. One is extracted in series with 1 liter of methylene chloride. Dry, add 25 g of activated charcoal, suction, rinse, distill to dryness and obtain 481 g of crude product. This amount of substance is taken up in 2.1 liters of isopropyl ether. You suck and wash with 2 times 420 ml

- 37 - DK 161082 B

iaopropylether. Dry under vacuum to constant weight to give 424.6 g of the expected product.

The product is identical to that prepared in Step C of Example 3.

5 gr Gi Diethylamine salt of 3-aethoxyinethyl-7- (2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetamido) -oeph-3-em-4-carboxylic acid. s.vn-is omer.

200 g of 2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetic acid from step P above is placed in a flask and then 1200 ml of methylene chloride. The suspension is refluxed under stirring under argon and then 600 ml of methylene chloride is distilled off at atmospheric pressure. It is brought to 18-20 ° C and 54 g of dicyclohexylcarbodiimide are then introduced into 54 ml of methylene chloride while maintaining this temperature. Stir for 1 hour at 18-20 ° C under 15 argon and then add at this temperature a solution of 61.4 g of 7-aminocephalo-sporanoic acid in 900 ml of methylene chloride and 63 ml of triethylamine over 15 minutes at this temperature. Stir for 1 hour 30 minutes at 20 ° C (pH 6.5-7) # 50 ml of acetic acid is then added, allowed to stand for 15 minutes under stirring at 20 ° C and then suctioned to eliminate it as a starting product. used 7-aminocephalosporanoic acid. Rinse with 200 ml of methylene chloride. The organic solution is washed with 400 ml of demineralized water 3 times and then dried over magnesium sulfate.

It is suctioned, rinsed with 200 ml of methylene chloride twice and distilled to dryness under vacuum and under argon. Dissolve dry oily extract at 20-25 ° with stirring and under argon in 700 ml of dioxane. 300 ml of a mixture of dioxane and methylene chloride are distilled under vacuum and under argon at a temperature below 30 ° C.

Bring to 20 ° C + 2 ° C and then add 500 ml of ether. 30 ml of diethylamine are added. After approx. Ten minutes, the diethylamine salt of 2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetic acid crystallizes. Leave for 1 hour under argon at 20 ° C.

Suction and rinse with 3x 100 ml of a solution of dioxane and ether. The resulting diethylamine salt is dried and 113.6 g are obtained. The organic solution is precipitated over 30 minutes with stirring in 3.25 liters of isopropyl ether. Allow to stand for 15 minutes under stirring and then suction under vacuum. Rinse with 400 ml of isopropyl ether twice, dry under vacuum to obtain 182 g of product identical to that obtained in Example 5.

'38 ~ DK 161082 B

Example 1- 5-Acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamide o) -ceph-5-em-4-carboxylic acid «syn isomer 18 182 g of the The product obtained above is introduced with stirring and under argon at 28-30 ° C in 347 ml of formic acid and 87 ml of demineralized water. A complete dissolution and crystallization of triphenylcarbinol occurs. Stir under argon and under argon for 2 hours 30 minutes at 28-30 ° C and then precipitate over 15 minutes with stirring in 1740 ml of demineralized water and 847 g of ammonium sulfate. Allow to stand for 30 minutes with stirring. Suction, rinse twice with 174 ml of demineralized water, dry under vacuum at 25-30 ° C and obtain 147 g of a mixture of the expected product and triphenylcarbinol. The crude product is quenched for 1 hour with 735 ml of ether at 18-20 ° C. Suction is taken off, 15 rinses with 147 ml of ether twice, dried at 25 ~ 30 ° C to obtain 89 g of the expected product.

This product is precipitated under stirring and under nitrogen atmosphere with 445 ml of ethanol. The suspension is stirred at 45-50 ° C and stirred for 1 hour under these conditions. It is then stirred for 1 hour at 18-20 ° C. It is suctioned, rinsed with 45 ml of ethanol twice, dried under vacuum at 20 ° C to give 76.85 g of the expected product.

This product is contacted with 230 ml of acetic acid.

It is stirred for 15 minutes under a nitrogen atmosphere and then 77 ml of demineralized water is added, then about 25 ml of water are added to this solution. Allow to stand for 1 hour with stirring at 18-20 ° C, then add over approx. 10 minutes 269 g of ammonium sulphate, leave for 15 minutes and then add 3.85 g of activated charcoal. Allow to stand for 15 minutes, stirring, rinsing and rinsing with 77 ml of demineralized water with 25 $ acetic acid. 30 ml is added at 18-20 ° C and with stirring 154 ml of formic acid and a crystal of the final product is added and the crystallization is then promoted by scraping. It is allowed to stand for 2 hours with stirring at 18-20 ° C and then 2 hours at 0-5 ° C. Suction and wash with 4 times 77 ml of demineralised water with 5 $ formic acid. Dry at 20-25 DEG C. under vacuum. 49.45 g of product is obtained in the form of formate.

