NZ196551A - Cephalosporin derivatives and pahrmaceutical compositions; intermediate cephalosporin derivatives - Google Patents

Description

New Zealand Paient Spedficaiion for Paient Number 1 96551 196551 % j Priority J-'A1- • • • I Cemple&e Specification FiSsd: '£ h?L j Class: CpJ^or)f\b\K?Af^.... ■ Publication Date: ■ ■ - 5 ^ MAY* -1^84 /<Pb8 f*0. Joumsl No: Iraana***-- Jk * ft{| *r% gB JJwrajB n; %■'• /, "' '" ' '■ " i«J <4$? _ iiBIri *&r. I.fl No.: Date: NEW ZEALAND PATENTS ACT, 1953 x.

COMPLETE SPECIFICATION CEPHALOSPORIN DERIVATIVES ^We, F. HOFFMANN-LA ROCHE & CO. AKTIENGESELLSCHAFT, 124-184 Grenzacherstrasse, Basle, Switzerland, a Swiss company, hereby declare the invention for which X / we pray that a patent may be granted to Kie/us, and the method by which it is to be performed, to be particularly described in and by the following statement: - (followed by la) - la- 196551 MM! -VHP/14-i The present invention is concerned with novel cephalosporin derivatives of the general formula h h ch3on=c conh n 1 h- COOH in which X represents one of the groups (a) h3c N—N AX cooh (b) N wherein R represents lower alkyl or (together with the oxygen) a 10 readily hydrolysable ether group, as well as readily hydrolysable esters of the compounds of formula I wherein X represents the group (b) or (a) in which R^ represents lower alkyl and salts of these compounds and hydrates of the compounds of formula I or of esters and 15 salts thereof. ' *5 ■V/ ; .7 : 1 96 2"5 1 <> ^ - 2 -: V-.V R"*" in the group (a) can represent lower alkyl, ;i.-e".:ia :straight-chain-cbr.\; branched talkyl,i>grpup;twith ;-l-;7 carbon atoms such as ethyl, n-propyl, isopropyl, n-butyl,. isobutyl, sec-butyl, t-butyl, n-pentyl or, especially, 5 methyl. R"*" can (together with the oxygen) also. represent a readily hydrolysable ether group. Thus, R^" can signify, for example: lower alkanoyloxyalkyl , (e.g. acetoxymethyl, pivaloyloxymethyl, 1-acetoxyethyl and 1-pivaloyloxyethyl), lower alkoxycarbonyloxyalky1 (e.g. methoxycarbonyloxymethy1, 10 1-ethoxycarbonyloxyethyl and 1-isopropoxycarbonyloxyethyl), lactonyl groups (e.g. phthalidyl and thiophthalidyl), lower alkoxymethyl (e.g. niethoxymethyl) and lower alkanoylamino-methyl (e.g. acetamidomethyl) . The term "lower" in each case denotes a group containing up to and including 7 carbon atcins.

As readily hydrolysable esters of the compounds of 15 formula I there are to be understood compounds of foirmula I in which at least one carboxy group is present in the form of a readily hydrolysable ester group. Examples of such esters, which can be of the conventional type, are the lower alkanoyloxyalkyl esters (e.g. the acetoxymethyl, pivaloyloxy-20 methyl, 1-acetoxyethyl and 1-pivaloyloxyethyl ester), the lower alkoxycarbonyloxyalkyl esters (e.g. the methoxycarbonyi-oxymethyl, 1-ethoxycarbonyloxyethyl and l-isopropoxycazbonyl-cxyethyi ester), the lactonyl esters (e.g. the phthalidyl and thiophthalidyl ester) the lower alkoxymethyl esters (e.g. 25 the methoxymethyl ester) and the lower alkanoylaminomethy1 (e.g. the acetamidomethyl ester). Other esters (e.g. i I I 2 3 MAR WSJ •-... 196551 the benzyl and cyanomethyl esters) can also be used. The esters can be monoesters or diesters. The second ester formation can occur in the compound with the .carboxy group of the 3-carboxy-l-methyl-lH-l,2,4-triazol-5-yl group X [group (b)].

Examples of salts of the compounds of formula I are alkali metal salts such as the sodium and potassium salt, the ammonium salt, alkaline earth metal salts such as the calcium salt, salts with organic bases such as salts with amines (e.g. salts with N-ethyl-piperidine, procaine, dibenzylamine, N,N1-dibenzylethylenediamine, alkylamines or dialkylamines) as well as salts with amino acids such as, for example, salts with arginine or lysine. The salts can be mono-salts or also di-salts. The second salt formation can occur in compounds with the carboxy group of the 3--carboxy-l-methyl-lH-1,2,4-triazol-5-yl group X [group (b)].

The compounds of formula I also form addition salts with organic or inorganic acids. Examples of such salts are hydrohalides (e.g. hydrochlorides, hydrobromides and hydro-iodides) as well as other mineral acid salts such as sulphates, nitrates, phosphates and the like, alkylsulphonates and mono-arylsulphonates such as ethanesulphonates, toluenesulphonates, benzenesulphonates and the like and also other organic acid salts such as acetates, tartrates, maleates, citrates, benzoates, salicylates, ascorbates and the like.

The compounds of formula I as well as their salts and readily hydrolysable esters can be hydrated. The hydration can be effected in the course of the manufacturing process 5 or can occur gradually as a result of the hygroscopic properties of an initially anhydrous - product.

The products in accordance with the invention can be present in the syn-isomeric form or in the anti-isomeric form U \\ II -c CONH-—> \ or as a mixture of these twojforms. The syn-isomeric i form or mixtures of the syn-fand anti-isomeric forms in which the syn-isomeric form predominates is/are preferred. 7 PATgNTT GFF1CS 2 3 MAR 1983 cC£iVED Preferred products are (6R,7R)-7-/2-(2-amino-4-thiazolyl)-2-[(Z)-methoxy-imino]acetamido7-3-£[ (2,5-dihydro-6-methoxy-2-methyl--5-oxo-as-triazin-3-yl)thio]methyl7~8-oxo-5-thia-■5 -1-azabicyclo[4.2.0]oct-2-ene-carboxylic acid,- (6R,7R)-7-/2-(2-amino-4-thiazolyl)-2-[(Z)-methoxy-imino]acetamidQ7-3-ZX( 3-carboxy-l-methyl-lH-l,2,4--triazol-5-yl)thio]methyl7-8-oxo-5-thia-l-azabicyclo-[4.2.0]oct-2-ene-2-carboxylic acid, 10 ~ (6R,7R)-7-[2-(2-amino-4-thiazolyl)-2-((Z)-methoxy- imino)acetamido]-3-[[[2,5-dihydro-2-methyl-5-oxo--6-[(pivaloyloxy)methoxy]-as-triazin-3-yl]thio]methyl] -8-oxo-5-|thia--l-azabicyclo [4.2 .0] oct.-2-en«-2-carboxyl:K acid and their salts as well as the corresponding hydrates.

The above cephalosporin derivatives can be manufactured in accordance with the invention by (a) cleaving off the protecting group R, and, if necessary, carboxy protecting groups which may be.present, in a compound of the general formula h h. 2 3 MAR 1983 "cevEO CH3ON RHN —c—conh- -ch 2 s ^ COOH II or (b) in which X"*" has the same significance as X in formula I, whereby the carboxy group of group (b) can be protected, R represents a cleavable protecting group and the carboxy group in the 4--position of the cephalosporin moiety can be present in protected form, reacting a halide of the general formula h h CHaON= —s—X- III c h2v cooh in which X"*" has the significance given above and Y represents a halogen atom and the carboxy group in the 4-position of the cephalosporin 15 moiety can be present in protected form, with thiourea, or (c) for the manufacture of a readily hydrolysable ester of a compound of formula I, subjecting a carboxylic acid of 20 formula I to a corresponding esterification, or 19655 (d) for the manufacture of salts or hydrates of a compound of formula I or hydrates of these salts, converting a compound of formula I into a salt or hydrate or into a hydrate of said salt.

If desired, carboxy groups present in the starting materials of formulae II and III can' be protected; for example, by esterification to form a readily cleavable ester such as a silyl ester (e.g. the trimethylsilyl ester). The readily hydrolysable esters mentioned above also come into 10 consideration. The carboxy groups can also be protected by salt formation with an inorganic or tertiary organic base such as triethylamine. Possible R-protecting groups are, for example, protecting groups which are cleavable by acid hydrolysis such as, for example, t-butoxycarbonyl or 15 trityl, or protecting groups which are cleavable by basic hydrolysis such as, for example, trifluoroacetyl. Preferred R-protecting groups are chloroacetyl, bromoacetyl and iodoacetyl, especially chloroacetyl. The last-mentioned protecting groups can be cleaved off by treatment with 20 thiourea.

