SE438508B - NEW CEPHALOSPORIN ANTIBIOTICS AND PROCEDURES FOR PRODUCING THEREOF - Google Patents

Description

7904578 7 a new class of cephalosporin antibiotics, containing one 7β-he-etherified oximino) -acylamido group, the oximino group has sight configuration. Donna class of antibiotic compounds is characterized by high antibacterial activity with respect to a diversity of gram-positive and gram-negative organisms as well particularly high stability against β-lactenases, which are generated by various gram-negative organisms.

The discovery of this class of compounds has stimulated further research in the same field in order to find compounds, which have improved properties, for example with with regard to specific classes of organisms and in particular 1 gram-negative organisms.

British Patent Specification 1,496,757 discloses cephalopods sporin antibiotics, which contain a 7f ~ acylamido group with the formula Flcwcovna- A R II N i! , ~ \, q '\ I CO, ontn fl mc (c_12) n cm (A) have R wherein R is a thienyl or furyl group, RA and RB may have many different meanings and for example are C 1-4 alkyl groups or together with the carbon atom to which they are attached form a C 3-7 cycloalkylidene group and m and n are each 0 or 1 with the proviso that the sum of m and n is 0 or 1, wherein the compounds are syn-isomers or mixtures visual and anti-isomers containing at least 90% of its isomers. The 3-position of the cephalosporin molecule may be unsubstituted or may contain a variety of possible substituents. These compounds have been shown to be special good activity with respect to gram-negative organisms. ._.______: ...-- J ..- ___- iår- _, »Ar - rw- fl -... a- ...___ V 7904578 7 other compounds with a similar structure have been developed going from these associations in order to further find antibiotics with improved broad-spectrum antibiotic activity and / or high activity with respect to gram-negative organisms more. Such lines of development have not included variations only in the 7β-acylamido group of the above formula but also the introduction of special groups into the 3- of the cephalosporin molecule position.

Thus, for example, it is disclosed in the Belgian patent specification 852,427 cephalosporin antibiotic compounds, which fall within within the framework of British Patent Specification 1, 399,086 and wherein the group R in the above formula (A) may be replaced by a diversity of organic groups including 2-aminothiazole 4-yl and the sulfur atom in the oxyimino group is attached to an ali- phatic hydrocarbon group¿ which may itself be a substituted example- show with carboxy. In such compounds, the substituent in 3- the position is limited to an acyioxymethyl, hydroxymethyl, formyl- or optionally substituted heterocyclic thiomethyl- QICUQP.

Furthermore, Belgian Patent 836.83 discloses cephalopods trace compounds, wherein the group R in formula (A) above can be obtained with, for example, 2-aminothiazol-4-yl and the oxyimino group is a hydroxyimino or blocked hydroxyimino group, e.g. pelvis and methoxyimino group. In such compounds, cephalo- the 3-position of the sporin molecule substituted by a methyl group, which itself may be substituted by groups, which derive from a large number of nucleophilic compounds described therein.

In the above-mentioned patent specification such compounds are not attributed no antibiotic activity but is mentioned only as an intermediate products for the production of those described in the patent specification antibiotics.

Belgian Patent 853,545 discloses cephalosporin antibiotics, wherein the vf-acylamiaosidokeajan is primarily a 2- (2- aminothiazol-4-yl} -2- (syn) -methoxyimino-acetamido group and 1904578 7 substituentone in the 3 ~ position is defined in the broad way that disclosed in the above-mentioned Belgian patent specification 836.8l3.

It has now been shown that by an appropriate choice of one small number of special groups in the 7ß position in combination with a methyl group in the 3-position, the cephalosporin compounds with particularly advantageous activity (as described in more detail below) with respect to a plurality usually existing pathogenic organisms.

The present invention provides cephalosporin antibodies. biotics of the general formula ma? x F., H h * LJ c. <: O.NH S in; Ra §_.a »--- N / m3 (i) “I \, -__ f: ¿_¿0w coon Oh wherein Ra and Rp, which may be the same or different, are each a C 1-4 alkyl group (preferably a straight chain alkyl group, i.e. a methyl, ethyl, n-propyl or n-butyl group and i in particular a methyl or ethyl group) or Ra and Rb together with the carbon atom to which they are attached form a C3-7 cycloalkylidene group (preferably a C3-5 cycloalkylidene group) alkylidene group), and non-toxic salts and non-toxic bolically labile esters thereof.

The compounds of the invention are syn-isomers. The iso- the more visual form is defined by the configuration of the group a R - ~ O.C.COÛH Oh .,.....--_......._in. ._. _... ._, 7904578 with respect to the carboxamido group. In the present context the following structure is attributed to the syn configuration fff- c.c.o.rrf ~: - u 1 'J -uçj More / Jov- IN Rb It will be appreciated that since the compounds of the invention gene are geometric isomers, some mixture with the corresponding anti-isomer may occur.

The solvates also fall within the scope of the present invention (especially the hydrates) of the compounds of formula (I). Also esters of the compounds of formula (I) fall within the scope of present invention.

The compounds of the present invention may exist in tautomeric forms (with respect to the 2-aminothiazolyl group) and it will be appreciated that such tautomeric forms, e.g. the iminothiazolinyl form, falls within the scope of the present invention invention.

It should also be understood that when Ra and kb in the above different C 1-4 alkyl groups, the carbon atom to which they are bound includes an asymmetry center. Such compounds are diastereoisomers and the present invention include separate diastereoisomers of these compounds as well as mixtures thereof.

