NL8702295A - CEPHALOSPORINE ANTIBIOTICS AND PREPARATIONS CONTAINING THESE ANTIBIOTICS. - Google Patents

Description

- Cephalosporin antibiotics and preparations containing these antibiotics -

The invention relates to cephalosporins and improvements thereof, in particular to new cephalosporin compounds and derivatives thereof with valuable antibiotic properties.

The cephalosporin compounds of the invention are referred to with reference to "cefam" according to J. Amer. Chem. Soc., 1962, 84, 3400, wherein the term "cefemT" denotes the double bond cefamba base structure.

Cephalosporin antibiotics are widely used in the treatment of diseases or conditions caused by pathogenic bacteria in humans and animals, and are particularly useful in the treatment of diseases caused by bacteria resistant to other antibiotics such as penicillin compounds, and in the treatment of patients sensitive to penicillin. It is often desirable to use a cephalosporin antibiotic which has activity against both Gram-positive and Gram-negative microorganisms, and considerable research has been conducted on the development of different types of broad spectrum cephalosporin antibiotics.

For example, Applicant's British Patent No. 1,399,086 describes a new class of cephalosporin antibiotics containing a 7 ft (¾-etherified oxyimino) acyl amido side chain, which oxyimino group has the synon configuration. This class of antibiotic compounds has been marketed by a powerful antibacterial action against a range of Gram-positive and Gram-negative organisms, coupled with particularly high stability against lactamases produced by various Gram-negative organisms.

The discovery of this class of compounds has stimulated further research in the same field, seeking compounds that have improved properties, for example against certain classes of organisms, especially Gram-negative organisms. This interest is evident from the very large ^ 70 2 25? * Λ - 2 - number of patent applications pertaining to cephalosporin anti-biotics with certain substituents both in the 7β-acylamido side chain and in the 3-position of the cephalosporin nucleus.

For example, British Pat. No. 1,567,625 contains a general description of cephalosporin antibiotics with a 7β- (ole-etherified oxyimino) acetamido side chain wherein the etherifying group is an aliphatic hydrocarbon group which may optionally have suitable substituents (in addition to a wide variety of possibilities, for example, a carboxy group or a protected carboxy group and a phenyl group substituted by up to three hydroxy groups), which side chain is further substituted by a group which may include an aminothiazolyl group. However, none of the examples mentioned specifically contain a carboxyphenylalkyl etherifying group. The group 15 in the 3-position can also be chosen from a large number of alternatives, but a hydrogen atom is not included.

A large number of cephalosporin antibiotics are generally described in British Patent 1,604,971, the side chain in the 7-position being selectable from among others a 2- (2-aminothiazol-4-yl) -2- (etherified oxyimino) - acetamido group, wherein the etherifying group, in addition to many other meanings, may be an alkyl group (for example methyl) substituted by both phenyl and carboxy, although there is no specific example of compounds having a carboxyphenylalkyl group and the etherifying group preferably an unsubstituted methyl group. The group in the 3-position can also be chosen from a large number of alternatives and the possible 3 substituents also include a hydrogen atom.

In British Pat. No. 2,017,702, the oxyimino etherifying group, according to the general definition, may include a Krcarboxyphenylmethyl group. In the named compounds, the carboxyphenylmethyl group is always combined with an acetoxymethyl group in the 3-position.

British Patent 2,104,888 generally discloses cephalosporin antibiotics wherein the side chain at the γ1 position is a 2- (2-aminothiazol-4-yl) -2- (etherified oxyimino) acetamido group. The oxyimino etherifying group may include, inter alia, 870220ύ

V

- 3 - be a carboxyphenylmethyl group, where the phenyl group may be substituted by a hydroxy group. The 3-position group is an isothiazolylthiomethyl or iminoalkylidenedithithane group.

European Patent 197,409 5 generally describes cephalosporin antibiotics wherein the side chain in the 7 position is a 2- (2-aminothiazol-r4-yl) -2- (etherified oxyimino) acetamido group. The oxyimino etherifying group may include catecholcarboxymethyloxyimino group.

The substituent at the 3 position can be selected from a number of 10 alternatives, but this does not include a hydrogen atom.

It has now been found that by choosing a (Z) -2- (2-aminothiazol-4-yl) -2— (etherified oxyimino) acetamido group in the 7-position in combination with a hydrogen atom in the 3-position, and by choosing a ty-carboxy substituted phenylmethoxyimino group as the etherified oxyimino group, cephalosporin compounds can be obtained, which have a particularly favorable activity profile (described in greater detail below) against a wide variety of commonly occurring pathogenic organisms and / or as intermediates can be used in the preparation of other active compounds.

Accordingly, cephalosporin compounds of the general formula 1 are provided, wherein and R 2, which may be the same or different, each represent a hydrogen atom or a carboxyl group blocking group,

Rg represents a hydrogen atom or an amino-protecting group, R 1 and R 2 each represent a hydroxy or substituted hydroxy group or R 1 and together represent a cyclic protected diol group, ZS or SH) (0C- or $ -), and / / · The dotted line bridging the 2, 3 and 4 positions indicates that the compound is a cef-2-em or cef-3-em compound, and non-toxic salts and solvates (in particular hydrates) 35 thereof.

When and / or R 2 represent carboxyl group blocking groups, the blocking group may be, for example, the remainder of an ester-forming aliphatic or araliphatic alcohol "70": ♦ - 4 - or of an ester-forming phenol, silanol or stannanol (where the alcohol, phenol, silanol or stannanol preferably contains 1 to 20 carbon atoms) or a symmetrical or mixed anhydride blocking group derived from a suitable acid.

Preferred examples of or are t-butyl, diphenylmethyl and p-nitrobenzyl.

When R 1 is an amino protecting group, the protecting group may, for example, be an aralkyl group having 7-20 carbon atoms (e.g. a triphenylmethyl or 4-1Ω methoxybenzyl group), an acyl group, such as an optionally substituted alkanoyl group having 1 to 6 carbon atoms (e.g. a formyl) or chloroacetyl group) or an optionally substituted alkoxycarbonyl group (e.g. a t-butoxycarbonyl or 2,2,2-trichloroethoxycarbonyl group), or an aralkyloxycarbonyl group (e.g. a benzyloxycarbonyl group) or a silyl group (e.g. a trimethylsilyl group).

In general, compounds of formula 1 in which R1 is a hydrogen atom are preferred.

2Q When R ^ or R ^ represents a substituted hydroxy group, it may be, for example, an acyloxy group / for example a formyloxy group or a group of formula -OCORg (where Rg is an alkyl group with 1-8 carbon atoms, for example a methyl group) 7, a carbonate group -OCC ^ Rg (where R & has the meaning defined above) a silyloxy group / e.g. a (C 1 -alkyl) silyloxy group such as a trimethyl-silyloxy group or a t-butyldimethylsilyloxy group 7, or a borate / -0B (0R7) or phosphate [-0Έ (0 ) (OR ^) group (where R ^ represents an alkyl group of 1-4 carbon atoms).