The resulting formate is stirred for 1 hour at 45-50 ° C in 25Ο ml of ethanol and then left for 1 hour at 18-20 ° 0. Suction and rinse twice with 50 ml of ethanol. Dry at 20 DEG C. under vacuum and then 10-15 hours at 35 DEG-40 DEG C. 45.45 g of the expected product are obtained, ap = + 65.5 ° (0.5 $, water with 0.5 $ NaHCO3).

-39- DK 161082 B

The product is identical to that of Examples 4, 6 and 14.

Example 17

5 The sodium salt of 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido) -ceph-5-em-4-earboxylic acid, crystallized. ayn isomer.

Dissolve 19.8 g of 3-acetoxymethyl-7- (2- (2-amino-4- * thiazolyl) -2-methoxyiminoacetamido) -ceph-3β-4-carboxylic acid, syn-isomer, prepared according to Examples 4, 6, 14 or 15 in 65 ml of 1 M sodium acetate solution in methanol. Crystallize 35 minutes at room temperature, add 1 ml of ethanol over 1 hour, stir in ice bath for 2 hours 30 minutes, suction, wash twice with 10 ml of a mixture of methanol and 15 ethanol (1: 1), twice with 10 ml of ethanol and then twice with 20 ml of ether. After drying for 2 hours at 45 ° C under vacuum and then 48 hours in the desiccator under vacuum with sulfuric acid, 16.191 g of the crystalline product is obtained.

By avoiding any contact with the humidity of the atmosphere, you get a product whose physical constants are as follows:

HgO (Karl Fischer) = 0.2 $ methanol under 0.1 $) determination by vapor phase chromatography ethanol 0.45 $ Analysis: G- ^ gH ^ gOyN ^ SgNa = 477.5 calculated: C $ 40.24 H $ 3.38 N $ 14.67 S $ 13.43 Na $ 4.81 Found: 39.9 3.5 14.5 13.1 4.8

When left in the air, the product is rehydrated. The X-ray spectrum (Debye-Scherrer) has made it possible to confirm the crystalline nature of the obtained product.

It is also possible to use isopropanol instead of ethanol to obtain the crystallization of the expected product.

Example 18 The sodium salt of 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido) -ceph-3-em-4-carboxylic acid, crystallized, syn isomer.

Step A: Solvate between 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamide o) -ceph-3-em-4-carboxylic acid, syn-4aomer. and formic acid.

In small portions and with stirring, 87.2 g of diethyl-amine salt of 3-acetoxymethyl-7 - '(2- (2-tritylamino-4-thiazolyl) -2- | -methoxyiminoacetamido) -ceph-3 are added. em-4-carboxylic acid, prepared according to Example 5, for the ep mixture of 220 ml of pure formic acid and 220 ml of water.

Stir for 30 minutes at 50 ° C, cool and eliminate by filtration 30.1 g of triphenylcarbinol. 450 ml of water is poured into the filtrate, eliminates a weak activated charcoal precipitate and evaporates at 40 ° C under vacuum until a precipitate is formed. 200 ml of anhydrous ethanol are added, cooled with ice, filtered, washed with ethanol and with ether and dried under vacuum.

10 31 »1 g of the expected product is obtained.

Analysis; C ^ gHH j ^ I ^₂S₂, HCO₂H, H₂O = 541.5 calculated: C $ 39.3 H $ 4.08 13.48 S # 12.34 H2 O $ 3.46.

found: 39.2 4.1 13.2 12.8 4.15 p

Step B: Crystallized sodium salt.

Dissolve 15 g of freshly prepared solvate from step A in 75 ml of methanol and treat the solution with 4.5 g of potassium acetate and 3 g of activated charcoal. After filtration, 5 ml of isopropanol is added with stirring. After 16 hours at 0 ° C, the crystals are isolated, washed with ethanol and ether and dried for 2 hours under extreme vacuum at 50 ° 0.

7.95 g of the expected product is obtained.

The product then briefly stays in the open air.

