NL193284C - Antibacterial preparation and a 7 - [(Z) -2- (etherified oxyimino) -2- (2-aminothiazol-4-yl) -acetamido] -3-ammoniomethyl-3-cephem-4-carboxylic acid suitable for use in this preparation derivative. - Google Patents

Description

1 193284

Antibacterial preparation and a 7 - [(Z) -2- (etherified oxyimino) -2- (2-aminothiazol-4-yl) acetamido] -3-ammoniomethyl-3-cephem-4-carboxylic acid derivative suitable for use in this preparation

The invention relates to an antibacterial composition comprising a 7 - [(Z) -2- (etherified oxyimino) -2- (2-5 aminothiazol-4-yl) acetamido] -3-ammoniomethyl-3-cephem-4-carboxylic acid derivative and an inert carrier.

Such a preparation is known from British patent application 2,040,921. In the 7 - [(Z) -2- (etherified oxyimino) -2- (2-aminothiazol-4-yl) acetamido] -3-ammoniomethyl-3-cephem-4-carboxylic acid derivatives to be used according to this British patent application the oxyimino moiety etherified with a 1-carboxy-1,1-dialkylmethyl group, in which each of the alkyl radicals has 1-4 carbon atoms, or with a 10-1-carboxycycloalkyl group, in which the cycloalkyl radical has 3-7 carbon atoms, while the ammonio methyl group can be represented as -CH2NR1R2R3 (+), wherein R 1, R 2, and R 3 are each an alkyl group of 1-4 carbon atoms. The compounds have a broad spectrum of antibacterial activity, with high activity against strains of Pseudomonas organisms and various Enterobacteriaceae, according to the British patent application. Test results are not disclosed in British patent application 2,040,921.

It has now been found that by using certain, hereinafter described, 7 - [{Z) -2- (etherified oxyimino) -2- (2-aminothiazol-4-yl) acetamido] -3-ammoniomethyl-3-cephem -4-carboxylic acid derivatives provide an antibacterial preparation which, in addition to having excellent activity against Enterobacteriaceae and acceptable to strong activity against Pseudomonas strains, also has good activity against Staphylococcus strains.

The invention therefore relates to the antibacterial preparation described in the preamble, characterized in that it is a compound of the formula 1 of the formula sheet, wherein R 2 is a straight or branched chain alkyl group with 1-4 carbon atoms or an allyl, 2- butenyl or 3-butenyl group, or it contains a non-toxic pharmaceutically acceptable salt or solvate or a non-toxic physiologically hydrolyzable ester thereof.

It should also be noted that the antibacterial activities of 7 - [(Z) -2- (etherified oxyimino) -2- (2-25 aminothiazol-4-yl) acetamido] -3- (substituted or unsubstituted methyl) -3-cephem -4-carboxylic acid derivatives have attracted much attention. The reason for investigating such compounds is evident from a publication by M. Ochiai et al. In Chem. Pharm. Bump. 25, 3115-3117 (1977) combining the good properties of 7 - [(Z) -2-methoxyimino-2- (2-furyl) acetamido] -3-aminocarbonyloxymethyl-3-cephem-4-carboxylic acid (cefuroxime) with the good properties of 7- [2- (2-aminothiazol-4-yl) acetamido] -3- (substituted or unsubstituted methyl) -3-cephem-4-carboxylic acid derivatives. The compounds described by Ochiai et al., Including 7 - [(Z) -2- (methoxyimino) -2- (2-aminothiazol-4-yl) acetamido] -3-acetoxymethyl-3-cephem-4-carboxylic acid (cefotaxime) with formula 19 of the present formula sheet, have good activity against various types of bacteria, including B-lactamase producing bacterial strains.

R. Bucourt et al., In Tetrahedron 34, 2233-2243 (1978) start from in a 7- [2- (2-aminothiazol-4-35 yl) acetamido] -3- (substituted or unsubstituted methyl) -3 -cephem-4-carboxylic acid incorporating a substituent at position 2 of the acetamido side chain. The 7 - [(Z) -2- (whether or not improved oxyimino-2- (2-aminothiazol-4-yl) acetamido] -3-acetoxymethyl-3-cefem-4-carboxylic acid derivatives, including cefotaxime, prepared by Bucourt et al. , have: a) a degree of activity relative to gram-positive bacteria that is comparable to that of the best 40 commercially available cephalosporins; b.with regard to gram-negative bacteria a degree of activity that far exceeds the activity of comparative compounds; c. high resistance to 3-lactamases, similar to that of cefuroxime and related compounds.

45 U.S. Patent 4,258,041 describes that 7 - [(Z) -2- (etherified oxyimino) -2- (2-aminothiazol-4-yl) acetamido] -3 - [(1-pyridinium) methyl] - 3-cephem-4-carboxylates exhibit a broad spectrum of antibacterial activity, with very high activity against Pseudomonas organisms. The oxyimino moiety etherifying group is the same 1-carboxy-1,1-dialkylmethyl group or 1-carboxycycloalkyl group as described above in British Patent Application 2,040,921. Preferred are the compound 7 - [(Z) -2- (2-carboxyprop-2-oxyimino) -2- (2-aminothiazol-4-yl) acetamido] -3 - [(1-pyridinium) methyl ] -3-cephem-4-carboxylate (ceftazidime) of the formula 20 of the present formula sheet.

U.S. Patent 4,278,671 describes 7 - [(Z) -2-methoxyimino-2- (2-aminothiazol-4-yl) acetamido] -3- (substituted or unsubstituted methyl) -3-cephem-4-carboxylic acid derivatives which exhibit a broad spectrum of antibacterial activity, including activity against gram-negative bacteria, and are resistant to β-lactamase. The compounds satisfy the formula 21 of the present formula sheet, wherein R 2 represents a hydrogen atom or an amino group protecting group and R 3 is a hydrogen atom or the rest of a nucleophilic compound. R3 may be a quaternary ammonium 193284 2 group, in which context pyridinium, substituted pyridinium, quinolinium, picolinium and lutidinium are mentioned. There is no suggestion that the quaternary ammonium group would be a fully saturated five-membered heterocyclic ring quaternized to the ring nitrogen atom. Neither alone nor read in conjunction with the literature discussed earlier in this specification, U.S. Patent 5 4,298,671 gives an expert reason to believe that a 7 - [(Z) -2- (etherified oxyimino) -2- {2 -aminothiazol-4-yl) acetamido] -3 - [(1-methyl-1-pyrrolidinium) methyl] -3-cephem-4-carboxylate of the formula 1, wherein R2 has the aforementioned meaning, in addition to excellent activity against Enterobacteriaceae and an acceptable to strong activity against Pseudomonas strains would also have a good activity against Staphylococcus strains. US Patent No. 4,168,309 describes cephalosporin derivatives of formula 22 of the present formula sheet wherein R is phenyl, thienyl or furyl; Ra and Rb are independently hydrogen, alkyl, cycloalkyl, phenyl, naphthyl, thienyl, furyl, carboxy, alkoxycarbonyl or cyano, or Ra and Rb taken together with the nitrogen to which they are attached form a cycloalkylidene or cycloalkenylidene ring; and m and n are each 0 or 1 such that the sum of m and n is 0 or 1; and R1, together with the nitrogen atom to which it is attached, is broadly defined and may be inter alia a saturated 5-ring. The compound of formula 19 of the present formula sheet is a preferred compound and is described in Example 5 of this U.S. Patent. The compounds of the present formula 22 have a broad spectrum of antibacterial activity, with high activity against β-lactamase producing gram negative organisms. Moreover, when at least one of Ra and Rb has a meaning other than hydrogen, the compounds of the present formula 22 have a high activity relative to Pseudomonas aeruginosa. Neither alone nor read in conjunction with the literature discussed earlier in this specification, U.S. Patent 4,168,309 gives an expert reason to believe that the 2-aminothiazol-4-yl group and another etherified oxyimino group containing 7- [ (Z) -2- (etherified oxyimino) -2- (2-aminothiazol-4-yl) acetamido] -3 - [(1-methyl-1 -25 pyrrolidinium) methyl] -3-cephem-4-carboxylates with the formula 1, wherein R2 has the aforementioned meaning, would have both an excellent activity against Enterobacteriaceae and an acceptable to strong activity against Pseudomonas strains as well as a good activity against Staphylococcus strains.