2q When R ^ and R ,. together forming a cyclic protected diol group, it may be an alkylidene dioxy group, preferably having from 1 to 20 carbon atoms, for example, a methylenedioxy, ethylenedioxy, or isopropylidenedioxy group, optionally having one or more substituents such as phenyl, C 1-4 alkoxy- or oxo substituents, for example methoxymethylenedioxy, diphenylmethylenedioxy or carbonyldioxy groups; a cyclic borate group, for example, -0B (0H) O-; a cyclic phosphate group, for example -0P (0) (0H) 0- or -0P (0) (0R7) 0- (in which the meaning defined above ¢ 702295 * - 5 - s> -) or a cyclic silyl ether group, for example duck (C 1-4 alkyl) silyldioxy group, such as a dimethylsilyldioxy group.

In general, such silyl-5 oxy, borate or phosphate groups represent protected hydroxy groups that can be cleaved to give a compound of formula 1 with free hydroxy groups.

In general, R ^ and R1. each preferably an acetoxy group or especially a hydroxy group.

^ In the compounds of formula 1, Z is preferably S.

Cef-3 em compounds of the invention are particularly preferred.

If the compound is to be used in medicine ^ 5, any ester of the carboxyl groups in the molecule must be a non-toxic, metabolically labile ester function. Examples of non-toxic metabolically labile ester derivatives are acyloxyalkyl esters, for example low alkanoyl-oxy-methyl or ethyl esters such as acetoxy-methyl or ethyl-20 or pivaloyloxymethyl esters, and alkoxyearbonyloxyethyl esters, for example low alkoxyearbonyloxyethyl esters such as the ethoxy-carbonyloxyethyl ester.

It will be understood that, in addition to the aforementioned ester derivatives, the invention also includes the active compounds of formula 1 in the form of other physiologically acceptable equivalents, ie physiologically acceptable compounds which, as well as the metabolically labile esters are converted in vivo to the parent antibiotic compounds of the invention.

The non-toxic salt derivatives which can be formed by reaction of the carboxyl group present in the compounds of formula 1 include inorganic base salts, such as alkali metal salts (eg sodium and potassium salts) and alkaline earth metal salts (eg calcium salts), amino acid salts (e.g. lysine and arginine salts), organic base salts (e.g. procaine, phenylethylbenzylamine, dibenzylethylenediamine, ethanolamine, diethanolamine and N-methylglucosamine salts).

6 70 2 29 - 6 -

Non-toxic salt derivatives include acid addition salts, e.g., formed with hydrochloric, hydrobromic, sulfuric, nitric, phosphoric, formic and trifluoroacetic acids. The salts may also be in the form of resin products, for example, formed with a polystyrene resin or a cross-linked polystyrene-divinylbenzene copolymer resin containing amino or quaternary amino groups or sulfonic acid groups, or with a resin containing carboxyl groups, for example a polyacrylic acid resin.

Soluble base salts (e.g., alkali metal salts such as the sodium salt) of the compounds of formula 1, due to the rapid distribution of such salts in the body after administration, can be easily used for therapeutic applications. If in a particular application, for example for use in depot preparations, insoluble salts of compounds of formula 1 are desired, then such salts can be prepared in a usual manner, for example with suitable organic amines.

The compounds of the invention are syn isomers. The syn isomeric form is defined by the configuration of the group of formula 10 relative to the carboxamido group. In this description, the syn-configuration is presented as in formula 1. It will be understood that since the compounds of the invention are geometric isomeric, some mixing with the corresponding anti-isomer may occur.

25 ......

Furthermore, it will be understood that the oxime etherifying group is the carbon atom adjacent to the oxy group chiral and may therefore exist in the R or S configuration.

The scope of the invention therefore includes the individual R and S forms of this eiriral carbon atom as well as mixtures (including diastereomeric mixtures) thereof. In general, compounds of formula 1 in which this ehiral carbon atom has the S configuration and R / S mixtures in which the S isomer predominates are preferred.

The compounds of the invention can prevent oc tautomeric forms (for example, with respect to the 2-aminothiazolyl group), and it will be appreciated that such "702295" -7-tautomeric forms, for example, the 2-iminothiazolinyl form, are within the scope of the invention.

As previously indicated, the compounds of the invention are active against a wide variety of commonly occurring pathogenic organisms and / or can be used as intermediates for the preparation of other active compounds. When the compounds of the invention are used as intermediates, the groups and R2 are often carboxyl group blocking groups; the R group is then often 10 * an ammoprotective group and the groups and R 4 are then often protected hydroxy groups such as silyloxy, borate or phosphate groups, or together form a cyclic protected diol group.

Non-toxic derivatives in which and / or R 1 represent acyloxy groups such as acetoxy groups can act as intermediates as well as active compounds.

In general, active compounds of the invention are cef-3-em compounds of formula 1 wherein

V

E |, R 2 and R j represent hydrogen atoms, Z represents S and R, and Re which may be the same or different, hydroxy or 20 y C | -C ^ -cyloxy groups such as acetoxy groups.

Important active compounds according to the invention have the formula 1a, in which R1a is a hydroxy or acetoxy group,

Re is a hydroxy or acetoxy group, 25. , 2 and their non-toxic salts, solvates, hydrates and metabolically labile esters.

A particularly preferred compound according to the invention is: (6R, 7R, 2'Z, S) -7- / 2- (2-aminothiazol-4-yl) -2- / "(carboxy) (3,4-dihydroxyphenyl) methoxyimino_ acetamidq7cef-3-em-30 · 4-carboxylic acid, and its non-toxic salts, solvates, hydrates and metabolically labile esters.

It has been found that (6R, 7R, 2'Z, S) -7- (2- (2-aminothiazol-4-yl) -2- (tcarboxy) (3,4-dihydroxyphenyl) methoxyiminq-acetamido / cef-3 -em-4-carboxylic acid can be prepared and recovered in an advantageous and reproducible manner with a high degree of purity in crystalline form, and this is a further aspect of the invention. In particular, crystalline hydrated (RR ^ R ^ 1- Z, S) -7- ^ 2- (2-aminothiazol-4-yl) -2- / f (carboxy) (3,4-dihydroxyphenyl) - 870 2 - - - 8 - Methoxyiminoj / -acetamidojcef-3-em-4-carboxylic acid prepared and recovered. This new substance is not only excellent in crystallinity and high purity, but is also obtained in high yield and has great storage stability even at high temperature for long periods. Because of these properties, the crystalline substance is particularly valuable for pharmaceutical use.

The above hydrated crystalline material can be characterized by the infrared-10 spectrum and / or by the X-ray diffraction pattern of the powder. IR spectrum (Nujol) V 3700-2100 (wide 1760, 1720, 1660, 1555, 1520, 1350, 1300 max 1290, 1240, 1215, 1170, 1155, 1120, 1030, 1000, 920, 860, and 755 cm ” 1.

15 X-ray diffraction pattern (given as d-distances in AngstrÖm units and percent intensities I).

12.5 (21); 9.97 (37); 9.51 (22); 6.79 (13); 6.33 (35); 5.29 (10); 4.76 (30); 4.63 (100); 4.39 (54); 4.32 (5); 4.18 (15); 4.09 (31), 3.91 (24); 3.78 (69); 3.69 (17); 3.61 (9); 3.52 (12); 3.48 (21); 3.42 (6); 3.33 (37); 3.15 (36); 3.07 (22); 3.02 (4); 2.96 (36); 2.85 (9); 2.78 (3); 2.75 (3); 2.65 (11); 2.59 (49); 2.50 (6); 2.44 (9); 2.38 (6); 2.31 (3); 2.27 (4); 2.20 (5); 2.16 (9); 2.10 (10); 2.05 (4), 2.01 (9).