The starting materials of formula II can be prepared, for example , by N-acylating the corresponding 7-amino compound, namely by reacting a compound of the general formula 1 £ r o' h h2n— cooh in which X"*" has the significance given above and the carboxy group and/or the amino group can be present in protected form, with an acid of the general formula in which R has the significance given above, or with a reactive functional derivative of this acid and, if desired, cleaving off carboxy protecting groups which may be present.

If desired, the carboxy groups present in the 7-amino compound of formula IV can be protected in the manner h 196551 mentioned above for the starting material of .formula II to be prepared. The amino group of the compound of formula IV can be protected, for example, by a silyl protecting group such as trimethylsilyl.

As reactive functional derivatives of acids of formula V there come into consideration, for.example, halides, (i.e. chlorides, bromides and fluorides), azides, anhydrides, especially mixed anhydrides with strong acids, reactive esters (e.g. N-hydroxysuccinimide esters) and amides (e.g. 10 imidazolides).

The reaction of the 7-amino compound of formula IV with an acid of formula V or a reactive functional derivative thereof can be carried out in a manner known per se. Thus, for example, a free acid of formula V can be 15 condensed with one of the aforementioned esters corresponding to formula IV by means of a carbodiimide such as dicyclo-hexylcarbodiimide in an inert solvent such as ethyl acetate, acetonitrile, dioxan, chloroform, methylene chloride, benzene or dimethylformamide and subsequently the ester group 20 can be cleaved off. Oxazolium salts (e.g. N-ethyl-5-phenyl--isoxazolium-31-sulphonate) can also be used as the • condensation agent in place of carbodiimides.

According to another embodiment, a salt of an acid of formula IV (e.g. a trialkylammonium salt such as the triethy1- - io - 196551 ammonium salt) is reacted with a reactive functional derivative of an acid of formula V as mentioned above in an inert solvent (e.g. one of the solvents named above).

According to a further embodiment, an acid halide, preferably the chloride, of an acid of formula V is reacted with the amine of formula IV. The reaction is preferably carried out in the presence of an acid-binding agent, for example, in the presence of aqueous alkali, preferably sodium hydroxide, or in the presence of an alkali metal carbonate such as potassium carbonate or in the presence of a lower alkylated amine such as triethylamine. As the solvent there is preferably used water, optionally in admixture. with an inert organic solvent such as tetrahydro-furan or dioxan. The reaction can also be carried out in an aprotic organic solvent such as, for example, dimethyl-formamide, dimethyl sulphoxide or hexamethylphosphoric acid triamide. When silylated starting materials of formula IV are used, the reaction is carried out in an anhydrous medium.

The reaction of the 7-amino compound of formula IV with the acid of formula V or a reactive functional derivative thereof can conveniently be carried out at temperatures between about -40°C and room temperature, for example at about 0-10°C. 1 0) £ ^ c I wherein R is wot\J)qjj.qs Starting materials of formula Ii wnerein k is nt5tqja qj monohalogenated (as in bromo-, chloro and iodoacetyl) can also be prepared by thiolation, namely by reacting a compound of the general formula H H CHoON:— O FfHN •CONH VI COOH in which R°. is as R but cannot be monohalogenated, Z represents a leaving group and the carboxy group can be protected by salt formation with an inorganic or tertiary organic base, with a thiol of the general formula HS— X" VII in which X has the significance given above, and, if desired, cleaving off a carboxy protecting group which may be present.

As the leaving group Z in a compound of formula VI there come into consideration, for example, halogens (e.g. chlorine, bromine or iodine), acyloxy groups (e.g. lower 12 1 0)(^ ; alkanoyloxy groups such as acetoxy), lower alkylsulphonyloxy or arylsulphonyloxy groups such as mesyloxy or tosyloxy, or the azido group.

The reaction of a compound of formula VI with a thiol of formula VII can be carried out in a manner known per se; for example, at a temperature between about 40 °C and 80°C, conveniently at about 60°C, preferably in water or in a buffer solution with a pH of about 6 to 7, preferably 6.5.

The carboxy group of the resulting compounds of formula II can subsequently be protected if desired (e.g. by salt formation or esterification).

The thiols of formula VII are in tautomeric equilibrium with the corresponding thiones as shown in the following formulae h Vila 1 Vila 2 1965 wherein R"'" has the above significance or HoC 3 \ N—N n — n hs cooh h Vllb1 Vllb 2 wherein the carboxy group can be protected.

The preparation of these compounds is described in Examples 1,2 and 3. For the preparation of thiols (thiones) corresponding to group (a) which are etherified in the 6--position, the R"*"- group is generally introduced by reacting a S-protected thiol (e.g. by benzhydryl) with the halide containing the R"*"- group, preferably the iodide, in an inert organic solvent in the presence of an acid-binding agent (e.g. potassium carbonate), preferably at about 10-50°C, and cleaving off the protecting group (benzhydryl can be cleaved off with anisole and trifluoroacetic acid at room temperature).

In accordance with variant (a) of the process in accordance with the invention, the amino protecting group R 19655 in a starting material of formula II is cleaved off. Protecting groups which are cleavable by acid hydrolysis are preferably removed with the aid of a lower alkane-carboxylic acid which may be halogenated. In particular, formic acid or trifluoroacetic acid is used. This cleavage is generally carried out at room temperature, although it can be carried out at a slightly higher or slightly lower temperature, for example, a temperature in the range of about 0°C to +40°C. Protecting groups which are cleavable under alkaline conditions are generally hydrolysed with dilute aqueous caustic alkali at 0°C to 30°C. The chloroacetyl, bromoacetyl and iodoacetyl protecting groups can be cleaved off by means of thiourea in acidic, neutral or alkaline medium at about 0-30°C. Hydrogenolytic cleavage (e.g. cleavage of benzyl) is unsuitable in this case, since the oxime function would be reduced to the amino group during the hydrogenolysis.

After carrying out process variant (a), carboxy protecting groups which may be present in the reaction product can be cleaved off if desired. When the protecting group is a silyl group (silyl ester), this group can be cleaved off especially readily by treating the reaction product with water. Lower alkanoyloxyalkyl, alkoxycarbonyl-oxyalkyl, lactonyl, alkoxymethyl and alkanoylaminomethyl esters are preferably cleaved enzymatically with the aid of a suitable esterase (at about 20-40°C). When a carboxy 196551 group is protected by salt formation (e.g. with triethylamine,) then the cleavage of this salt-forming .protecting group can be carried out by treatment with acid. The acid which can be used for this purpose can be, for example, hydrochloric acid, sulphuric acid, phosphoric acid or citric acid.

The carboxy protecting groups can be cleaved off in the same manner as just described also prior to the cleavage of the protecting group R.

The halide of formula III used in accordance with the invention can be prepared, for example, by reacting a compound of formula IV with a halogenated carboxylic acid of the general formula CH3on=c—cooh co j VIII ch2y in which Y represents a halogen atom, or with a reactive derivative of this compound. The halogenated carboxylic acid of formula VIII is used either in free form in the presence of a condensation agent (e.g. a N, N1-disubstituted carbodiimide such as N,N1-dicyclohexyl-carbodiimide or an azolide compound such as N,N'-carbonyl- 196551 diimidazole or N,N'-thionyldiimidazole) or in the form of an acid haiide such as the acid chloride or acid bromide, in the form of an acid anhydride such as an acid anhydride with a carbonic acid monoester (e.g. with monomethyl carbonate or monoisopropyl carbonate) or in the form of an activated ester such as the p-nitrophenyl ester, 2,4--dinitrophenyl ester, N-h'ydroxysuccinimide ester or N-hydroxyphthalimide ester. The reaction is generally carried out in an inert organic solvent, for example in a halogenated hydrocarbon such as chloroform, dichloromethane or carbon tetrachloride, in an ether (e.g. tetrahydrofuran or dioxan), in dimethylformamide, dimethylacetamide, water or mixtures thereof. The reaction temperature mainly, lies in the range of about -50°C to +40°C, preferably at about -10°C to +10°C.