As shown on the formula sheet, the compounds of formula 1 have the "syn" or "Z" configuration relative to the -OR2 defined alkoxyimino or alkenyloxyimino group. The compounds of the formula 1 to be used in the antibacterial preparation according to the invention consist of at least 90% of the syn isomer. Preferably, the compounds of formula 1 are "syn" isomers that are essentially free of the corresponding "anti" isomers.

The pharmaceutically acceptable salts of the compounds of the formula I include the inorganic basic salts, such as the alkali salts (eg, the sodium and potassium salts) and the alkaline earth salts (eg, the calcium salts), ammonium salts, as well as the organic alkaline salts (eg, with triethylamine, procaine, phenethylbenzylamine, dibenzylethylenediamine) and the acid addition salts (for example, the salts with hydrogen chloride, hydrogen bromide, formic acid, nitric acid, sulfuric acid, methanesulfonic acid, phosphoric acid, acetic acid or trifluoroacetic acid). The physiologically hydrolyzable esters include the acyloxyalkyl esters, for example, (lower) alkanoyl (lower) alkyl esters, such as acetoxymethyl, acetoxyethyl and pivaloyloxymethyl.

The compounds of the formula 1 exhibit high antibacterial activity against various gram-positive and gram-negative bacteria and are effective in the treatment of bacterial infections of animals and humans. The compounds of formula 1 can be formulated for parenteral use in conventional manner using known pharmaceutical carriers and diluents and can be formed into unit dosage forms or multi-dose containers. The formulation may be in the form of solutions, suspensions or emulsions in oily or aqueous carriers and these may contain conventional dispersing, suspending or stabilizing agents. The compositions may also be in the form of a dry powder which is prepared before use with, for example, sterile pyrogen-free water. The compounds of formula 1 can also be formulated as 50 suppositories using a conventional suppository base, such as cocoa butter or other glycerides. The compounds of this invention may be administered in combination with other antibiotics such as penicillin or other cephalosporins, if desired.

When supplied in unit dosage forms, the compositions will preferably comprise about 50 to about 1500 mg of the active ingredient of the formula 1. The adult human dose will preferably be in the range of from about 500 to about 5000 mg per day, depending on the frequency and route of administration. When administered intramuscularly or intravenously to adults, a total dose of about 750 to about 3000 mg per day, in divided doses, will normally suffice, although in the case of Pseudomonas infections higher daily doses of some of the compounds may be desired. to be.

In the primary evaluation of the compounds of this invention, the minimal inhibitory concentrations (MRCs) of these compounds and two reference compounds (cefotaxime of the formula 19 and ceftazidime of the formula 20) were determined by the two-way dilution method in Mueller-Hinton agars compared to 32 strains of test organisms in six groups. The geometric mean of the MRCs determined in this test is given in Table A.

It can be seen that all test compounds were more active than cefotaxime against the (G -) - 11 and (G -) - 111 groups of test organisms, ie Enterobacteriaceae and Pseudomonas strains, with the most preferred compound 1a being clearly more active. All test compounds were more active than ceftazidime against the (G +) - 1a and (G +) - 1b groups of test organisms, ie Staphylococcus strains, the most preferred compound 1a being clearly more active than ceftazidime against all groups of test organisms except (G-) ) -lll, which was somewhat more sensitive to ceftazidime.

The absorption of the most preferred compound 1a and reference compounds (cefotaxime and ceftazidime) in mice was determined after a single intramuscular injection of the test compound (dissolved in 0.1 M phosphate buffer; pH 7) at a dose of 20 mg / kg. Blood samples were collected from the orbital cavity in heparinized capillary tubes and analyzed in Mueller-Hinton medium using Morganella Morganii A9695 as a test organism. The blood levels at different time intervals, the half-life values (ty2) and the areas under the curve (AUC) are listed in Table B.

Tests to identify organisms resistant to the preferred compound 1a were also performed with cefotaxime and ceftazidime. The MRC of these three compounds against 240 strains of Enterobacteriaceae were determined in a Mueller-Hinton medium and an MRC equal to or greater than 8 for at least one of the test compounds was chosen at random as an indication of a resistant organism. Of the 240 strains, 27 were found to be resistant to at least one of 25 test compounds. The results, which indicate 3 strains resistant to compound 1a, ie 7 - [(Z) -2-methoxyimino-2- (2-aminothiazol-4-yl) acetamido] -3 - [(1-methyl-1- pyrrolidinium) methyl] -3-cefem-4-carboxylate, 22 strains of organisms resistant to ceftazidime and 26 strains resistant to cefotaxime are listed in Table C.

30 TABLE A

Minimal Inhibitory Concentrations

Geometric mean of MRC (pg / ml) against bacterial strains 35 Compound with the (G +) - la (G +) - lb (G -) - la (G -) - lb (G -) - ll (G -) - lll formula T.

la; R2 = methyl 1.2 3.1 0.025 0.13 0.33 2.8 lb, R2 = ethyl 1.4 3.1 0.087 0.32 1.0 5.6 40 lc; R2 = isopropyl 1.4 3.6 0.35 1.3 3.2 11 1d; R2 = allyl 1.8 3.6 0.53 1.1 2.4 13

For comparison:

Cefotaxime (a) 1.0 2.2 0.015 0.35 4.1 22

Ceftazidime (a) 5.1 12 0.070 1.7 2.6 1.8 45 - (G +) - 1a: penicillin-sensitive S. aureus (5 strains) (G +) - lb: penicillin-resistant S. aureus (5 strains) (G -) - 1a: cephalothin-sensitive E. coli (2 strains), KI. pneumoniae (1 strain) and Pr. mirabilis (2 strains) 50 (G -) - lb: cephalothin resistant E. coli (3 strains) and KI. pneumoniae (3 strains) (G -) - ll: Pr. Morganii (1 strain), Ent. cloacae (2 strains) and Ser. marcescens (2 tribes) (G -) - lll: Ps. aeruginosa (6 strains) (a) average of five trials 193 284 4