Compounds of the invention exert a broad spectrum antibiotic activity against both Gram positive and Gram negative organisms, including many β-lactamase producing strains. The compounds also have high stability to lactamases produced by a variety of Gram negative and Gram positive organisms.

It has been found that compounds of the invention have a potent activity against strains (including penicillinase-forming strains) of Gram-positive bacteria such as Staphylococcus aureus, Staphylococcus epidermidis and Streptococcus species, coupled with excellent activity against Pseudomonas species .and also a strong action against various members of the Enterobacteriaceae (eg strains of Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, 8702295 «- 9 -

Serratia mareescens, Proteus mirabilis and indole-positive Proteus organisms such as Proteus vulgaris, Proteus morganii and Providence species), and strains of Haemophilus influenzae and Acinetobacter calcoaceticus. This combination of a strong action against 5 Gram positive organisms with a strong action against Gram negative organisms, in particular against Pseudomonas, of the compounds according to the invention is unusual and particularly advantageous.

The IQ cephalosporin derivatives described herein have been found to have a desirable long serum elimination half-life in vivo.

Compounds of the invention can be used to treat a variety of diseases or conditions caused by pathogenic bacteria in humans and animals, such as respiratory tract infections and urinary tract infections.

The antibiotic active compounds of the invention can be processed in any suitable manner for administration purposes, in analogy to other anti-biotics, and the invention therefore also includes pharmaceutical compositions comprising one or a number of antibiotic active compounds of the invention, suitable for for use in human or veterinary medicine. Such formulations can be presented for a conventional application, using any necessary pharmaceutical carriers or excipients.

The antibiotic active compounds of the invention may be processed for injection purposes and presented in unit dosage form, in ampoules or in multiple dose containers, if necessary with an added preservative. The compositions may also be suspensions, solutions or emulsions in oily or aqueous carriers, and may contain processing agents such as suspending agents, stabilizing agents and / or dispersing agents.

It is also possible that the active ingredient is in a powder form for processing with a suitable carrier, for example sterile, pyrogen-free water, before use.

If desired, such powdery preparations may contain a suitable, non-toxic base, in order to improve the water solubility of the active ingredient and / or ensure that when the powder is processed with water, the pH of the resulting aqueous base is improved. preparation is physiologically acceptable. It is also possible that the base is present in the water with which the powder is processed. For example, the base may be an inorganic base such as sodium carbonate, sodium bicarbonate or sodium acetate, or an organic base such as lysine or lysine acetate.

The composition may also be presented in a form suitable for absorption by the gastrointestinal tract, for example, tablets, capsules, syrups or suspensions for oral administration and suppositories.

For example, veterinary formulations may be formulated into breast formulations in long-acting or rapid release bases.

The compositions may contain from 0.1%, e.g. 0.1-99%, of active material, depending on the mode of administration. When the compositions are in unit doses, each unit will preferably contain 100-3000 mg of the active ingredient, for example 200-2000 mg.

The daily dosage for treating adult humans will preferably range from 200-12000 mg, for example 1000-9000 mg, per day, depending inter alia on the nature of the infection and the manner and frequency of administration. Generally, intravenous or intramuscular administration will be used, for example, with 400 to 4000 mg per day of the active ingredient in the treatment of adult humans. It will be understood that in some circumstances, for example in the treatment of newborns, smaller unit doses and daily doses may be desirable.

The antibiotic active compounds according to the invention can be administered in combination with other therapeutic agents, such as antibiotics, for example penicillins, cephalosporins or other (Vlactam compounds).

The compounds of the invention can be prepared by a number of methods discussed below 1702295-11.

Thus, the invention provides a process for the preparation of an antibiotic active compound of the general formula T as defined above or a non-toxic salt, solvate or hydrate thereof, by acylation of a compound of the formula 2, wherein, Z and the dotted line have the meaning defined for the formula 1, or a salt, for example an acid addition salt, formed with for example an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid or phosphoric acid or an organic acid such as methanesulfonic acid or toluene-p-sulfonic acid, or 7-N-silyl derivative thereof, with an acid of formula 3, wherein R 2, R 2, R 2 and the above defined mean, or a salt thereof, or with an acylating agent corresponding thereto, after which, whenever the necessary and / or desirable, each of the following reactions, in any suitable order, is carried out: 2 i) conversion of a Δ isomer to .3.

a desired Δ isomer,

ii) reduction of a compound in which Z

20 is S- * 0 to a compound in which there is ZS, iii) conversion of a carboxyl group to a non-toxic metabolically labile ester function, iv) formation of a non-toxic salt or solvate, v) removal of any carboxyl group blocking and / or N-protecting groups, and vi) removal of any hydroxy group blocking groups present.

The above reactions i) to vi) can be carried out in the usual manner.

In acylation, the starting material of formula 2 is preferably a compound wherein Z is S and the dotted line represents a cef-3-em compound.

Acylating agents which can be used in the preparation of compounds of formula 1 include acid halides, in particular acid chlorides or bromides. Such acylating agents can be prepared by reaction 8702295-12 of an acid of formula 3 or a salt thereof, with a halogenating agent, for example, phosphorus oxychloride, thionyl chloride or oxalyl chloride.

Acylations using acid halides can be carried out in aqueous and non-aqueous reaction media, suitably at temperatures from -50 to + 50 ° C, preferably at temperatures between -40 and + 30 ° C, optionally in the presence of an acid-binding agent. Substantial reaction media include aqueous ketones, such as aqueous acetone, aqueous alcohols, such as aqueous ethanol, esters such as ethyl acetate, halohydrocarbons such as methylene chloride, amides such as dimethyl acetamide, nitriles such as acetonitrile, or mixtures of two or more such solvents. Suitable acid-binding agents are, for example, tertiary amines (eg triethylamine or dimethyl-aniline), inorganic bases (eg calcium carbonate or sodium bicarbonate) and oxiranes, such as low 1,2-alkylene oxides (eg ethylene oxide or propylene oxide) which are released in the acylation reaction hydrogen halide bond.

Acids of formula 3 can themselves be used as acylating agents in the preparation of compounds of formula 1. It is desirable to carry out acylations using acids of formula III in the presence of a condensing agent, for example a carbodiimide such as N, N'-dicyclohexylcarbodiimide or N-ethyl-N'-Y-dimethylaminopropyl-25 carbodiimide, a carbonyl compound such as carbonyldiimidazole, or an isoxazolium salt such as N-ethyl-5-phenylisoxazolium perchlorate, or N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline.

The acylation can also be carried out with other amide-forming derivatives of acids of the formula 3, such as, for example, an activated ester, a symmetrical anhydride or a mixed anhydride, for example formed with pivalic acid or with a haloformate, such as a lower alkyl haloformate.

Mixed anhydrides with phosphoric acids, for example phosphoric acid or phosphorous acid, sulfuric acid or aliphatic or aromatic sulfonic acids, for example toluene-p-sulfonic acid, can also be formed.