The compounds of the invention exhibit antibiotic broadband spectrum activity. Activity is unusually high with respect IN" / _.... § ..___.__.._._._..._- ..._. 7904578 7 on gram-negative organisms. This high activity includes also many β-lactamase-producing gram-negative strains.

The compounds also show high stability with respect to β-lactamases, which are produced by a variety of gram-negative organisms nismer.

The compounds of the invention have been found to be high activity with respect to different strains of Enterobac teriaceae (e.g., strains of Escherichia coli, Klebsiella pneumoniae, Salmonella typhimurium, Shigella sonnei, Enterobacter cloacae, Serratia marcescens, Providence- species, Proteus mirabilis and especially indole-positive Proteus organisms such as Proteus vulgaris and Proteus morganii) as well as with respect to strains of Haemophilus influenzae and good activity with respect to strains of Pseudomonas organisms, for example strains of Pseudomonas aeruginosa. This activity with respect to strains of indole-positive Proteus and Pseudomonas organisms are if one considers the antibacterial activity of known 3-methylcephalosporin antibiotics.

In addition to exhibiting high antibiotic activity, 3- the methyl compounds of the present invention more advantageous that they in comparison with analogous compounds which exhibit more sophisticated 3-substituents can be produced relatively relatively and economically on an industrial scale based on readily available starting materials, i.e. peni- cillin G or penicillin V.

Undigested salt derivatives, which can be prepared by turnover of one or both of the carboxyl groups of the compounds with the general formula (I), comprises salts of inorganic bases such as alkali metal salts (e.g. sodium and potassium salts) and alkaline earth metal salts (for example calcium salts), amino acid salts (eg lysine and arginine salts) and salts of organic bases (for example procaine, phenylethyl benzylamine, dibenzylethylenediamine, ethanolamine, diethanol 7904578 7 ænin and N-methylglucosamine salts). Other non-toxic salt derivatives includes acid addition salts, which are prepared, for example, with hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid acid, formic acid or trifluoroacetic acid. The salts may also come in the form of resinates, which are prepared, for example, with a polystyrene resin or a crosslinked polystyrene-divinyl- benzene copolymer resin containing amino or quaternary near amino groups or sulfonic acid groups, or with one resin containing carboxyl groups, for example a polyacrylic acid resin. Soluble base salts (eg alkali metal salts such as the sodium salt) of the compounds of formula (I) may be used for therapeutic use due to the rapid resorption tion of such salts in the body upon administration. About exem- pelvis insoluble salts of the compounds (I) are desirable for a special application, for example for use in depot prepared, such salts can be prepared in a conventional manner, for example with suitable organic amines.

These and other salt derivatives, such as the salts with toluene-p- sulfonic acid and methanesulfonic acid, can be used as intermediates products in the manufacture and / or purification of the present compounds of formula (I), for example in the following described procedures.

Oqiftiqa metabolically labile ester derivatives, which can be prepared by esterification of one or both of the carboxyl groups in the parent compound of formula (I), comprises acyloxyalkyl- esters, for example lower alkanoyloxy-methyl or ethyl esters such as acetoxymethyl or ethyl or pivaloyloxymethyl estrar. In addition to the above ester derivatives, the present invention comprises invention compounds of formula (I) in the form of other logically acceptable equivalents, i.e. physiologically acceptable only compounds which, like the metabolically labile ones the esters are converted in vivo to the antibiotic parent the compound of formula (I). _, _...._ ~ __._..- ..,.-..... _... w--. ..._- n 7904578 7 Preferred compounds of the present invention are below listed compounds of formula (I) and their non-toxic salts and non-toxic metabolically labile esters, namely (aR, 7R) -7 - {(z) -2- (2-aminothiazol-4-yl) -2- (2-carboxy- prop-2-oxyimino) acetamido: -3-methylcef-3-em-4-carboxylic acid; (6R, 7R) -7-yl) -2- (2-aminathiazol-4-yl) -2- (1-carboxy- cyclobut-1-oxyimino) acetamidoi-3-methylcef-3-em-4-carboxylic acid acid, and (en, vn) -7-β- (z) -2- (z-aminothiazol-α-yl) -2- (i-carboxy- cyclopent-1-yloxyimino) acetamido] -3-methylcephyl-3-em-4-carboxylic acid acid.

Other examples of compounds of formula (I) according to The present invention includes those wherein the groups Ra and Rb has the meanings given in the table below.

CHART Ra Rb a) Alkyl groups CH3 C2H5 Cz C s Cz fi s ; b) Cycloalkylidene groups z l the cyclopropylidene cyclohexylidene The compounds of formula (I) may be used for the treatment of a variety of diseases caused by pathogenic bacteria in humans and animals, such as respiratory tract infections and in the urinary tract. 7904578 7 According to another embodiment of the invention is provided a process for the preparation of an antibiotic compound of the general formula (I) or a non-toxic salt or a non-toxic metabolically labile ester thereof, which process characterized by a man acylates a compound of the formula H7: ____ L_ ~ _¿f 'H (11) 1 O? NY J | .

CGORL ; ïf _ '"“ _ ÛH., wherein B is ï> S or j> S -: O (u- or fi -) and R1 is hydrogen or a carboxyl blocking group, for example the residue of an ester-forming aliphatic or araliphatic alcohol or an ester-forming phenol, silanol or stannanol, said alcohol, phenol, silanol or stannanol preferably contain holds 1 to 20 carbon atoms, and the dashed line bridges 2-, 3- and 4 ~ positions indicate that the association is a cef-2-em or cef-3-em compound, or a salt, for example an acid addition salt, which is prepared, for example, with a mineral acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid or phosphoric acid or an organic acid such as methanesulfonic or toluene-p-sulfonic acid, or an N-silyl- derivatives thereof with an acid of the formula 5 N \ I -your .