The reaction in accordance with the invention of the ha-lide of formula III with thiourea, variant (b) of the process in accordance with the invention, is preferably carried out in an inert solvent such as, for example, in a lower alkanol (e.g. ethanol), in a lower ketone such as acetone, in an ether such as tetrahydrofuran or dioxan, in dimethylformamide, dimethylacetamide, in water or in mixtures thereof. The reaction temperature generally lies in the range of about 0°C to 60°C, preferably at room temperature. The chloride, bromide, fluoride or iodide can be used as the halide of formula III, the chloride or the bromide is preferably used. The free acid of formula III or', if desired, also a salt thereof can be used, whereby the 19S5ba same salts as the salts of the compounds of formula I mentioned above come into consideration.

In order to manufacture the readily hydrolysable esters of the carboxylic acids of formula I in accordance 5 with variant (c), the carboxylic acid is preferably reacted with the corresponding halide, preferably with the iodide, containing the ester group. The reaction can be accelerated with the aid of a base, for example an alkali metal hydroxide or carbonate or an inorganic amine such 10 as triethylamine. When group (b) is present the carboxy function thereof is also esterified, a readily hydrolysable diester being obtained. In this case an excess of the corresponding halide is preferably used. The esterification is preferably carried out in an inert organic solvent such 15 as dimethylacetamide, hexamethylphosphoric acid triamide, dimethyl sulphoxide or, preferably, dimethylformamide. The temperature preferably lies in the range of about 0-40°C.

The manufacture of the salts and hydrates of the compounds of formula I or the hydrates of these salts can 20 be carried out in a manner known per se; for example, by reacting the carboxylic acid of formula I with an equivalent amount of the desired base, conveniently in a solvent such as water or in an organic solvent such as ethanol, methanol, acetone etc. When a second equivalent of base is used 25 salt formation is also effected on the carboxy function of a 18 group (b) which may be present, a di-salt resulting. The temperature at which the salt formation is carried out is not critical. It is generally carried out at room temperature, but it can also be carried out at a temperature slightly above or below room temperature (e.g. in the range of 0°C to +50°C). automatically in the course of the manufacturing process or as a result of the hygroscopic properties of an initially anhydrous product. For the controlled manufacture of a hydrate, a completely or partially anhydrous product (carboxylic acid of formula I or ester, ether or salt thereof) can be exposed to a moist atmosphere (e.g. at about +10°C to +40°C).

The 7-amino compounds of formula IV used above can be prepared starting from a compound of the formula The manufacture of the hydrates usually takes place H H S- IX cooh ( 19 19 in which Z represents a leaving group and the carboxy group can be protected by salt formation with an inorganic or tertiary organic base, with a thiol of formula VII. The reaction can be carried out under the same conditions as those which have been described for the reaction of the starting materials of formulae VI and VII. On the other hand, the compounds of formula VI can be prepared starting from a compound of 10 formula IX and an acid of formula V or a reactive functional derivative thereof under the same conditions as have been described for the reaction of the compounds of formulae IV and V. resulting compound of formula IV can subsequently be protected if desired, for example by esterifying the carboxy group or forming a salt thereon or by silylating the amino group.

A syn/anti mixture of a compound of formula I which 20 may be. obtained can be separated into the corresponding syn- and anti-forms in the customary manner, for example by recrystallisation or by chromatographical methods using a suitable solvent or solvent mixture.

The carboxy group and/or the amino group of the ISSPri The compounds of formulae I and II as well as the corresponding readily hydrolysable esters and salts or the hydrates of these products have antibiotic, especially bactericidal, activity. They possess a broad spectrum of activity against gram-positive bacteria (e.g. Staphylococci) and against gram-negative bacteria such as, for example, Haemophilus influenzae and Neisseria, gonorrhoeae, as well as against various 3-lactamase-forming gram-negative organisms such as Escherichia coli, Serratia marcescens, Enterobacter cloacae, Pseudomonas aeruginosa, Proteus mirabilis and Proteus vulgaris.

The compounds of formula I and II as well as the corresponding readily hydrolysable esters and salts or the hydrates of these products can be used for the treatment and prophylaxis of infectious diseases. A daily dosage of about 0.1 g to about 2 g comes into consideration for adults. The parenteral administration of the compounds in accordance with the invention is especially preferred.

In order to demonstrate the antimicrobial activity of the aforementioned products, the following representative members were tested: Product A: (6R,7R)-7-/2-(2-amino-4-thiazolyl)-2-[(Z)- -methoxyimino] acetamidq7-3-/~t (2,5-dihydro-6--methoxy-2-methyl-5-oxo-as-triazin-3-yl)thio]-methyl7-8-oxo-5-thia-l-azabicyclo[4.2.0]oct--2-ene-carboxylic acid sodium salt..

Product B: (6R,7R)(2-amino-4-thiazolyl)-2-[(Z)--methoxyimino] acetamido7-3-/~[ (3-carboxy-l--methyl-lH-1,2,4-triazol-5-yl)thio]methy17--8-oxo-5-th±a-l-azabicyclo[4.2.0]oct-2-ene--2-carboxylic acid disodium salt.

Product C: (6R,7R)-7-[2-(2-amino-4-thiazolyl)-2-((Z)- -methoxyimino)acetaraido]-3-[[[2,5-dihydro-2" -methyl-5-oxo-6-[(pivaloyloxy)methoxy]-as--triazin-3-yl] thio]methyl] -8-oxo-5-thia-l--azabicyclo[4.2.0]oct-2-ene-2-carboxylie acid sodium salt.

Activity in vitro: Minimum inhibitory concentration (ng/ml) Pi paten"1- offta: 2 3 MAR 1983 DECEIVED L 22 - Pathogenic organism A B C .

Escherichia cola *) 0.04 - 0.08 : 0.10 Serratia marcescens *) 0.16 0.16 3 • 1 Enterobacter cloacae *) 1.2 Proteus mirabilis ..*) 0.-04 0.01 0.4 Proteus vulgaris *) 0.02 0.01 0.2 Pseudomonas aeruginosa strain 1 *) 40 40 50 strain 2 *) 80 40 50 Haemophilus influenzae 0.01 0.01 0.04 Neisseria gonorrhoeae 0.01 0.0025 0.006 Staphylococcus aureus 2.5 1.6 *) 0-lactamase-forming strain Compounds of formula I in which X represents group (b) as well as the corresponding esters, salts and hydrates are distinguished by a long half-life, i.e. they remain in the organism for a long time and can accordingly more effectively exert their germicidal function, Thus, for example, the disodium salt of (6R,7R)(2-amino-4-thiazolyl)-2-[(Z)- -methoxyimino]acetamidg7-3-/~[ (3-carboxy-l-methyl-lH-l,2,4- -triazol-5-yl)thio]methyl7-8-oxo-5-thia-l-azabicyclo[4.2.0]- oct-2-ene-2-carboxylie acid has a half-life in rats of 22 — PATENT 0FFfr_ _v«lutes after the administration of 20 mg of substance per 2 3 MAR J983 Vf'T) 4 196551 kg body weight.

Toxicity Test substance A B c LD100 m9/k9 i-v- > 500 > 500 s. c. >4000 >4000 p.o. >5000 >5000 >5000 The products in accordance with the invention can be used as medicaments, for example in the form of pharmaceutical preparations which contain them in admixture with a pharmaceutical, organic or inorganic inert carrier material which is suitable for enteral or parenteral administration such as, for example, water, gelatin, gum arabic, lactose, starch, magnesium stearate, talc, vegetable oils, polyalkylene glycols, Vaseline etc. The pharmaceutical preparations can be made up in solid form (e.g. as tablets, dragees, suppositories or capsules) or in liquid form (e.g. as solution suspensions or emulsions). If necessary, they can be sterilised and/or can contain adjuvants such as preserving, stabilising, wetting or emulsifying agents, salts for varying the osmotic pressure, anaesthetics or buffers. They can also contain still other therapeutically valuable substances. 196551 The compounds of formula I and their salts or hydrates are preferably administered parenterally and for this purpose are preferably prepared as lyophilisates or dry powders for dilution with customary agents such as water or isotonic sodium chloride solution. The readily hydrolysable esters of the compounds of formula I and their salts or hydrates also come into consideration for enteral administration.

