DE2760287C2 - - Google Patents

Description

The invention relates to the production of aminothiazolyl-syn-oxyiminoacetic acid derivatives and your Use for the preparation of corresponding 7-aminothiazolyl syn-oxyiminoacetamidocephalosporanic acids.

The process according to the invention relates to the preparation of aminothiazolyl-syn-oxyiminoacetic acid derivatives of the general formula I,

in which mean:

R₂H or a protective group which can be split off by acid hydrolysis or hydrogenolysis, R₁H, a protective group which can be split off by acid hydrolysis or hydrogenolysis or unsaturated C _2-4 -alkyl,
where R₁ is not H if R₂ is a protective group which can be split off by acid hydrolysis or hydrogenolysis,
and AH or C _1-4 alkyl.

From these compounds of formula I z. B. corresponding 7-aminothiazolyl-syn-oxyiminoacetamidocephalosporanic acids the general formula

wherein

R₁und R₂ the same as above
and AH, an alkali metal atom or an equivalent of an alkaline earth metal including Mg or an ammonium group derived from an organic amine base

mean, accessible, as explained in more detail in the experimental section is.

By acid hydrolysis or hydrogenolysis Removable protecting groups R₁ and R₂ are for example Benzyl, formyl, trichloroethoxycarbonyl and 2-tetrahydropyranyl and especially t-butoxycarbonyl, Trityl, benzhydryl, trichlorethyl and carbobenzyloxy.  

R₁ also means z. B. vinyl, propenyl, butenyl, Ethynyl and propargyl.

Compounds of general formula I are special the compounds described in the examples and especially 2-amino-4-thiazolyl-syn- 2-propenyloxyiminoacetic acid.

The compounds used according to the invention can be present either in the structure described by formula I or in a structure according to formula I _Z :

in the R₁, R₂ and A the above meaning have.

The inventive method for Preparation of the compounds of general formula I. is characterized by Reaction of a compound of formula II,

in which mean:  

XCl or Br, R₁H or a protective group which can be split off by acid hydrolysis or hydrogenolysis or unsaturated C _2-4 -alkyl,
where R₁ is H when X is Br,
and alk = C _1-4 alkyl,

with thiourea in an aqueous solvent and / or at room temperature in the presence of an approximately stoichiometric amount of thiourea and for a reaction time of a few hours and
Treatment with a base to give the corresponding syn compound of the formula III

with R₁ and alk as above and if necessary Implementation of the compound of formula III with a functional derivative can be split off by acid hydrolysis or hydrogenolysis Protecting group R'₂ for syn compound of formula IV  

with R₁, R'₂ and alk as above as well as if necessary Implementation of the compound of formula IV by treatment with a base and then with an acid to the compound of formula I.

When converting compound III to compound IV is preferably R'₂ chloroacetyl used.

For the functional derivative of the chloroacetyl group it is preferably chloroacetic anhydride or a halide, such as Monochloroacetyl chloride.

If the reaction with a chloroacetyl halide is carried out, it is preferred to work in the presence one of the basic specified below Medium.

In a preferred embodiment of the method is used as the base for the preparation of the compound Formula III uses potassium acetate, however also carbonates and bicarbonates of alkali metals, dilute sodium hydroxide or potassium hydroxide can be used.  

The functional derivative of acid Hydrolysis or hydrogenolysis removable protective group is preferably trityl chloride, which in Presence of triethylamine or other tertiary Nitrogen bases, like other trialkylamines, N-methylmorpholine or pyridine is used.

Other functional derivatives are also available usable with groups that are easily acidic Hydrolysis or can be split off by hydrogenolysis are. Such derivatives are e.g. B. t-butyl chloroformate, made in situ or t-butylazidoformate, Trichloroethyl chloroformate or benzyl chloroformate, the mixed anhydride from formic acid and acetic acid made in situ Benzyl or benzhydryl chloride or other benzyl or benzhydryl halides, phthalic anhydride or about N-carbethoxyphthalimide.

For saponification of the compound of formula IV sodium hydroxide is preferably used, however are other bases, such as Potassium hydroxide or barium oxide can be used.

To isolate the acid of formula I (A = H) preferably dilute hydrochloric acid is used, however, acetic acid or formic acid can also be used be used.

Acidic hydrolysis agents with which the Compounds of formula I optionally implemented are formic acid, trifluoroacetic acid and acetic acid. These acids can be anhydrous  or used in aqueous solution. As This is particularly suitable for hydrolysis agents Zinc / acetic acid system.

To split off t-butoxycarbonyl or Trityl groups are preferably used acidic hydrolysis agent, such as anhydrous trifluoroacetic acid, aqueous formic acid or acetic acid.

The zinc / acetic acid system is preferred for Cleavage of the trichloroethyl group, the catalytic Hydrogenation to split off benzyl, benzhydryl and carbobenzyloxy groups applied.

As a means for hydrogenolysis z. B. the catalytic Hydrogenation used.

The compounds of formula II are accessible by reacting a compound of formula VI

with alk as above with an alkylating agent to a compound of formula VII

with R₁ as above  and reacting this compound with a brominating agent to a compound of formula VIII

with R₁ and alk as above.

That for the transfer of the compounds of formula VI alkylating agents used in compounds of formula VII is preferably an alkyl halide, such as. B. a Alkyl chloride, bromide or iodide, or a C₁- bis C₄ alkyl sulfate.

As a brominating agent for transferring the compounds of formula VII in compounds of formula VIII bromine is preferably used.

The preparation of compounds of the formula IV,

in which mean:

R'₂eine protective group which can be split off by acid hydrolysis or hydrogenolysis, R₁H, a protective group which can be split off by acid hydrolysis or hydrogenolysis or unsaturated C _2-4 -alkyl
and alk = C _1-4 alkyl,

can in a manner known per se Implementation of a compound of formula III ′

with alk as above with 1 equivalent of a functional derivative one can be split off by acid hydrolysis or hydrogenolysis Protecting group R'₂ to a compound of Formula V

with R'₂ and alk as above and Reaction of this compound with an alkylating agent made to the corresponding compound of formula IV will.  