** ~ "- ~ C, LÛ" * ala; a 3 u \\ F, - 7 0.C-CORB lair f » QIII) 7904578 7 10 wherein Ra and Eb have the meanings given above, R2 is a carboxyl ~ blocking group, for example as described for R1, and R 3 is an amino or protected amino group, or with an opposite corresponding acylating agent, after which, if necessary and / or desired, perform any of the following reactions in lig scheme; i) conversion of an A2 isomer to the desired 33-isomer, ii) reduction of a compound, wherein B is j ~ S-ë = 0, to a compound wherein B is;> S, iii) conversion of a carboxyl group to a non-toxic salt or a metabolically unstable ester function and iv) removal of any carboxyl blockers and / or N-protecting groups. eg / In the above procedure / is the starting material of the formula (II) preferably a compound wherein B is EVS and the dashed line that line represents a cef-3-em compound.

Acylating agents which can be used in the preparation of compounds of formula (I) include acid halides, in particular acid chlorides or bromides. Such acylating agents can be produced by reacting an acid (III) or a salt thereof with a halogenating agent, for example phosphorus pentaclo- rid, thionyl chloride or oxalyl chloride.

Acylations, which use acid halides, can be performed in aqueous aqueous and anhydrous reaction media, preferably at temperature turns of from -5o ° to + 5o ° c, preferably at from -2o ° to + 30 ° C, if desired in the presence of an acid scavenger.

Suitable reaction media include aqueous ketones such as as aqueous acetone, esters such as ethyl acetate, halogenated hydrocarbons such as methylene chloride, amides such as dimethylacetamide, nitriles such as acetonitrile or mixtures of two or several of such solvents. Suitable acid-binding agents include tertiary amines (for example triethylamine or di- methylaniline), inorganic bases (for example calcium carbonate) or sodium bicarbonate) and oxiranes such as lower 1,2-alkylene 7904578 7 ll oxides (for example ethylene oxide or propylene oxide) which bind the hydrogen halide liberated in the acylation reaction.

Acids of formula (III) may themselves be used as acylating agents. agents in the preparation of compounds of formula (I).

Acylations utilizing acids (III) are conveniently carried out in the presence of a condensing agent, for example a carbodiimide as N, N'-dicyclohexylcarbodiimide or N-ethyl-N'-γ-dimethyl- aminopropylcarbodiimide, a carbonyl compound such as carbonyl diimidazole or an isoxazolium salt such as N-ethyl-5-phenyl- isoxazolium perxlorate.

Acylation can also be performed with other amide-forming derivatives of acids of formula (III), such as, for example, an activated ester, a symmetrical anhydride or a mixed anhydride (produced for example with pivalic acid or with a haloformate such as a lower alkyl haloformate). Mixture anhydrides can also with phosphorus-based acids (for example phosphoric acid or phosphoric acidity), sulfuric acid or aliphatic or aromatic sulfonic acids (for example toluene-p-sulfonic acid). An active ester can be conveniently prepared in situ during use for example 1-hydroxybenzotriazole in the presence of a condensing agent, as indicated above. Alternatively, it can activated ester is prepared in advance.

Acylation reactions involving the free acids or theirs the above-mentioned amide-forming derivatives are conveniently carried out in an aqueous free reaction medium, for example in methylene chloride, tetra- hydrofuran, dimethylformamide or acetonitrile.

If desired, the acylation reactions can be carried out in the presence of a catalyst such as 4-dimethylaminopyridine.

The acids of formula (III) and the corresponding acylating agent can mn so desired, prepared and utilized in the form of their acid addition salts. Acid chlorides can thus be suitably used such as the hydrochloride salts and acid bromides such as hydrobromide 7904578 7 the salts.

The reaction product can be separated from the reaction mixture, which may, for example, contain unreacted cephalosporin starting materials and other substances, through a variety of processes including recrystallization, ionphoresis, column chromatography and use of ion exchangers (for example by chromatography engraving on ion exchange resins) or macroreticular resins.

The L2 cephalosporin ester derivatives obtained according to the process according to the invention can be converted into the corresponding A3 derivatives, for example by treating the A2 ester with a base such as pyridine or triethylamine.

A cef-2-em reaction product can also be oxidized to obtain of the corresponding cef-3-em-1-oxide, for example by reaction with a peracid such as peracetic acid or m-chloroperbenzoic acid; the resulting sulfoxide can, if desired, then be reduced in the manner described below to obtain the corresponding cef-3- em-sulfide.

If a compound is obtained in which B is j> S <> O, this compound can be converted to the corresponding sulphide by, for example, reduction of the corresponding acyloxisulfonium or alkoxysulfonium salt, produced in situ through turnover with, for example, tyl chloride in the case of an acetoxysulfonium salt, thereby reducing is achieved, for example, with the aid of sodium additionite or with iodide ions as in a solution of potassium iodide in one water-miscible solvent, for example acetic acid, acetone, tetrahydrofuran, dioxane, dimethylformamide or di- methylacetamide. The reaction can be carried out at a temperature of from -2 ° to + 50 ° C.

Metabolically labile ester derivatives of the compounds of the formula (I) can be prepared by reacting a compound with formula (I) or a salt or protected derivative thereof with the appropriate esterifying agent in question, such as an acyloxyalkyl _....? __.__.._._.> .. - - ..-..-... »-._. .-- V 7904578 7 13 nalid (for example -iodide), suitably in an inert organic solvents such as dimethylformamide or acetone, followed by removal of any protecting groups, if required.