The following Examples illustrate the present invention: Example 1 Preparation of the sodium salt of (6R,7R)(2--amino-4-thiazolyl)-2-[(Z)-methoxyimino]acetamido7-3-/^ (2,5--dihydro-6-methoxy-2-methyl-5-oxo-as-triazin-3-yl)thio]methyl7--8-oxo-5-thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid. 34.5 g of the sodium salt of (6R, 7R)-7-/2-[2-(2--chloroacetamido)-4-thiazolyl]-2-[(Z)-methoxyimino]-acetamido7--3-£[ (2,5-dihydro-6-methoxy-2-methyl-5-oxo-as-triazin-3-yl) — thio]methyl7-8-oxo-5-thia-l-azabicyclo[4.2.0]oct-2-ene-2--carboxylic acid are dissolved in 700 ml of water together with 20 g of thiourea and stirred overnight at 25°C while gassing with nitrogen, the pH-value of the reaction solution being held at 7 by adding IN sodium hydroxide. The pH-value is re-adjusted to 4 by adding IN hydrochloric acid while stirring vigorously. The material which thereby precipitates is filtered off under suction and discarded. The yellow mother liquor is adjusted to pH 3 with IN hydrochloric acid. The precipitated product is filtered off under suction, washed with water, ethanol and low-boiling petroleum ether and dried at 40°C in vacuo. There is obtained beige coloured (6R,7R)--7-/2-(2-amino-4-thiazolyl)-2-[(Z)-methoxyimino]acetamidq7--3-/CI (2,5-dihydro-6-methoxy-2-methyl-5-oxo-as-triazin-3--yl)-thio]methyl7-8-oxo-5-thia-l-azabicyclo[4.2.0]oct-2-ene- ' «. ':c. 19655 f -2-carboxylic acid which, for conversion into the sodium salt, is suspended in 350 ml of methanol and treated with 20 ml of a 2N solution of sodium 2-ethylcaproate in ethyl acetate. A solution has resulted after ca 15 minutes and 5 this solution is diluted with 300.ml ofethanol. . Thereby, ■ ■■ ■ i V there precipitates a small amount of amorphous material which is filtered off under suction and discarded. The filtrate is concentrated strongly at 40°C in vacuo. The material which thereby precipitates is filtered off under suction, washed with ethanol and low-boiling petroleum ether, and dried overnight at 40°C in a high vacuum. There is obtained pure sodium salt of (6R,7R)(2-amino-4- -thiazolyl)-2-[(Z)-methoxyimino]acetamido7-3-/~[(2,5-dihydro- -6-methoxy-2-methyl-5-oxo-as-triazin-3-yl)thio]methyl7-8- ^ -oxo-5-thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid as a yellowish powder with [a]D = -140.9° (c = 1 in water).

The compound used as the starting material in the above process can be prepared as follows: (a) Preparation of tetrahydro-2-methyl-5,6-dioxo-3-thioxo-20 -as-triazine-l(2H)-carboxylic acid benzyl ester. 39.79 g of 1,2,5,6-tetrahydro-5,6-dioxo-3-mercapto--2-methyl-as-triazine are taken up in 500 ml of dimethylformamide and treated with 35 ml of triethylamine. 39 ml 112. PATENT 01 2 3 MAR 1983 of benzyloxycarbonyl chloride are added dropwise during -— srtOFHCl 196551 minutes, whereby the temperature of the reaction mixture rises to 45°C, triethylamine hydrochloride precipitates and the suspension becomes yellow in colour. The mixture is stirred at 25°C for 2 1/2 hours and subsequently 5 evaporated at 70°C in vacuo. The oily evaporation residue is stirred with 500 ml of water for 1 hour, whereby it becomes solid. The solid is filtered off under suction and washed with 50 ml of water. The yellow filter material is stirred well with 250 ml of ethanol, filtered off under 10 suction, washed with ethanol and dried at 40°C in vacuo. There is obtained a colourless crude crystallisate which is recrystallised from methanol and yields colourless tetrahydro-2-methyl-5,6-dioxo-3-thioxo-as-triazine-l(2H)--carboxylic acid benzyl ester of melting point 150-153°C. (b) Preparation of 3-[(diphenylmethyl)thio]-5,6-dihydro--2-methyl-5,6-dioxo-a"s-triazine-l (2H) -carboxylic acid benzyl ester. 13.2 g of tetrahydro-2-methyl-5,6-dioxo-3-thioxo-as--triazine-1(2H)-carboxylic acid benzyl ester are dissolved 20 in 500 ml of ethyl acetate and treated with a solution of diphenyldiazomethane in 90 ml of low-boiling petroleum ether. The initially violet reaction solution is left to stand at 25°C for 40 hours, the colour becoming pink. 3 ml of glacial acetic acid are added and, after 1 hour, 196551 the mixture is evaporated at 40°C in vacuo. A yellow oil is obtained as the evaporation residue. This is separated by means of column chromatography on silica gel with the elution agents benzene, benzene/ethyl acetate 95:5 5 and benzene/ethyl acetate 90:10. The fractions containing the desired substance are combined and evaporated at 40°C in vacuo. After recrystallisation from ethanol, there is obtained colourless 3-[(diphenylmethyl)thio]-5,6-dihydro-2--methyl-5,6-dioxo-as-triazine-l(2H)-carboxylic acid benzyl 10 ester of melting point 90-92°C. (c) Preparation of 3-[(diphenylmethyl)thio]-1,2-dihydro--2-methyl-as-triazine-5,6-dione. 6.9 g of 3-[(diphenylmethyl)thio]-5,6-dihydro-2--methyl-5,6-dioxo-as-triazine-l(2H)-carboxylic acid benzyl 15 ester are dissolved in 100 ml of ethyl acetate and stirred well with 30 ml of an aqueous, 7% ammonia solution at 25°C for 15 minutes. The ethyl acetate phase is separated and discarded. The aqueous phase is treated with 7 ml of concentrated hydrochloric acid and stirred in an ice-bath 20 for 30 minutes. The substance which thereby precipitates is filtered off under suction, washed with water and immediately recrystallised from ethanol. There is obtained colourless 3-[(diphenylmethyl)thio]-1,2-dihydro-2-methyl--as-triazine-5,6-dione of melting point 180-182°C. 1S 6 J- (d) Preparation of 3-[(diphenylmethyl)thio]-6-methoxy--2-methyl-as-triazin-5(2H)-one. 3.9 g of 3-[(diphenylmethyl)thio]-1,2-dihydro-2--methyl-as-triazine-5,6-dione are dissolved in 40 ml of dimethylformamide and treated with 1.6 8 ml of triethylamine as well as 4 ml of methyl iodide. After stirring for 2 hours at 25°C, 1.38 g of potassium carbonate are added to the mixture in order to accelerate the reaction. After stirring for a further 2 hours at 25°C, the starting material has reacted completely. The resulting red suspension is poured into water and extracted three times with ethyl acetate. The combined ethyl acetate extracts are washed with water, dried over sodium sulphate and evaporated at 40°C in vacuo. The red evaporation residue is purified by means of column chromatography on silica gel with ethyl acetate as the elution agent. Recrystallisation from ethyl acetate/ether/ petroleum ether yields yellowish 3-[(diphenylmethyl)thio]--6-methoxy-2-methyl-as-triazin-5(2H)-one of melting point 135-138°C. (e) Preparation of 3s4-dihydro-6-methoxy-2-methyl-3--thioxo- as-triazin-5(2H)-one. 2.6 g of 3-[(diphenylmethyl)thio]-6-methoxy-2-methyl--as-triazin-5(2H)-one-are dissolved in 26 ml of anisole and 50 ml of trifluoroacetic acid and stirred at;25°C for 3 hours. The reaction solution is evaporated at 40°C in vacuo. The evaporation residue is stirred with a small amount of ethyl acetate and a large amount of low-boiling petroleum 5 ether, filtered off under suction and recrystallised from methanol... There is obtained 3,4-dihydro-6-methoxy-2-methyl--3~thioxo-as-triazin-5(2H)-one as colourless, lustrous crystals of melting point 215-220°C. (f) Preparation of (6R,7R)-7-amino-3-/l(2,5-dihydro-10 -6-methoxy-2-methyl-5-oxo-as-triazin-3-yl)thio]- methyl7~8-oxo-5-thia-l-azabicyclo[4.2.0]oct-2-ene--2-carboxylic acid. 13.6 g of 7-aminocephalosporanic acid and 9.5 3 g of 3,4-dihydro-6-methoxy-2-methyl-3-thioxo-as-triazin-5(2H)-15 -one are suspended in 250 ml of water. The pH-value of the suspension is adjusted to 6.5 at 25°C with IN sodium hydroxide. The .suspension is heated to 60°C and stirred at 60°C for 4 hours while gassing with nitrogen, the pH-value being held at 6.5 with IN sodium hydroxide. .After 20 1 hour a solution results, after 2 hours some material begins to precipitate, and sodium hydroxide is no longer * consumed. The mixture is cooled to 25°C and separated . from a small amount of insoluble material by filtration. The orange filtrate is adjusted to pH 3.5 at 25°C with 25 concentrated hydrochloric acid. The precipitated substance HJL PATENT omce 2 3 MAR 1983" is filtered off under suction, washed successively with 100 ml of water, 300 ml of acetone and 300 ml of low-boiling petroleum ether and dried at 40°C overnight in vacuo.. There is obtained a beige coloured substance which, for purification, is suspended in 150 ml of water/methanol 1:1 and treated dropwise with 3.6 ml of triethylamine for 1 hour, whereby ^t pH 8.5 there results a dark solution.which still contains a small amount of insoluble material. After the addition of 3 g of decolourising charcoal, the mixture is stirred at 25°C for 15 minutes, filtered off from the decolourising charcoal over a hard filter and back-washed with 50 ml of water/methanol 1:1. The orange filtrate is adjusted to pH 3 with ca 2 ml of concentrated hydrochloric acid. The substance which thereby precipitates is filtered off under suction, washed successively with 50 ml of water/methanol 1:1, 100 ml of methanol, 50 ml of ether and 50 ml of low-boiling petroleum ether and dried at 40°C overnight in vacuo. There is obtained pure beige coloured (6R,7R)--7-amino-3-/~[ (2 ,5-dihydro-6-methoxy-2-methyl-5-oxo-as--triazin-3-yl)thio]methyl7-8-oxo-5-thia-l-azabicyclo(4.2.0]-oct-2-ene-2-carboxylic acid. g) »L2LPATEFVT0mCg 2 3 MAR 1983 * Preparation of the sodium salt'of- (6R,7R)-7-/2--[2-(2-chloroacetamido)-4-thiazolyl]-2-[(Z)-methoxyimino] acetamido7-3-/T(2,5-dihydro-6-methoxy-2-methyl--5-oxo-as-triazin-3-yl)thio]methyl7-8-oxo-5-thia-l-—azabicyclo[4.2.0]oct~2-ene-2-carboxylic acid. -cD g of phosporus pentachloride are added to 200 ml of methylene chloride dried over calcium chloride. The suspension is cooled to -10°C while stirring and 46 ml of N,N-dimethylacetamide are added dropwise for ca 5 minutes, the temperature rising to -5°C. The suspension is cooled to -15°C and stirred for 15 minutes. After cooling to -20°C, 33.3 g of 2-(2-chloroacetamido-4-thiazolyl)-2-(Z)- -methoxyimino-acetic acid are added and the mixture is stirred at -15°C for 45 minutes, a yellow-orange solution resulting. This solution is cooled to -30°C, treated with 50 g of ice and stirred at -50°C for 10 minutes. The methylene chloride phase containing the acid chloride formed is separated and stored initially at ca -15°C. This acid chloride solution is added-dropwise during 30 minutes while stirring vigorously to a solution, cooled to 0-5°C, which has been prepared by treating (6R,7R)-7-amino-3-^ft2,5-dihydro -6-methoxy-2-methyl-5-oxo-as-triazin-3-yl)thio]methyl7~8-oxo- -5-thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, prepared under (f), with 3N sodium hydroxide to pH 8.2 in 500 ml of water, the pH-value being held at 8.0-8.2 with 3N sodium hydroxide with the aid of an autotitrator. The mixture is stirred at 25°C for 2 hours. Thereafter, 300 ml of methylene chloride and 2 1 of n-butanol are added. The pH is re-adjusted to 2 with 3N hydrochloric acid while stirring vigorously. The organic phase is separated,. washed three times with 1 1 of water each time, stirred for 15__iainutes with 20 g of decolourising charcoal and filtered.