As a functional derivative one by acid Hydrolysis or hydrogenolysis easily split off Protecting group, trityl chloride is preferably used. Then in the presence of a base, preferably Triethylamine. However, there are other bases can also be used, for example others Trialkylamines, methylmorpholine or pyridine.

Other functional ones can also be used Derivatives of acidic hydrolysis or hydrogenolysis easily removable protective group, such as t-butyl chloroformate or azidoformate, trichloroethyl or dibenzyl chloroformate, mixed in situ Anhydride from formic acid and acetic acid, benzyl or dibenzyl chloride or another benzyl or Dibenzyl halide, phthalic anhydride or N-carbethoxyphthalimide can be used.

As an alkylating agent for the production of Compounds of formula IV is preferably a Alkyl halide, e.g. B. an alkyl iodide or an alkyl sulfate, used.

The invention is also the Use of aminothiazolyl-syn-oxyiminoacetic acid derivatives Formula I,

in which mean:

R₂H or a protective group which can be split off by acid hydrolysis or hydrogenolysis, R₁unsaturated C _2-4 -alkyl
and AH or C _1-4 alkyl,

for the preparation of corresponding 7-aminothiazolyl-syn-oxyiminoacetamidocephalosporanic acids.

For the preparation of corresponding 7-aminothiazolyl syn-oxyiminoacetamidocephalosporanic acids are the invention usable intermediates of formula I. with protective groups, if necessary after transfer into a functional derivative on the carboxyl group, e.g. B. with 3-acetoxymethyl-7-aminocephalosporanic acid of the formula  

implemented.

Preferred functional derivatives of the intermediates of the formula I are e.g. B. the anhydride and the acid chloride, with the anhydride passing through in situ Exposure to isobutyl chloroformate or dicyclohexylcarbodiimide to be produced on the acid can; other acid halides can also be used and other anhydrides used in situ by the action of other alkyl chloroformates, dialkyl carbodiimides or other dicycloalkylcarbodiimides can be produced. There are also others Acid derivatives can be used, for example the acid azide, activated acid amides or activated esters, the z. B. with hydroxysuccinimide, p-nitrophenol or 2,4-Dinitrophenol are accessible. If the implementation 3-acetoxymethyl-7-aminocephalosporanic acid with an acid halide of the formula I or with an anhydride generated with isobutyl chloroformate is carried out, one preferably works in the presence a basic agent.

Alkali metal carbonates, for example, can be used as basic agents or tertiary organic bases are used are, for example N-methylmorpholine, Pyridine and trialkylamines such as triethylamine.  

It was found that the syn configuration the accessible from the intermediates of formula I. End products on the configuration of the connections Formula III goes back because of the configuration of the connections resulting from the connections of formula III can be obtained in the course of production preserved.

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

It was thus found that the syn isomers were obtained when the implementation of the thiourea with the compounds of formula II in an aqueous Solvents such as aqueous acetone or aqueous Ethanol, or carried out at room temperature being an approximately stoichiometric amount Thiourea for a relatively short time from about 1 to 3 h, or if all both of the above measures are applied at the same time.

The compounds obtained in the examples Formula III have syn configuration.

The from the compounds of general formula I have available end products with syn structure particularly high antibiotic effectiveness both against gram positive bacteria such as staphylococci and Streptococci and especially penicillin resistant Staphylococci, as well as against gram-negative Bacteria, especially coli-like bacteria, Klebsiella, Salmonella and Proteus, and are suitable  therefore particularly useful as pharmaceutically Active substances for the treatment of corresponding infectious diseases.

The compounds of formula II, which are not yet known are from 4-chloro-2-hydroxyiminoacetoacetic acid ethyl ester (described in J. of Medicinal Chemistry 1973, Volume 16, No. 9) as explained below.

The compounds in which R₁ is an acid Hydrolysis or hydrogenolysis removable protective group are accessible in a manner known per se, being functional derivatives of these groups be used.

The compounds in which R₁ represents unsaturated C _2-4 alkyl are prepared by reacting ethyl 4-chloro-2-hydroxyiminoacetoacetate with corresponding alkyl halides or sulfates.

In addition to the compounds described in the examples, e.g. B. the following compounds are accessible from the intermediates used according to the invention:
3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2- (syn-2-butenyloxyimino) -acetamido] -ceph- (3) -em-4-carboxylic acid,
3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2- (syn-3-butenyloxyimino) acetamido] -ceph- (3) -em-4-carboxylic acid,
3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2- (syn-2-butinyl-oxyimino) -acetamido] -ceph- (3) -em-4-carboxylic acid,
3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2- (syn-3-butynyloxyimino) -acetamido] -ceph- (3) -em-4-carboxylic acid,
3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2- (syn-vinyloxyimino) acetamido] -ceph- (3) -em-4-carboxylic acid,
3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2- (syn-1-propenyl-oxyimino) -acetamido] -ceph- (3) -em-4-carboxylic acid,
3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2- (syn-1-propynyloxyimino) acetamido] -ceph- (3) -em-4-carboxylic acid,
3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2- (syn-1-butenyloxyimino) -acetamido] -ceph- (3) -em-4-carboxylic acid,
3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2- (syn-2-propynyloxyimino) acetamido] -ceph- (3) -em-4-carboxylic acid and
3-Acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2- (syn-1-butynyloxyimino) acetamido] -ceph- (3) -em-4-carboxylic acid.

The following examples illustrate manufacturing and Further processing of compounds which can be used according to the invention.