You get the following

Analysis: Ο ^ Η- ^ Ν ^ Ο ^, ΙΙ ^ Ο = 495.5 Calculated: C $ 38.78 H $ 3.66 B $ 14.14 Ba $ 4.64 S $ 12.94 Found: 38 , 6 3.7 13.8 4.6 13.2

Example 19.

The sodium salt of 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2 ~ 30-methoxyiminoacetamido) -ceph-3-em-4-carboxylic acid. amorphous, syn-iso.mer. Step A: Solvate between 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamide o) -oeph-3-em-4-carboxylic acid, syn isomer, and ethanol.

Dissolve 52 g of solvate with formic acid from step Å of Example 35 18 in a mixture of 3 liters of 96 $ ethanol and 350 ml of water. Evaporate in vacuo until a volume of approx. 300 ml, The solvate begins to crystallize during evaporation. One hour is cooled in an ice bath, filtered, washed with a little ethanol and dried under vacuum at room temperature in the presence of concentrated

DK 1610 8 2 B

sulfuric acid.

44 g of the expected product is obtained.

Analyzes ^ 16 ^ 17 ^ 5 ^ 7½9 0.8 mo1 G2H5OH s 492.3 calcd: C £ 42.94 H # 4.46 N # 14.23 S # 13.02 Found: 43.0 4.4 14 , 1 12.9

Step B; Amorphous sodium salt.

Step 3 of solvate with ethanol from step A is introduced into 60 ml of water at 0 ° C and 0.504 g of sodium bicarbonate dissolved in 6 ml of water are added with stirring. The neutral solution is filtered and lyophilized immediately. The product stays in the open air for a short time.

Analysis: ^ 16 ^ 16½ ^ 8½½919 ^ ½0 = 504.47 calculated: <# 38.09 3.8 N% 13.88 found: 38.2 3.9 13.6 15

Example 20

The sodium salt of 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2-methoxyiminoaoetamido) -ceph-3-em-4-oreboxylic acid. crystallized. syn isomer.

To 4.95 g of 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2-O-methoxyiminoacetamide o) -ceph-3-em-4-carboxylic acid, syn isomer, prepared according to Example 4, 6, 14 or 16, 5 ml of ethanol is added and then with stirring in an ice bath 10 ml of a 1 M aqueous solution of sodium bicarbonate. After dissolving, 15 ml of ethanol is added, evaporated at 30 DEG C. in vacuo, taken up in ethanol and dried to constant weight. Per a powder is obtained which is taken up in 15 ml of methanol. Crystallization is initiated and left to stand overnight in the refrigerator. Per 3.407 g of crystallized product is isolated.

Example 21.

O Π

The sodium salt of 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2-methoxyiminoaoetamido) -oeph-5-em-4-oreboxylic acid. crystallized. syn isomer.

0.5 g of amorphous sodium salt of Example 19 is dissolved in 2 ml of methanol, slowly added with stirring 0.25 ml of n-butanol 35 and cooled for 48 hours in a refrigerator at ca. The crystals are washed with slightly cold methanol and dried for 3 hours under vacuum at 40 ° 0 in the presence of concentrated sulfuric acid,

0.2 g of crystallized product are obtained.

The product then stays in the open air for a short time.

- «- DK 161082 B

Analysis: ^ 6% 6 ^ 5 ^ a ^ 7S2, '*', '' H2 ^ = 504.47 calculated: C fo 38.09 Efo 3.8 Ef »13.88 O fo 26.96 found: 38, 4 3.8 13.8 .27.1

Working under analogous conditions gives rise to 5 different crystalline forms, e.g. contains 0.5 mole of water or 1 mole of water and 1 mole of methanol. j The energy spectra (Debye-Scherrer) of the j products obtained above confirm the crystalline nature of the products obtained. j 10 'i

Example 22.

3-Acetoxymethyl-7- (2- (2-triethylamino-4-thiazolyl) -2-vinyloxyimino-acetamido) -ceph-5-em-4-carboxylic acid syn isomer.

Step A: Ethyl ester of 2- (2-bromethoxyimino) -2- (2-tritylamino-4- I

15-thiazolyl) -acetic acid syn syn isomer.