NJL FATWT OFRCS 2 3 MAR 1983 j The light yellow filtrate is concentrated strongly at 60°C in vacuo, the reaction product precipitating. The pre- . cipitated product is filtered off under suction and washed successively with n-butanol, ether and low-boiling petroleum ether and dried at 40°C overnight in vacuo. There is obtained crude, beige (6R,7R)-7-/2-[2-(2-chloroacetamido)-4--thiazolyl] -2- [ (Z) -methoxyimino] acetamido7-3-/~[ (2 ,5-dihydro--6-methoxy-2-methyl-5-oxo-as-triazin-3-yl)thio]methy17-8--oxo-5-thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid. For conversion into the sodium salt and purification, the acid is dissolved in a mixture of 300 ml of acetone and 50 ml'of methanol and treated with 35 ml of a 2N solution of sodium 2-ethylcaproate in ethyl acetate. The precipitated sodium salt is filtered off under suction, washed successively with acetone and low-boiling petroleum ether and dried at 25°C in vacuo. There is obtained the beige coloured sodium salt of (6R,7R)-7-/2-[2-(2-chloroacetamido)-4-thiazolyl]-2--[(Z)-methoxyimino]acetamido7-3-/"[ (2,5-dihydro-6-methoxy-2--methyl-5-oxo-as-triazin-3-yl)thio]methy!7-8-oxo-5-thia -1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid with [a]^ = -131.5° (c = 1 in water).

Example 2 © Preparation of the disodium salt of (6R,7R)-7-/2-—(2-amino-4-thiazolyl)-2-[(Z)-methoxyimino]acetamido7~3-~£X (3-carboxy-l-methyl-lH-l,2,4-triazol-5-yl)thio]methy!7--8-oxo-5-thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid. 53 g of (6R, 7R)-3-/TT ( 3-carboxy-l-methyl-lH-l, 2 , 4--triazol-5-yl)thio]methy17-7-/2-[2-(2-chloroacetamido)-4--thiazolyl]-2-[(Z)-methoxyimino]acetamido7-8-oxo-5-thia--1-azahicyclo[4.2.0]oct-2-ene-2-carboxylic acid are suspended in 1 1 of water together with 34. g of thiourea. . The pH is adjusted to 7 by the dropwise addition of IN sodium hydroxide, a dark brown solution resulting. This solution is stirred at pH 7 overnight while gassing with nitrogen, the pH being held constant with IN sodium hydroxide with the aid of an autotitrator. The pH of the reaction solution is re-adjusted to 3.5 with IN hydrochloric acid while stirring. The precipitated substance (fraction I) is filtered off under suction, washed with 250 ml of water and dried at 40°C in vacuo. Fraction I is brown in colour, and very much impure; it is discarded. The orange mother liquor is re-adjusted.to pH 2.65 with IN hydrochloric acid while stirring. The precipitated material is filtered off under suction and washed with 250 ml of water. . The beige coloured filter material is azeotropically distilled with ethanol under reduced pressure, filtered off under suction, washed with ethanol and low-boiling petroleum ether and dried at 40°C in vacuo. There is thus obtained a beige-brown fraction II which, for purification and conversion into the disodium salt, is suspended in 2 1 of methanol, treated with 50 ml of a 2N solution of sodium 2-ethylcaptroate in ethyl acetate and subsequently stirred with 200 ml of.water for 30 minutes. A small amount of insoluble, brown material is filtered off and discarded. Ca 1 1 is removed from the orange coloured filtrate by evaporation at 40°C in vacuo, then 1 1 of ethanol is added and the mixture is concentrated in vacuo to a volume of ca 500 ml. The solution is decanted off fxom the viscous brown resin, . a heavy mixture, which thereby separates. The decanted-off yellow-orange solution is treated with 11.of ethanol and concentrated strongly at 40°C in vacuo, substance partially precipitating. After adding 2.5 1 of methanol, there is obtained a 'yellow solution which is concentrated strongly at 40°C in vacuo. The thereby precipitated disodium salt is filtered off under suction, washed with methanol and low-boiling petroleum ether and dried at 35°C in a high vacuum. There is obtained the pure, beige coloured disodium salt of (6R,7R)-7-/2-(2-amino-4-thiazolyl)-2-[(Z)-methoxyimino] acetamidc7-3-ZT(3-carboxy-l-methyl-lH-l,2,4-triazol- -5-yl)thio]methyl7-8-oxo-5-thia-l-azabicyclo[4.2.0]oct-2- -ene-2-carboxylic acid with [a]D = -38.7° (c = 1 in water).