Example 1 3-acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl) -2-syn- trityloxyiminoacetamido] -ceph- (3) -em-4-carboxylic acid Step A: Ethyl 2- (2-amino-4-thiazolyl) -2-syn-hydroxyiminoacetate

2 g of ethyl 4-chloro-2-hydroxyiminoacetoacetate in 5 ml of ethanol and 0.76 g of thiourea brought together and at room temperature overall Stirred for 16 h. After the crystallization of the Hydrochloride was diluted with 5 ml ether, suction filtered and with ethanol / ether (1: 1) and then with Ether washed; 1.55 g of hydrochloride were obtained.

The 1.55 g of hydrochloride obtained were at 40 to 50 ° C dissolved in 8 ml of water; after that by adding sodium acetate to a pH neutralized from 5 to 6. The free amine crystallized out. After cooling with ice, suction, washing with water and drying were initially 1.22 g anti-isomer obtained; F. 154 ° C.

The mother liquors and washing liquids of the various Experiments were combined and evaporated. After resuming in water, washed with ether, mixed with sodium hydrogen carbonate, suction filtered and washed with water; 1.9 g fell of a product that  Thin layer chromatography revealed two spots. It was chromatographically on silicon dioxide with elution Ether cleaned. The cleaned fractions of the syn- Isomers were evaporated, stirred with ether, centrifuged and dried; 50 mg of syn isomer were obtained.

Step B: 2- (2-tritylamino-4-thiazolyl) -2-syn-trityloxyiminoacetic acid ethyl ester

To a solution of 5.4 g of that prepared in stage A. Product in 54 ml chloroform and 7.5 ml triethylamine was a solution of 15 g of trityl chloride in at 10 ° C. Given 30 ml of chloroform. After standing for 1 h with 40 ml of water and then with 20 ml of water containing 4 ml 1N hydrochloric acid contained, washed, decanted, dried and evaporated to dryness.

The residue was taken up with 10 ml ether, mixed with 50 ml of methanol, stirred, suction filtered and washed with methanol; two factions fell from a total of 14.2 g of the expected product.

Step C: 2- (2-tritylamino-4-thiazolyl) -2-syn-trityloxyiminoacetic acid

10.5 g of the ester obtained in Step B were approached suspended in reflux in 55 ml of dioxane, then slowly 17 ml of 2 N NaOH solution were added. After maintaining a slight reflux and cooling the salt is centrifuged off and mixed with 60 ml of methylene chloride, Resume 20 ml of water and 2 ml of acetic acid. The Acid was suctioned off and washed with water; be the first  Fraction was obtained 7 g of acid.

After evaporation of the dioxane from the mother liquor were again 20 ml of methylene chloride, 10 ml of water and 1 ml of acetic acid added, followed by a second fraction 1.5 g of the same product was isolated. A total of 8.5 g of product were obtained.

Analysis C₄₃H₃₃O₃N₃S0.5 H₂O:
Calculated C 75.85, H 5.03, N 6.17, S 4.7%. Found C 75.8, H 4.9, N 5.9, S 4.6%.

Step D: 3-acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl) - 2-syn-trityloxyiminoacetamido] -ceph- (3) -em-4-carboxylic acid

A suspension of 8.5 g of that prepared in Step C. Intermediate was stirred in 50 ml of methanol and mixed with 5 ml of N-methylmorpholine. After stirring for 10 min at 30 ° C. Add 30 ml methylene chloride, evaporate, add 100 ml ether, digesting, suction, washing with ether and drying a first fraction of 7.2 g salt was obtained. After evaporation to dryness and resume with ether was a second fraction of the same product receive.

4.24 g of the above morpholine salt were stirred and inert gas suspended in 60 ml of methylene chloride.

After stirring for 5 min and cooling to -5 ° C 6 ml of a 1 m solution of isobutyl chloroformate in methylene chloride added. The mixture was stirred at -5 ° C. for 5 min,  then cooled to -20 ° C and with a solution of 1.36 g of 7-aminocephalosporanic acid in 25 ml Methylene chloride and 1.4 ml of triethylamine were added. After standing at room temperature for 1 h, 50 ml Washed water containing 10 ml of 1N hydrochloric acid, aspirated, decanted, washed with water and evaporated to dryness. The product was in Triturated ether, centrifuged and washed with ether. 4.5 g of crude product were obtained.

The crude product was 1 h at + 10 ° C in 10 ml Stirred methylene chloride. The insoluble part was centrifuged off and washed with methylene chloride. After adding 50 ml ether to the filtrate, Stirring, centrifuging the precipitate and Washing with ether was 2.29 g of the expected Get final product.

Furthermore, a second fraction of 0.856 g obtained, so that a total of 3.146 g of the expected Product.

Example 2 3-acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl) -2-syn- (2-propenyloxyimino) acetamido] -ceph- (3) -em-4- carboxylic acid Step A: 2- (2-amino-4-thiazolyl) -2-syn- (2-propenyloxyimino) - ethyl acetate

• a) 9.7 g of 4-chloro-2-hydroxyiminoacetoacetic acid ethyl ester, 30 ml of acetone and 9.15 ml of 3-iodopropene were added  added 27.3 ml of 2N NaOH solution while cooling with ice; the mixture was then left to stand at room temperature for 1.5 h.
• b) The reaction mixture was treated with 3.8 g of thiourea added and 15 minutes at 60 ° C and then Kept at room temperature for 45 min; after eviction of acetone, addition of methylene chloride, water and potassium carbonate, stirring, decanting, extraction with methylene chloride, drying and evaporation 9.75 g of residue were obtained to dryness chromatographed on silica eluting with ether has been.

2.7 g of products were isolated using isopropyl ether was resumed; the crystals were suction filtered, washed and dried.

783 mg of the expected product were obtained. F. 100 ° C.