An activated ester can conveniently be formed in situ using, for example, 1-hydroxybenzotriazole in the presence of a condensing agent, as indicated above. Also, the activated ester can be pre-formed.

It is desirable to conduct acylation reactions using the free acids or their aforementioned amide-forming derivatives in an anhydrous reaction medium, such as methylene chloride, tetrahydrofuran, dimethylformamide, acetonitrile, dimethyl acetamide, or dimethyl sulfoxide.

Another activation method is, for example, the reaction of an acid of formula 3 with a solution or suspension previously formed by adding a carbonyl halide, especially oxalyl chloride or phosgene, or a phosphoryl halide, such as phosphorus oxychloride, to a solvent such as a halohydrocarbon, for example methylene chloride, containing a (low acyl) tert-amide such as N, N-dimethylformamide. The activated form of the acid of formula 3 can then be reacted with a 7-amino compound of formula 2 in a suitable solvent or mixture of solvents, for example halohydrocarbons such as dichloromethane, alcohols such as an alkanol, for example ethanol or technical denatured alcohol esters such as ethyl acetate, ethers such as tetrahydrofuran or dioxane, ketones such as acetone, amides such as dimethylacetamide, acetonitrile, water and mixtures thereof.

The acylation reaction can conveniently be carried out at a temperature between -50 to + 50 ° C, preferably between -40 and + 30 ° C, optionally in the presence of an acid binding agent, for example as mentioned above, for example dimethylaniline, triethylamine or sodium bicarbonate.

If desired, the above described 30. . .

acylation reactions are carried out in the presence of a catalyst such as 4-dimethylaminopyridine.

The acids of formula 3 and acylating agents corresponding thereto, if desired and suitable, may be prepared and used in the form of their acid addition salts. For example, acid chlorides can suitably be used as their hydrochloride salts and acid bromides as their hydrobromide salts.

For use as starting materials for 8702295-14 - the preparation of compounds of the general formula 1 according to the invention, compounds of the general formula 3 and the amide-forming derivatives thereof, such as corresponding acid halides and anhydrides, are preferably used in their - * syn isomeric form or in the form of mixtures of the syn isomers and the corresponding anti isomers containing at least 90% of the syn isomer.

Acids of formula 3 and their derivatives can be prepared by etherification of a compound of formula 4, wherein Rg has the meaning defined above, and Rg represents a hydrogen atom or a carboxyl group blocking group, or a salt thereof, by selective reaction with a compound of the general formula 5, wherein T is a chlorine, bromine or iodine atom, a sulfate group, or a sulfonate group such as the tosylate group, followed by removal of any carboxyl group blocking group Rg.

The etherification reaction is expediently carried out in the presence of a base, for example potassium carbonate or sodium hydride, and preferably in an organic solvent, for example dimethyl sulfoxide, a cyclic ether such as tetrahydrofuran or dioxane or a Ν, Ν-disubstituted amide such as dimethylformamide.

The isomers can be separated before or after such etherification. Under the conditions described above, the configuration of the oxyimino group is substantially unaltered by the etherification reaction.

When the compound of formula 4 is used in the form of a free acid or a salt with a base, the etherification is generally carried out in the presence of a strong base, for example potassium t-butoxide, whereby sufficient base is added to form a dianion. In addition, the reaction must be carried out in the presence of a base, if an acid addition salt of a compound of formula 4 is used, the amount of base being sufficient to rapidly neutralize the acid in question.

Acids of formula 3 can also be prepared by reacting a compound of formula 6, wherein Rg and Rg have the meaning defined above, with a compound 8702295? Of formula 7, wherein, R 4 and the have the meaning defined above, followed by removal of any carboxyl group blocking group R 9 present, and separation of syn and anti isomers if necessary.

The reaction is expediently carried out in a solvent such as dimethylformamide, dimethylacetamide, dimethyl sulfoxide, tetrahydrofuran or methanol, all optionally in the presence of water, at a temperature of -20 to + 50 ° C, preferably 0 to 30 ° C.

The acids of formula 3 can be converted into the corresponding acid halides and anhydrides and acid addition salts by conventional methods, for example as described above.

The intermediates of formula 7 can be prepared by treating compounds of formula 8, wherein Y is an imido group, for example a phthalimido, suecin imido or maleimido group, with a hydrazine reagent such as hydrazine hydrate or an alkyl hydrazine such as methylhydrazine. The reaction is generally carried out in an inert solvent, for example a halohydrocarbon such as methylene chloride at a low temperature, for example -70 to + 30 ° C.

The intermediates of formula 8 can be prepared by alkylation with compounds of formula 9, wherein Hal is a halogen atom such as a chlorine or bromine atom of a suitable N-hydroxyimide, for example N-hydroxyphthalimide, N-hydroxysuccinimide or N-hydroxymaleimide in the presence of a base such as triethylamine in a solvent such as acetonitrile at, for example, -10 to + 30 ° C.

The intermediates of formula 9 are either known compounds or can be prepared using methods analogous to those used for the preparation of the known compounds.

The compounds of formula II which are used as starting materials in acylation are known from the literature, for example from British patent application 2,159,817, or can be prepared according to information from the literature.

2.

An Δ-cephalosporm master derivative obtained according to the method of the invention can be converted into e 7 0 2 2 0 o ft 3 - 16 - the corresponding, desired Δ derivative by, for example, bonding the A ester with a base, such as pyridine or triethyl amine.

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

If a compound is obtained in which ZS is -►O, it can be converted to the corresponding sulfide by, for example, reduction of the corresponding acyloxysulfonium or alkoxysulfonium salt, prepared in situ by reaction with, for example, acetyl chloride in the case of an acetoxysulfonium-1 - * salt, the reduction being carried out, for example, with sodium dithionite or iodide ions, such as in a solution of potassium iodide in a solvent such as acetic acid, acetone, tetrahydrofuran, dioxane, dimethylformamide or dimethyl acetamide. The reaction can be carried out at a temperature from -20 to + 50 ° C.

90

In the oxidations and reductions described above, the groups R 1 and in the starting materials are preferably not hydroxy groups.

Metabolically labile ester derivatives of the carboxyl groups in the compounds of formula 1 can be obtained by reacting a compound of formula 1 or a salt or protected derivative thereof with the suitable esterifying agent, such as an acyloxyalkyl halide or alkoxycarbonyloxyalkyl halide (eg iodide) suitable in an inert organic solvent, such as dimethylformamide or acetone, optionally followed by removal of any protecting groups.

Base salts of the compounds of formula 1 can be obtained by reacting an acid of formula 1 with a suitable base. For example, the sodium or potassium salts can be obtained using the respective acetate, 2-ethylhexanoate or hydrogen carbonate salt.

Acid addition salts can be prepared by reacting a compound of formula 1 or a metabolically labile ester derivative thereof with the appropriate acid.

8702295 - 17 -

a

For example, if a compound of formula 1 is obtained as a mixture of isomers, the syn isomer can be obtained by conventional methods such as crystallization or chromatography.

It will be understood that in some of the conversions described above it may be necessary to protect any sensitive groups in the molecule of the compound in question, in order to avoid undesired side reactions. Examples of suitable protecting groups are given in "Protective Groups in Organic Synthesis" by Theodora W. Greene (John Wiley and Sons, 1981).