Under argon, a mixture of 4.94 g of hydrochloride of the ethyl ester of 2-hydroxyimino-2- (2-tritylamino-4-thiazclyl) acetic acid, syn isomer, is added to 10 ml of dimethylformamide and introduced at room temperature. temperature over 3 minutes 4.14 g of potassium carbonate. Stirring | 20 20 minutes at 20 ° 0 and adding 8.65 ml of 1,2-dibromoethane. Stir for 30 hours and pour into an environment comprising ICO ml of distilled water and 20 ml of methylene chloride, then decant, re-extract with methylene chloride, wash. with distilled water, re-extract, dry the organic solutions, suction, rinse and distill to dryness. The crude product is obtained which is chromatographed on silica gel eluting with benzene with 5 $ ether. A first fraction is collected which is recrystallized from methanol after dissolution at 50-60 ° 0 and suction at 0-5 ° C. 1.16 g of cream-white product are obtained, mp 117 ° 0.30 A homogeneous fraction of 1.258 g is then obtained.

H.M.R. spectrum, ppm (CDCl3):

Triplet = 3.55 J = 7 Hz CH2Br triplet = 4.51 J = 6 Hz H-O-CH2 singlet 6.55, proton in thiazole ring.

Step B; Ethyl ester of 2- (2-tritylamino-4-thiazolyl) -2-vinyloxyimino acetic acid, syn isomer.

6.9 g of ethyl ester of 2- (bromethoxyimino) -2- (2-tritylamino-4-thiazolyl) acetic acid, syn isomer, are prepared in step A in 35 ml of dimethyl sulfoxide, then introduced at 20-25 ° C

DK 161082 B

over 5 minutes 16 ml of 1 M solution of potassium tert.-butylate in tetrahydrofuran. Stir for 30 minutes and 3 ml of 1 M potassium tert-butylate solution in tetrahydrofuran are stirred at room temperature. Stir again for 30 minutes, again introduce 3 ml of the same potassium-5-tert-butylate solution and after 30 minutes a third time 3 ml of the solution.

After the last introduction, stir for 30 minutes and pour into a mixture of 350 ml of water and ice and 90 ml of methylene chloride.

Acidify to a pH of 2 with 28 ml of 1 l of hydrochloric acid, decant and re-extract with 3 times 50 ml of methylene chloride.

Wash with distilled water, dry, suction, rinse and distill to dryness to obtain a black resin which is purified by chromatography on silica gel column eluting with methylene chloride. Thus, there are obtained 3.227 g of the expected product, which is immediately used in the next step.

I.M.R, Spectrum, ppm (CDCl3):

Solid from 4.16 to 4.83 ppm = CHg solid from 6.66 to 7.33 ppm = CH singlet at 6.65, proton in thiazole ring.

After recrystallization from absolute ethanol, a product is obtained, mp 114 ° 0,

Analysis: ^ 28 ^ 25 ^ 3 ^ 3 ^ Calculated: C1 * 69.54 H # 5.21 W ° 8.69 S # 6.36 Found: 69.2 5.2 8.3 6.6 2 5

Step G: 2- (2-Tritylamino-4-thia2; olyl) -2-vinyloxyiminoacetic acid. syn isomer.

3.227 g of ethyl ester of 2- (2-tritylamino-4-thiazolyl) -2-vinyloxyimino acetic acid, syn isomer, are prepared in step B in 3.2 ml of dioxane. At room temperature, 20 ml of 30 M of potassium hydroxide in absolute ethanol and 12 ml of absolute ethanol are added. It is heated to 50 ° C with stirring and argon atmosphere and maintained for 1 hour. The potassium salt crystallizes out. It is brought to 20 ° C, suctioned, rinsed with ethanol and taken up in a mixture of 50 ml of ethyl acetate and 50 ml of distilled water.

35

Acidify with 6 ml of 2 L hydrochloric acid and stir until two homogeneous phases are obtained. Decant, wash with 3 x 20 ml distilled water, re-extract with ethyl acetate, dry, suction and rinse with ethyl acetate. Evaporate to 15 ml and ice-cool at 0-5 ° C. You suck and rinse with ethyl acetate. Dry and give 2.53 g of the expected product. The product is used

'44' DK 161082 B

immediately in the next step.

After recrystallization, a purified sample is obtained with m.p.

150 ° C.

Step D: 3-Acetoxymethyl-7- (2- (2-tritylamino-4-thiazolyl) -2-vinyl-5- (oxyiminoacetamido) -ceph-3-em-4-carboxylic acid, syn isomer.

2.53 g of 2- (2-tritylamino-4-thiazolyl) -2-vinyl oxyimino acetic acid, syn isomer, are prepared in step C in 20 ml of methylene chloride. It is cooled to 5 ° C and 0.632 g of di-1 are introduced at one time eyclohexylcarbodiimid. You let recharge room temperature. Stir under argon for 1 hour to obtain suspension A. Separately, 0.718 g of 7-aminocephalosporanic acid is introduced into 10 ml of methylene chloride under argon atmosphere. It is cooled to 10 ° C and 1.1 ml of triethylamine is added to obtain solution B.