Stage B: 2- (2-tritylamino-4-thiazolyl) -2-syn- (2-propenyloxyimino) - ethyl acetate

511 mg of the product made in Step A, 0.92 ml Dimethylformamide, 1.8 ml methylene chloride and 0.29 ml triethylamine were mixed; after cooling to -15 ° C 615 mg of trityl chloride added. After standing for 1.5 hours 2 ml of 1 N hydrochloric acid and then 5 ml were at room temperature Water added, then decanted, dried and was evaporated to dryness.

1.28 g of crude products were obtained.  

Step C: 2- (2-tritylamino-4-thiazolyl) -2-syn- (2-propenyloxyimino) - acetic acid

1.28 g of the product obtained in step B, 6.2 ml Dioxane and 3 ml of 2N NaOH solution were combined at 120 ° C. The mixture was refluxed for 1 hour; the crystallized sodium salt was filtered off with washed with an ether-dioxane mixture and after drying isolated in an amount of 805 mg.

The product was dissolved in 10 ml of methylene chloride and 3 ml of 1N hydrochloric acid resumed and until Dissolution stirred; after decanting, drying, Evaporate to dryness, resume with ether and aspirating was 715 mg of the expected product receive. F. 170 ° C.

Step D: 3-acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl) - 2-syn- (2-propenyloxyimino) acetamido] -ceph- (3) -em- 4-carboxylic acid

470 mg of that prepared in Step C. Intermediate 5 ml of methylene chloride and 130 mg of dicyclohexylcarbodiimide were mixed. After that was rinsed with some methylene chloride and the mixture with stirring for 1 h at room temperature ditched. After sucking off the dicyclohexylurea formed  the filtrate was cooled and under intergas with a solution of 136 mg of 7-aminocephalosporanic acid in 2.4 ml of methylene chloride and 0.14 ml of triethylamine. After standing at room temperature for 1.5 h, addition of 2 ml 1 N hydrochloric acid and water, stirring, decanting, Washing with water, drying and evaporation were 610 mg Get raw product.

Example 3 3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-syn- (2-propenyloxyimino -) - acetamido] -ceph- (3) -em-4-carboxylic acid

610 mg of the product obtained in Example 2 were at 60 ° C for 15 min with 3 ml of 50% aqueous formic acid implemented; then 4 ml of water was added. After stirring, Centrifuge the triphenylcarbinol, wash with Water, evaporate to dryness in vacuum, resume with water, digestion, suction and washing Obtained 120 mg of the acid. F. ∼160 ° C.

UV spectrum
in ethanol:
Maximum 236 nm, = 375, ε = 18,000
Turning point 252 nm, = 316
Turning point 295 nm, = 138, ε = 6600;
in ethanol +0.1 N hydrochloric acid:
Maximum 263 nm, = 380, ε = 18 300
Turning point 280 nm, = 317.

NMR (90 MHz, DMSO), ppm:
2.02 -O-CO-CH₃
6.68 proton of the thiazole ring.

Example 4 3-acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl) -2-syn-vinyloxyimino-acetamido] - ceph-3-em-4-carboxylic acid Stage A: 2-syn- (2-bromoethoxyimino) -2- (2-tritylamino-4- ethyl thiazolyl) acetate

A mixture of 4.94 g of 2-syn-hydroxyimino-2- (2-tritylamino- 4-thiazolyl) ethyl acetate hydrochloride in 10 ml of dimethylformamide was submitted, after which at room temperature 4.14 g of potassium carbonate were added in 3 minutes. After stirring for 20 min at 20 ° C, 8.65 ml of 1,2 dibromoethane admitted. After stirring for a further 30 h Medium from 100 ml of distilled water and 20 ml of methylene chloride poured, then decanted, with methylene chloride extracted and after washing again with  distilled water extracted again; after this Drying of the organic solutions was suctioned off, washed and evaporated to dryness. The received Crude product was made on silica using Chromatographed benzene with 5% ether as eluent. A first fraction was obtained, the one after dissolving was recrystallized from 50 to 60 ° C from methanol, wherein was suctioned off at 0 or + 5 ° C; in this way 1.16 g of a creamy white product were obtained; F. 117 ° C.

Thereafter, a homogeneous fraction was obtained in an amount of 1.258 g.
NMR (CDCl₃), ppm:
3.55 triplet, J = 7 Hz -CH₂Br
4.51 triplet, J = 6 Hz NO-CH₂
6.55 singlet, proton of the thiazole ring.

Step B: 2- (2-tritylamino-4-thiazolyl) -2-syn-vinyloxyimino ethyl acetate

6.9 g of the 2-syn- (bromoethoxyimino) - prepared in stage A - 2- (2-tritylamino-4-thiazolyl) acetic acid ethyl ester were introduced into 35 ml of dimethyl sulfoxide. After that were 16 ml of a 1 M solution at 20 to 25 ° C in 5 min of potassium t-butoxide in tetrahydrofuran added. After stirring for 30 min, a further 3 ml the 1 M solution of potassium t-butoxide in tetrahydrofuran admitted. The mixture was then stirred for a further 30 min, followed by another 3 ml of the above potassium t-butoxide solution and then again after a further 30 min 3 ml Solution were added.  

After the last addition, stirring was continued for 30 min, whereupon in a mixture of 350 ml of water and ice and 90 ml Methylene chloride was poured in.

After acidification with 28 ml of 1 N hydrochloric acid to pH 2, Decant and extract three times with 50 ml Methylene chloride, washing with distilled water, drying, Suction, washing and evaporation to dryness obtained a black, resinous product, which by Column chromatography on silica using was cleaned of methylene chloride as the eluent.

In this way, 3.227 g of the aimed product get that unchanged for the next stage was used.

NMR (CDCl₃), ppm:
4.16 to 4.83 solid CH₂
6.66 to 7.33 solid CH
6.65 singlet, proton of the thiazole ring.

After recrystallization from absolute ethanol received a product with F. = 114 ° C.

Analysis (C₂₈H₂₅O₃N₃S):
Calculated C 69.54, H 5.21, N 8.69, S 6.36%. Found C 69.2, H 5.2, N 8.3, S 6.6%.