TABLE B

Blood levels after intramuscular administration to mice (20 mg / kg)

5 Compound with Blood Levels (pg / ml) minutes after administration * 1/2 AUC

formula 1: (minutes) (µg hour / ml) 10 20 30 45 60 90 10 -1a; R2 = methyl (a) 20.7 19.6 13.6 8.8 4 -0.9 17 13.5

For comparison: 27.8 19.3 13 9.1 4.6 1.2 15 14.9

Cefotaxime (b)

Ceftazidime (c) 21.5 18.4 14.9 8.7 4.4 -0.8 17 13.8 15 - (a) average of 2 trials (b) 1 trial (c) average of 3 trials

20 TABLE C

Resistance (MRC = ^ 8 μ / ml to one or more test compounds of 240 strains Enterobacteriaceae

Mueller-Hinton medium

Geometric mean MRC (pg / ml) of 25 test compound)

Organism Number of Ceftazidime Cefotaxime strains 30 Escherichia coli 1 0.25 32 8

Escherichia coli 1 4 0.5 8

Klebsiella pneumoniae 1 2 16 0.13

Enterobacter aerogenes 3 0.25 32 13

Enterobacter aerogenes 1 4 8 32 35 Enterobacter cloacae 1 0.13 4 8

Enterobacter cloacae 3 0.5 40 50

Enterobacter cloacae 3 1.6> 63> 63

Enterobacter cloacae 1> 32> 63> 63

Citrobacter freundii 1 0.35 45 32 40 Citrobacter species 2 0.03> 63 32

Proteus vulgaris 1 0.06 8 8

Morganella Morganii 1 0.06 32 32

Serratia marcescens 1 1 1 16

Serratia marcescens 1 2 8 16 45 Serratia marcescens 2 2.8 2 11

Serratia marcescens 1 4 8 63

Serratia marcescens 1 8 16 8

Serratia marcescens 1 32> 63> 63 50 Total number of resistant strains 27 3 22 26 1a: compound of the formula 1; R2 = methyl

In view of the test results given in Tables A, B and C, in the antibacterial preparation 55 according to the invention, as the compound of the formula 1, it is preferred to use 7 - [(Z) -2-methoxyimino-2- (2 -aminothiazol-4-yl) acetamido] -3 - [(1-methyl-1-pyrrolidinium) methyl] -3-cephem-4-carboxylate or the 7 - [(Z) -2-ethoxyimino-2- (2- aminothiazol-4-yl) acetamido] -3 - ((1-methyl-1-pyrrolidinium) methyl] -3-cephem-4-5 193284 carboxylate, optionally in the form of a non-toxic pharmaceutically acceptable salt or solvate or a non toxic physiologically hydrolyzable ester thereof.

The invention further relates to a 7 - [(Z) -2- (etherified oxyimino) -2- (2-aminothiazol-4-yl) acetamido] -3-ammoniomethyl-3-cephem-4-carboxylic acid derivative, suitable for use in the anti-bacterial preparation according to the invention, characterized in that it is a compound of the formula 1 of the formula sheet, wherein R 2 is a straight or branched chain alkyl group with 1 to 4 carbon atoms or an allyl, 2-butenyl- or is 3-butenyl group, or a non-toxic pharmaceutically acceptable salt or solvate or a non-toxic physiologically hydrolyzable ester thereof.

For the preparation of the compounds of the formula I it is preferred to start from an easily available other cephalosporin. There are two basic procedures for converting an easily available starting cephalosporin into another cephalosprin with different substituents at the 7 and 3 positions. One can first remove the 7 substituent and replace it with the desired 7 substituent, and then insert the desired 3 substituent. On the other hand, the desired 3 substituent can be introduced first and subsequently the 7 substituent can be exchanged. The compounds of formula 1 can be prepared by either procedure, and both are within the scope of this invention, but it is preferred to first introduce the desired 7 substituent and then the desired 3 substituent. The preferred procedure is shown in reaction scheme A and the alternative procedure in reaction scheme B of the formula sheet. The abbreviation "Tr" represents the trityl (triphenylmethyl) group which is a preferred amino protecting group. Abbreviation "Ph" represents the phenyl group. Thus, the -CH (pH) 2 unit is the benzhydryl group, which is a preferred carboxyl protecting group.

Although the above reaction schemes indicate preferred multistage procedures for the preparation of the compounds of formula 1, it is clear that other starting materials and procedures may also be used to prepare the intermediates used in the key step of each reaction scheme. Thus, the key step in reaction scheme A is the reaction of compound 7 with N-methylpyrrolidine. Compound 7 can be prepared by itself by other procedures. Similarly, the key step in reaction scheme B is the acylation of compound 12 with compound 4. Both compounds, 12 and 4, can be prepared by other procedures.

Thus, there is provided a process for the preparation of compounds of formula 1, wherein R 2 is a straight or branched chain alkyl group of 1 to 4 carbon atoms, allyl, 2-butenyl or 3-butenyl, as well as 30 non-toxic pharmaceutically acceptable salts, physiologically hydrolyzable esters and solvates thereof, according to which a compound of formula 14 of the formula sheet, wherein R2 has the aforementioned meanings, B1 is a known carboxyl protecting group and B2 is a known amino protecting group, with N-methylpyrrolidine in reaction is brought to form a compound of formula 15 of the formula sheet, and then all protecting groups are removed by conventional means.

The reaction is carried out in a non-aqueous organic solvent, such as methylene chloride, chloroform, diethyl ether, hexane, ethyl acetate, tetrahydrofuran, acetonitrile, or mixtures of such solvents. The reaction is conveniently conducted at a temperature from about -10 to about + 50 ° C; with preference being given to conducting the reaction at room temperature.

At least one mole of N-methylpyrrolidine is used per mole of compound 14; normally it is preferred to use a 50-100% excess of N-methylpyrrolidine.

Carboxyl protecting groups suitable for use as B1 in the aforementioned reaction are known to those skilled in the art and include aralkyl groups such as benzyl, p-methoxybenzyl, p-nitrobenzyl and diphenylmethyl (benzhydryl); alkyl groups such as t-butyl; haloalkyl groups such as 2,2,2-trichloroethyl and other 45 carboxyl protecting groups as described in the literature, for example in British Pat. No. 1,399,086. It is preferred to use carboxyl protecting groups, which are easily removed by acid treatment. Particularly preferred carboxyl protecting groups are the benzhydryl and t-butyl units.

Amino protecting groups suitable for use as B2 are also known in the art and include the trityl group and acyl groups such as chloroacetyl. Preferred are amino protecting groups that can be easily removed by acid treatment, for example a trityl group.

