IE45600B1 - New oxime derivatives of 7-amino-thiazolyl-acetamido-cephalosporanic acid, processes for their preparation and pharmaceutical compositions containing them - Google Patents

Description

This Application relates to new oxinte derivatives of 7-amino-thiazolyl-acetamido-cephalosporanic acid, processes for their preparation and pharmaceutical compositions containing them.

In our Patent Specification No.Ij ζϋϊ$ there are described and 5 claimed new oxime derivatives of 7 - amino - thiazolyl - acetamido cephalosporanlc acid of the general formula: NH — R wherein R represents a hydrogen atom or a group removable by acid hydrolysis or hydrogenolysis, R‘ represents a hydrogen atom, a group removable by acid hydrolysis or by hydrogenolysis or a saturated or unsaturated hydrocarbyl radical having from 1 to 4 carbon atoms, A represents a hydrogen· atom, an alkali metal atom, an equivalent of an alkaline-earth metal atom, an equivalent of a magnesium atom or a quaternary ammonium group, with the provisos that when R1 represents a group removable by acid hydrolysis or by hydrogenolysis R represents the same or a different group removable by acid hydrolysis or by hydrogenolysis and that when R‘ represents a hydrogen atom R also represents a hydrogen atom, the compound being in the form of the syn isomer.

The aforesaid specification also describes and claims processes for the preparation of the compounds of general formula I and pharmaceutical compositions containing them, as well as new antibiotic intermediates of general formula VI employed in the process described and claimed.

The present invention concerns certain new compounds falling within general formula I, but not disclosed in the aforementioned specification, which have been found to have exceptional antibiotic activity.

Accordingly, this invention provides 7-/2-(2- amino - 4 15 thiazolyl) - 2 - hydroxyimino - acetamid£] - 3 - acetoxy - methyl ceph - 3 - em - 4 - carboxylic acid, syn isomer, and salts thereof formed with alkali metals, alkaline-earth metals, magnesium or organic amines.

Amongst these compounds 7-/.2- (2 - amino - 4 - thiazolyl) - 2 hydroxyimino - acetamido] - 3 - acetoxy - methyl - ceph - 3 - em - 4 20 carboxylic acid, syn isomer, and the sodium salt of 7-/2-(2- amino - 4 - thiazolyl) - 2 - hydroxyimino - acetamido] - 3 acetoxy - methyl - ceph - 3 - em - 4 - carboxylic acid, syn isomer, have been found to display particularly remarkable activity.

Furthermore, the invention provides 7-/2- (2 - tritylamino - 4 25 thiazolyl) - 2 - trityl - hydroxyimino - acetamidoj - 3 - acetoxy - methyl - ceph - 3 - erne - 4 - carboxylic acid, syn isomer. 45®θ° The above compounds may all be prepared by the processes described and claimed in the aforementioned specification. However, this invention also provides alternative processes for preparing the new compounds and other compounds of general formula I.

Thus, in another aspect this invention provides a process for the preparation of compounds of the general formula . CH,- 0 — CH, |( 3 (Ib) syn isomerj (wherein R^ represents a hydrogen atom or a saturated or unsaturated hydrocarbyl radical having from 1 to 4 carbon atoms) - that is to say, a compound of general formula I wherein R and A each represent a hydrogen atom and R' represents a hydrogen atom or a hydrocarbyl radical having from 1 to 4 carbon atoms - in which process a derivative of 7 - amino - cephalosporanic acid of the general formula: COgAl '3 (wherein A^ represents a group removable by acid hydrolysis or by hydrogenolysis) is reacted with an acid of the general formula: (Π) OR 1 syn isomer, (wherein R) represents a group removable by acid hydrolysis or by hydrogenolysis or a chloracetyl radical, and R'^ represents a group removable by acid hydrolysis or by hydrogenolysis, a chloracetyl radical or a saturated or unsaturated hydrocarbyl radical having from 1 to 4 carbon atoms) or a functional derivative thereof, to obtain the corresponding compound of the general formula: ',45600 syn isomers (wherein A-j, R^ and R’-j are as defined hereinbefore) which compound XI is treated with one or more of acid hydrolysis agents, hydrogenolysis agents and thiourea to obtain the desired product of general formula lb.

The preferred groups removable by acid hydrolysis or by hydrogenolysis, that may be represented by R^ and R'p include the tert-butoxycarbonyl, trityl, benzyl, benzhydryl, tri chloroethyl, carbobenzyloxy and formyl groups. Other such groups include the trichloroethoxycarbonyl and 2 - tetrahydropyranyl groups.

Preferred groups A^ include the benzhydryl, tert-butyl, benzyl, paramethoxybenzyl and tri chloroethyl radicals.

In the preferred method of carrying out the above process, a 15 compound of the general formula: CHg—0 ί02Α1 ·—0—— CFL II 3 0 (wherein A^ is as defined hereinbefore) is reacted with a functional derivative of an acid of general formula II such as an anhydride or the acid chloride. Preferably the anhydride is formed in situ by the action of isobutyl chloroformate or dicyclohexylcarbodiimide on the acid. Other acid halides or other anhydrides - formed in situ by the action of another alkyl chloroformate, a dialkylcarbodiimide or another dicycloalkylcarbodiimide - may also be employed. It is also possible to employ other derivatives of the acid, such as the acid azide, the activated acid amide or an activated acid ester, for example, the esters formed with hydroxysuccinitiride, j>-nitrophenol and 2,4-di nitrophenol.