Base salts of the compounds of formula (I) may be prepared by reacting an acid of formalin (I) with the appropriate the base in question. So can, for example, sodium or potassium salts produced using resp. 2-ethylhexanoate or bicarbonate salt. Acid addition salts can be prepared by reacting a compound of formula (I) or a bolic labile ester derivative thereof with the appropriate acid in question.

If a compound of formula (I) is obtained as a mixture isomers, the syn isomer can be obtained, for example, by conventional methods such as crystallization or chromatography engraving.

For use as a starting material for the production of compounds of the general formula (I) according to the invention preferably use compounds with the general formula flour (III) and the corresponding acid halides and anhydrides in their isomeric visual form or in the form of mixtures of isomers and corresponding anti-isomers, which mixtures contains at least 90% of the syn isomer.

Acids of formula (III) (provided that Ra and Rb together with the carbon atom to which they are attached does not constitute a propylidene group) is prepared by etherification of a compound of the formula 'IN _: . “__. ,. fl ívänzr , n_ \ ........_....._..__. \ '. 7904578 7 14 wherein R 3 is as defined above and R 4 is a carboxyl blocking group, by reaction with a compound thereof general formula e.g. '-11 xx wherein Ra and RD have the meanings given above and T is halogen such as chlorine, bromine or iodine, sulfate or sulfonate such as tosilate, after which the carboxyl blocking group R4 is removed nas. Separation of isomers can be performed either before or after such a change. The preference reaction is carried out in generally in the presence of a base, for example potassium carbonate or sodium hydride, and is preferably carried out in an organic solution such as dimethyl sulfoxide, a cyclic ether such as tetrahydrofuran or dioxane or an N, N-disubstituted amide such as dimethylformamide. Under these conditions, the the configuration of the imino groups is essentially unchanged during the etherification reaction. The reaction should be carried out in the presence of a base if using an acid addition salt of a compound with formula (IV). The base should be used in an amount sufficient to to rapidly neutralize the acid in question.

Acids of the general formula (III) can also be prepared by reacting a compound of the formula .. 'w ... švï) .Hr f * q fi lie t ..-......_....._.._ VJQL * VL * - wherein R 3 and R 4 have the meanings given above, with a compound ..__.._.__, .__,,. , 7904578 7 with the formula wherein Ra, Rb and R2 have the meanings given above, after which removes the carboxyl blocking group R 4 and, if so, required, separates the visual and anti-isomers.

The latter reaction is particularly applicable to the formation of acids of formula (III), wherein Ra and Rb together with the carbon atom to which they are attached constitute a pylidene group. In this case, the relevant associations can join formula (VII) is prepared in a conventional manner, for example by means of the synthesis described in the Belgian patent specification 866,422 for the preparation of t-butyl-1-amino-oxycyclo- propane carboxylate.

The acids of formula (III) can be converted to the corresponding acid halides and anhydrides and acid addition salts by means of conventional methods.

It should be noted that in some of the above conversions it may be necessary to protect any sensitive groups per in the molecule of the compound in question in order to avoid non desirable side reactions. This can happen, for example, during any of above reaction sequences may be necessary to protect The NH 2 group in the aminothiazolyl radical, for example by trityl ring, acylation (for example chloroacetylation), protonation or other suitable method. The protection group can then removed in any suitable manner which does not cause degradation of the desired compound, for example by using 7904578 7 16 a possibly halogenated carboxylic acid in the case of a trityl- group, for example acetic acid, formic acid, chloroacetic acid or trifluoroacetic acid, or using a mineral acid, for example hydrochloric acid or mixtures of such acids, preferably in the presence of a protic solvent such as water, or by treatment with thiourea in question a chloroacotyl group.

Carboxyl blocking groups used in the preparation of compounds of formula (I) or in the preparation of required starting materials, are suitably groups, as light can be cleaved at an appropriate stage of the reaction sequence, preferably in the last step. However, it may be appropriate to use in certain cases non-toxic, metabolically labile boxyl blocking groups such as acyloxymethyl or ethyl (for example acetoxymethyl or ethyl) and pivaloyloxymethyl and maintaining these in the final product to obtain one suitable ester derivative for a compound of formula (I).

Suitable carboxyl blocking groups are well known in the art. and a list of representative blocked carboxylates groups are found in British Patent Specification 1,399,086.

Preferred blocked carboxyl groups include aryl (lower) alkoxy) carbonyl groups such as p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl and diphenylmethoxycarbonyl, lower alkoxycarbonyl such as t-butoxycarbonyl and lower haloalkoxyl carbonyl groups such as 2,2,2-trichloroethoxycarbonyl. Carboxyl- blocked groups can then be removed by any means preferably by the appropriate methods disclosed in the literature; for example, acid or base catalyzed hydrolysis is suitable in many cases as well as enzymatically catalyzed lights up.

The antibiotic compounds of the invention may be prepared for administration by any conventional means by analogy with other antibiotics, and in the context of the present therefore, pharmaceutical compositions »X 7904578 7 17 comprising an antibiotic compound according to the invention and suitable for use in human or veterinary medicine.

Such compositions may be provided for use on conventionally by means of the required pharmaceuticals carriers or excipients.

The antibiotic compounds of the invention may be prepared for injection purposes and may be provided in unit in ampoules or in multiple dose containers, even possibly with the addition of preservative medcl. The compositions may also be in the form of suspensions, solutions or emulsions in oily or aqueous vehicles and can contain additives such as suspending agents, stabilizers dispersants and / or dispersants. Alternatively can the active ingredient is present in powder form for situation with a suitable carrier, for example sterile pyrotropic gene-free water, before use.