The present invention also provides a process for preparing compounds of the formula 1, wherein R 2 is a straight or branched chain alkyl group of 1 to 4 carbon atoms, allyl, 2-butenyl or 3-butenyl, as well as non-toxic pharmaceutically acceptable salts, physiologically hydrolysable esters and solvates 55 thereof, according to which method a compound of the formula 16 of the formula sheet, or an N-silyl derivative thereof, wherein B1 is hydrogen or a known carboxyl protecting group, with an acylation derivative of an acid of the formula 17 of the formula sheet, wherein B2 is a known 193284 6 amino protecting group, and R2 has the aforementioned meanings, is acylated to form a compound of the formula 15 of the formula sheet, and then all protecting groups are removed.

The acylation derivatives of the acid of formula 17 include the acid halides, {and in particular the acid chloride), mixed acid anhydrides (such as the acid anhydrides formed with pivalic acid or a haloformate such as ethyl chloroformate), and activated esters (such as can be formed with N- hydroxybenzthiazole in the presence of a condensing agent such as dicyclohexylcarbodiimide). The acylation can also be carried out using the free acid of formula 17 in the presence of a condensing agent, such as dicyclohexylcarbodiimide, carbonyldimidazole or an isoxazolium-10 salt. As used herein, the term "acylation derivative" of the acid of formula 17 means the free acid itself in the presence of a condensing agent as described above. The preferred acylation derivative of the acid of formula 17 is the acid chloride used, for example, in the presence of a acid binding agent (in particular a tertiary amine, such as triethylamine, dimethylaniline or pyridine).

When the acylation is carried out with an acid halide, it is possible to use an aqueous reaction medium, although a non-present medium is preferred. When acid anhydrides, activated esters or the free acid are used for the acylation in the presence of a condensing agent, the reaction medium should be non-aqueous. Particularly preferred solvents for the acylation reaction are halogenated hydrocarbons, such as methylene chloride and chloroform, but tertiary amides such as dimethylacetamide or dimethylformamide can also be used as well as other conventional solvents such as tetrahydrofuran or acetonitrile.

The acylation reaction can be carried out at a temperature from about -50 to about + 50 ° C. However, it is preferred to carry it out at or below room temperature and most preferably at about -30 to about 0 ° C. It is usually preferred to acylate the compound of Formula 16 with about a stoichiometric amount of the acylating agent of Formula 17, although a small excess (e.g. 5-25%) of the acylating agent may also be used.

It is preferred that the compound of formula 16 be acylated in the form of its N-silyl derivative (if a non-aqueous reaction medium is used). This is expediently performed in situ by simply adding a suitable silylating agent (eg, Ν, Ο-bistrimethylsilylacetamide) to the solution of compound 16 before the addition of the acylating agent of the formula 17. It is preferred about 3 moles of silylating agent per mole of connection 16, although this is not critical. The silyl compound is easily removed after addition of water by acylation.

The acylating acids of formula 17 including carboxyl and amino-protected derivatives thereof are known in the art or can be prepared according to known procedures.

The following Preparations 1-4 show the preparation of compounds of formulas 3, 4, 5 and 8 shown in reaction schemes A and B. Six exemplary embodiments follow, which lead to obtaining compounds of formula 1.

Preparation No. 1 40 See formula 3 of the formula sheet.

Ethyl (Z) -2-methoxyimino-2- (2- [tritylamino] thiazol-4-yl) acetate (3a)

A mixture of ethyl (Z) -2-hydroxyimino-2- (2- [tritylamino] thiazol-4-yl) acetate (Formula 2; 5.00 g, 10.9 mmol), CH3I (2.04 mL, 32 , 8mmol) and K2CO3 (4.54g, 32.8mmol) in dry dimethyl sulfoxide (DMSO) (100ml) was stirred at room temperature for 16 hours and then poured into water (250ml).

45 The precipitate formed was collected by filtration, washed with water and then dried. The title compound (5.15 g) was obtained in quantitative yield. Melting point 115 ° C (decomposition).

NMR in CDCl3; maxima in ppm (δ values): 1.32 (3H, t), 3.98 (3H, s), 4.30 (2H, q), 6.42 (1H, s), 7.2 (1H , m), 7.25 (15H, s).

Compounds 3b, 3c and 3d were prepared according to the general procedure described above, but 50 replacing the methyl iodide with the appropriate iodide.

Compound of Formula 3 R2 Yield Melting Point Literature <1) (%) (° C) Mp. (° C) 55 3a methyl 100 115 {dec.) Ca. 120 (dec.) 3b ethyl 67 97-98 7 193284 (continued)

Compound of Formula 3 R2 Yield Melting Point Literature <1> (%) (° C) Mp. (° C) 5 -------- 3c isopropyl 26 51-55 3d allyl * The ester was hydrolyzed without isolation.

10 (1) Tetrahedron 34, 2233-2243 (1978).

Preparation No. 2

See formula 4 of the formula sheet.

(Z) -2-Methoxyimino-2- (2- [tritylamino] thiazol-4-yl) acetic acid (4a) 15 The ethyl ester 3a, (6.00 g, 12.7 mmol) prepared in Preparation No. 1 in ethanol (120 ml) was treated with 2N NaOH (12.7 ml) at room temperature for 16 hours. The reaction mixture was then adjusted to pH 8 by addition of powdery solid carbon dioxide and the solvent was then evaporated under reduced pressure. The residue was dissolved in water (100 ml) and the solution was acidified to pH 2 with 1N HCl and then with ethyl acetate (3 x 50 ml) extracted. The combined extracts were washed with a saturated aqueous NaCl solution, dried and evaporated. The residue was crystallized from ethyl acetate-hexane to obtain 5.56 g (yield 98%) of the title product.

Melting point 138-143 ° C (decomposition).

NMR in cdci3; maxima in ppm (δ values): 3.89 (3H, s), 6.52 (1H, s), 7.2 (15H, s).

Compounds 4b, 4c and 4d were similarly prepared according to the general procedure described above.

Compound of formula 4 R2 Yield Melting point Literature (1) (%) (° C, dec.) Mp. (° C, dec.) 30 4a methyl 98 138-143 ca. 140 4b ethyl 85 140-145 not listed 4c isopropyl 85 166-169 ca. 170 4d allyl 66 170-178 ca. 170 35 (1) Tetrahedron 34, 2233-2243 (1978).

Preparation No. 3

Benzhydryl 3-hydroxymethyl-7-phenylacetamido-3-cephem-4-carboxylate (8)

To a stirred suspension of phosphate buffer (pH 7, 162.5 ml) and wheat bran (20 g, dry) at room temperature was added 7-phenylacetamidocephalosporanic acid sodium salt (5 g, 12.1 mmol) at one time. The course of the reaction was followed by HPLC until the hydrolysis was completed (5 hours). The suspension was filtered to remove the wheat bran and the filtrate was cooled to 5-10 ° C for extractive esterification. To the cooled solution, methylene chloride (82 ml) was added followed by a 0.5 M solution of diphenyldiazomethane in methylene chloride (24 ml). The pH was then adjusted to 3.0 with 28% phosphoric acid. After 1 hour, the temperature of the reaction mixture was allowed to rise to 20 ° C. Heptane (56 ml) was added slowly and the resulting crystalline title product recovered by filtration. The yield of the title product was 3.0 g (50%).