In the case where the reaction is effected using a halide of the acid of general formula II or with an anhydride thereof formed with isobutyl chloroformate, it is preferably carried out in the presence of a basic agent. Preferred basic agents include alkali metal carbonates and tertiary organic bases, such as N-methyl-morpholine, pyridine or a trialkylamine such as triethylamine.

The transformation of the products of general formula XI to the products of general formula lb involves replacement of the substituents R^ and A.| by hydrogen atoms, if necessary, and in the case when R'l represents a group removable by acid hydrolysis or by hydrogenolysis or the chloracetyl group, the transformation also involves replacement of this group by a hydrogen atom. The choice of the appropriate agent from those set out hereinbefore to effect this transformation in a particular case is believed to be within the competence of one skilled in the art. However, it is pointed out that the compound of V. 4560 0 general formula XI is treated with one or more acid hydrolysis agents when the substituents R-j and A^ represent a group removable by acid hydrolysis and R1^ represents a group removable by acid hydrolysis or a hydrocarbyl radical. The compound of general formula XI is treated with one or more hydrogenolysis agents when the substituents Rl and A^ represent a group removable by hydrogenolysis and R‘l represents a group removable by hydrogenolysis or a hydrocarbyl radical. The compound of general formula XI is treated with one or more acid hydrolysis agents and by one or more hydrogenolysis agents when at least one of the substituents R^, A^ and R1^ represents a group removable by acid hydrolysis and at least one of these substituents represents a group removable by hydrogenolysis. Finally, the compound of general formula XI is treated with thiourea, and optionally thereafter with one or more acid hydrolysis agents or hydrogenolysis agents, in the case when at least one of the substituents R-| and R'^ represents a chloracetyl group.

Preferred acid hydrolysis agents are formic acid, trifluoroacetic acid or acetic acid. These acids can be employed in anhydrous form or in aqueous solution. An example of a hydrogenolysis agent that may be employed is the zinc-acetic acid system.

An acid hydrolysis agent such as an anhydrous trifluoroacetic acid or aqueous fonrn'c or acetic acid is preferably used to eliminate butoxycarbonyl or trityl groups as substituents R^ and R’p or to eliminate benzhydryl, tert-butyl, or jj-methoxybenzyl groups as the substituent Ap The zinc-acetic acid system is preferably used to eliminate the trichloroethyl group as substituents Rp R'^ and A.

A hydrogenolysis agent such as hydrogen in the presence of a catalyst is preferably used to eliminate the benzhydryl and carbobenzyloxy groups as substituents R1 and R'p and to eliminate the benzyl group as substituents Rp R1^ and Ap The reaction of thiourea with a compound of general formula XI containing at least one chloroacetyl group is preferably carried out in a neutral or acidic medium. . This type of reaction is described for other compounds by MASAKI (JACS, 90, 4508/1968).

The compounds of general formula II employed in the above process or in the process described and claimed in the aforementioned specification may be prepared by an alternative process in which an appropriate compound of the general formula C02alk (IV) syn isomer, I (wherein R represents a hydrogen atom, a group removable by acid hydrolysis or by hydrogenolysis or a saturated or unsaturated alkyl radical having from 1 to 4 carbon atoms and alk represents an alkyl radical having from 1 to 4 carbon atoms) is reacted firstly with a base and then with an acid to obtain a product of the general formula: 4SQG0 NH, COgH OR (XII) syn isomer, (wherein R1 is as defined hereinbefore) which product is treated with a compound capable of introducing a group removable by acid hydrolysis or by hydrogenolysis or a compound capable of introducing the chloracetyl group to obtain the desired product of general formula II.

The base which is used to saponify the product of formula IV is preferably sodium hydroxide but other bases such as potassium hydroxide or barium hydroxide can also be used.

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

The compound capable of introducing a group removable by acid hydrolysis or by hydrogenolysis is preferably trityl chloride, and this is conveniently used in the presence of triethylamine or another tertiary amine such as other tri alkyl amines, N - methyl - morpholine or pyridine.

Other compounds capable of introducing groups removable by acid hydrolysis or by hydrogenolysis include tert-butyl ehloroformate, tert20 butyl azidoformate, tri chloroethyl and benzyl chloroformates, the mixed formyl acetic anhydride, benzyl and benzhydryl halides and particularly benzyl chloride and benzhydryl chloride, phthalic anhydride and N-carbethoxyphthalimide.

The compound capable of introducing the chloracetyl group is preferably chloracetic anhydride or a chloracetyl halide such as monochloracetyl chloride. In the case where the reaction is effected with a chloracetyl halide, the reaction is preferably carried out with a basic agent such as one of those basic agents set out hereinbefore.

In addition the compounds falling within the general formula: NH-R1 syn isomer, (wherein R-| and alk are as defined hereinbefore and Rc, which is different to R,, represents a group removable by acid hydrolysis or by hydrogenolysis or a chloracetyl group) which are employed in the processes for the preparation of compounds falling within general formula I as described and claimed herein or in our aforementioned specification (general formula Vc above encompassing compounds falling within general formula III and Illb of that earlier specification), may be prepared by treating an appropriate compound of the general formula: ., 45500 I OH syn isomer, (wherein R-j and alk are as defined hereinbefore) with a compound capable of introducing a group removable by acid hydrolysis or by hydrogenolysis (Rc) or a compound capable of introducing the chloracetyl group (Rc) so as to obtain the desired compound of general formula Vc.

The group removable by acid hydrolysis or by hydrogenolysis which may be represented by Rj is preferably the trityl group.

The compound capable of introducing a group removable by acid hydrolysis or by hydrogenolysis which is reacted with the product X is preferably dihydropyran.

One can however use any of the functional derivatives mentioned hereinbefore. Accordingly, in a preferred process Rc represents a tetra hydropyranyl radical.