If desired, such powder formulations may contain a suitable non-toxic base in order to improve the active ingredient water solubility and / or to ensure that the pH value of the aqueous preparation obtained when the powder is constituted with water is physiologically acceptable. Alternative the base may be present in the water with which the powder is present reconstituted. The base may be, for example, an inorganic base such as sodium carbonate, sodium bicarbonate or sodium acetate or an organic base such as lysine or lysine acetate.

The antibiotic compounds can also be prepared in the form of suppositories, which include, for example: conventional suppository bases such as cocoa butter or other glycerides.

Compositions for veterinary use may, for example prepared as intramammary preparations in either long-term active or fast-acting bases.

The compositions may contain from 0.1% and up, for example 7904578 7 18 0.1 - 99%, of the active substance, depending on the administration method. When the compositions comprise dosage units each unit preferably contains 50 - 1500 mg of the active va component. The dosage used in treatment of adults preferably amounts to 500 - 6000 mg per day, depending on the mode and frequency of administration. So for example, it is normally sufficient that in the treatment of adult administering l000 - 3000 mg per day intravenously nostril or intramuscularly. In the treatment of Pseudomonas infections, higher daily doses may be required.

The antibiotic compounds of the invention may be administered administered in combination with other therapeutic agents such as antibiotics, such as penicillins or other cephalopods sporinerÅ The invention is further illustrated by the following examples, wherein the temperature indications refer to degrees Celsius and "Petrol" refers to petroleum ether with a boiling point range of 40 - 60oC.

PRODUCTION OF INITIALS Try l Ethyl (Z) -2- (2-aminothiazol-4-yl) -2- (hydroxyimino) acetate To a stirred and ice-cooled solution of 292 g of ethyl acetoacetate in 296 ml of glacial acetic acid was added a solution of 180 g of sodium nitrite in 400 ml of water at such a rate that the reaction temperature the tour was kept below 100. Stirring and cooling were continued for about 30 minutes, after which a solution of 160 g of potassium chloride in 800 ml of water was added. Down obtained the mixture stirred for 1 hour. The lower oil phase was separated and the aqueous phase was extracted with diethyl ether. Combined extract was washed with the oil and washed successively with water 7904578 7 19 and saturated brine, dried and evaporated. The return standing oil, which solidified when allowed to stand, was washed with "Petrol" and dried in vacuo over potassium hydroxide, 309 g of ethyl (Z) -2- (hydroxyimino) -3-oxobutyrate were obtained.

A stirred and ice-cooled solution of 150 g of ethyl (Z) -2- (hydroxy- imino) -3-oxobutyrate in 400 ml of dichloromethane was treated dropwise ¿With 140 g sulfuryl chloride. The resulting solution was maintained room temperature for 3 days and then evaporated. Re- the column was dissolved in diethyl ether, washed with water until then the washings were practically neutral, dried and fumes. The residual oil (177 g) was dissolved in 500 ml ethanol and 77 ml of dimethylaniline and 42 g of thiourea were added while stirring. After 2 hours, the product was recovered through filtration, washed with ethanol and dried to give of 73 g of the title compound of the melting point 1ss ° (decomposition).

Experiment 2 Ethyl (Z) -2-hydroxyimino-2- (2-tritylaminothiazol-4-yl) acetate hydrochloride 16.75 g of trityl chloride was added portionwise over an additional 2 hours a stirred and cooled (-300) solution of 12.91 g of the product from Experiment 1 in 28 ml of dimethylformamide containing 8.4 ml triethylamine. The mixture was allowed to warm to 150 ° C 1 hour, stirred for a further 2 hours and distributed then between 500 ml of water and 500 ml of ethyl acetate. The or- The organic phase was separated, washed with 2 x 50 ml of water and was then shaken with 500 ml of 1N HCl. Precipitation was taken, washed successively with 100 ml of water, 200 ml ethyl acetate and 200 ml of ether and dried in vacuo to give holding 16.4 g of the title compound such as a white solid with melting point l84 - l86O (decomposed sharing). 7904578 7, 20 Experiment 3 Ethyl (Z) -2- (2-t-butoxycarbonylprop-2-oxyimino) -2- (2-tri- tylaminothiazcl-4-yl) acetate 34.6 g of potassium carbonate and 24.5 g of t-butyl-2-bromo-2-methyl- propionate in 25 ml of dimethyl sulfoxide was added to a stirred solution 49.4 g of the product from Experiment 2 i 200 ml of dimethyl sulfoxide and the mixture was stirred at room temperature temperature for 6 hours. The mixture was poured into 2 l of water, was stirred for 10 minutes and filtered. The solid material was washed with water and dissolved in 600 ml of ethyl acetate. Loose- The mixture was washed successively with water, 2N hydrogen chloride acid, water and saturated brine, dried and evaporated. des. The residue was recrystallized from petroleum ether (boiling point range 60 - 800) to obtain 34 g of the i title compound with melting point 123.5 - 1250. < Experiment 4 (Z) -2- (2-t-butoxycarbonylprop-2-oxyimino) -2- (2-tritylamino- thiazol-4-yl) acetic acid 2 g of the product from Experiment 3 were dissolved in 20 ml of methanol and 3.3 ml of ZN sodium hydroxide was added. The mixture is refluxed. was boiled for 1.5 hours and then concentrated. Eating The residue was taken up in a mixture of 50 ml of water, 7 ml of hydrochloric acid and 50 ml of ethyl acetate. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic solvents The compounds were combined, washed successively with water and saturated brine, dried and evaporated. the remainder was recrystallized from a mixture of carbon tetrachloride and "Petrol" to give 1 g of the title compound with melting point 152 - 1560 (decomposition). 7904578 7 21 Experiment 5 Ethyl- (Z) -2- (2-tritylaminothiazol-4-yl) -2- (1-t-butoxycarbonyl- cyclobut-1-oxyimino) acetate 55.8 g of the product from Experiment 2 were stirred under nitrogen for 400 ml dimethyl sulfoxide with 31.2 g finely powdered potassium carbonate at room temperature. After 39 minutes, 29.2 g of t butyl 1-bromocyclobutanecarboxylate. After 8 hours was added an additional 31.2 g of potassium carbonate. Additional potassium carbonate (6 portions of 16 g each) were added during the following 3 days and an additional 3.45 g of t-butyl-1-bromocyclobutane carboxylate was added after 3 days. After a total of 4 days the mixture was poured into about 3 l of ice-water and the solid The material was collected by filtration and washed well water and "Petrol". The solid was dissolved in ethyl acetate and the solution was washed twice with brine, dried over magnesium sulphate and evaporated to a foam.