Suspension A is introduced over 10 minutes at 5-10 ° C

15 in solution B. Stir, allowing to re-enter room temperature (over 35 minutes). Then stir for 3 hours at 20-25 ° C.

0.6 ml of acetic acid is then added, stirred for 10 minutes and then the dicyclohexylurea formed is suctioned off. The organic solution is washed with water, dried and distilled under vacuum.

A yellow resin is obtained, which is taken up in ethyl acetate, filtering out some insoluble material. Evaporates to dryness to obtain 3.475 g of the expected product.

The hydrochloride of the ethyl ester of 2-hydroxyimino-2- (2-triethylamino-4-thiazolyl) acetic acid, syn isomer used as the starting product of Examples 1-Y, is prepared as follows: 2- (2-amino-4 -thiazolyl) -2-hydroxyiminoacetic acid ethyl ester, syn-isomer.

0.8 g of thiourea is dissolved in 2.4 ml of ethanol and 4.8 ml of water. The solution of 2 g of 4-chloro-2-hydroxyiminoacetylacetic acid ethyl ester is added over 5 minutes and stirred for 1 hour at room temperature. Most of the ethanol is evaporated under partial vacuum and neutralized to a pH of 6, adding solid sodium bicarbonate. One is cooled, suctioned, washed with water and dried under vacuum at 40 ° C. 1.32 g of the expected product, m.p. 232 ° C.

Analysis: Calculated: C $ 6 39.06 W ° 4.21 Wfo 19.52 S% 14.9 Found: 38.9 4.4 19.7 14.6

DK 161082 B

Hydrochloric acid of 2- (2-trit, ylamino-4-thiazolyl) - 2-h, hydroxyiminoacetic acid ethyl ester, syn isomer.

43 * 2 g of 2- (2-amino-4-thiazolyl) -2-hydroxyimino-acetic acid ethyl ester, syn isomer, prepared above in 120 ml of dry dimethylformamide are introduced.

It is cooled to -35 ° C, introduced with 32 ml of triethylamine and then 60 g of trityl chloride in portions over 30 minutes. The temperature is raised, a complete solution is observed and then a heating to 30 ° C. After 1 hour, pour 10 into 1.2 liters of ice-cold water containing 40 ml of 22 ° Be hydrochloric acid.

Stir in an ice-water bath, suction, rinse with 1N hydrochloric acid and elute with ether to obtain 69.3 g of hydrochloride.

Example 23

The sodium salt of 3-aethoxymethyl-7- (2- (2-tritlamino-4-thiazolyl) -2-vinyloxyiminoaoetamido) -ceph-3-em-4-carboxylic acid, syn isomer .

At room temperature 2.628 g of product prepared in Example 22 are dissolved in the form of a resin in 5.2 ml of acetone. 10.4 ml of 1 M solution of sodium bicarbonate in water are added at once.

The sodium salt is suctioned off after 1 hour of contact at 20 ° 0 and rinsed twice with 2 ml of a mixture of water and acetone (1: 1) and with 2 ml of distilled water. Dry under vacuum at 40 ° C to give 1.612 g of the expected crude product.

Example 24.

3-acetoxymethyl-7- (2- (2-amlno-4-thiazolyl) -2-vinylox.viminoacetamido) - ceph-3-em-4-oarboxylsyre. syn isomer.

1.612 g of sodium salt of 3-acetoxymethyl ~ 7- (2- (2-tritylamino-4-thiazolyl) -2-vinyloxyiminoacetamido) -oeph-3-30-em-4-carboxylic acid, syn isomer, are prepared in Example 1. 23 in 4 ml 98 $ formic acid. Immediately after 4 ml of distilled water is added.

The mixture is stirred at 40 ° C for 15 minutes. The heterogeneous suspension is ground for 30 minutes at room temperature. Extract at 20 ° C and rinse twice with 1.6 ml of 50 $ formic acid. It is distilled to 35 dryness and taken up in 5 ml of distilled water. It is extracted at room temperature, rinsed with distilled water and then with ether. Dry and obtain 300 mg of the expected product. The insoluble material is separated in the same way as above using 50 $ formic acid. Thus, 0.214 g of additional acid is obtained.