The new compounds described and claimed herein, in common with those of the aforementioned specification, possess a very good antibiotic activity, on the one hand against the Gram (+) bacteria such as the staphylococci and the streptococci and especially against the penicillin-resistant staphylococci, and on the other hand against the Gram (r) bacteria, especially the coliform bacteria, the Klebsiella, the Salmonella and the Proteus.

These properties may make the compounds of the invention useful as medicaments in the treatment of diseases caused by sensitive microorganisms, and notably in the treatment of staphylococcal infections such as staphylococcal septicaemia, malignant facial or skin staphylococcal infections, pyodermatitis, septic or suppurating sores, anthrax, phlegmons, erysipelas, acute primary or post-influenza staphylococcal infections, bronchopneumonia and pulmonary suppurations.

The compounds of the invention may also be useful as medicaments in the treatment of colibacillosis and associated infections, in infections caused by Proteus, Klebsiella and Salmonella, and in other diseases caused by Gram (-) bacteria.

However, before any of the compounds of this invention are used in medicine, they are preferably formed into pharmaceutical compositions by association with suitable pharmaceutical vehicles.

The term “pharmaceutical is used herein to exclude any possibility that the vehicle or the active material, as appropriate, considered in relation to the route by which the composition is intended to be administered, could result in the composition being harmful rather than beneficial. The choice of a suitable mode of presentation, together With an appropriate vehicle, is believed to be within the competence of those accustomed to the preparation of pharmaceutical formulations.

Accordingly, in yet another aspect this invention provides pharmaceutical compositions containing, as active principle, one or more of the compounds of the invention, in association with a suitable pharmaceutical vehicle.

Preferred active materials for use in the compositions are: 4δ® OO -Q2 - (2 - amino - 4 - thiazolyl) - 2 - hydroxyimino - acetamidj - 3 - acetoxy -methyl - ceph - 3 - em - 4 - carboxylic acid, syn isomer, and the salts thereof formed with alkali metals, alkaline-earth metals, magnesium and organic amines, and more particularly: 7-(2-(2- amino - 4 - thiazolyl) - 2 - hydroxyimino - acetamido] - 3 - acetoxy - methyl - ceph - 3 - em - 4 - carboxylic acid, syn isomer, the activity of which in relation to a variety of germs appears to be particularly remarkable; and: the sodium salt of 7 -Γ2 -(2- amino - 4 - thiazolyl) - 2 hydroxyimino - acetamido] - 3 - acetoxy - methyl - ceph - 3 - em - 4 carboxylic acid, syn isomer.

The compositions of this invention may be administered by buccal, rectal or parenteral routes, or by local route by topical application to the skin or mucous membranes, and thus the pharmaceutical vehicle is preferably: a) the ingestible excipient of a tablet, coated tablet, sublingual tablet or pill, the ingestible container of a capsule or cachet, the ingestible pulverulent solid carrier of a powder or granules, or the investible liquid medium of a syrup , solution, suspension or elixir; ' b) the solid or liquid medium of a paste, lotion, salve, ointment, cream, gel or unguent; c) a sterile injectable liquid solution or suspension medium, or d) a base material or a suppository.

Whilst the modes of presentation and vehicles just listed represent those most likely to be employed, they do not necessarily exhaust the possibilities. ' 45680 Ihe compositions may thus be in the form of a solid or liquid and the preferred pharmaceutical forms currently used in human medicine in which they may be presented are as plain or sugar-coated compressed tablets, gelatin capsules, granules, suppositories, injectable preparations, ointments, creams or gels. The vehicle is typically one .or more of the excipients conventionally employed in these pharmaceutical compositions such as talc, gum arabic, lactose, starch, magnesium stearate, cocoa butter and aqueous or non-aqueous liquid vehicles including fatty substances of animal or vegetable origin, paraffin derivatives and glycols. The compositions may also include one or more wetting, dispersing or emulsifying agents, and/or one or more preservatives.

The dose of active material administered will vary according to the complaint treated, the person concerned, the route of administration and the product under consideration. However, for the compounds of Examples 1 and 2 it may be, for example, between 0.500 g and 1 g. taken three times daily, for administration to man by intramuscular route.

Certain of the intermediate compounds used in the preparation of the compounds of general formula I and related compounds are new, and in another aspect this invention provides these useful intermediates per se.

Thus, this invention also provides the compounds of the general formula: /45600 syn isomerο wherein R-j R’-j and are as defined hereinbefore.

These compounds, in common with the compounds of general formula I, possess an antibiotic activity.

The following Examples are now given, though only by way of illustration, to show certain preferred aspects of the invention.

EXAMPLE 1 - /2-(2- amino - 4 - thiazolyl) - 2 - hydroxyimino acetamido] 3 - acetoxymethyl - ceph - 3 - em - 4 - carboxylic acid, syn isomer Stage A: Ethyl 2 - (2 - amino - 4 - thiazolyl) - 2 - hydroxyimino - acetate, syn isomer. q 3 One dissolves 0.8 g of thiourea in 2.4 cm ethanol and 4.8 cm water. 15 Over 5 minutes one adds a solution of 2 g of ethyl 4 - chloro - 2 hydroxyimino - acetylacetate and agitates for 1 hour at ambient temperature. One drives off the majority of the ethanol under partial vacuum and neutralizes to pH6 by adding solid sodium bicarbonate.