Preparation No. 4 50 Benzhydryl 7-amino-3-chloromethyl-3-cephem-4-carboxylate (5)

Pyridine (3.2g, 40mmol) was added to a suspension of PCI5 (8.3g, 40mmol) in CH2Cl2 (100ml) and the mixture was stirred at 20 ° C for 20 minutes. Benzhydryl 3-hydroxymethyl-7-phenylacetamido-3-cephem-4-carboxylate (prepared in Preparation No. 3; 5.1 g, 10 mmol) was added all at once to the mixture under stirring at -40 ° C. The mixture was stirred at -10 ° C 55 for 15 minutes and then left at -10 to -15 ° C for 7 hours. Propane-1,3-diol (10 ml) was added to the cooled solution (~ 20 ° C) and the mixture was stored at -20 ° C for 6 hours, after which it was stirred at room temperature for 20 minutes. The resulting solution was washed 193284 with ice water (2 x 20 ml) and saturated aqueous NaCl (10 ml), then dried with MgSO 4 and then concentrated under reduced pressure. The gummy residue (12 g) was dissolved in a mixture of CHCl 3 and n-hexane (2: 1), and subjected to chromatography using a silica gel column (200 g) and the same solvent as eluent. Fractions containing the title compound were evaporated under reduced pressure and the residue triturated with n-hexane to afford the title product (2.1 g, 51%). Melting point at> 110 ° C (dec.).

IR in KBr; maxima at: 3400, 2800, 1785, 1725 cm -1.

UV in ethanol; maximum at 265 nm (E70 cm 160).

NMR in DMSO-d6 plus CDCI3; maxima in ppm (δ values): 3.69 (2H, s), 4.43 (2H, s), 5.09 (1H, d, J = 4.5 10 Hz), 5.24 (1H, d, J = 4.5 Hz), 6.87 (1H, s) 7.3 (10, m).

Example I

7 - [(Z) -2-Methoxyimino-2- (2-aminothiazol-4-yl) acetamido] -3 - [(1-methyl-1-pyrrolidinium) methyl] -3-cephem-4-carboxylate (1a) 15 A. Benzhydryl 3-chloromethyl-7 - [(Z) -2-methoxyimino-2- (2- (tritylamino) thiazol-4-yl) acetamido] -3-cephem-4-carboxylate (6a ')

Benzhydryl 7-amino-3-chloromethyl-3-cephem-4-carboxylate, prepared in Preparation No. 4, (2.29 g, 5.52 mmol) in CH 3 CN (57 ml) was mixed with bis (trimethylsilyl) acetamide (BSA ; 4.09 ml, 16.6 mmol) at room temperature for 50 minutes to give a clear solution. To the solution was added an acid chloride solution, which was prepared from (Z) -2-methoxyimino-2- (2- (tritylamino) thiazol-4-yl) acetic acid, prepared in Preparation No. 2, (2.04 g, 4.60 mmol) and PCI5 (1.15 g, 5.52 mmol) in methylene chloride (20 ml). The mixture was stirred at room temperature for 30 minutes, then poured into cold water (200ml) and extracted with ethyl acetate (3 x 100ml). The combined extracts were washed with aqueous NaCl, dried and evaporated. The residue syrup (4 g) was chromatographed on silica gel 25 (150 g) by successively eluting with 10: 1 and 3: 1 mixtures of toluene and ethyl acetate. The fractions containing the desired title compound were combined and evaporated to yield 2.61 g (68%) of 6a 'as an amorphous powder.

NMR in CDCl3; maxima in ppm (δ values): 3.50 (2H, s), 4.02 (3H, s), 4.33 (2H, s), 4.98 (1H, d), 5.87 (1H , q), 6.65 (1H, s), 6.90 (1H, s), 7.3 (25H, m).

B. Benzhydryl 3-iodomethyl-7 - [(Z) -2-methoxyimino-2- (2- (tritylamino) thiazol-4-yl) acetamido] -3-cephem-4-carboxylate (7a ')

A mixture of the 3-chloromethyl derivative (6a ') (1.50g, 1.79mmol) and Nal (1.34g, 8.93mmol) in methyl ethyl ketone (30ml) was stirred at room temperature for 1 hour. After evaporation of the solvent, the residue was dissolved in ethyl acetate (100ml) and washed with water, aqueous Na2S2O3 and aqueous NaCl, then dried and then evaporated to give the title compound 7a '(1.47g, 89%) as an amorphous powder.

NMR in CDCl3; maxima in ppm (δ values): 3.55 (2H, ABq), 4.00 (3H, s), 4.25 (2H, s), 4.97 (1H, d), 5.80 (1H , q), 6.65 (1H, s), 6.90 (1H, s), 7.3 (25H, m).

C. 7 - [(Z) -2-Methoxyimino-2- (2-aminothiazol-4-yl) acetamido] -3 - [(1-methyl-1-pyrrolidinium) methyl] -3-40 cephem-4-carboxylate (1a)

A mixture of 7a '(4.5g, 4.83mmol) and N-methylpyrrolidine (0.65ml, 6.28mmol) in CH 2 Cl 2 (45ml) was stirred at room temperature for 20 minutes. Ether (400 ml) was added to the mixture to separate the quaternary salt from the blocked cephalosporin which was collected by filtration and treated with 90% trifluoroacetic acid (TFA) at room temperature for 1 hour. The mixture was then evaporated under reduced pressure below 20 ° C. The residue was triturated with ether to give the TFA salt of 1a (2.40 g), which was dissolved in methanol (5 ml) and treated with a 1M solution of sodium 2-ethyl hexanoate (SEH) at room temperature for 30 minutes. ) in ethyl acetate (8 ml). After addition of ethyl acetate (100 ml), the precipitate formed (1.94 g) was collected by filtration. HPLC analysis showed that the crude product was 7% pure 50, with a 1: 8 ratio of the Δ isomer to the A2 isomer. Purification of the product by HPLC was repeated three times (Lichrosorb RP-18, 8 x 300 mm) eluted with 5% aqueous CH3OH or 0.01 M ammonium phosphate buffer (pH 7.2) containing 5% CH3OH, containing 35 mg ( 1.5%) of the title product was obtained as a colorless powder. Estimated purity (by HPLC) 90%. Melting point 150 ° C (decomposition).

55 IR in KBr; maxima at: 1770, 1660, 1620 cm'1.

UV in phosphate buffer, pH 7; maxima (with molar extinction coefficient) at: 235 (16200), 258 (15400) nm.