For example, during any of the above reactions, it may be necessary to protect the M 2 group of the amino-thiazolyl moiety, for example, by tritylation, acylation (eg, chloroacetylation or formylation), protonation, or other conventional method. The protecting group can then be removed in any suitable manner that does not degrade the desired compound, for example in the case of a trityl group by using an optionally halogenated carboxylic acid, for example acetic acid, formic acid, chloroacetic acid or trifluoroacetic acid, or using an inorganic acid, for example hydrochloric acid or mixtures of such acids, preferably in the presence of a protic solvent such as water, or in the case of a chloroacetyl group by treatment with thiourea.

Likewise, it may be necessary to protect the hydroxy groups of the catechol moiety during any of the above reactions. Hydroxy protecting groups which can be removed under mild conditions will generally be suitable, for example, acetyl or silyl groups. Such groups can be introduced in the usual manner and, if desired, removed such that the product is not degraded. Thus, in the case of an acetyl group, the group can be removed by solvolysis with an aqueous solvent such as aqueous methanol or aqueous ethanol in the presence of a base such as sodium bicarbonate, ammonium hydroxide, ammonium carbonate or ammonium carbamate. For example, a trimethylsilyl group can be split off 8702295 s-18 by treatment with a dilute aqueous acid.

It is desirable that groups that block carboxyl groups and that are used in the preparation of compounds of formula 1 or in the preparation of necessary starting materials are groups that can be readily cleaved off at a suitable step in the reaction sequence, suitably in the latter step. However, it may sometimes be convenient to use non-toxic, metabolically labile, carboxyl group blocking groups, such as acyloxymethyl or ethyl groups (eg, acetoxymethyl or ethyl or pivaloyloxymethyl) and retain them in the final product in order to obtain a suitable ester derivative of a compound with formula 1.

Suitable groups that block the carboxyl group are known in the art; a list of 15 representative blocked carboxyl groups is included in UK patent 1 399 086. Recommended blocked carboxyl groups include aryl (lower alkoxy) carbonyl groups such as p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl and diphenyl-methoxycarbonyl, lower alkoxycarbonyl groups such as t-butoxycarbonylf 20 and low haloalkoxycarbonyl groups such as 2,2,2-trichloroethoxy-carbonyl. The carboxyl group blocking group can then be removed by any of the suitable methods described in the literature; for example, an acid-catalyzed hydrolysis or reduction can often be used as well as an enzyme-catalyzed hydrolysis.

If a certain enantiomer of a compound of formula 1 is required, starting materials of the desired stereochemical configuration must be used in the above processes. Such intermediates can be obtained by conventional separation methods. For example, enantiomeric intermediates of formula 8, wherein R.2 is a hydrogen atom and wherein the chiral carbon atom has the (R) or (S) configuration, can be obtained by reacting a mixture of enantiomers with a separating agent such as 35 a chiral organic base C-for example R - (+) - - methyl-benzylamine / in a solvent such as acetone or acetonitrile under formation of the corresponding diastereomeric salts. The salts can then be separated by known methods, and the desired chiral intermediate of formula 8 can then be obtained by treatment with an aqueous acid such as hydrochloric acid at, for example, room temperature.

The crystalline (6R, 7R, 2'Z, S) -7- / - - (2-aminothiazol-4-yl) -2- / (carboxy) (3,4-dihydroxypheny1) with hoxy imino / acetamido7cef-3 -em-4-carboxylic acid as the hydrate can conveniently be prepared from an aqueous solution of a salt of the above cephalosporin or a solvate thereof.

The invention therefore further provides a process for preparing crystalline hydrated (6R, 7R, 2 'Z, S) -7- / 2- (2-aminothiazol-4-yl) -2- / T (carboxy) (3} 4-dihydroxy-phenyl) methoxyiminoacetamido / cef-3-em-4-carboxylic acid by crystallization of the acid from an aqueous solution, preferably an aqueous solution of an acid addition salt or base salt thereof or a solvate of those salts, to form the crystalline hydrated product. Crystallization is normally followed by recovery and drying of the product.

Suitable base salts are, for example, inorganic base salts such as alkali metal salts and alkaline earth metal salts and organic base salts as described above. A particularly suitable base salt is the sodium salt.

The acid addition salts used as starting material can be formed with an organic or inorganic acid. Organic acids that can be used are, for example, carboxylic acids and sulfonic acids, for example formic acid, trifluoroacetic acid and methanesulfonic acid. Inorganic acids that can be used are, for example, hydrochloric acid, hydrobromic acid or sulfuric acid.

According to another aspect of the invention, the desired material can be crystallized from a solution of an acid or base salt of (6R, 7R, 2'Z, S) -7- / 2- (2-aminothiazol-4-yl) - 2 - / (carboxy) (3,4-dihydroxyphenyl) methoxyiminq7-35 acetamido7ceph-3-em-4-carboxylic acid or a solvate thereof, in an aqueous medium by adjusting the pH to 1.0 to 4.0.

87 0 2? .S; j i - 20 -

For example, a hydrate can be crystallized from an aqueous solution of a base salt by adding an organic or inorganic acid to a pH in the above range. Suitable acids which can be used in the crystallization are, for example, hydrochloric acid and sulfuric acid. Also, the desired hydrate can be crystallized from an aqueous solution of an acid addition salt by adding an organic or inorganic base of a suitable pH in the above range. Bases that can be used are, for example, alkali metal or alkaline earth metal hydroxides, carbonates, bicarbonates or acetate, for example sodium hydroxide, sodium carbonate, sodium bicarbonate or sodium acetate.

The aqueous medium can contain a water-miscible organic solvent such as an alcohol, for example ethanol or isopropanol, an ether, for example tetrahydrofuran or dioxane, an amide, for example Ν, Ν-dimethylformamide or Ν, Ν-dimethyl acetamide, a ketone, for example acetone, or a nitrile, for example acetonitrile.

It is desirable to carry out the crystallization at ambient temperature, for example from 0 to 30 ° C, preferably from 15 to 25 ° C. The crystalline product can be obtained by filtration and washed and dried in the usual manner, for example by air drying or by careful drying under reduced pressure.

In the following examples, Nujol and

Sorbsil trademarks. Sorbsil U30 is a silica gel manufactured by Joseph Crosfield and Son of Warrington, Cheshire, England. Unless otherwise stated, NMR spectra were included in DMSO-dg. The X-ray diffraction measurements were made on a 50 Siemens D-500 diffractometer using CuK2 radiation.

The X-ray intensity was measured in 0.02 ° increments of 2Θ for 5 sec. , with a scintillation counter, between values of 2 and 90 °. The results were processed by a computer.

35 6 702 295% - 21 -

Intermediate 1 2- (3,4-Dioxycarbonylphenyl) -2- (phthalimidooxy) acetic acid

A solution of N-hydroxyphthalimide (14.6 g) and triethylamine (25 ml) in acetonitrile (50 ml) was added to a suspension of 2- (3.4) under stirring at -2 ° C over 20 min. -dioxycarbonylphenyl) -2-chloroacetic acid (20.5 g) in acetonitrile. After an additional hour at about 0 ° C, a solution of concentrated hydrochloric acid (7.5 ml) in water (100 ml) was added rapidly. Water (100 ml) was added slowly and the initially formed oil crystallized, aided by seeding.