One chills, vacuum-filters, washes with water and dries under vacuum at 40°C. One obtains 1.32 g of desired product. M.Pt 232°C Analysis: CgHgOgNgS Calculated: C% 39.06 Found 38.9 NMR (CDClg, 60 MHz) HSS 4.21 NS6 19.52 4.4 19.7 S% 14.9 14.6 (b) (a) CH2-CHg (a) triplet centred on 1.25 ppm d = 7 Hz (b) quadruplet centred on 4.27 ppm J 7 Hz (c) singlet at 6.83 ppm (d) singlet at 7.11 ppm (e) singlet at 11.4 ppm Stage B: 2-(2- amino - 4 - thiazolyl) - 2 - hydroxyimino-acetic acid, syn isomer.

One introduces 21.5 g of the product prepared in stage A into 200 cm3 of absolute ethanol and 55 cm0 of 2N sodium hydroxide. One agitates in a bath of water at 45°C. After 30 minutes one places in a bath of iced water, then adjusts to pH 6 with acetic acid. One observes a precipitation. -45600 One vacuum filters and rinses with 50% aqueous ethanol, then with ether. After drying, one obtains 16.9 g of the desired product.

Rf = 0.05 (eluant: ethyl acetate-ethanol-water 70-20-10).

Stage C: Sodium salt of 2 - (2 - tritylamino - 4 - thiazolyl) - 2 - trityl hydroxyamino - acetic acid, syn isomer. o One mixes 16.9 g of the acid prepared in stage B, 50 cm of dimethyl 0 formamide and 42 cm of triethylamine. One agitates for 15 minutes at ambient temperature and observes a total dissolution.

One re-cools to -20°C, and the triethylamine salt partially crystallizes. One introduces over 15 minutes at -20°C 54 g of trityl chloride and 100 cm3 of chloroform. One agitates for 1 hour while leaving to return to ambient temperature. One pours into 3 3 200 cm of water containing 40 cm of 2N hydrochloric acid.

One decants, washes the organic phase twice with 200 cm3 of water, dries, vacuum-filters, drives off the solvent under reduced pressure, and takes up the product in ethyl acetate.

One adds 100 cm3 of a saturated solution of sodium bicarbonate, agitates and decants. The sodium salt crystallizes out. One chills for 30 minutes, vacuum-filters and rinses with ethyl acetate.

One obtains 27 g of the desired sodium salt. Rf = 0.33 (ether).

Stage D: Tert-butyl 7 - [2 - (2 - tritylamino - 4 - thiazolyl) - 2 - trityl hydroxyimino - acetamidq] - 3 - acetoxymethyl - ceph - 3 - em 25 4 - earboxylate, syn isomer.

One mixes 17.2 g of the sodium salt of 2 - (2 - trityl - amino 18 6 0 0 - thiazolyl) - 2 - trityl oxyimino - acetic acid, syn isomer, obtained in stage C, in 170 cm of chloroform, and 170 cm of N hydrochloric acid. One decants and washes 5 times with water. One dries, vacuum-filters and drives off the solvent.

The residue is taken up with 170 cm of methylene chloride. One adds 2.8 g of dicyclohexylcarbodiimide. One agitates for 1 hour, then vacuum-filters off 1.9 g of di cyclohexyl urea.

One adds to the filtrate 3.56 g of tert-butyl 7 - amino - 3 acetoxymethyl - ceph - 3 - em - 4 - carboxylate.

One agitates for 2 hours at ambient temperature, and washes with normal hydrochloric acid, then with water, with a 5% aqueous solution of sodium bicarbonate, then with water.

One dries the organic phase, vacuum-filters, drives off the solvent, takes up the product with methylene chloride and elutes on a silica column with methylene chloride containing 55! of ether.

The fractions of interest (Rf = 0.78 in ether) are reunited. One drives off the solvent under reduced pressure and takes up the product in isopropyl ether.

One disintegrates the solid matter, vacuum-filters and rinses with isopropyl ether.

Analysis: θ57^5ΐθ7^5^2 Calculated: C% 69.7 HSS 5.2 N« 7.1 S% 6.5 Found: 70.4 5.6 6.5 5.9 NMR (CDC13, 60 MHz) (a) (b) (c) (d) $3 singlet at 2.06 ppm singlet at 6.45 ppm singlet at 7.31 pp, Stage E: - [^2-(2- amino - 4 - thiazolyl) - 2 - hydroxyimino-acetamido] - acetoxymethyl - ceph - 3 - em - 4 - carboxylic acid, syn isomer.

One places 1 g of the product obtained in stage D in 3 cm of 10 trifluoroacetic acid. One agitates for 30 minutes at ambient temperature, then adds 30 cm3 of isopropyl ether. The salt precipitates. One vacuum-filters and rinses with isopropyl ether. One obtains in this manner 0.652 g of the trifluoroacetic acid salt of 7 -[^2 -(2- amino 4 - thiazolyl) - 2 - trityl oxy imi no - acetamido] - 3 - acetoxymethyl - ceph - 3 - em - 4 - carboxylic acid.

This product is dissolved in 6 cm of tetrahydrofuran, and one adds cm3 of 50% aqueous formic acid. One agitates for IS minutes at 50°C, drives off the solvents, takes up with ether and vacuum-filters, rinsing with ether. One obtains in this manner 0.441 g of the formate of the 56 00 desired product. The salt is triturated in 2 cm of water containing 3 drops of pyridine (ρΗ^β). One vacuum-filters, rinsing with water, dries and thus obtains 0.136 mg of the desired product. The filtrate is taken to dryness, then taken up with ethanol. One vacuum-filters, rinsing with ethanol, and obtains a supplementary 0.04 g of the product. (a) : singlet at 2.01 ppm (b) : singlet at 6.67 ppm (c) : singlet at 7.08 ppm (d) : singlet at 11.3 pp EXAMPLE 2 7-/2-(2 - amino - 4 - thiazolyl) - 2 - hydroxyimino - acetamidoj - 3 - acetoxymethyl - ceph - 3 - em - 4 - carboxylic acid, syn isomer.