NMR in D20; maxima in ppm (δ values): 2.31 (4H, m), 3.08 (3H, s), 3.63 (4H, m), 4.09 (3H, s), 5.43 (1H , 193284 d, J = 4.8 Hz), 5.93 (1H, d), 7.08 (1H, s).

Example II

7 - [(Z) -2-Methoxyimino-2- (2-aminothiazol-4-yl) acetamido] -3 - [(1-methyl-1-pyrrolidinium) methyl] -3-cephem-4-5 carboxylate (1 a)

To a stirred solution of 20.4 g (21.9 mmol) of 7a ', prepared according to Example IB, in 150 ml of dry methylene chloride, 2.242 g (28.5 mmol) of N-methylpyrrolidine were added all at once at room temperature. The mixture was stirred for 5 minutes and then poured into 1000 ml ether with vigorous stirring to form a precipitate which was filtered, washed with ether (5 x 20 ml) and then dried under reduced pressure, 19.3 g product as a light yellow powder was obtained.

IR in KBr; maxima at: 3400, 1780 (s), 1740, 1675, 1530 cm1.

TLC, solvent ethanol-CHCl 3 (1: 3): Rf = 0.30.

The solid was dissolved in 185 ml of trifluoroacetic acid water (99: 1), and this solution was stirred at room temperature for 1 hour and then concentrated to about 30 ml below 10 ° C. The concentrate was poured into 1000 ml of ether with vigorous stirring to form a precipitate, which was filtered, washed with ether (5 x 50 ml) and dried under reduced pressure to give 10.6 g of a pale yellow powder . The powder was dissolved in 20 ml of methanol and the solution was filtered. 45 ml of 0.8M SEH ethyl acetate was added to the filtrate. The resulting suspension was poured into 400 ml of ethyl acetate and filtered to give 8.08 g of a solid, which was a mixture of the title compound and the corresponding A2 isomer (Δ3 / Δ2 = 1: 8) as shown by HPLC analysis (Lichrosorb RP-18, 10, 15% methanol in 0.01 M phosphate buffer pH 7). A second test, starting from 28.9 g (31.0 mmol) of 7a ', gave 16.0 g of the crude product (Δ3 / Δ2 = 1: 8). Isolation of the desired A3 isomer from the combined crude product (24.08 g) using preparative HPLC 25 (System 500, Waters Associates, PrePAK 500 / Cia, 5-10% CH 3 OH) gave 769 mg of compound 1a.

Example III

7 - [(Z) -2-Methoxyimino-2- (2- (2-aminothiazol-4-yl) acetamido] -3 - [(1-methyl-1-pyrrolidinium) methyl] -3-cephem-4-carboxylate (1a) A series of tests was conducted to determine the effect of solvent, amount of solvent and reaction period on the yield of the compound 1 and the Δ3 / Δ2 ratio in the reaction product The general procedure was, referring to Table D, as follows:

To a suspension of the 3-iodomethyl derivative 7a '(prepared according to example IB; 45 mg, 0.048 mmol) in the indicated amount of the indicated solvent was added a solution of 35 N-methylpyrrolidine (0.01 ml, 0.097 mmol) in ether ( 0.1 ml) and the mixture was stirred at room temperature for the indicated period. The reaction mixture was diluted with ether (5 ml) and the resulting precipitate was collected by filtration and mixed with 90% TFA. The mixture was stirred for 1 hour and then evaporated to dryness under reduced pressure below 20 ° C to yield the product. The ratio of Δ3 / Δ2 in the product was determined by HPLC 40 (Lichrosorb RP-18; mobile phase: 0.01 M ammonium phosphate buffer (pH 7.2) with 15% CH3OH; retention time: Δ3 6.60 minutes, Δ2 5, 56 minutes). Product yield and the ratio of Δ3 / Δ2 isomers for each test are given in Table D below.

TABLE D

45 -

Experiment No. Solvent Ratio of Reaction Yield Ratio 7a '(in g) to time (min) (%) Δ3 / Δ2 Solvent (in ml) 50 - 1 CH2CI2 1:20 15 73 1/8 2 CH2CI2 Ether 1: 100 15 25 4/1 (1/10) 3 Ethyl acetate ether 1: 100 15 27 4/1 55 (1/10) 193 284 10 TABLE D (continued)

Experiment No. Solvent Ratio of Reaction Yield Ratio 7a '(in g) to Time (min) (%) Δ3 / Δ2 5 Solvent (in ml) 4 Ethyl Acetate Ether 1: 100 60 64 2/1 (1/10) 10 5 Ether 1: 100 15 31 6/1 6 Ether 1: 100 60 62 3/1 7 Ether 1:60 15 55 3.5 / 1 8 Ether 1:60 60 82 1/1 15

Example IV

7 - [(Z) -2-Ethoxyimino-2- (2-aminothiazol-4-yl) acetamido] -3 - [(1-methyl-1-pyrrolidinium) methyl] -3-cephem-4-carboxylate (1b) A. Benzhydryl 3-chloromethyl-7 - [(Z) -2-ethoxyimino-2- (2- (tritylamino) thiazol-4-yl) acetamido] -3-cephem-4-20 carboxylate (6b)

To a solution of (Z) -2-ethoxyimino-2- (2- (tritylamino) thiazol-4-yl) acetic acid (4b) (1.095 g, 2.4 mmol) in dichloromethane (20 ml) was added phosphorus pentachloride (500 mg ) added. After stirring for 1 hour at room temperature, the mixture was added all at once to an ice-cooled solution of compound 5 (prepared in Preparation No. 4; 1.083 g, 2.4 mmol) and BSA (1 ml) in dichloromethane (20 ml). After stirring for 0.5 hours, the reaction mixture was poured into 10% aqueous NaHCO 3 (200 ml) and extracted with CHCl 3 (100 ml). The extract was washed with water, dried over MgSO 4 and evaporated under reduced pressure.

The residue was chromatographed on a silica gel column. Elution with CHCI3 gave 6b as an amorphous powder (1.76 g) (86%).

30 NMR in CDCI3: maximums in ppm (δ values): 1.40 (3H, t), 3.53 (2H, ABq), 4.37 (2H, s), 4.60 (2H, q), 4.90 (1H, d), 5.89 (1H, d), 6.88 (1H, s), 6.91 (1H, s).

B. Diphenylmethyl 7 - [(Z) -2-ethoxyimino-2- (2- (tritylamino) thiazol-4-yl) acetamido] -3-iodomethyl-3-cephem-4-carboxylate (7b)

A mixture of 6b (1.07g, 1.25mmol) and Nal (562mg, 2.75mmol) in acetone (20ml) was stirred for 1 hour. The mixture was filtered and the filtrate poured into water and extracted with ethyl acetate. The organic layer was washed successively with 5% aqueous Na2S2O3, water and saturated NaCl, then dried with MgSo4 and then evaporated to yield 1.04 g (89%) of compound 7b.

NMR in CDCI3: Maximums in ppm (δ values): 3.55 (2H, q), 4.27 (2H, s), 5.02 (1H, d), 5.87 (1H, d), 6 .68 (1H, 40s), 6.93 (1H, s).