The compound used as the starting material in the above process can be prepared as follows: (a) Preparation of 4,5-dihydro-l-methyl-5-thioxo-lH--O.^2,4-triazol-3-carboxylic acid methyl ester. 196551 12.9 g of l-methoxyoxalyl-2-methyl-thiosemicarbazide are added to a solution of 1.6 g of sodium in 200 ml of methanol. The mixture is boiled under reflux for 4 hours. The substance which precipitates after 1/2 hour is 5 separated. The mother liquor is evaporated at 40°C in vacuo. The evaporation residue is dissolved in 100 ml of water and acidified with concentrated hydrochloric acid. The crystallisate which thereby separates out yields, after recrystallisation from water, pure, colourless 4,5-dihydro-10 -l-methyl-5-thioxo-lH-l,2,4-thiazole-3-carboxylic acid methyl ester of melting point 188-190°C (decomposition). (b) Preparation of 4,5-dihydro-l-methyl-5-thioxo-lH--1,2,4-triazole-3-carboxylic acid. 3.46 g of 4f5-dihydro-l-methyl-5-thioxo-lH-l,2,4-15 -triazole-3-carboxylic acid methyl ester are dissolved in 50 ml of IN sodium hydroxide. After stirring for 1/2 hour at 25°C, the reaction solution is made acid with 25 ml of 2N hydrochloric acid and concentrated at 40°C in vacuo, the desired acid crystallising. This acid is filtered 20 off under suction, washed with ice/water and dried at 45°C in a high vacuum. There is obtained colourless 4,5-dihydro--l-methyl-5-thioxo-lH-l,2,4-triazole-3-carboxylic acid of melting point 177-178°C (decomposition). (c) Preparation of (6R,7R)-7-amino-3-,£I (3-carboxy-l--methyl-1,2,4-triazol-5-yl)thio]methyl7-8-oxo-5--thi'a-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid. 34 g of 7-aminocephalosporanic acid are suspended -5 together with 25.5 g of 4,5-dihydro-l-methyl-5-thioxo--1H-1,2,4-triazole-3-carboxylic acid in 500 ml of water. The pH is adjusted to 6.5 with 3N sodium hydroxide. The mixture is stirred at 55°C for 4 hours while gassing with nitrogen, the pH being held constant at 6.5 with 3N sodium 10 hydroxide with the aid of an autotitrat'or. The dark solution is cooled to 25°C and adjusted to pH 3.5 with concentrated hydrochloric acid. After 15 minutes, the precipitated material is filtered off under suction, suspended in 500 ml of water and brought into solution at 15 pH 7 with 3N sodium hydroxide. The orange-red solution is adjusted to pH 3 with concentrated hydrochloric acid. The precipitated substance is filtered off under suction and washed successively with 250 ml of water, 500 ml of acetone, 500 ml of low-boiling petroleum ether and dried 20 at 40-45°C overnight in vacuo. There is obtained (6R,7R)--7-amino-3-/T(3-carboxy-l-methyl-l,2 f 4-triazol~5-yl)thio] ~ methyl7-8-oxo-5-thia-l-azabicyclo[4.2.0]oct-2-ene-2--carboxylic acid as a beige,coloured powder.

J&Z. PATENT GPRCl i < 2 3 MAR 1983 I ! (d) Preparation of (6R,7R)-3-/T(3-carboxy-l-methyl-lH--1,2,4-triazol-5-yl)thio]methyl7-7-/2~[2~(2-,-chloroacetamido)-4-thiazolyl]-2-[(Z)-methoxyimino]-acetamido7-8-oxo-5-thia-l-azabicyclo[4.2.0]oct--2-ene-2-carboxylic acid.

This compound is prepared in accordance with paragraph (g) of Example 1 starting from 100 g of 2-(2-chloroacetamido--4-thiazolyl)-2-(Z)-methoxyimino-acetic acid and 111.5 g of (6R,7R)-7-amino-3-/Tt (3-carboxy-l-methyl-l,2,4-triazol--5-yl)thio]methyl7-8-oxo-5-thia-l-azabicyclo[4.2.0]oct-2--ene-2-carboxylic acid. There is obtained beige coloured (6R,7R)-3-/Tt (3-carboxy-l-methyl-lH-l,2,4-triazol-5-yl)thio]-methyl7~7-/2- (2-chloroacetamido)-4-thiasolyl] -2-[ (Z) - -:.-; -methoxyimino]'acetamidg7-8-oxo-5-thia~l-azabicyclo[4.2.0]-oct-2-ene-2-carboxylic acid.

Example 3 Preparation of the sodium salt of (6R,7R)-7-[2-(2--amino-4-thiazolyI)-2- ( (Z) -methoxyimino) acetamido] -3- [ [ [2 ,5--dihydro-2-methyl-5-oxo-6-[(pivaloyloxy)methoxy]-as-triazin--3-yl] thio] methyl] -8- oxo-5-thia-l-azabicyclo [4 .2 .0] oct-2--■ene-2-carboxylic acid. 13.5g of(6R,7R)-7-[4-bromo-l-[(Z)-methoxyimino]aceto-acetamido]-3-[[[2 f 5-dihydro-2-methyl-5-oxo-6-[(pivaloyloxy)- 196551 methoxy]-as-triazin-3-yl]thio]methyl]-8-oxo-5-thia-l--azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid and 2.96 g of thiourea arfe dissolved in 80 ml of ethanol. The solution is stirred at 25°C for 2 hours and then treated with 22 ml of a 2N solution of sodium 2-ethylcaproate in ethyl acetate, the sodium salt precipitating immediately. The mixture is now diluted to 1000 ml with ethanol. 500 ml of methanol are added, a yellow-brownish solution resulting. Ca. 600 ml of solvent are> distilled off at 40°C in vacuo. The material which thereby precipitates is filtered off under suction, washed with ethanol and low-boiling petroleum ether and dried at 40°C in a high vacuum. There is obtained a brownish fraction I which is discarded.

A further 250 ml of solvent are distilled off from the mother liquor at 40°C in vacuo. The material which thereby precipitates if filtered off under suction, washed with ethanol and low-boiling petroleum ether and dried at 40°C overnight in a high vacuum. A beige fraction II is obtained. For purification, this substance is dissolved in 100 ml of water and treated with 2 1 of ethanol. This solution is strongly concentrated at 40°C in vacuo. The precipitated material is filtered off under suction, washed with ethanol and low-boiling petroleum ether and dried at 40 in a high vacuum. There is obtained a fraction III which is discarded. The mother liquor is, for the removal of water, 196551 treated twice with 1 1 of ethanol each time and strongly concentrated at 40°C in vacuo. Finally, acetone is added. The substance which thereby precipitates is filtered off under suction, washed with acetone and low-boiling petroleum ether and dried at 40°C overnight in a high vacuum.

There is thus obtained pure, non-crystalline title substance 25 with [a]D = 124.6° (c = 1 in water). The nuclear resonance spectrum and the microanalysis correspond to the given structure.