Stage A: Ethyl 2-(2- tritylamino - 4 - thiazolyl) - 2 - hydroxyimino acetate, syn isomer.

One introduces 43.2 g of ethyl 2-(2- amino - 4 - thiazolyl) - 2 hydroxyimino - acetate, syn isomer, prepared in stage A of Example 1, into 120 cm3 of dry dimethyl formamide.

One chills to -35°C and introduces 32 cm3 of triethylamine, then 60 g of trityl chloride in fractions over 30 minutes. One allows the temperature to rise, observes a total dissolution, then heats to 30°C. After one hour one pours into 1.2 litres of iced water containing 40 cm of 22° Be hydrochloric acid.

One agitates in a bath of iced water, vacuum-filters, rinsing with normal hydrochloric acid, and forms into a paste· with ether.

One obtains 69.3 g of hydrochloride.

The free base is obtained by dissolving the product in 5 volumes of 10 methanol treated with 120% of triethylamine, then causing precipitation gently with 5 volumes of water.

Analysis: CggHggOgNgS; 1/4 HgO Calculated: C% 67.6 H% 5.1 67.5 5.1 N% 9.1 S% 6.9 8.8 6.8 NMR (CDClg, MHz) (a) : Triplet centred on 1.31'ppm J = 7 Hz (b) : Quadruplet centred on 4.37 ppm J = 7 Hz (c) : Singlet at 6.37 ppm (d) : Singlet at 7.28 ppm Stage Β: Ethyl 2-(2- trityl ami no - 4 - thiazolyl) - 2 tetrahydropyranyloxyimino - acetate, syn isomer.

One places 5.6 g of the product prepared in stage A in 56 cm of 5 redistilled dihydropyran. One places in a bath of iced water, then adds 2.4 g of para-toluenesulphonic acid.

One agitates for one hour, thirty minutes while leaving the temperature to return to ambient. One pours into a mixture of 100 cm3 of benzene, 100 cm3 of water and 2 cm3 of triethylamine.

One decants, washes with water, dries, vacuum-filters, rinses with benzene, then drives off the solvent. One takes up with isopropyl ether, initiates crystallisation, leaves overnight in a refrigerator, and vacuum-filters, rinsing with isopropyl ether. One obtains 4.42 g of the product - M.Pt . = 184°C.

Analysis: para-toluenesulphonic acid salt C3gH3g07N3S2 Calculated: C% 63.9 Hit 5.5 Found: 63.7 5.5 NMR (CDClg, 60 MHz) Nit 5.9 5.8 Sit 9.0 8.9 (e)^ 3~==c~NH (d) H (?) X LJ (b) (a) , CHg—CHg (a) b) (c) d e Triplet centred on 1,36 ppm Quadruplet centred on 4.39 ppm Singlet at 6.60 ppm Singlet at 6.91 ppm Singlet at 7.28 ppm 4560° Stage C: 2-(2- tri tyl ami no - 4 - thiazolyl) - 2 - tetrahydropyranyl oxyimino - acetic acid, syn isomer.

One places 4.56 g of the product prepared in stage B in 45 cm of dioxane and 8.4 cm of 2N sodium hydroxide. One takes to reflux for one hour thirty minutes. One chills in a bath of iced water, and the salt precipitates. One vacuum-filters, rinsing with aqueous dioxane, then with ether, and obtains 4.6 g of the sodium salt.

One obtains the acid by dissolving the product in 50 cm of dioxane, acidifies with formic acid (pH 5) and precipitates with 90 cm3 of water. M.Pt. = 180°C.

NMR (CDClg), 60 MHz) 6.69 ppm (proton of the thiazolic ring); 7.31 (aromatic).

Stage D: Tert-butyl 1 -{2. - (2 - tri tyl ami no - 4 - thiazolyl) - 2 tetrahydropyranyl - oxyimino - acetamido]- 3 - acetoxymethyl - ceph - 3 - em - 4 - carboxylate, syn isomer.

One introduces 0.362 g of the product obtained in stage C, together with 0.244 g of tert-butyl 7 - amino - 3 - acetoxymethyl - ceph - 3 20 em - 4 - carboxylate and 0.280 g of dicyclohexylcarbodiimide, into 4 cm of dry chloroform. One agitates for 2 hours at ambient temperature. One vacuum-filters the di cyclohexyl urea which is formed and rinses it with chloroform.

One drives off the solvent from the filtrate under reduced pressure dissolved in 1 cm3 of ether, then chromatographs on a column of silica, eluting with ether.

One combines the fractions having Rf = 0.38, drives off the solvent under reduced pressure, takes up in isopropyl ether, disintegrates the solid matter, and vacuum-filters, rinsing with, isopropyl ether. One obtains 0.184 g of the desired product.

Analysis:C43H45°8N5S2 Calculated: C% 62.7 Found: 62.8 NMR (CDC13, , 60 MHz) H% 5.5 N% 8.5 S% 7.8 5.S 8.1 7.5 (b) CH--O-C—CH, 2 i, 3 (b) : 2.07 ppm (c) : 5.46 ppm (d) : 6.76 ppm (e) : 7.28 ppm 4,5θ θθ Stage Ε: - ^2-(2- amino - 4 - thiazolyl ) - 2 - hydroxyimino - acetamido] 3 -.acetoxymethyl. - ceph - 3 - em - 4 - carboxylic acid, syn isomer. 638 mg of the product obtained according to stage D are agitated for 15 minut.es at ambient temperature in 1.8 cm of trifluoroacetic acid, q One. adds 18 cm of isopropyl ether and vacuum-filters off 404 mg of the product which is precipitated. These 404 mg of product are taken up and agitated for 15 minutes at 50°C with 2 cm3 of 50% aqueous formic acid. One concentrates to dryness under vacuum at 30°C, takes up with 1 cm of ethanol, adds a drop of pyridine and vacuum-filters off the desired product.