Step C: 2- (2-tritylamino-4-thiazolyl) -2-syn-vinyloxyimino acetic acid

3.227 g of the 2- (2-tritylamino-  4-thiazolyl) -2-syn-vinyloxyiminoacetic acid ethyl ester were placed in 3.2 ml of dioxane. At room temperature 20 ml of a 0.5 M solution of KOH in absolute ethanol and 12 ml absolute ethanol. The mixture was then stirred and in an argon atmosphere heated to 50 ° C and kept at this temperature for 1 h, the potassium salt crystallized out. After cooling at 20 ° C was suction filtered and washed with ethanol; the Product was in a mixture of 50 ml of ethyl acetate and Resumed 50 ml of distilled water.

It was then acidified with 6 ml of 2N hydrochloric acid, where was stirred until two homogeneous phases were achieved. It was then decanted off, distilled three times with 20 ml Washed water, extracted with ethyl acetate, dried, suction filtered and washed with ethyl acetate. After constriction to 15 ml, cooling to 0 or + 5 ° C, suction, Washing with ethyl acetate and drying were 2.53 g of the get the desired product, unchanged for the subsequent reaction stage was used.

After recrystallization, a purified product receive; F. 150 ° C.

Step D: 3-acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl) - 2-syn-vinyloxyimino-acetamido] -ceph- (3) - em-4-carboxylic acid

2.53 g of the 2- (2-tritylamino- 4-thiazolyl) -2-syn-vinyloxyiminoacetic acid introduced into 20 ml of methylene chloride. After that was on Cooled to + 5 ° C, all at once with 0.632 g of dicyclohexylcarbodiimide added and allowed to warm to room temperature.  To obtain a suspension, the mixture was stirred under argon for 1 h (Suspension A).

0.718 g of 7-aminocephalosporanic acid were separated from this under argon introduced into 10 ml of methylene chloride. After cooling to + 10 ° C, 1.1 ml of triethylamine were added (Solution B).

The suspension A was in 10 min at +5 to + 10 ° C. added to solution B, whereupon with stirring in about 35 min was allowed to warm to room temperature. After that was Stirred at 20 to 25 ° C for 3 h.

After adding 0.6 ml of acetic acid, the mixture was stirred for 10 min, whereupon the dicyclohexylurea formed is suctioned off has been. The organic solution was washed with water, dried and distilled in vacuo. Hereby a yellow, resinous product was obtained, which in Ethyl acetate was resumed; an easy, insoluble The residue was filtered off. After evaporation 3.475 g of the desired product were obtained to dryness.

The one used as starting material in Example 4 2-syn-hydroxyimino-2- (2-tritylamino-4-thiazolyl) -acetic acid ethyl ester-r- Hydrochloride was made as follows:

2- (2-Amino-4-thiazolyl) -2-syn-hydroxyiminoacetic acid ethyl ester

0.8 g of thiourea were dissolved in 2.4 ml of ethanol and 4.8 ml of water dissolved. Then a solution became in 5 min  of 2 g of 4-chloro-2-hydroxyiminoacetoacetic acid ethyl ester added and stirred for 1 h at room temperature. Subsequently most of the ethanol was evaporated in a slight vacuum, whereupon by adding solid sodium hydrogen carbonate was neutralized to pH 6. After cooling with ice water, suction, washing with water and drying in vacuo at 40 ° C were 1.32 g of the desired product receive; F. 232 ° C.

Analysis (C₅H₉O₃N₃S):
Calculated C 39.06, H 4.21, N 19.52, S 14.9%. Found C 38.9, H 4.4, N 19.7, S 14.6%.

2- (2-Tritylamino-4-thiazolyl) -2-syn-hydroxyiminoacetic acid ethyl ester-- hydrochloride

43.2 g of the 2- (2-amino-4-thiazolyl) - prepared as above 2-syn-hydroxyiminoacetic acid ethyl ester were placed in 120 ml of dry dimethylformamide.

After cooling to -35 ° C, 32 ml of triethylamine and then 60 g of trityl chloride in portions in 30 minutes admitted. After reheating became complete Dissolution and further warming up to 30 ° C were determined.

After 1 h was poured onto 1.2 l of ice water, the Contained 40 ml of hydrochloric acid (22 ° B´).

The mixture was then stirred in an ice-water bath, suction filtered, washed with 1 N hydrochloric acid and with ether inclined. Thereafter, 69.3 g of hydrochloride were obtained.  

Example 5 Sodium salt of 3-acetoxymethyl-7- [2- (2-tritylamino-4- thiazolyl) -2-syn-vinyloxyimino-acetamido] -ceph- (3) -em-4- carboxylic acid

At room temperature, 2.628 g of the in Example 4 Product obtained in resin form dissolved in 5.2 ml of acetone and all at once with 10.4 ml of a 1 M aqueous sodium hydrogen carbonate Solution added. After 1 h at 20 ° C the sodium salt was suctioned off and twice with 2 ml a water-acetone mixture (1: 1) and 2 ml distilled Washed water. After drying in a vacuum 40 ° C, 1.612 g of the desired crude product were obtained.

Example 6 3-acetoxymethyl 7- [2- (2-amino-4-thiazolyl) -2-syn-vinyloxyiminoacetamido] - ceph- (3) -em-4-carboxylic acid

1.612 g of the sodium salt obtained in Example 5 the 3-acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl) -2-syn- vinyloxyiminoacetamido] -ceph- (3) -em-4-carboxylic acid were dissolved in 4 ml of 98% formic acid. Right away then 4 ml of distilled water were added, whereupon the mixture was stirred at 40 ° C. for 15 min. The heterogeneous Suspension was triturated for 30 minutes at room temperature. It was then suctioned off at 20 ° C and twice with 1.6 ml Washed 50% hydrochloric acid, evaporated to dryness and resumed with 5 ml of distilled water. After suction at room temperature, wash with  distilled water and then with diethyl ether and drying were 300 mg of the target Product received. The separation of the insoluble Residue was made in the same way as above 50% formic acid.