■ The (6R,7R)-7-[4-bromo-2-[(Z)-methoxyimino]aceto- acetamido]-3-[[[2,5-dihydro-2-methyl-5-oxo-6-[(pivaloyloxy)-me.thoxy]-as-triazin-3-yl]thio]methyl]-8-oxo-5-thia-l--azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid used as the starting material in the above process can be prepared 15 as follows: (a) Preparation of tetrahydro-2-methyl-5,6-dioxo-3--thioxo-as-triazine-1(2H)-carboxylic acid benzyl ester. 39.79 g of 1,2,5,6-tetrahydro-5,6-dioxo-3-mercapto-2-20 -methyl-as-triazine are taken-up in 500 ml of dimethylformamide and treated with 35 ml of triethylamine. 39 ml of benzyloxycarbonyl chloride are added dropwise during 15 minutes, whereby the temperature of the reaction mixture rises to 45°C, triethylamine hydrochloride precipitates and the -41- - 19 6 & £> 1 suspension becomes yellow in colour. The mixture is stirred at 25°C for 2 1/2 hours and subsequently evaporated at 70°C in vacuo. The oily evaporation residue is stirred' with 500 ml of water for 1 hour, whereby it becomes solid. The solid obtained is filtered off under suction and washed with 50 ml of water. The yellow filter material is stirred well with 250 ml of ethanol, filtered off under suction, washed with ethanol and dried at 40°C in vacuo. There is obtained a colourless crude crystallisate which is recrystallised from methanol and yields colourless pure substance of m.p. 150-15 3°C. (b) Preparation of 3-[(diphenylmethyl)thio]-5,6-dihydro- -2-methyl-5,6-dioxo-as-triazine-l(2H)-carboxylic acid benzyl ester. 13.2 g of tetrahydro-2-methyl-5 , 6 -dioxo-3-thioxo--as-triazine-1(2H)-carboxylic acid benzyl ester are dissolved in 500 ml of ethyl acetate and treated with a solution of diphenyldiazomethane in 90 ml of low-boiling petroleum ether. The initially violet reaction solution is left to stand at 25°C for 40 hours, the colour becoming pinko 3 ml of glacial acetic acid are added and, after 1 houre the mixture is evaporated at 40°C in vacuo. A yellow oil is obtained as the evaporation residue. This is separated by means of column chromatography on silica gel with the elution agents benzene, benzene/ethyl acetate 95:5 and benzene/ethyl acetate 1965^1 90:10. The fractions containing the desired substance are combined and Evaporated at 40°C in vacuo. After recrystallisation from ethanol, there is obtained colourless pure substance of m.p. 90-92°C. (c) Preparation of 3-[(diphenylmethyl)thio]-1,2-dihydro--2-methyl-as-triazine-5,6-dione. 6.9 g of the substance prepared under (b) are dissolved in 100 ml of ethyl acetate and stirred well with 30 ml of an aqueous, 7% ammonia solution at 25°C for 15 minutes. 10 The ethyl acetate phase is separated and discarded. The aqueous phase is treated with 7 ml of concentrated hydrochloric acid and stirred in an ice-bath for 30 minutes. The substance which thereby precipitates is filtered off under suction, washed with water and immediately recrystallised from ethanol. 15 There is obtained colourless pure substance of m.p. 180--182 °C. (d) Preparation of 3-[(diphenylmethyl)thio]-6-pivaloyloxy-methoxy-2-methyl-as-triazin-5(2H)-one. 3.-25 g of the compound prepared under (c) are dissolved 20 in 40 ml of dimethylformamide and treated with 1.52 g of potassium carbonate. 2.66 g of pivaloyloxymethyl iodide are added to this mixture in one portion, a pale yellow solution 186551 resulting. The reaction mixture is stirred at 25°C for 4 hours and once again treated with 2.66 g of pivaloyloxy-methyl iodide. This mixture is stirred at 25°C for 15 hours and subsequently evaporated at 35°C in a high vacuum. The 5 resulting evaporation residue is partitioned between 100 ml of water and 100 ml of ethyl acetate. The aqueous phase is extracted twice with 50 ml of ethyl acetate each time. The combined ethyl acetate phases are dried over sodium sulphate and evaporated at 35°C in vacuo. The residual yellow oil 10 is chromatographed on a silica gel column with ethyl acetate as the elution agent. The fractions containing the desired product are combined and evaporated at 35°C in vacuo. The residual pale blue oil is dried at 25°C for 1 hour in a high vacuum, the pure substance being obtained as a pale blue 15 ' resin. (e) Preparation -of [ (2, 3,4,5-tetrahydro-2-methyl-5-oxo-3--thioxo-as-triazin-6-yl)oxy]methyl pivalate. 3.6 g of the compound prepared under (d) are stirred at 25°C for 2 1/2 hours in 18 ml of anisole and 18 ml of trifluoro 20 acetic acid. The solution is then evaporated at 50°C in vacuo. The oily residue is stirred with 50 ml of low-boiling petroleum ether, crystallisation occurring. The crystallisate is filtered off under suction and washed with low-boiling petroleum ether. There are obtained white crystals which, 1tT> F ^ "* A after recrystallisation from ether/low-boiling petroleum ether, yield white, crystalline pure substance of m.p. 112-113°C. The nuclear resonance spectrum and the microanalysis correspond to the given structure. (f) Preparation of (6R,7R)-7-amino-3-[[[2,5-dihydro-2--methyl-5-oxo-6- [ (pivaloyloxy.) methoxy]-as-triazin--3-yl]thio]methyl]-8-oxo-5-thia-l-azabicyclo[4.2.0]-oct-2-ene-2-carboxylic acid. 3.0 g of the compound prepared under (e) are suspended in 50 ml of water together with 2.72 g of 7-aminocephalo-sporanic acid. While gassing with nitrogen the suspension is adjusted to pH 6.5 with IN sodium hydroxide and stirred for 4 hours at pH 6.5-7 and at 55-60°C. The desired compound precipitates out and is filtered off under suction after cooling the mixture to 25°C. After washing with water, acetone and low-boiling petroleum ether and drying at 40°C overnight in a high vacuum, there is obtained the pure substance, the microanalysis and nuclear resonance spectrum of which correspond to the given structure. (g) Silylation of (6R,7R)-7-amino~3-[[[2,5-dihydro--2-methyl-5-oxo-6-[(pivaloyloxy)methoxy]-as-triazin--3-yl]thio]methyl]-8-oxo-5-thia-l-azabicyclo[4.2.0]-oct-2-ene-2-carboxylic acid. ^,96551 14.56 g of the compound prepared under (f) are suspended in 300 ml of ethyl acetate and treated with 30 ml of N,0-bis-(trimethylsilyl)-acetamide. The mixture is stirred at 35-40°C for 30 minutes, a dark solution resulting. 5 This solution is cooled to -5°C to -10°C and stirred with the exclusion of moisture until the acylation has taken place. (h) Preparation of (Z)-2-hydroxyimino-3-oxo-butyric acid tert.-butyl ester. 592.1 g of acetoacetic acid tert.-butyl ester are 10 dissolved in 560 ml of glacial acetic acid. A solution of 290.6 g of sodium nitrite in 655 ml of water is added drop-wise to this solution at 5-lO°C during 2 1/2 hours. The resulting yellow suspension is stirred at 20°C for 30 minutes, treated with 940 ml of water and stirred for a 15 further 2 hours. The mixture is treated with 900 ml of water and 900 g of ice and extracted in a stirrer vessel three times with 1 1 of ethyl acetate each time. The combined ethyl acetate extracts are washed three times with 1 1 of water each time, then treated with 5 1 of water and 20 the pH-value is adjusted to 6C8 with sodium hydrogen carbonate After separating the aqueous phase, the organic phase is washed once with water. Thereafter, the ethyl acetate solution is dried over sodium sulphate and evaporated at 40°C in vacuo. There is obtained (Z)-2-hydroxyimino-3-oxo-25 -butyric acid tert.-butyl ester as a yellow oil which is 19655* dried at 40°C for a further 9 hours in a high vacuum. (i) Preparation of (Z)-2-methoxyimino-3-oxo-butyric acid tert.-butyl ester. 626.65 g of (Z)-2-hydroxyimino-3-oxo-butyric acid 5 tert.-butyl ester are dissolved in 2.86 1 of acetone. The solution is cooled to 5°C and treated portionwise with 703.5 g of potassium carbonate. 322 ml of dimethyl sulphate are then added dropwise to the yellow suspension without cooling during 1 hour, whereby the temperature of the mixture should not rise 10 above 25°C. The light beige suspension is stirred at 20-25°C for ca. 4.hours until starting material is no longer detected by thin-layer chromatography. Thereafter, the mixture is poured into 7 1 of water and extracted three times with 1 1 of ethyl acetate each time. The combined ethyl acetate 15 extracts are washed three times with 1 1 of water each time, dried over sodium sulphate and evaporated at 40°C in vacuo. The residual, yellow oil is dried at 40°C for a further 6 hours in a high vacuum. There is obtained (Z)-2-methoxy-imino-3-oxo-butyric acid tert.-butyl ester \ . • - as a yellow oil with a boiling point of 57°C at 0.02 mm Hg. 1 (j) Preparation of (Z)-2-methoxyimino-3-oxo-butyric acid. 86 g of (Z)-2-methoxyimino-3-oxo-butyric acid tert.--butyl ester are dissolved in 400 ml of trifluoroacetic acid, The solution is left to stand at 25°C for 1 hour and thereafter evaporated at 35°C in vacuo. The oily residue is crystallised from ether/petroleum ether. There is obtained yellowish, water-soluble (Z)-2-methoxyimino-3-oxo-butyric „ i acid of m.p. 80-85 C. ' (k) Preparation of (Z)-4-bromo-2-methoxyimino-3-oxo- -butyric acid. 145 g of (Z)-2-methoxyimino-3-oxo-butyric acid are dissolved in 1000 ml of alcohol-free anhydrous dichloro-methane. 10 ml of 30% hydrobromic acid in glacial acetic acid are added to this solution. Then, a solution of 37.5 ml of bromine in 112.5 ml of dichloromethane is added dropwise during ca. 2 hours, the temperature of the reaction mixture being held at 20-25°C by means of slight cooling. Now, in order to remove hydrogen bromide from the reaction mixture, nitrogen is vigorously blown through. Subsequently, 250 g of ice, 250 ml of water and 2 1 of ether are successively 196551 added. The aqueous phase is separated and discarded. The organic phase is washed with 250 ml of water and 250 ml of saturated sodium chloride solution, dried with sodium sulphate and evaporated in vacuo. There remain behind 170 g 5 of a brown oil which is recrystallised from carbon tetrachloride. There is obtained almost colourless (Z)-4-bromo--2-methoxyimino-3-oxo-butyric acid. * I (1) Preparation of the acid chloride of (Z)-4-bromo--2-methoxyimino-3-oxo-butyric acid. 6.72 g of (Z)-4-bromo-2-methoxyimino-3-oxo-butyric acid are dissolved in 70 ml of anhydrous, alcohol-free 15 methylene chloride and treated at 0-5°C with 6.24 g of phosphorus pentachloride. The reaction solution is stirred at 0-5°C for 15 minutes and without cooling for 45 minutes. (m) Preparation of (6R,7R)-7-[4-bromo-2-[(Z)-methoxyimino]-acetoacetamido]-3-[[[2,5-dihydro-2-methyl-5-oxo-6-20 -[(pivaloyloxy)methoxy]-as-triazin-3-yl]thio]methyl]- -8-oxo-5-thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid.