This product is identical to that obtained in Example 1.

EXAMPLE 3 7-^2-(2- tritylamino - 4 - thiazolyl) - 2 .- trityloxyimino 15 acetamido] - 3 - acetoxymethyl - ceph - 3 em - 4 - carboxylic acid, syn isomer.

Stage A: Ethyl 2-(2- tritylamino - 4 - thiazolyl) - 2 - trityloxyimino acetate, syn isomer.

One dissolves 1.08 g of ethyl 2 -(2- amino - 4 - thiazolyl) 2 - hydroxyimino - acetate, syn isomer, prepared in stage A of example 1, in 8 cm of chloroform. One adds 1.5 cm of triethylamine, then adds at +5°C over 25 minutes, a solution of 3 g of trityl chloride in 6 cm3 of chloroform, and agitates for one hour at ambient temperature. One washes 3 with 18 cm of water, 6 cm of normal hydrochloric acid and 3 times with cm of water. One dries, vacuum-filters and concentrates to dryness. The product is taken up with isopropanol in which it crystallises. One obtains 2.3 g of the desired product. M.Pt. = 140°C.

Stage B: Sodium salt of 2-(2- tritylamino - 4 - thiazolyl) - 2 trityl oxy imino-acetic acid, syn isomer. q One dissolved 0.7 g of the product obtained in stage A in 3.5 cm of warm dioxane. One heats to 110°C and adds 1 cm3 of 2N sodium hydroxide, drop by drop under agitation. One continues the agitation for 1 hour 50 minutes at a temperature close to reflux, cools and vacuum-filters the sodium salt which has precipitated.

This product is identical to that obtained in stage C of Example 1.

Stage C: 7-/2-(2- tritylamino - 4 - thiazolyl) - 2 - trityloxyimino - acetamido] - 3 - acetoxymethyl - ceph - 3 em - 4 - carboxylic acid, syn isomer.

One suspends 5.12 g of the sodium salt obtained in stage B in 3 cm of chloroform. One adds 50 cm of normal hydrochloric acid.

One agitates, decants and washes three times with 50 cm of water. One dries, vacuum-filters and concentrates to dryness. One obtains in this way 2-(2- tritylamino - 4 - thiazolyl) - 2 - trityloxyimino acetic acid, syn isomer.

The acid obtained is dissolved in 50 cm3 of methylene chloride.

One adds 1.6 g of dicyclohexylcarbodiimide, agitates for one hour at 4S.6 0 0 ambient temperature, vacuum-filters off the dicyclohexylurea, cools the solution to -J0°C and adds 1.1 g of 7 - amino - cephalosporanic acid 3 3 in solution in 10 cm of methylene chloride and 1.2 cm of triethylamine. One agitates for 2 hours at ambient temperature, adds 50 cm of normal hydrochloric o acid, agitates, decants, washes 3 times with 50 cm of water, dries, vacuumfill ters and concentrates to dryness.

One disintegrates the solid product in ethanol, and vacuum-filters off 2.36 g of the crude condensate. This product is transformed into the diethylamine salt of 7-(2- (2 - tritylamino - 4 - thiazolyl) - 2 10 trityl - hydroxyimino - acetamido] - 3 - acetoxymethyl - ceph - 3 - em 4 - carboxylic acid, syn isomer, by the action of diethylamine in ether.

The salt obtained is displaced by normal hydrochloric acid in the presence of methylene chloride until an acidic pH is obtained. The solution is washed with water, dried and taken to dryness.

One obtains 0.75 g of the desired product, in purified form.

EXAMPLE 4 7-(2- (2 - amino - 4 - thiazolyl )-2- hydroxyimino - acetamido] - acetoxymethyl - ceph - 3 - em - 4 - carboxylic acid, syn isomer.

The ditritylated product obtained in example 3 is dissolved in 5 cm 20 of 50% aqueous formic acid. One agitates for 15 minutes at 50°C, drives off the solvents, takes up in ether and vacuum-filters, rinsing with ether. One obtains in this way the product as its formate.

This formate is triturated in 3 cm of water containing several drops of pyridine (pH±6). One vacuum-filters, rinsing with water, dries and so obtains the desired product which is identical to that obtained in Examples 1 and 2.

EXAMPLE 5 Sodium salt of 7 - [2 - (2 - amino - 4 - thiazolyl) - 2 - hydroxyimino acetamido] - 3 - acetoxymethyl - ceph - 3 - em - 4 - carboxylic acid, syn isomer.

One dissolves 0.613 g of the acid obtained in Example 1, 2 or 4 in a mixture of 2 cm3 of distilled water and 2 cm3 of a molar solution of sodium acetate in methanol. One adds 60 mg of active carbon, vacuumfilters, rinsing with 2 cm of methanol, concentrates to dryness under vacuum at 30°C and takes up in ethanol to obtain, after vacuum-filtering and drying, 0.432 g of the desired sodium salt.

EXAMPLE 6 Crystalline salt of 7 - - (2 - amino - 4 - thiazolyl) - 2 hydroxyimino - acetamido] - 3 - acetoxymethyl - ceph - 3 - em 4 - carboxylic acid, syn isomer.