In this way, an additional 0.214 g of acid receive.

Analysis (C₁₇H₁₇O₇N₅S₂):
Calculated: C 43.68, H 3.66, N 14.98, S 13.72%. Found C 44.2, H 3.9, N 15.1, S 13.0%.

NMR (DMSO), ppm:
6.91 proton of the thiazole ring
7.28 free amino group NH₂.

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

To 5.08 g of the 3-acetoxymethyl 7- [2- (2-amino-4-thiazolyl) -2-syn-vinyloxyiminoacetamido] - ceph- (3) -em-4-carboxylic acid was 5 ml of ethanol and then with stirring in an ice-water bath 10 ml a 1 M aqueous sodium hydrogen carbonate solution was added. After dissolving and adding 15 ml of ethanol was in vacuo Evaporated at 30 ° C, then taken up again with ethanol  and dried to constant weight.

Example 8 3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-syn-hydroxyiminoacetami-do] - ceph- (3) -em-4-carboxylic acid Step A: Ethyl 2- (2-amino-4-thiazolyl) -2-syn-hydroxyiminoacetate

0.8 g of thiourea was dissolved in 2.4 ml of ethanol and 4.8 ml of water dissolved within 5 min with a Solution of 2 g of ethyl 4-chloro-2-hydroxyiminoacetoacetate added and stirred for 1 h at room temperature. Then most of the ethanol was evaporated in vacuo and by adding solid sodium hydrogen carbonate neutralized to pH 6. After cooling with ice, Vacuuming, washing with water and drying in a vacuum at 40 ° C 1.32 g of the expected product was obtained. F. 232 ° C.

Analysis (C₅H₉O₃N₃S):
Calculated C 39.06, H 4.21, N 19.52, S 14.9%. Found C 38.9, H 4.4, N 19.7, S 14.6%.

NMR (60 MHz, DMSO), ppm:

• (a) Triplet with center at 1.25, J = 7 Hz
• (b) Quadruplet centered at 4.27, J = 7 Hz
• (c) Singlet 6.83
• (d) Singlet 7.11
• (e) Singlet 11.4.

Step B: 2- (2-amino-4-thiazolyl) -2-syn-hydroxyiminoacetic acid

21.5 g of the product obtained in step A were in 200 ml of absolute ethanol and 55 ml of 2 N NaOH solution were added and stirred in a water bath at 45 ° C. After 30 min cooled in an ice-water bath and with acetic acid brought to pH 6, whereupon a precipitate precipitated, the aspirated and with 50% aqueous ethanol and then was washed with ether. After drying were Obtained 16.9 g of the expected product.

Rf = 0.05 (eluent ethyl acetate / ethanol / Water 70: 20: 10).

Step C: sodium salt of 2- (2-tritylamino-4-thiazolyl) - 2-syn-trityloxyiminoacetic acid

16.9 g of the acid obtained in stage B were mixed with 50 ml of dimethylformamide and 42 ml of triethylamine mixed. While stirring at room temperature for 15 min, was complete Resolution found.

After cooling to -20 ° C, the triethylammonium salt crystallized partially out. Then within 15 g at -20 ° C 54 g trityl chloride in 100 ml chloroform added, stirred for 1 h and warmed to room temperature  calmly. Then it was poured into 200 ml of water, which contained 40 ml of 2N hydrochloric acid.

After decanting, wash the organic twice Phase with 200 ml water, drying, suction, Evaporate the solvents in vacuo, resume with ethyl acetate, adding 100 ml of one saturated sodium bicarbonate solution, stirring and Decanting crystallized the sodium salt. After that was cooled with ice for 30 min, suction filtered and with Washed ethyl acetate. There were 27 g of the expected Obtained sodium salt. Rf = 0.33 (ether).

Step D: 3-acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl) - 2-syn-trityloxyiminoacetamido] -ceph- (3) -em-4-carboxylic acid- t-butyl ester

17.2 g of the sodium salt of 2- (2-tritylamino- 4-thiazolyl) -2-syn-trityloxyiminoacetic acid were mixed in 170 ml of chloroform with 170 ml of 1N hydrochloric acid. Then it was decanted, washed five times with water, dried and vacuumed. After evaporation of the solvent and resuming the residue with 170 ml of methylene chloride, 2.8 g of dicyclohexylcarbodiimide admitted. After stirring for 1 h, 1.9 g Aspirated dicyclohexylurea.

3.66 g of 3-acetoxymethyl-7-amino were added to the filtrate. ceph- (3) -em-4-carboxylic acid t-butyl ester added.

The mixture was then stirred at room temperature for 2 h and with 1 N hydrochloric acid, then with water, with a 5% aqueous solution Sodium bicarbonate solution and again with water washed.  

After drying the organic phase, suction, Evaporation of the solvent, resume with Methylene chloride and column chromatography on silica using a mixture of methylene chloride with 5% ether those of interest Fractions (Rf = 0.78 in ether) combined, whereupon the solvent evaporated in vacuo and with Isopropyl ether was resumed.

After digesting and aspirating with isopropyl ether washed.

5.8 g of the expected product were obtained.

Analysis C₅₇G₅₁O₇N₅S₂:
Calculated: O 69.7, H 5.2, N 7.1, S 6.5%. Found: O 70.4, H 5.6, N 6.5, S 5.9%.

NMR (60 MHz, CDCl₃), ppm:

• (a) Singlet 1.55
• (b) Singlet 2.06
• (c) Singlet 6.45
• (d) Singlet 7.31.