The mixture was filtered and the filter cake was washed with water and ethyl acetate: petroleum ether (bp 40-60 ° C) (1: 2) and dried to give the title compound (30 g), mp.

183 to 185 ° C; £ 5.84 (s, OCH), 7.53 (s, catechol 5-H and 6-H), 7.69 (s, catechol 2-H), and 7.85 (s-phthalimido protons).

Intermediate 2 20 (a) (R) - (+) - & - Methylbenzylamine salt of (S) —2— (3,4-dioxycarbonylphenyl) -2- (phthalimidooxy) acetic acid

A solution of R - (+) - 0 (-methylbenzyl amine (16.3 ml) in acetone (100 ml) was rapidly added at 21 ° C under nitrogen to a magnetically stirred solution of Intermediate 1 (45 g). ) in acetone (1.25 L) After 30 min, the mixture was filtered and the precipitate was washed thoroughly with acetone to give the title compound (16.57 g) + 242 ° (c 1.07, EtOH); 6 1.46 (d, J 7Hz, CH, CH), 5.48 (s, 0CH), 7.3 to 7.6 (m, aromatic

30 J

protons) and 7.80 (s, phthalimidoprotons) (b) Diphenylmethyl- (S) -2- (3,4-dioxycarbonylphenyl) -2- (phthalimidooxy) acetate 2M Hydrochloric acid (10ml) under nitrogen at 21 ° C and added with stirring to a suspension of Intermediate 2 (a) (5.0 g) in water (30 ml). After 2 min, a solution of diphenyldiazomethane with one equivalent in methylene-870 2 295-22-chloride (11 ml) was added. After vigorous stirring for 35 min, the organic layer was separated and added dropwise to ethanol (150 ml) with stirring. The mixture was stirred at 21 ° C for 10 min and then stored at 4 ° C for 1 h. The crystals were collected by filtration, washed with ethanol and dried to give the title compound (3.59 g), mp. 135-135.5 ° C; + 112.6 ° (c, 1.18, ethyl acetate).

Intermediate 3 10 (S) - (3,4-Dihydroxypheny1) (diphenylmethoxycarbony1) methoxyamine

Intermediate 2 (b) (5.71 g) was stirred at about 40 ° C in methanol (390 ml) with M hydrochloric acid (5.5 ml) for 4.5 hours. The solution was concentrated and mixed with methylene chloride. After washing twice with water, the solution was dried over magnesium sulfate and evaporated to a foam. A solution of this foam in methylene chloride (176 ml) was cooled to -50 ° C with stirring and under nitrogen, after which hydrazine hydrate (1.5 ml) was added. The mixture was allowed to warm slowly to 21 ° C with stirring. After 5.75 hours, the mixture was filtered and the filter cake was leached with methylene chloride. The combined filtrates were diluted with ethyl acetate and washed with citric acid solution and brine. After drying over magnesium sulfate and evaporation, the title compound 25 (4.35 g) was obtained as a foam; Γ% Ί ^ λ + 17.8 ° (c, 1.03, methanol); 6 5.02 (s, OCHCO), 6.68 and 6.81 (m, thiazole H, OCH and catechol protons) 7.1 to 7.4 (m, aromatic protons) 9.04 (s, 0H).

Intermediate 4 (Z, S) -2- (2-Aminothiazol-4-yl) -2- (3,4-dihydroxyphenyl) (diphenylmethoxycarbonyl) methoxyiminoacetic acid 2- (2-Aminothiazol-4-yl) glyoxylic acid (1, 91 g) was added to a solution of Intermediate 3 (4.20 g) in N, N-dimethylformamide (22 ml) at 3 ° C with stirring over 3 min. After an additional 30 min with cooling by ice water, 8702295 i-23 - the solution was allowed to warm to 21 ° C over 1.5 h, after which it was added dropwise to an ice-water mixture (110 g) over 20 min with stirring. . The precipitate was collected by filtration, washed with water and dried. The precipitate was resuspended in methylene chloride (45 ml), stirred for 15 min and filtered.

The filter cake was washed with methylene chloride and dried to give the title compound (3.76 g); / o7D21 + 25.4 ° (c, 1.02, methanol); b 5.59 (s, OCHCO), 6.6 to 6.9 (m, thiazole H, OCH and catechol 10 protons), 7.0 to 7.5 (m, N ^ and phenyl protons) and 9.06 and 9.13 (2s, 0H).

Intermediate 5 (Z, S) -2- (2-Formamidothiazol-4-yl) -2- / (3,15-dihydroxyphenyl) (diphenylmethoxycarbonyl) methoxyimino7acetic acid 2- (2-Formamidothiazol-4-yl) glyoxylic acid (6, 96 g) was added at room temperature with stirring to a solution of / /, 4-dihydroxyphenyl) (diphenylmethoxycarbonyl) ^ -methoxyamine (12.73 g) in Ν, Ν-dimethylformamide (100 ml). After 1.5 hours more Ν, Ν-dimethylformamide (40 ml) was added, and after 2.5 hours the resulting aqueous solution (700 ml) was poured. The products were extracted with ethyl acetate (3 x 250 ml). The combined extracts were washed with water (2x250ml), dried over magnesium sulfate and evaporated to a solid. This solid was treated with ethyl acetate (20ml) and then with dichloromethane (200ml) to give a suspension. After cooling, the solid was collected by filtration, washed with dichloromethane and dried to give the title compound (11.38 g), mp. 125-127 ° C; CKJjj + 22.2 ° (c, 0.99, methanol).

Example I

Diphenylmethyl- (6R? 7R, 2 TZ, S) -7- / 2- (2- 35-aminothiazol-4-yl) -2-yl (3,4-dihydroxyphenyl) (diphenylmethoxycarbonyl) -methoxyiminq acetamidoy cef-3-em -4-carboxylate.

The tosylate salt of diphenylmethyl- (6R, 7R) -7-aminocef-3-em-4-carboxylate (1.24 g) was converted into the free 8702295-24-amine by partition between methylene chloride and aqueous sodium bicarbonate solution. The organic layer was washed with water and dried over magnesium sulfate. Evaporation yielded a foam which was mixed with intermediate 4 (1.0 g) 5 in THF (25 ml) with stirring, then Ν, Ν'-dicyclohexylcarbodiimide (830 mg) and hydroxybenzotriazole hydrate (290 mg) were added. After stirring at 21 ° C for 16 hours, the mixture was filtered and the filter cake was leached with ethyl acetate. The combined organic solutions were evaporated to a foam, which was redissolved in ethyl acetate and filtered through a column of Sorbsil U30 (100 g) in ethyl acetate. Evaporation of the appropriate eluate gave the crude product, which was redissolved in chloroform and applied to a column of Sorbsil U30 (100 g), poured with chloroform. The column was eluted with chloroform and then with 1%, 2% and 2% methanol in chloroform. The title compound (800 mg) was obtained as a foam by evaporation of the appropriate eluate;

Zoc7d2 1 + 36.25 ° (c, 1.17, DMSO); λ inf1 (EtOH) 230.8 (e]% 303), 290 (e]% 117) and 300.4 nm (e]% 84).