This foam was dissolved in a 1: 2 mixture of ethyl acetate and "Petrol" and filtered through 500 g of silica gel. Evaporation of 60 g of the title compound as a foam, Vmax (CHBr 3) 3400 (NH) and 1730 cm -1 (ester).

Experiment 6 (Z) -2- (1-t-butoxycarbonylcyclobut-1-oxyimino) -2- (2-trityl- aminothiazol-4-yl) -acetic acid A mixture of 3.2 g of the product from Experiment 5 and 1.65 g potassium carbonate was refluxed in 180 ml of methanol and 20 ml water for 9 hours, after which the mixture was cooled to room temperature. temperature. The landing was concentrated and the remainder * was placed between ethyl acetate and water, to which was added 1.2 ml ZN HCl. The organic phase was separated and the aqueous phase was extracted. cured with ethyl acetate. The combined organic extracts washed with saturated brine, dried and evaporated to obtain 2.3 g of the title compound V, ia Imax (ethanol) 20 nm (nlcm 243). 7904578 7 22 Experiment 7 Ethyl (Z) -2- (1-t-butoxycarbonylcyclopent-1-yloxyimino) -2- - (2-tritylaminothiazol-4-yl) acetate 10 g of the product from Experiment 2 was stirred with 7 g of t-butyl-2- bromocyclopentanecarboxylate in 40 ml of dimethyl sulfoxide containing holding 10 g of potassium carbonate under nitrogen at 210 under 21 hours. The mixture was poured into 500 ml of ice-water and the gray the solid was collected by filtration, washed with water and air dried. Recrystallization of this solids from 500 ml of methanol gave 11.7 g of the indicated compound with melting point 179 - 1800; Omax (CHBr3) 3410 (NH), 1735 (ester), 1275 (ester) and 1 755 cm -1 (phenyl).

Experiment 8 (Z) -2- (1-t-butoxycarbonylcyclopent-1-yloxyimino) -2- (2- tritylaminothiazol-4-yl) acetic acid 625 mg of the product from Experiment 7 was refluxed with 0.5 ml of a 2N sodium hydroxide solution and 1 ml of water in 12 ml of methanol for 7 hours. The mixture was allowed to cool over nattem. After dilution with water, orthophosphorus was added. acid for setting the pH of the solution to 2. Precipitating The mixture was extracted with ether and the combined extracts washed with brine. After drying with magnesium sulphate The solvent was evaporated to give 493 mg of one rubber material. Recrystallization from diisopropyl ether gave 356 mg of the title compound, m.p. 171 - 1730; Max (CHBI3) 2500-3500 (OH and NH), 1755 (ester), 1692 (acid) and 755 and 170 cm -1 (phenyl). 7904578 7 PREFERRED EMBODIMENTS ACCORDING TO THE HPPTINHINGEN Example 1 a) diphenylmethyl- (6R, 7R) -EXX) -2- (2-t-butoxycarbonylprop-2- oxyimino) -2- (2-tritylaminothiazol-4-yl) acetamino} 3-methyl- cef-3-em-4-carboxylate A stirred solution of 2.86 g of the product from Experiment 4 and 2.09 g of diphenylmethyl- (6R, 7R) -7-amino-3-methylceph-3-em-4- carboxylate in 50 ml of dimethylformamide was cooled to 0 ° C and treated was added with 745 mg of hydroxybenzotriazole and 1.14 g of dicyclo- hexylcarbodiimide. The mixture was warmed to room temperature and stirred overnight. The mixture was filtered and white the solid was washed with a small amount of ethyl acetate.