Step E: 3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-syn- hydroxyiminoacetamido] -ceph- (3) -em-4-carboxylic acid

1 g of the product obtained in stage D was dissolved in 3 ml Trifluoroacetic acid. After stirring for 30 min at room temperature 30 ml of isopropyl ether were added, followed by the salt precipitated, which was filtered off with isopropyl ether was washed. In this way, 0.652 g of the Trifluoroacetate of 3-acetoxymethyl-7- [2- (2-amino-4- thiazolyl) -2-syn-trityloxyiminoacetamido] -ceph- (3) -em-4- receive carboxylic acid.

The product was dissolved in 6 ml of tetrahydrofuran and mixed with 3 ml of 50% aqueous formic acid. After stirring for 15 min at 50 ° C., evaporating off the solvents, Resume with ether, aspirate and wash with ether became 0.441 g of formate of the expected product receive.

The salt was in 2 ml of water, the 3 drops of pyridine contained (pH ∼ 6), triturated. After suction, wash with Water and drying became 0.136 mg of the expected product receive. By evaporating the filtrate to dryness, Resume with ethanol, aspirate and wash with A further 0.04 g of product was obtained from ethanol.  

NMR (60 MHz, DMSO), ppm:

• (a) Singlet 2.01
• (b) Singlet 6.67
• (c) Singlet 7.08
• (d) Singlet 11.3.

Example 9 3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-syn-hydroxyimino acetamido] -ceph- (3) -em-4-carboxylic acid Step A: 2- (2-tritylamino-4-thiazolyl) -2-syn-hydroxyimino ethyl acetate

43.2 g of that prepared in Step A of Example 8 2- (2-Amino-4-thiazolyl) -2-syn-hydroxyiminoacetic acid ethyl ester were in 120 ml of dry dimethylformamide brought in.  

After cooling to -35 ° C, 32 ml of triethylamine and then 60 g of trityl chloride in portions within 30 minutes added. Then the temperature was allowed to rise again being full resolution and after that Warming up to 30 ° C were found. After 1 h was poured onto 1.2 l of ice water containing 40 ml of hydrochloric acid (22 ° B´) contained.

After stirring in an ice-water bath, suction, washing with 1 N hydrochloric acid and digest with ether Obtained 69.3 g of hydrochloride.

The free base was obtained by dissolving the product in 5 volumes of methanol, to which 120% thriethylamine was added and then slow precipitation obtained with 5 volumes of water.

Analysis C₂₆H₂₃O₃N₃S · ¼ H₂O:
Calculated C 67.6, H 5.1, N 9.1, S 6.9%. Found C 67.5, H 5.1, N 8.8, S 6.8%.

NMR (60 MHz, CDCl₃), ppm:

• (a) Triplet centered at 1.31, J = 7 Hz
• (b) Quadruplet centered at 4.37, J = 7 Hz
• (c) Singlet 6.37
• (d) Singlet 7.28.

Step B: 2- (2-tritylamino-4-thiazolyl) -2-syn-tetrahydropyranyloxyiminoacetic acid, ethyl ester

5.6 g of the product obtained in step A were in 56 ml of distilled dihydropyran added. Then in cooled in an ice-water bath and with 2.4 g of p-toluenesulfonic acid transferred.

After stirring for 1.5 h, the temperature rose again Allow room temperature to rise, then mix of 100 ml of benzene, 100 ml of water and 2 ml of triethylamine was poured in.

Then it was decanted off, washed with water, dried, suctioned off and washed with benzene, whereupon the solvent was evaporated. After resuming with isopropyl ether, seeding for crystallization, leaving to stand overnight in the refrigerator, suction and washing 4.42 g of product were obtained with isopropyl ether; F. 184 ° C.

Analysis of p-toluenesulfonate C₃₈H₃₉O₇N₃S₂:
Calculated: C 63.9, H 5.5, N 5.9, S 9.0%. Found C 63.7, H 5.5, N 5.8, S 8.9%.

NMR (60 MHz, CDCl₃), ppm:  

• (a) Triplet centered at 1.36
• (b) Quadruplet centered at 4.39
• (c) Singlet 6.60
• (d) Singlet 6.91
• (e) Singlet 7.28.

Step C: 2- (2-tritylamino-4-thiazolyl) -2-syn-tetrahydropyranyloxyiminoacetic acid

4.56 g of the product obtained in stage B was dissolved in 45 ml Dioxane and 8.4 ml of 2N NaOH solution added; thereafter became 1.5 h held at reflux. After cooling in the ice-water bath, this fell Sodium salt, which after suction and washing with aqueous Dioxane and then with ether in an amount of 4.66 g.

The free acid was obtained by dissolving the sodium salt in 50 ml of dioxane, acidifying to pH 5 with formic acid and precipitating with 90 ml of water. F. 180 ° C.
NMR (60 MHz, CDCl₃), ppm:
6.69 proton of the thiazole ring
7.31 aromatic H.

Step D: 3-acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl) - 2-syn-tetrahydropyranyloxyiminoacetamido] -ceph- (3) - em-4-carboxylic acid t-butyl ester

0.362 g of the product obtained in Step C, 0.244 g 3-acetoxymethyl-7-amino-ceph- (3) -em-4-carboxylic acid-t- butyl ester and 0.280 g dicyclohexylcarbodiimide placed in 4 ml of dry chloroform. After stirring for 2 h at room temperature the dicyclohexylurea formed suction filtered and washed with chloroform.

The solvents of the filtrate were evaporated in vacuo; the residue was dissolved in 1 ml ether and then on a silica column with elution Chromatographed ether.

The fractions with an Rf value of 0.38 were cleaned; after evaporation of the solvent in vacuo, Resume with isopropyl ether, digest, aspirate and washing with isopropyl ether were 0.184 g of the expected Product received.

Analysis C₄₃H₄₅O₈N₅S₂:
Calculated: C 62.7, H 5.5, N 8.5, S 7.8%. Found C 62.8, H 5.9, N 8.1, S 7.5%.