The solution prepared according to (1) is added dropwise at -10°C to 0°C during ca. 30 minutes to the solution prepared according to (g) . The mixture is stirred at -10°C to 0°C for 30 minutes and without cooling for 1 hour. 300 ml of ethyl acetate and 200. ml of water are then added. The precipitated material is filtered off under suction and discarded. The aqueous phase of the filtrate is separated and discarded. The organic phase is washed 6 times with 250 ml of water each time, dried with sodium sulphate, concentrated strongly at 40°C in vacuo and finally poured into 500 ml of ether. The precipitated substance is filtered off under suction, washed with ether and low--boiling petroleum ether and dried at 40°C overnight in a high vacuum. There is obtained beige, amorphous pure substance The nuclear resonance spectrum and the infrared spectrum correspond to the given structure.

Example 4 Production of dry ampoules for intramuscular administration: .

/A lyophilisate of 1 g of the disodium salt of (6R,7R)-(2-amino-4-thiazolyl)-2-[(Z)-methoxyimino)acetamido7-3--ZTl (3-carboxy-l-methyl-lH-l,2,4-triazol-5-yl)thio]methyl7-8-

Claims (3)

196551 -oxo-5-thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid is prepared in the customary manner and filled into an ampoule. Prior to the administration, the lyophi'lisate is treated with 2.5 ml of a 2% aqueous lidocaine hydrochloride solution. eg ij illci/143 - 51 - what fwg cmn CLAIMS: 1

1.

Cephalosporin derivatives of the general formula h h

CH30n=c conh n [

COOH

in which X represents one of the groups

o

(a)

h-jc

N—N

AX

cooh

(b)

wherein R represents lower alkyl or (together with the oxygen) a readily hydrolysable ether group,

10 as well as readily hydrolysable esters of the compounds of formula I wherein X represents the group (b) or (a) in which R"*" represents lower alkyl and salts of these compounds and hydrates of the compounds of formula I or of esters and salts thereof.

- 52 -

''96 55

2. Cephalosporin derivatives according to Claim 1,

wherein X represents the group (a) in which R1" represents methyl or the group (b).

3. Cephalosporin derivatives in accordance with Claim 1,

wherein R represents the group (a) in which R"*" represents pivaloyloxymethy1.

4.

(6R, 7R) -7-/2- (2-Amino-4-thiazolyl) -2 - [ (Z) -methoxyimino] acetamido7-3-/l(2,5-dihydro-6-methoxy-2-methyl-5--oxo-as-triazin-3-yl)thio]methyl]-8-oxo-5-thia-1-azabicyclo-[4.2.0]oct-2-ene-2-carboxylic acid as well as salts of this compound and hydrates of this compound or salts.

5 . (6R,7R)-7-[2-(2-Amino-4-thiazolyl)-2-[(Z)-methoxy imino] acetamido7~3-/I(3-carboxy-l-methyl-lH-l,2,4-triazol~ -5-yl) thio]methy 1,7-8-0X0-5-thia- 1-azabicyclo [4.2.0] oct-™2-ene-2-carboxylic acid as well as salts of this compound and hydrates of this compound or salts.

6. (6R,7R)-7-[2-(2-Amino-4-thiazolyl)-2-((Z)-methoxy imino) acetamido]-3-[[[2,5-dihydro-2-methyl-5-oxo-6-[(pivaloyloxy) methoxy]-as-triazin-3~y1]thio]methy13-8-oxo-5-thia-l---azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid as well as salts of this compound and hydrates of this compound or salts.

- 53 -

133551

Cephalosporin derivatives of the general formula

H H

CH30N===C CONH

n s

■CH'2 ^'

•COOH

II

in which X represents one of the groups h3c

N—N n

cooh

(a)

(b)

10

wherein R represents lower alkyl or (together with the oxygen) a readily hydrolysable ether group, whereby the carboxy group of group (b) can be protected, R represents a cleavable protecting group and the carboxy group in the 4-position or the cephalosporin moiety can be present in protected form.

- 54 -

196551

Compounds of the general formula h h ch2—s cooh

IV

in which X represents one of the groups

h3c

S;N^ ^0

N-~ N

L W

'cooh

(a)

(b)

10

wherein R represents lower alkyl, whereby the carboxy group of group (b) can be protected, and the carboxy group in the 4-position of the cephalosporin moiety and/or the amino group can be present in protected form.

Halides of the general formula

H H

CH3ON:

c conh-

I

CO ^

CH2V

CH2 s x III

COOH

in which X represents one of the groups

H3C

N—N

N ° • COOH

(a) (b)

wherein Y represents a halogen atom and R"*" represents lower alkyl or (together with the oxygen) a readily hydrolysable ether group, whereby the carboxy group of group (b) can be protected, and the carboxy group in the 4-position or the cephalosporin moiety can be present in protected form.

- 56 -

10. Compounds in accordance with one of claims 1-6 in a form suitable for use as pharmaceutically active substances.

11. . Compounds in accordance with one of claims 1-6 in a form suitable for use as pharmaceutically active substances for the treatment and prophylaxis of infectious diseases.

HJL PATENT <Y:'

j 2 3 MAR 1983 I

deceived t

12. J Pharmaceutical preparations, characterised in that they contain a compound in accordance with one of claims 1

13.| Pharmaceutical preparations for the treatment and prophylaxis of infectious diseases, characterised in that they contain a compound in accordance with one of claims 1-6. •

N.Z. PATBvj

2 3 MAR /983

196551

- 58 -

14. A process for the manufacture of the compounds in accordance with one of claims 1-6, which process comprises (a) cleaving off the protecting group R, and, if necessary, carboxy protecting groups which may be present, in a compound of the general formula h h*

ch3on=c conh

II

-c h 2 s x cooh or (b)

in which X has the same significance as X in claim 1, whereby the carboxy group of group (b) can be protected, R represents a cleavable protecting group and the carboxy group in the 4--position of the cephalosporin moiety can be present in protected form,

reacting a halide of the general formula

h h ch2 s x'

cooh

III

19655

. . -1

in which X"*" has the significance given above and Y represents a halogen atom and the carboxy group in the 4-position of the cephalosporin moiety can be present in protected form,

with thiourea,

or . ..

(c) for the manufacture.of a readily hydrolysable ester of a compound of formula I, subjecting a carboxylic acid of formula I to a corresponding esterification,

or

(d) for the manufacture of salts or hydrates of a compound of formula I or hydrates of these salts,converting a compound of formula I into a salt or hydrate or into a "^ hydrate of said salt. 1

15. A process according to claim 14,' characterised in that there is used a starting material of formula II or III wherein X represents the group (a) in which R^" represents methyl or the group (b) .

196551

- 60 -

16. , A process for the manufacture of the compounds in accordance with one of claims 1-6, substantially as hereinbefore described with particular reference to any one of the foregoing Examples 1 to 3.

17. Compounds as claimed in any one of Claims 1-6, whenever prepared according to the process as claimed in Claim 14, 15 or 16.

18. Cephalosporin derivatives according to any one of claims 1-6 in the syn-isomer form or mixtures of the syn-and : anti-isomers in which the syn-isomer form predominates.

son

PPLICANTS

PATENT QFFiCE

3 f JAN I984