One suspends 1.78 g of the acid obtained in Example 1, 2 or 4 in 8 cm3 of methanol, one adds 8 cm3 of a molar solution of sodium acetate in methanol, and the desired sodium salt crystallises out immediately.

One agitates for 15 minutes in order to complete the transformation, vacuum-filters, and washes with ethanol and then with ether. One obtains 1.53 g of a white crystalline product.

Analysis: C^H^OyNgS^Na Calculated M 38.88 H% 3.04 N% 15.11 SSS 13.84 Na% 4.96 38.8 3.1 15.1 13.8 4.85 Infra-red (Nojol Trade Mark ) > C * 0 1753, 1724, 1683 cm1 NH, OH, 3597 cm1 NMR (DMSO, 60 MHz) Singlet at 2.01 ppm — C— CH, II Singlet at 6.65 ppm (proton of the thiazole ring) Singlet at 7.16 ppm (NHg amine group) Example 7 One made up a preparation for injection of the formula: -7 - [l· - (2 - amino - 4 - thiazolyl) - 2 - hydroxyimino - acetamido] - 3 acetoxymethyl - ceph - 3 - em - 4 - carboxylic acid, syn isomer 500 mg - Sterile aqueous excipient qsv. 5 cm EXAMPLE 8 One made up gelatin capsules corresponding to the formula: -7 - J2 - (2 - amino - 4 - thiazolyl) - 2 - hydroxyimino - acetamido] - 3 - acetoxymethyl - ceph - 3 - em - 4 - carboxylic acid. syn isomer 250 mg -Excipient q.s. for a gelatin capsule up to 400 mg Pharmacological study of the product of the invention.. Activity in vitro.

Dilution method in liquid medium: One prepares a series of tubes between which one divides a sterile nutrient medium in equal quantities. One places in each tube increasing 20 quantities of the product to be studied, then each tube is inoculated with a bacterial strain. 6 0 0 After incubation for 24 or 48 hours in an oven at 37°C, the inhibition of growth is assessed by transillumination, so as to permit the determination of the minimum inhibiting concentration (M.I.C.

Q expressed in pg/cnr).

The following results have been obtained.

Product of Examples 1, 2 and 4 STRAINS M.I.C. in ug/ml ΣΤΗ” 48 H Staphylococcus aureus ATCC 6 538 Penicillin-Sensitive 0.5 0.5 Staphylococcus aureus UC 1 128 Penicillin-resistant 1 1 Staphylococcus aureus exp. n° 54 146 1 1 Streptococcus pyogenes A 561 0.02 0.02 Streptococcus faecal is 5 432 5 5 Streptococcus faecal is 99 F 74 10 10 Bacillus subtilis ATCC 6 633 0.5 1 Escherichia Coli Sensitive to Tetracyclin ATCC 9 637 0.05 1 Escherichia Coli Resistant to Tetracyclin ATCC 11 303 0.1 0.1 Escherichia Coli Exp. TOggBg 0.05 0.1 Escherichia Coli Resistant to Gentamicin Tobramycin R 55 123 D 0.2 0.2 Klebsiella pneumoniae Exp. 52 145 0.05 0.1 Klebsiella pneumoniae 2 536 Resistant to Gentamicin 0.2 0,5 Proteus mirabilis (indol-) A 235 0.1 0.2 Salmonella typhimurium 420 0.1 0.1 Enterobacter clocae 681 5 10 Providencia Du 48 2 2 Serratia Resistant to Gentamicin 2 532 5 10 »45600 .

Claims (29)

1. 7 - |_2 - (2 - amino - 4 - thiazolyl) - 2 - hydroxyimino - acetamido] 3 - acetoxy - methyl - ceph - 3 - em - 4 - carboxylic acid, syn isomer, and salts thereof formed with alkali metals, alkaline-earth metals, 5 magnesium or organic amines.

2. 7-(2-(2- amino - 4 - thiazolyl) - 2 - hydroxyimino - acetamido] 3 - acetoxy - methyl - ceph - 3 - em - 4 - carboxylic acid, syn isomer.

3. The sodium salt of 7-(2-(2- amino - 4 - thiazolyl) - 2 hydroxyimino - acetamido] - 3 - acetoxymethyl-ceph - 3 - em - 4 10 carboxylic acid, syn isomer.

4. 7-(2-(2- tritylamino - 4 - thiazolyl) - 2 - tri tyl oxy imino acetamido] - 3 - acetoxy - methyl - ceph - 3 - em - 4 carboxylic acid, syn isomer.

5. A process for the preparation of compounds of the general formula syn isomer» (wherein Ri represents a hydrogen atom or a saturated or unsaturated hydrocarbyl radical having from 1 to 4 carbon atoms), in which process a derivative of 7 - amino - cephalosporanic acid of the general formula: (wherein represents a group removable by acid hydrolysis or by hydrogenolysis) is reacted with an acid of the general formula: syn isomer, (wherein R 1 represents a group removable by acid hydrolysis or by hydrogenolysis or a chloracetyl radical, and R 1 ^ represents a group removable by acid hydrolysis or by hydrogenolysis , a chloracetyl radical 10 or a saturated or unsaturated hydrocarbyl radical having from 1 to 4 carbon atoms) or a functional derivative thereof, to obtain the corresponding compound of the general formula: syn isomer, (wherein Aj, Rj and R'j are as defined hereinbefore) which compound XI is treated with one or more of acid hydrolysis agents, hydrogenolysis agents and thiourea to obtain the desired product of general formula lb.