DK 161082

- 4 «- I

_ - i * |

Analysis: C17% 7®7 ^ 5 ^ j Calculated: Ofo 43 * 68 3 * 66 W $> 14 * 98 13 * 72 Found: 44 * 2 3.9 15.1 13.0 j EMR Spectrum, ppm ( DMSO): δ 6.91 (proton in thiazole ring) 7.28 (free amine IH2)

Example 25

The Hatrium salt of 5-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2-vinyl-oxyiminoacetamido) -ceph-3-em-4-carboxylic acid.

To 5.08 g of 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2-vinyloxyiminoacetamido) -ceph-3-em-4-carboxylic acid, syn isomer, prepared by example are added. ml of ethanol and then stirred in ice-water bath 10 ml of 1 L aqueous solution of sodium bicarbonate. After dissolving, 15 ml of ethanol * is then evaporated to 30 ° C under vacuum, taken up in ethanol and dried at constant weight «The expected product is obtained, 20

Example 26

An injection preparation according to the recipe is prepared: - 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) - 2-hydroxyiminoacetamido) -ceph-3-em-4-carboxylic acid 500 mg - sterile aqueous additive in 5 ml

Example 27

An injection preparation according to the prescription is prepared: 5 ml

Example 28.

An injection preparation according to the recipe is prepared: - The sodium salt of 3-acetoxymethyl-7 - (. 2- (2- -amino-4-thiazolyl) -2-methoxyiminoacetamide o) mg - sterile aqueous additive in 5 ml

. 47. DK 161082 B

Example 29.

An injection preparation according to the recipe was prepared: - 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) - 2- (1-methylethoxyimino) -acetamido) -ceph-3- 5 -era-4-carboxylic acid 500 mg - sterile aqueous additive in 5 ml

Example 30.

Gelatin capsules are prepared according to the recipe: 10-3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2- -methoxyiminoacetamido) -ceph-3-em-4-carboxylic acid 250 mg - additive to form 1 400 mg gelatin capsule

Example 31.

Gelatin capsules are prepared according to the prescription: forming 1 400mg gelatin capsule 20

Example 32.

Gelatin capsules are prepared according to the recipe: - 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2- (1-methylethoxyimino) -acetamide o) -ceph-3-em-4- 25-carboxylic acid 250 mg - additive to form 1 400 mg gelatin capsule

Example 33

30 -K-—

An injection preparation according to the prescription is prepared: ad 5 ml 35

Example 34.

Gelatin capsules are prepared following the prescription: - 3-acetoxymethyl-7- (2- (2-amino-4-thiazolyl) -2- -vinyloxyiminoacetamido) -ceph-3-em-4-carboxylic acid, syn isomer 250 mg - additive for forming a gelatin capsule of 400 mg

DK 161082B

Pharmacological study.

In vitro activity.

Method with dilutions in orphan chemistry.

You make a series of glasses in which you distribute the same amount of sterile nutrient environment. In each glass, growing amounts of the product to be examined are distributed, and then each glass is seeded with a bacterial strain.

After incubation for 24 or 48 hours in a heating cabinet at 37 ° C, growth inhibition is assessed by illumination, which makes it possible to determine the minimum inhibitory concentrations of MIC expressed in μg / ml.

Results achieved, 15 20 25 30 35 - 49 -

DK 161082 B

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'52' DK 161082 B

Product of Example 9 Stems ME in itg / ml ____ 24 h 48 h 5 Staphylococcus aureus ATCC + 638 penicillin sensitive 1 2

Staphylococcus aureus UC 1 128 penicillin resistant 2 2

Staphylococcus aureus exp, No. 54 146. 2 2 10 Staphylococcus aureus Co 15 cephalexin resistant 10 20

Streptococcus pyogenes A 561 0.02 0.02

Streptococcus faecalis 5 432 2 10 15 Bacillus subtilis ATCC 6 633 1. 1

Escherichia coli tetracycline sensitive ATCC 9 637 1 1

Escherichia coli tetracycline resistant ATCC 11 303 0.1 0.1 20

Escherichia coli exp. TOggBg 1 1

Escherichia coli gentamicinresis ^ 6 ^ tobramycin B. 55 123 D 11

Klebsiella pneumoniae exp. 52 145 0.05 0.1 25 Klebsiella pneumoniae 2,536 gentamicin resistant 2 5

Proteus mirabilis (indole) A 235 0.2 0.2 30 35

-53- DK 161082 B

Product of Example 9 Strains MIO in µg / ml 24 h 48 h 5 Proteus vulgaris (indole +) A 232 2 10

Salmonella typhimurium 420 1 10

Providencia Du 48 35 10

Pseudomonas 3 935 exp. SG 20 20

Serratia gentamicin resistant 2,532 0.4 1 15 20 25 30 35

. 54. DK 161082 B

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- 58 - DK 161082 B

Experimental infection with Escherichia coli (T) ° 26B6 * A) The effect of the product of Examples j 4, 6, 14 and 16 on an experimental infection with Escherichia coli j is investigated in mice. Teams of 10 male mice with an average weight of 5 21.5 g were infected by intraperitoneal injection of 0.5 ml of one 24 hour old culture in nutrient broth of the Escherichia coli strain (!)