C. 7 - [(Z) -2-Ethoxyimino-2- (2-aminothiazol-4-yl) acetamido] -3 - [(1-methyl-1-pyrrolidinium) methyl] -3-cephem-4-carboxylate ( 1b)

A mixture of 7b (333mg, 0.85mmol) and N-methylpyrrolidine (60mg, 0.7mmol) in CH 2 Cl 2 (5ml) was stirred at room temperature for 0.5h then evaporated under reduced pressure.

45 The residue was washed with ether and then dissolved in 90% aqueous TFA. After being stored at room temperature for 0.5 hours, the mixture was concentrated under reduced pressure. Ether was added to the concentrate to separate the quaternized product which was then collected by filtration and dissolved in a small amount of methanol. The solution was chromatographed on an HP-2 column (40 ml). Elution with 30% aqueous CH3OH followed by freeze drying gave 50 0.062 g of a mixture of the Δ2 and A3 isomers (Δ2: Δ3 = 5: 1). The mixture was purified by HPLC (Lichrosorb RP-18, 8 x 300 mm, 15% methanol) and the desired A3 isomer (1b) was isolated as a pale yellow powder (4.9 mg, 2.7%).

UV in phosphate buffer, pH 7; maxima (with molar extinction coefficient) at: 235 (15000), 258 (14000) nm.

NMR in D20; maxima in ppm (δ values): 1.43 (3H, t), 2.33 (4H, m), 3.10 (3H, s), 3.64 (4H, m), 4.36 (2H .55 q), 5.44 (1H, d), 5.95 (1H, d), 7.08 (1H, s).

11 193284

Example V

7 - [(Z) -2- (lsopropoxyimino) -2- (2-aminothiazol-4-yl) acetamido] -3 - [(1-methyl-1-pyrrolidinium) methyl] -3-cephem-4-carboxylate ( 1c) A. Diphenylmethyl 3-chloromethyl-7 - [(Z) -2- (2-isopropoxyimino) -2- (2- (tritylamino) thiazol-4-yl) acetamidoj-3-5 cephem-4-carboxylate (6c )

A mixture of (Z) -2- (2-isopropoxyimino) -2- (2- (tritylamino) thiazol-4-yl) acetic acid (4c) (707 mg, 1.5 mmol) and phosphorus pentachloride (344 mg, 1, 65 mmol) in dichloromethane (14 ml) was stirred at room temperature for 1 hour and then in a solution of compound 5 (prepared in Preparation No. 4; 677 mg, 1.5 mmol) and BSA (1.1 ml, 4, 5 mmol) in dichloromethane (15 ml). The reaction mixture was stirred at room temperature for 30 minutes, then diluted with ethyl acetate (200 ml). The whole was washed with aqueous NaHCO 3 (100 ml) and water (3 x 100 ml), then dried with sodium sulfate and then evaporated to give 1.4 g (100%) of compound 6c.

IR in KBr; maxima at: 3360, 3020, 3060, 2960, 1785, 1725, 1680, 1520, 1500, 1450, 1375, 1300, 1250, 1160, 1090, 1060, 1010, 990, 840, 740, 700 cm'1.

UV in EtOH; maxima (with molar extinction coefficient) at: 240 (24600), 260 (20700) nm.

NMR in CDCl3; maxima in ppm (δ values): 1.35 (6H, d, J = 6 Hz), 3.50 (2H, s), 4.35 (2H, s), 4.58 (1H, m, J = 6 Hz), 5.00 (1H, d, J = 4.5 Hz), 5.91 (1H, dd, J = 4.5 &9Hz; d of D20, J = 4.5 Hz), 6 .68 (1H, s), 6.88 (1H, s), 7.25 (25H, s).

B. Diphenylmethyl 3-iodomethyl-7 - [(Z) -2- (2-isopropoxyimino) -2- (2- (tritylamino) thiazol-4-yl) acetamido] -3-20 cephem-4-carboxylate (7c)

A mixture of compound 6c (500mg, 0.55mmol) and sodium iodide (248mg, 1.66mmol) in acetone (10ml) was stirred at room temperature for 50 minutes. After evaporation, the residue was dissolved in ethyl acetate (15 ml), washed successively with 10% aqueous sodium thiosulfate (10 ml), water (10 ml) and aqueous NaCl (10 ml), then dried with sodium sulfate and then evaporated to yield of 494 mg (90%) of the title compound 7c.

IR in KBr; maximums at: 3360, 3040, 3020, 2960, 1785, 1720, 1680, 1600, 1520, 1500, 1450, 1370, 1300, 1230, 1150, 1115, 1080, 990, 900, 840, 750, 700 cm'1.

UV in EtOH; maxima (with molar extinction coefficient) at: 240 (24900), 260 (19400) nm.

NMR in CDCl3; maxima in ppm (δ values): 1.30 (6H, d, J = 6 Hz), 3.37 & 3.70 (1H each, d, J = 16 Hz), 4.22 30 (2H, s ), 4.55 (1H, m, J = 6 Hz), 4.95 (1H, d, J = 4.5 Hz), 5.83 (1H, dd, J = 4.5 & 9 Hz; d of D20), 6.66 (1H, s), 6.87 (1H, s), 7.25 (25H, s).

C. 7 - [(Z) -2- (2-Osp [rp [pxuo, omp] -2- (2-aminothiazol-4-yl) acetamido] -3 - [(1-methyl-1-pyrrolidinium) methyl ] -3-cefem-4-carboxylate (1c)

A mixture of compound 7c (545mg, 0.55mmol) and 1-methylpyrrolidine (70mg, 0.82mmol) in 35 dichloromethane (10ml) was stirred at room temperature for 30 minutes and then diluted with ether (100ml) . The resulting precipitate was collected by filtration. A solution of the precipitate in 90% TFA (4.5 ml) was stirred at room temperature for 10 minutes and then evaporated under reduced pressure. The residue was triturated with ether to give 317 mg of the crude product which was chromatographed on an HP-20 column (50 ml) eluting with water (500 ml) and 30% CH 3 OH (500 40 ml). The 30% CH3OH eluate was concentrated and lyophilized to give 109 mg of a mixture of the Δ2 and A3 isomers (Δ2 / Δ3 = 6/1), 100 mg of which were purified by HPLC (Lichrosorb RP-18, 15% MeOH) to form 5 mg (3%) of the desired title compound 1c.

UV in pH 7 phosphate buffer; maxima (with molar extinction coefficient) at: 236 (15100), 252 (14600) nm.

NMR in D20; maxima in ppm (δ values): 1.42 (6H, d, J = 6 Hz), 2.33 (4H, s), 3.10 (3H, s), 3.65 (4H, s), 45 3.83 & 4.23 (1H each d, J = 17 Hz), 5.45 (1H, d, J = 4.5 Hz), 5.95 (1H, d, J = 4.5 Hz) , 7.05 (1H, s).