The filtrate and washings were diluted with 100 ml of water and was extracted with ethyl acetate. The organic extracts combine was washed successively with ZN hydrochloric acid, water, sodium bicarbonate solution and saturated brine, dried and evaporated. The residue was eluted through a silica gel column with ethyl acetate. The product-containing eluate was recovered and concentrated to give 3.6 g of the title compound. given compound; .kmax (ethanol) 238 nm (Eââm 283), Åinf 263.5 mn (Eiâm 186); [Lålšo + 6,5 '(c 1,0, masa). b) (6R, 7R) -7 - [(Z) -2- (2-aminothiazol-4-yl) -2- (2-carboxyprop-2- oxyimino) acetamido--3-methylceph-3-em-4-carboxylic acid 20 ml of trifluoroacetic acid was added to a solution of 2.8 g of the product from step a) in 20 ml of anisole at 0 DEG. The mixture was stirred at room temperature for 2 hours and concentrated. The residue was dissolved in ether and re-evaporated. The new The residue was dissolved in ethyl acetate and extracted with saturated sodium bicarbonate solution. The aqueous extracts The pH was adjusted to 6 and the solution was washed with ethyl acetate. The aqueous phase was acidified to pH 1.5 under ethyl acetate and extracted with ethyl acetate. The combined bodies 7904578 7 The extracts were washed with saturated brine, dried and evaporated. The residue was dissolved in 40 ml of a warm 75% aqueous solution of formic acid, allowed to stand for 1 hour. The mixture diluted with water and filtered. The filtrate was concentrated. The residue was taken up in water, re-filtered and lyophilized. was obtained to obtain 900 mg of the title (pH 6 buffer) 236 nm (287) 261 nm 1% lcm l Association; Ä l% _ lcm 1720 (co, H), 1765 cm -1 inf (E 254), 296 nm (E 115; Vm (Nujol) 1530, 1665 (CONH), ear (ß-lactam).

Example 2 a) diphenylmethyl- (6R, 7R) -§] Z) -2- (1-t-butoxycarbonylcyclobut-1- oxyimino) -2- (2-tritylaminothiazol-4-yl) -acetamido] -3-methyl- cef-3-em-4-carboxylate A stirred solution of 2.2 g of the product from Experiment 6 and 2.2 g of diphenylmethyl- (6R, 7R) -amino-3-methylceph-3-em-4-carboxy- lat in 45 ml of dimethylformamide was cooled to 0 DEG C. and 655 mg of hydroxybenzotriazole was added, followed by 982 mg of dicyclohexyl carbodiimide. The mixture was warmed to room temperature and stirred. was stirred overnight. The mixture was filtered. The filtrate was diluted with 300 ml of water and extracted with ethyl acetate. The organic ones the extracts were combined, washed successively with 2N hydrochloric acid, sodium bicarbonate solution and saturated brine, dried and evaporated. The residue was eluted through a silica cone column with a 3: 1 mixture of ether and "Petrol". The the appropriate fractions were concentrated to give 2.1 g of the title compound; inf (ethanol). 1% - W lcm 67) '\ 244 mn (Eíäm 238), 302 nm (E (cHBr3) 1520, 1682 max (conn), 1722 (coza), 1788 cm_1 (β-lactam). 7904578 7 25 b) (6R, 7R) -7- [(Z) -2- (2-aminothiazo] -4-yl) -2- (1-carboxy] cyclo- but-1-oxyimino) acetamido-13-methylccc-3-em-4-carboxylic acid 8 ml of trifluoroacetic acid was added to a solution of 1.9 q of the product from xteq a) in 2 ml of anisole at 00. The mixture was stirred for 5 minutes and 32 ml of trifluoroacetic acid were added. Among- the mixture was stirred at room temperature for 30 minutes and centered. The residue was dissolved in ethyl acetate and fumes. The residue now obtained was dissolved in ethyl acetate and extracted with saturated sodium bicarbonate solution. Water- extracts with a pH of 7 - 7.5 were washed with ethyl acetate, acidified to pH 1.5 under ethyl acetate and extracted into des with ethyl acetate. The combined organic extracts are dried was evaporated and evaporated. The residue was dissolved in 30 ml of formic acid an addition of 9 ml of water and the mixture was stirred at room temperature. temperature for 2 hours. The mixture was diluted with 200 ml of aqueous and filtered. The filtrate was concentrated. The remainder taken up in water, refiltered and lyophilized to give holding 730 mg of the title compound; 1% 1% lmax (pm 6 btffert) 241 nm (slcm 272), linfzsz nm 291 nm (E1% 129); Ed 20+ 62.5 '(C 1.0, Dmso). lcm D 269), , §Example 3 a) Alphenylmethyl- (6R, 7R) -7-f (z) -2- (1-t-but® x-carbonylcyclo- pent-1-yloxyimino) -2- (2-tritylaminothiazol-4-yl) acetamido] -3- methylcef-3-em-4-carboxylate 1 g of the product from Experiment 8 was dissolved in 25 ml of tetrahydro furan and 760 mg of diphenylmethyl- (6R, 7R) -7-amino-3-methylceph-3-em- 4-carboxylate and 380 mg of 1-hydroxybenzotriazole hydrate was stirred. Once a clear solution had been formed, 520 mg of dicyclohexylcarbodiimide were added and the mixture was stirred. 24 hours at 210 DEG C. The solution was filtered and the filtrate concentrated. This solution was percolated through a card column of 30 mg of neutral alumina eluting with a 1: 3 mixture of ethyl acetate and petroleum ether with boiling point 7904578 7 20 the range 60 - 800. The product was applied to a column of silica (silica gel 60G with a particle size of meter; 50 g). The column was eluted with a 1: 3 mixture of ethyl acetate and petroleum ether with a boiling point range of 60 -800 at a pressure of 8 Jb / sq. in. and evaporation of the appropriate the fractions in question gave 510 mg of the title š9 + 25.2 '(c 0.95, cnc13), v (cHßr3) 3405, 3275 (NH), 1790 (β-lactam), 1727 (esters) 1683 1 the compound as a foam; max and 1527 cm -1 (amia). b) (sR, 7R) -7-η (z) -2- (2-aminothiazol-4-yl) -2- <1-Carb021cyc pent-1-yloxyimino) -acetamidof-3-methylceph-3-em-4-carboxylic acid, trifluoroacetate salt 408 mg of the product from step a) were mixed with 0.3 ml of anisole and 2 ml of trifluoroacetic acid were added. After 2 hours at 210 The solution was concentrated and the residue was poured into 20 ml of water. ten. This mixture was washed with ether (3 times), whereupon each time the ether was re-extracted with water. They combined the water layers were evaporated to dryness and the residue triturated. was treated with ether to give 88 mg of the title (PH e buffer) 239.5 nm (z1% 304), given compound; Ä lem “Max 290 (ßíâm 131), Jmax (Nujo1) 3700 - 2100 (wH *, NH och infl.