1 cm 1 cm 1cm 20

Example II

Trifluoroacetic acid salt of (6R, 7R, 2'Z, S) - 7- / 2- (2-Aminothiazol-4-yl) -2 - / (carboxy) (3,4-dihydroxyphenyl) methoxyimino7acetamido / cef-3 -em-4-carboxylic acid 25

The compound of Example I (740 mg) was stirred with anisole (1.5 ml) and trifluoroacetic acid (7.5 ml) was added. After 1 min, diisopropyl ether (50 ml) was added.

The precipitate was collected by filtration, washed with diisopropyl ether and dried in vacuo to give the title compound (450mg); / DC? / 1 + 102.8 ° (c 0.88, DMSO); ^ max (pH6 buffer) 236.8 nm (e! ^ 335), ^ infl 250.0 (e | ^ 257), 277.4

Am 17 cm 1 cm (E / o 173) and 296.0 nm (E A 139).

1 cm 1 cm

Example III

Diphenylmethyl- (6R, 7R, 2 * Z, S) -7 ~ Z2- (2-formamido-thiazol-4-yl) -2 - [(3,4-dihydroxyphenyl) (diphenylmethoxycarbonyl) methoxyimino-acetamido2cef-3- em-4-carboxylate 8702295 - 25 - i

Intermediate 5 (1.2 g) was suspended in methylene chloride (20 ml) with stirring and under nitrogen, and Ν, Ν-dimethylacetamide (4 ml) was added. The solution was cooled to -25 ° C and phosphorus oxychloride (0.28 ml) was added. After stirring at -25 to -20 ° C for 1 hour, a solution of diphenylmethyl- (6R, 7R) -7-aminocef-3-em-4-carboxylate (prepared from the tosylate salt (1.35 g) was reacted with sodium bicarbonate solution) in methyl chloride (20 ml) with N, N-dimethylaniline

(2 ml) added. The solution now had a temperature of -10 ° C

10 and was then surrounded by ice water and stirred for 1.75 hours.

The reaction mixture was then washed successively with sodium bicarbonate solution, dilute hydrochloric acid and three times with water, the water layers each being back extracted with methylene chloride. The combined organic layers were dried over magnesium sulfate and concentrated. The crude product in methylene chloride was applied to a column of Sorbsil U30 (50 g) poured into 40% ethyl acetate in petroleum ether (bp 40-60 ° C) with 2% acetic acid. The title compound (1.56 g) was obtained by evaporation of the appropriate eluate; 20 WD21 + 36.2 ° <c> 1> 05> DMSO); Λ max (EtOH) 269.2 nm (e]% 184) λinf1 232.0 (e!% 285) 260.4

Cm 1cm '(e; 179) and 266.8 nm (El% 183).

Icm Icm

Example IV

25 - r

Diphenylmethyl- (6R, 7R, 2, Z, S) -7- / 2- (2-amino-thiazol-4-yl) -2 - ^ (3,4-dihydroxyphenyl) (diphenylmethoxycarbonyl) -methoxyimino7acetamido7ceph-3-em -4-carboxylate

The compound of Example III (1.5 g) was stirred at 21 ° C with methanol (15 ml), and 60% perchloric acid (1 ml) was added. After 2.5 hours, the solution was partitioned between water and ethyl acetate, after which sodium bicarbonate solution in excess was carefully added. The water layer was extracted with more ethyl acetate and the combined organic layers were washed twice with brine, dried over magnesium sulfate and evaporated to give the title compound (1.34 g); Z ^ 2d21 + 36.5 ° (c 0.96, DMSO); β 7 0 Z i C: - 26 - O (Nujol) 3600 to 3100 (wide), 1785, 1730 and 1680 cm- ^.

Ill 3.X

Example V

Sodium salt of (6R, 7R, 2'Z, S) -7-ji (2 -5-aminothiazol-4-yl) -2- / Xcarboxy) (3,4-dihydroxypheny1) methoxyimino / -acetamidoj cef-3- em-4-carboxylic acid

The compound of Example II (11 g) was dissolved in ethanol (110 ml) with stirring at 21 ° C and saturated sodium acetate solution (4.8 ml) was added. After 10 min, the mixture was filtered. The filter cake was washed well, 4 times with ethanol and 4 times with ether, and dried under vacuum to give the title compound (5.45 g).

Another yield (2.73 g) was obtained by adding more saturated sodium acetate solution (2 ml) to the mother liquors; WD21 + 185.0 ° (c 0.78, DMS0); Λ max (pH6 buffer) 234.4 (e] )386); "λίηίΐ 250.4 (E276), 276.8 to 07 17 cm Ί cm (E 184) and 294.4 nm (E / o 148).

1cm 1 cm

Example VI

20 (6R, 7R, 21Z, S) -7-Z2- (2-Aminothiazol-4-yl) - 2- / f (carboxy) (3,4-dihydroxyphenyl) methoxyiminq7 acetamido7cef-3-em- 4-carboxylic acid

The compound of Example V (1.0 g) was mixed with water (4 ml), and when a solution was formed, sodium acetate (41 mg) was added. The solution was filtered and crystallization was started by scratching. The mixture was stored at 21 ° C for 14.5 hours, after which the crystals were collected by filtration. These crystals were washed with water and dried under vacuum at 21 ° C to give the title compound (250 mg). By addition of 2N hydrochloric acid (0.9 ml), further crystalline product (360 mg) was obtained; A; d21 + 136.27 ° (c 0.54, DMSO); 234 max 234.4 nm (e!% 408), ληί1 249.6 (e]% 295), 277.8 (e]% 192) and. «1 cm 1 cm 1 cm 295.0 nm (E 156) .

Icm 35

Example VII

(6R, 7R, 2'Z, S) -7- / 2- (2-Aminothiazol-4-yl) -2- & u29 5 - 27 - / “(carboxy) (3,4-dihydroxyphenyl) methoxyimino7 acetamido7cef -3-em-4-carboxylic acid, hydrochloride

The compound of Example IV (27.4 g) was dissolved in formic acid (110 ml) with stirring at 20 ° C, after which 3 equivalents of concentrated hydrochloric acid (8.2 ml) were added. The mixture was stirred for an additional 15 min, then added dropwise over 6 min with stirring at 20 ° C over diisopropyl ether (1.1 L). The precipitate was collected by filtration, washed with diisopropyl ether (500 ml). and dried under vacuum to give the title compound (18.02 g); ^ max (Nujol) 3600-2800 (wide) and en 1780 cm ^; S 3.3-3.7 (m, H-2), 5.05 (d, H-6), 5.4 (s, CHON), 5.8 (m, H-7), 6.46 (m, H-3) and 9.6 (d, NHCO).