Example VI

7 - [(Z) -2-Allyloxyimino-2- (2-aminothiazol-4-yl) acetamidoj-3 - [(1-methyl-1-pyrrolidinium) methyl] -3-cefern-4-carboxylate (1 d) 50 A. Benzhydryl 7 - [(Z) -2-allyloxyimino-2- (2- (tritylamino) thiazol-4-yl) acetamido] -3-chloromethyl-3-cephem-4-carboxylate (6d)

To a suspension of compound 5 (prepared in Preparation No. 4; 1.35 g, 3 mmol) in methylene chloride (20 ml), BSA (1.1 ml, 4.5 mmol) was added and the mixture was stirred for 30 minutes at stirred at room temperature to give a clear solution. A mixture of (Z) -2-allyloxyimino-2- (2-55 (tritylamino) thiazol-4-yl) acetic acid (4d) (1.40 g, 3.0 mmol) and phosphorus pentachloride (690 mg, 3.3 mmol) in methylene chloride (20 ml) was stirred at room temperature for 15 minutes and poured into the solution of the trimethylsilylated compound 5 all at once. The resulting mixture was kept for 20

Claims (4)

193284 Stirred at room temperature for 12 minutes and then diluted with ethyl acetate (200 ml), washed with aqueous sodium hydrogen carbonate and water, dried and evaporated under reduced pressure. The oily residue was purified by silica gel column chromatography (Wako gel, C-200, 30 g). The column was eluted with chloroform and the fractions containing the desired product were combined. After evaporation under reduced pressure, the title compound (6d) was obtained as an amorphous powder, yield 2.32 g (89%). Melting point 100-115 ° C (decomposition). IR in KBr; maximums at: 3990, 1790, 1730, 1680, 1530, 1380, 1250, 1160, 1020 cm -1. NMR in CDCl3; maxima in ppm (δ values): 3.50 (2H), 4.32 (2H, s), 4.6-6.1 (7H, m), 6.70 (1H, s), 6.90 (1H, s), 7.1-7.6 (30H, m). Elemental analysis calculated for C 4 H 40 N 5 O 5 S 2 Cl • 1 / 3CHCl 3: C, 64.05; H, 4.45; N, 7.73; S, 7.08; Cl, 7.82%. found: C, 64.13; 63.99; H, 4.61, 4.64; N, 7.50, 7.30; S, 6.85, 6.85; Cl, 7.55, 7.46%. B. Benzhydryl 7 - [(Z) -2-allyloxyimino-2- (2- (tritylamino) thiazol-4-yl) acetamido] -3-iodomethyl-3-cephem-4-carboxylate (7d) A mixture of compound 6d (2.03g, 2.65mmol) and sodium iodide (2g, 13.3mmol) in acetone (15ml) were stirred at room temperature for 1 hour and then evaporated under reduced pressure. A solution of the oily residue in ethyl acetate (200 ml) was washed with 10% sodium thiosulfate and water, then evaporated under reduced pressure to afford compound 7d as an amorphous powder, which was used in the next step without further purification. Yield 2.52 g (99%). 20 C. 7 - [(Z) -2-Allyloxyimino-2- (2-aminothiazol-4-yl) acetamido] -3 - [(1-methyl-1-pyrrolidinium) methyl] -3-cephem-4-carboxylate (1 d) A mixture of compound 7d (478 mg, 0.5 mmol) and N-methylpyrrolidine (0.05 ml, 0.5 mmol) in methylene chloride (5 ml) was stirred at room temperature for 20 minutes and then with ether (50 ml) diluted precipitating the quaternized product (yield 500 mg). A mixture of the quaternized product and TFA (2ml) was stored at room temperature for 1Vfe hours and then diluted with ether to precipitate the crude TFA salt of the product (yield 265g) which was chromatographed on a column of HP-20 (1.8 x 18 cm). The column was eluted with water and 30% aqueous methanol. The methanolic eluate was evaporated under reduced pressure and the residue was lyophilized to form an amorphous powder, (yield 124 mg) containing the desired product (17%) and the corresponding A2 isomer (83%). The mixture was purified by HPLC / Lichrosorb RP-18; 0.01 M NH 4 H 2 PO 4 (pH 7: CH 3 OH = 87:15). The eluate was acidified to pH 3 with dilute HCl and chromatographed on a column of HP-20 (1.8 x 10 cm). The column was eluted with water and then with 30% aqueous methanol. The methanolic eluate was evaporated under reduced pressure and the residue was lyophilized to afford the title compound (1d) as an amorphous powder (yield 13 mg, 5.1%); mp 155 ° C (decomposition). IR in KBr; maxima at: 3600-2800, 1770, 1670, 1610, 1530, 1200 cm'1. UV in pH 7 phosphate buffer; maxima (with molar extinction coefficient) at: 235 (16600), 253 (15600) nm. NMR in 6D20; maxima in ppm (δ values): 2.1-2.5 (4H, m), 3.10 (3H, s), 3.4-3.8 (4H, m), 5.95 (1H, d, J = 4 Hz), 7.10 (1H, s). 40 An antibacterial preparation comprising a 7 - [(Z) -2- (etherified oxyimino) -2- (2-aminothiazol-4-yl) acetamido] -3-45 ammoniomethyl-3-cephem-4-carboxylic acid derivative and an inert carrier, characterized in that it is a compound of the formula 1 of the formula sheet, wherein R 2 is a straight or branched chain alkyl group with 1 to 4 carbon atoms or an allyl, 2-butenyl or 3-butenyl group, or a non- toxic pharmaceutically acceptable salt or solvate or a non-toxic physiologically hydrolyzable ester thereof. Preparation according to claim 1, characterized in that the compound of the formula 1 contains the 7 - [(Z) -2- 50 methoxyimino-2- (2-aminothiazol-4-yl) acetamido] -3 - [(1 -methyl-1-pyrrolidinium) methyl] -3-cephem-4-carboxylate, or a non-toxic pharmaceutically acceptable salt or solvate or a non-toxic physiologically hydrolyzable ester thereof. Preparation according to claim 1, characterized in that the compound of the formula 1 contains the 7 - [(Z) -2-ethoxyimino-2- (2-aminothiazol-4-yl) acetamido] -3 - [(1- methyl 1-pyrrolidinium) methyl] -3-cephem-4-carboxylate, 55 or a non-toxic pharmaceutically acceptable salt or solvate or a non-toxic physiologically hydrolyzable ester thereof. 4. A 7 - {(Z) -2- (etherified oxyimino) -2- (2-aminothiazol-4-yl) acetamido] -3-ammoniomethyl-3-cephem-4 -13 193284 carboxylic acid derivative, suitable for use in the antibacterial preparation according to claim 1, characterized in that it is a compound of the formula 1 of the formula sheet, wherein R 2 is a straight or branched chain alkyl group with 1 to 4 carbon atoms or an allyl, 2-butenyl or 3-butenyl group , or a non-toxic pharmaceutically acceptable salt or solvate or a non-toxic physiologically hydrolyzable ester thereof. Hereby 5 sheets drawing