Dilute OggBg from the Pasteur Unit to 1/6 with distilled water.

A certain amount of the product is administered by subcutaneous injection or orally 1 hour, 5 hours and 24 hours after injection.

Mortality is recorded over 8 days.

Achieved results: . Mortality after Survivors. 15 1) 08: 18_ 21 h 50 28h 50 52 h mus the 8 «da

Control 9 1 0/10 0.05 mg subcutaneous 10/10 0.1 mg subcutaneous 10/10 0.25 mg subcutaneous 10/10 20 o, l mg orally 1 9/10 0.25 mg orally 10/10 0, 5 mg orally 10/10 B) Working under similar conditions and infecting mice with an average weight of 22.5 g by intraperitoneal injection of a 24 hour old culture of the strain Escherichia coli (T) ° 26B6 diluted to 1/5, the following is obtained: Results with the product of Example 4. 6 «14 and 16; 30 35

- 59 - DK 161082 B

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Experimental infection with Salmonella typhimurium.

The effect of the product of Example 7 on experimental infection with Salmonella typhimurium on mice

Holds of 10 mice infected with an average weight of j 5 20.5 g by intraperitoneal injection of 0.5 ml of a 24 hour j in old culture in oxoid broth of the strain Salmonella typhimurium 52K - diluted to 1/75 with distilled water *

A specific amount of the product is administered by subcutaneous injection at 4 hours, 8 hours, 24 hours and 32 hours after injection. 10 Results achieved; 15 i

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. 61 _ DK 161082 B

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- 62 - DK 161082 B

Experimental infection with Proteus mirabilis.

The effect of Example 31 products on an experimental infection with Proteus mirabilis in mice is investigated.

Teams of 10 mice with an average weight of 22 g 5 were infected by intraperitoneal injection of 0.5 ml of a 24 h old culture culture in oxoid broth of the strain Proteus mirabilis A.23? Diluted 1 to 1/4. .

A certain amount of the product is administered 1 hour, 5 hours and 24 hours after injection. j 10 ^ 1

Results obtained: j i i i i i i 15 20 j 25 30 35

'63 -. DK 161082 B

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- 54 "DK 161082 B

Study of acute toxicity in mice by intravenous route.

The acute toxicity of the product of Example 7 "is determined on female mice of 19-21 g, Swiss CDI Specific Pathogen Free (breeding CH. Hiver - France), in holdings of 8-10 animals per dose of the compound"

The animals are kept in a protected cage (temperature 21 ° C + 1 ° C, constant humidity ,, air pressure room) during the 4 days of the observation period with free access to food and drinking water.

The product of Example 7 is dissolved at a concentration of 10 100 mg / m-1 in apyrogenic sterile distilled water. It is injected into a tail vein in a variable volume at a rate of 1 ml per ml. minute.

Dose mg / kg Deadly on day # Total mortality _ 1 2 5 4 _ 15 500 0/10 1000 0/10 2000 0/8

The lethal dose is over 2000mg / kg.

Symptomatology.

20 No poisoning symptom is observed during either the injection or the observation period.

25 30 35

Claims (13)