6. A process as claimed in claim 5, in which the first step comprises reacting a compound of the general formula: 10 (wherein Aj, is as defined in claim 5) with an anhydride or the acid chloride of an acid of general formula II.

7. A process as claimed in claim 6, in which the anhydride is that formed jn situ by the action of isobutyl chloroformate or di cycl ohexylcarbodi imide on the free acid.

8. A process as claimed in claim 6, in which the reaction of the 5 acid chloride or the anhydride formed with isobutyl chloroformate is carried out in the presence of a basic agent.

9. A process as claimed in claim 8, in which the basic agent is an alkali metal carbonate, N-methyl morpholine, pyridine or triethylamine.

10. 10. A process as claimed in any of claims 5 to 9, in which the compound of general formula XI is treated with formic acid, trifluoroacetic acid, acetic acid, or the zinc-acetic acid system.

11. A process as claimed in any of claims 5 to 10, in which the compound of general formula XI is treated with hydrogen in the 15 presence of a catalyst.

12. A process as claimed in any of claims 5 to 11, in which thiourea is reacted with a compound of general formula XI containing at least one chloroacetyl group in a neutral or acidic medium.

13. A process for the preparation of a compound of general 20 formula I (as defined hereinbefore) as claimed in any of claims 5 to 12 in which the starting material of general formula II is prepared by reacting an appropriate compound of the general formula: ' 45600 syn isomer, (wherein R' represents a hydrogen atom, a group removable by acid hydrolysis or by hydrogenolysis or a saturated or unsaturated alkyl radical having from 1 to 4 carbon atoms and alk represents an alkyl radical having from 1 to 4 carbon atoms) firstly with a base and then with an acid to obtain a product of the general formula: (XII) syn isomer t (wherein R' is as defined hereinbefore) which product is treated with a compound capable of introducing a group removable by acid hydrolysis or by hydrogenolysis or a compound capable of introducing the chloracetyl group to obtain the desired product of general formula II.

14. A process as claimed in claim 13, in which the base used to saponify the product of formula IV is sodium hydroxide, potassium hydroxide or barium hydroxide.

15. A process as claimed in claim 13 or claim 14, in which the 5 acid used to isolate the acid of formula XII is dilute hydrochloric acid, acetic acid or formic acid.

16. A process as claimed in any of claims 13 to 15, in which the compound capable of introducing a group removable by acid hydrolysis or by hydrogenolysis is trityl chloride. 10

17. A process as claimed in any of claims 13 to 15, in which the compound capable of introducing a group removable by acid hydrolysis or is by hydrogenolysis ^tert-butyl chloroformate, tert-butyl azidoformate, tri chloroethyl or benzyl chloroformate, the mixed formylacetic anhydride, a benzyl halide, a benzhydryl halide, phthalic anhydride 15 or N-carbethoxyphthalimide.

18. A process as claimed in any of claims 13 to 15, in which the compound capable of introducing the chloroacetyl group is chloracetic anhydride or a chloracetyl halide.

19. A process for the preparation of a compound of general formula I 20 (as defined hereinbefore) which employs a compound of the general formula: 456ΰθ C0 2 alk (Vc) R c syn isomer s (wherein Rj is as defined in claim 5, alk is as defined in claim 13 and RC, which is different to Rj represents a group removable by . and a acid hydrolysis or by hydrogenolysis or a chloracetyl group),/ in which the compound of general formula Vc is prepared by treating an appropriate, compound of the general formula: C0 2 alk (X) syn Isomer, 10 (wherein Rj is as defined in claim 5 and alk is as defined in claim 13) with a compound capable of introducing a group removable by acid hydrolysis or by hydrogenolysis, or a compound capable of introducing the chloracetyl group, as appropriate.

20. A process as claimed in claim 19, in which the group 15 removable by acid hydrolysis or by hydrogenolysis represented by Rj is the trityl group.

21. A process as claimed in claim 19, in which R c represents a tetrahydropyranyl radical.

22. A process as claimed in any of claims 5 to 21 and substantially as described herein with reference to any one of 5 Examples 1 to 6. (as defined hereinbefore)

23. A compound of general formula I/whenever prepared by a process as claimed in any of claims 5 to 22.

24. A pharmaceutical composition containing, as active principle, one or more of the compounds as claimed in any of claims 1 to 4, in 10 association with a suitable pharmaceutical vehicle.

25. A pharmaceutical composition as claimed in claim 24, in which the active principle comprises one or more of 7-0-(2- amino 4 - thiazolyl) - 2 - hydroxyimino - acetamid^ - 3 - acetoxy - methyl - ceph - 3 - em - 4 - carboxylic acid, syn isomer, and salts thereof 15 formed with alkali metals, alkaline-earth metals, magnesium and organic amines.

26. A pharmaceutical composition as claimed in claim 25, in which the active material is 7 -C2 -(2- amino - 4 - thiazolyl) - 2 hydroxyimino - acetamido] - 3 - acetoxymethyl - ceph - 3 - em - 4 20 carboxylic acid, syn isomer.

27. A pharmaceutical composition as claimed in claim 25, in which the active material is the sodium salt of 7-(2-(2- amino - 4 thiazolyl) - 2 - hydroxyimino - acetamido] - 3 - acetoxymethyl - ceph 3 - em - 4 - carboxylic acid, syn isomer. ‘· 45600

28. A pharmaceutical composition as claimed in claim 25 and substantially as described herein with reference to Example 7 or Example 8.

29. A compound of general formula: syn isomer, wherein R-j, R 1 ^ and A-j are as defined in claim 5. Dated this 1st day of April 1977,