OH), 1770 (β-lactam and 1680 cm -1 (amide and acid).

PHARMACEUTICAL COMPOSITIONS Example A - Dry powder for injection purposes Composition per ampoule (6R, 7R) -7 - [(Z) -2- (2-aminothiazol-4-yl) -2- (2-carboxypropyl) -2- oxyimino) acetamide-3-methylceph-3-em-4-carboxylic acid 500 mg Sodium carbonate, anhydrous ll3 mg ......-.._...... _.... _ _ 7904578-7 27 Method The sterile cephalosporin antibiotic was mixed with sterile sodium carbonate under aseptic conditions. The mixture is was aseptically placed on glass ampoules under a sterile atmosphere nitrogen. The ampoules were sealed using rubber discs or plugs held in place by aluminum covers, which prevented gas exchange or penetration of microorganisms.

The product was intended for reconstitution by dissolution in water for injection or other suitable sterile vehicle shortly before administration.

Example B - Dry powder for injection Sterile disodium salt of (6R, 7R) -7 - [(Z) -2- (2-aminothiazol-4-yl) - -2- (1-carboxycyclobut-1-oxyimino) acetamido-3-methylceph ~ 3-em ~ 4- carboxylic acid was filled into glass ampoules so that each ampoule came to contain an amount equivalent to 500 mg of antibiotic acid.

The filling was performed aseptically under an atmosphere of sterile nitrogen. The ampoules were closed with rubber discs or plugs, held in the position of aluminum covers, which prevented gas exchange or penetration of microorganisms. The product was intended for reconstitution by dissolving in water for in ~ injection or other suitable sterile vehicle, shortly before administration.

Claims (9)

7904578-7 28 PATENTKRAV _l. Cephalosporin antibiotics with the general formula NHZ / \ å! Eš / S (I) c.co.NH H Ra 4 N \ o.šë.coon 0 C93 Rb cooH wherein Ra and Rb, which may be the same or different, are each a C1-4 alkyl group, or Ra and Rb together with the carbon atom to which they are attached constitutes a C 3-7 cycloalkylidene group, as well as non-toxic salts and non-toxic esters thereof, in vivo Transferable to the corresponding acids. Compounds according to claim 1, characterized in that at least one of the substituents Ra and Rb is a methyl or ethyl group. Compounds according to claim 1, characterized in that Ra and Rb together with the carbon atom to which they are attached form a C 3-5 cycloalkylidene group. 4. A compound according to claim 1, wherein it is (6R, 7R) -7-I (Z) -2- (2-aminothiazol-4-yl) -2- (2-carboxypropyl). 2-oxyimino) acetamidol-3-methylcef-3-em-4-carboxylic acid and its non-toxic salts. A compound according to claim 1, characterized in that it is (6R, 7R) -7-I (Z) -2- (2-aminothiazol-4-yl) -2- (1-carboxycyclobut-1- oxyimino) acetamido [-3-methylcef-3-em-4-carboxylic acid and its non-toxic salts. 7904578-7 29 A compound according to claim 1, characterized in that it is (6R, 7R) -7-I (Z) -2- (2-aminothiazol-4-yl) -2- (1-carboxycyclopent-1- yloxyimino) acetamidol-3-methylceph-3-em-4-carboxylic acid and its non-toxic salts. A process for the preparation of an antibiotic compound of the general formula NH 2 // \\ v H. S (I) C .CO.NH Il Ra 4 N \ o.'f ~ coo fl ° (m3 Rb coon wherein Ra and Rb, which may be the same or different, are each a C1-4 alkyl group, or Ra and Rb together with the carbon atom to which they are attached form a C 3-7 cycloalkylidene group, or a non-toxic salt or a non-toxic ester thereof, in vivo transferable to the corresponding acid, characterized in that acylating a compound of the formula H ) (m3 cooul wherein B is S or SO, R1 is hydrogen or a carboxyl blocking group and the dashed line of formula (II) bridging the 2-, 3- and 4-positions indicates that the compound is a cef-2-em or cef-3-em compound, or an acid addition salt or N-silyl derivative thereof, with an acid of the formula 1904578-7% Å í = ;: zï-c.cooH (III) 3 y \ * o, cooR 2 R b wherein R a and R b are as defined above, R 2 is a carboxyl blocking group and R 3 is an amino or protected amino group, or with a corresponding acylating agent, after which, if required and / or desired in in each particular case, any of the following reactions are carried out in the appropriate order: i) conversion of an? 1 isomer to the desired [β 3 isomer, ii) reduction of a compound wherein B is: DS- # 0 to a compound wherein B is:> S, iii) conversion of a carboxyl group to a non-toxic salt or a non-toxic ester function, in vivo transferable to the corresponding acid, and iv) removal of any carboxyl blocking and / or N-protecting groups. 8. A process according to claim 7, characterized in that the starting material of formula (II) is a cef-3-em compound, wherein B is IIS. An antibiotic compound according to any one of claims 1 to 6, optionally in combination with a pharmaceutical carrier or excipient, for use in human or veterinary medicine.