1. Analogous Process for Preparation of Syn-Isomeric Cephalosporanoic Acid Derivatives of Formula I NH A. 10 'O co2a where R' represents a hydrogen atom or a saturated or unsaturated alkyl group of 1-4 carbon atoms, and A represents either a hydrogen atom or an alkali metal, alkaline earth metal, magnesium or a protonated organic amine, characterized in that a) 7-Amino-Cephalosporanoic Acid of Formula Ν-, Ν S 20 Ύ-Ί ^ y ^ - * ^^ ai2-oc-cH3 'o co2h is reacted with an acid of formula II NH-R, XS »(II) 30' - = Li c / CO 2 H II N OR '1 syn isomer 35 66. DK 161082 B j or a functional derivative of this acid in which | represents a group which can be readily eliminated by acidic hydrolysis or by hydrogenolysis, and R 'represents a | -L in a group which can be easily eliminated by acid hydrolysis or by hydrogenolysis, or a saturated or unsaturated alkyl group of 1-4 carbon atoms, to produce a product of the formula la jj NHR1] 10 1 SN \ - / s (la) ! CC-NH --- ^^ J LJ 1 I - n \ ^^ ch, -oc-ch OR, 1 T 2 o CO-H 15 syn isomers where R 2 and R 4 represent the same as above and the compound of formula Ia is optionally hydrolyzed in an acidic environment or hydrogenolysed to produce a compound of formula Ib NH_ 2S X Δ SN \ - / (Ib ·) ss-NH-i-τ'S r or 'υ γ ch oc-ch3 30 c ° 2 "° syn isomer wherein R 'represents a hydrogen atom or a saturated or unsaturated alkyl group of 1-4 carbon atoms corresponding to a compound of formula I wherein A represents a hydrogen atom and the compound having the formula Ib is converted 67. DK 161082 B desired for salt, or b) for the preparation of compounds of formula Ib NH, 5 1 S H O \ - (Ib) T .s? η-r ^ OR 'λ-Nv. 10 syn-salts and salts thereof, in which Formula Ib R 'represents a hydrogen atom or a saturated or unsaturated alkyl group of 1-4 carbon atoms, which Formula Ib corresponds to a compound of Formula I wherein A represents a hydrogen atom, either a salt of a compound of formula Ia is treated, wherein R R represents a group which can be easily eliminated by acid hydrolysis and R 'represents a group which can be easily eliminated by acid hydrolysis. or a saturated or unsaturated alkyl group of 1-4 carbon atoms, with an acid to prepare a compound of formula Ib, or also a salt of a compound of formula Ia, wherein 25 represents a group which can be readily eliminated by hydrogenolysis, and R * represents a group which can be readily eliminated by acid hydrolysis, or a saturated or unsaturated alkyl group of 1-4 carbon atoms, with a hydrogenolysing agent to prepare a salt of a compound of formula Ib. 30 Analogous process according to claim 1 characterized in that 3-acetoxy-methyl-7- (2- (2-amino-4-thiazolyl) -2-methoxy-iminoacetamido) -ceph-3-em-4-carboxylic acid is prepared, syn isomer. 35 An analogous process according to claim 1, characterized in that the sodium salt of 3-acetoxy 68. B-methyl-7- (2- (2-amino-4-thiazolyl) -2-methoxy-imino-acetamido) -ceph-3-em-4-carboxylic acid, syn-isomer. 4. An analogous process according to claim 1, characterized in that the crystalline sodium salt of 3-acetoxy-methyl-7- (2- (2-amino-4-thiazolyl) -2-methoxy-amino) is prepared. -acetamido) -ceph-3-em-4-carboxylic acid, syn-isomer. Analogous process according to claim 1, characterized in that the crystalline sodium salt of 3-acetoxy-methyl-7- (2- (2-amino-4-thiazolyl) -2-methoxy-imino-acetamido) -ceph is prepared. -3-em-4-carboxylic acid, syn-isomer, in hydrated or solvated form. 15 Analogous process according to claim 1, characterized in that compounds of the general formula I according to claim 1 are prepared, wherein A represents a hydrogen atom, a sodium atom or a protonated diethylamine base. 20 An analogous process according to claim 1, characterized in that 3-acetoxy-methyl-7- (2- (2-amino-4-thiazolyl) -2-ethoxy-imino-acetamido) -ceph-3-em. 4-. . -carboxylic acid, syn isomers, and salts thereof with alkali, alkaline earth metals, magnesium or protonated organic amines. An analogous method according to claim 1, characterized in that one. prepares 3-acetoxy-methyl-7- (2- (2- 30-amino-4-thiazolyl) -2 - ((2-propenyl) -oxymino) -acetamido) -ceph-3-em-4-carboxylic acid, syn isomers, and salts thereof with alkali, alkaline earth metals, magnesium or protonated organic amines. Analogous process according to claim 1, characterized in that 3-acetoxy-methyl-7- (2- (2-amino-4-thiazolyl) -2 - ((1-methyl-ethoxy) -imino) -acetamido is prepared ) - 69. B-ceph-3-em-4-carboxylic acid, syn-isomer, and the salts of this compound with alkali, alkaline earth metals, magnesium or a protonated organic amine. 5 10 15 20 25 30 35