NMR (60 MHz, CDCl₃), ppm:  

• (a) 1.53
• (b) 2.07
• (c) 5.46
• (d) 6.76
• (e) 7.28

Step E: 3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-syn- hydroxyiminoacetamido] -ceph- (3) -em-4-carboxylic acid

638 mg of the product obtained in Step D were 15 min at room temperature in 1.8 ml trifluoroacetic acid touched. After adding 18 ml of isopropyl ether Aspirated 404 mg of precipitated product, which with 2 ml 50% aqueous formic acid resumed and Stirred at 50 ° C for 15 min. After evaporation to Dry in vacuo at 30 ° C, reabsorb with 1 ml Ethanol and adding a drop of pyridine became that expected product suctioned off with the product of Example 8 was identical.  

Example 10 3-acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl) -2-syn-trityloxyimin-oacetamido] - ceph- (3) -em-4-carboxylic acid Step A: 2- (2-tritylamino-4-thiazolyl) -2-syn-trityloxyiminoacetic acid ethyl ester

1.08 g of that obtained in Step A of Example 8 2- (2-Amino-4-thiazolyl) -2-syn-hydroxyiminoacetic acid ethyl ester were dissolved in 8 ml of chloroform and with 1.5 ml triethylamine and then within 25 min at + 5 ° C with a solution of 3 g trityl chloride added in 6 ml of chloroform and for 1 h at room temperature touched. Then with 18 ml of water, 8 ml of 1 N hydrochloric acid and washed three times with 20 ml of water, dried, suction filtered and evaporated to dryness. The product was with isopropanol in which it crystallized. 2.3 g of the expected product were obtained. F. 140 ° C.

Stage B: sodium salt of 2- (2-tritylamino-4-thiazolyl) - 2-syn-trityloxyiminoacetic acid

0.7 g of the product obtained in step A was in Dissolved 3.5 ml of hot dioxane. After heating to 110 ° C 1 ml of 2N NaOH solution was added dropwise with stirring admitted. Then at a temperature close to Reflux continued to stir for 1 h 50 min. The fancy Sodium salt was suctioned off. The product was with the obtained in Step C of Example 8 identically.  

Step C: 3-acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl) - 2-syn-trityloxyiminoacetamido] -ceph- (3) -em-4- carboxylic acid

5.12 g of the sodium salt obtained in step B were suspended in 50 ml of chloroform and with 50 ml of 1N hydrochloric acid transferred. After stirring, decanting, three times Wash with 50 ml of water, dry, suction and concentration to dryness was 2- (2-tritylamino-4- obtained thiazolyl) -2-syn-trityloxyiminoacetic acid.

The acid obtained was dissolved in 50 ml of methylene chloride solved. After adding 1.6 g of dicyclohexylcarbodiimide and stirring for 1 hour at room temperature was formed Aspirated dicyclohexylurea, whereupon the solution Cooled -10 ° C and with 1.1 g of 7-aminocephalosporanic acid in solution in 10 ml methylene chloride and with 1.2 ml Triethylamine was added. After stirring for 2 h at room temperature, Add 50 ml of 1N hydrochloric acid, stir, decant, washing three times with 50 ml of water, drying, Suck off and evaporate to dryness using ethanol digested, whereupon 2.36 g of crude condensation product is suctioned off were. This product has been exposed of diethylamine in ether in the diethylammonium salt of 3-acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl) -2-syn- trityloxyiminoacetamido] -ceph- (3) -em-4-carboxylic acid transferred.

The salt obtained was until a acidic pH with 1 N hydrochloric acid in methylene chloride transferred. The solution was washed with water, dried and evaporated to dryness, whereupon 0.75 g of the expected  Pure product were obtained.

Example 11 3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-syn-hydroxyiminoacetami-do] - ceph- (3) -em-4-carboxylic acid

The double tritylated obtained in Example 10 Product was in 5 ml of 50% aqueous formic acid solved. After stirring at 50 ° C for 15 min, the solvents evaporated, whereupon resumed in ether, was suction filtered and washed with ether.

The formate was thus obtained.

The formate was triturated with 3 ml of water, the contained a few drops of pyridine (pH ∼ 6). After suction, Washing with water and drying was what was expected Obtain product that with that of Examples 8 and 9 was identical.

Claims (2)

1. Process for the preparation of aminothiazolyl-syn-oxyiminoacetic acid derivatives of the formula I, in which: R₂H or a protective group which can be split off by acid hydrolysis or hydrogenolysis, R₁H, a protective group which can be split off by acid hydrolysis or hydrogenolysis or unsaturated C _2-4 -alkyl,
where R₁ is not H if R₂ is a protective group which can be split off by acid hydrolysis or hydrogenolysis,
and AH or C _1-4 alkyl, characterized by reacting a compound of formula II in which: XCl or Br, R₁H or a protective group which can be split off by acid hydrolysis or hydrogenolysis or unsaturated C _2-4 -alkyl,
where R₁ is H when X is Br,
and alkC _1-4 alkyl, with thiourea in an aqueous solvent and / or at room temperature in the presence of an approximately stoichiometric amount of thiourea and for a reaction time of a few hours and treatment with a base to give the corresponding syn compound of the formula III with R₁ and alk as above and, if necessary, reaction of the compound of formula III with a functional derivative of a protective group R'₂ which can be split off by acid hydrolysis or hydrogenolysis to give the syn compound of formula IV with R₁, R'₂ and alk as above and, if necessary, reaction of the compound of formula IV by treatment with a base and then with an acid to give the compound of formula I. 2. Use of aminothiazolyl-syn-oxyiminoacetic acid derivatives of the formula I, in which: R₂H or a protective group which can be split off by acid hydrolysis or hydrogenolysis, R₁unsaturated C _2-4 -alkyl
and AH or C _1-4 alkyl, for the preparation of corresponding 7-aminothiazolyl-syn-oxyiminoacetamidocephalosporanic acids.