15

Example VIII

(6R, 7R, 2'Z, S) -7- ^ 2- (2-Aminothiazol-4-yl) -2- / f (carboxy) (3,4-dihydroxypheny1) methoxyiminqacetamido7 cef-3-em-4- carboxylic acid The compound of Example VII (2.65 g) was added to distilled water (10.6 ml) at 20 ° C with stirring, and the mixture was stirred for 30 min, during which time crystallization took place. The stirred suspension was cooled in ice / water for 15 min, after which the crystals were collected by filtration, washed with ice-cold distilled water (10 ml) and dried in an oven at 30 ° C to obtain the title compound (2.09 g); water content 12% (Karl Fischer); IR spectrum (Nujol) 30 ^ max 3700 "2100 (wide), 1760, 1720, 1660, 1555, 1520, 1350, 1300, 1290, 1240, 1215, 1170, 1155, 1120, 1030, 1000, 920, 860 and 755 cnf1; X-ray diffraction pattern (given as d-distances in Angstrom units and percent intensities I).

12.5 (21); 9.97 (37); 9.51 (22); 6.79 (11); 6.33 (35); 5.29 (10); 4.76 35 (30); 4.63 (1-0); 4.39 (54); 4.32 (5); 4.18 (15); 4.09 (31); 3.91 (24); 3.78 (69); 3.69 (17); 3.61 (9); 3.52 (12); 3.48 (21); 3.42 (6); 3.33 (37); 3.15 (36); 3.07 (22); 3.02 (4); 2.96 (36); 2.85 (9); 2.78 (3); 2.75 (3); 2.65 (11); 2.59 (49); 2.50 (6); 2.44 (9); 2.38 (6); 87 0 2 2 9 5 Μ - 28 - 2.31 (3); 2.27 (4); 2.20 (5); 2.16 (9); 2.10 (10); 2.05 (4); 2.01 (9).

Example IX

tert-Butyl- (6R, 7R, 21Z, S) -7- / 2- (2-formamido-thiazol-4-yl) -2- / j (3,4-dihydroxyphenyl) (diphenylmethoxycarbonyl) methoxyimine / acetamido / cef-3-em-4-carboxylate

Intermediate 5 (1.53 g) was dissolved with stirring in methylene chloride (15 ml) and N, N-dimethylacetamide (2.9 ml), the solution was cooled to -35 ° C, and phosphorus oxychloride (0 .41 ml) was added. The mixture was stirred at -25 to -20 ° C for 1 hour, after which a solution of tert-butyl (6R, 7R) -7-aminocef-3-em-4-carboxylate (prepared from the tosylate salt (1.0 g ) by reaction with aqueous sodium bicarbonate solution as described in Example 1 for the diphenylmethyl ester) in 15 methylene chloride (15 ml) with Ν, Ν-dimethylaniline (2.9 ml) was added. The reaction mixture was kept at about 0 ° C for 16 hours and then washed successively with water, dilute hydrochloric acid (2 times) and water; each time the water layer was back extracted with methylene chloride. The combined organic 20 layers were concentrated to a foam, which was redissolved in methylene chloride (15ml) and passed through a column of Sorbsil U30 silica (5g) with methylene chloride (30ml) as eluent. The eluate was concentrated to a small volume and treated with diisopropyl ether (15 ml) to precipitate the title compound as a white solid (1.2 g); 6 (CDCl3) 1.57 (s, (CH3) 3), 5.74 (s.OCHCO), 6.15 (m, H-7), 6.49 (d, H-3), 8.4 -8.7 (m, NHCO and -OH).

Example X.

30 1 tert-Butyl- (6R, 7R, 21Z, S) -7- / 2- (2-amino-thiazol-4-yl) -2- (3,4-dihydroxyphenyl) (diphenylmethoxycarbonyl) -methoxyimino / acetamido / cef-3-em-4-carboxylate

The compound of Example IX (6.29 g) was stirred at 20 ° C in methanol (64 ml) and 1,4-dioxane (16 ml), 35 and 60% perchloric acid (5.63 ml) was added. After 3.5 hours, the solution was added to sodium bicarbonate solution (240 ml) with vigorous stirring. The mixture was stirred for an additional hour 7 7 2: 5-29 and then cooled. The product was collected by filtration, washed with water and dried. The product was then dissolved in a mixture of ethyl acetate and methylene chloride (1: 1), and this solution was applied to a column of "Sorbsil" U3Q silica 5 (65 g), which column was eluted with more solvent. The title compound (4.0 g) was obtained by evaporation of the appropriate eluate fraction; 1.5 (s, (CH3) 3), 5.06 (d, H-6), 5.61 (s, CHON), 5.8 (m, H-7), 6.41 (m, H -3), 9.0 (broad, -OH), 9.5 (d, NHCO).

10 ~

Example XI

(6R, 7R, 2% S) -7-Z2- (2-Aminothiazol-4-yl) - 2- / T (carboxy) (3,4-dihydroxyphenyl) methoxyimino7 acet amido. 7 cef-3-em- 4 -carboxylic acid, hydrochloride 15. .

The compound of Example X (1.5 g) was dissolved in formic acid (6 ml) with stirring at 20 ° C, and concentrated hydrochloric acid (0.35 ml, 2.1 equivalents) was added. The mixture was stirred for 2 hours, filtered and then evaporated under reduced pressure. Ethyl acetate (6 ml) was added to the residue and the mixture was again evaporated to a foam which was triturated with ethyl acetate (6 ml). The resulting suspension was stirred at ambient temperature for 30 min. The solid was obtained by filtration, washed twice with ethyl acetate and dried in vacuo to give the title compound (1.33g) which was identical to the product of Example VII.

Dry powder for injection purposes (6R, 7R, 21Z, S) -7-i2- / 2-Aminothiazol-30 4-yl / -2 - / "(carboxy) (3,4-dihydroxy-phenyl) methoxyiminq / acetamido] · cef- 500 mg of 3-em-4-carboxylic acid

Sodium carbonate (anhydrous) 99 mg

The two sterile components were mixed aseptically and filled into sterile vials. The headspace of the vials 8 7 0 2 2 P 5 * - 30 - was purged with sterile nitrogen and the vials were sealed with rubber stoppers and a metal coating (applied by shrinking). The product can be processed by dissolving it shortly before administration in water for injection purposes or in another suitable sterile vehicle.

8? 01 29 5

Claims (9)

1. A compound of the general formula I, wherein R 1 and R 2, which may be the same or different, each represent a hydrogen atom or a carboxyl group blocking group, R 8 represents a hydrogen atom or an amino protecting group, R 1 and R 2 each represent a hydroxy or substituted hydroxy group or R 4 and R 4 together represent a cyclic protected diol group, Z represents -S- or -SO- (IX or β-), and the dotted line representing the 2, 3 and 4 bridges positions, indicates that the compound is a cef-2-em or cef-3-em compound, and its salts and solvates (including hydrates). 2. A compound according to claim 1 of formula 1a, wherein R 1 is a hydroxy or acetoxy group, R 3 a is a hydroxy or acetoxy group, and the non-toxic salts, solvates (including hydrates) and metabolic labile esters thereof. The compound of claim 2, wherein the oxime etherifying group has the S configuration. 4. (6R, 7R, 21Z, S) -7- / 2- (2-Aminothiazol-4-yl) -2- / tcarboxy) (3,4-dihydroxyphenyl) methoxyimino / acetamido7cef- 3-em-4- carboxylic acid and its non-toxic salts, solvates (including hydrates) and their metabolically labile esters. A compound according to claim 4 in crystalline hydrated form. Crystalline hydrated compound 30 according to claim 5, characterized by an infrared spectrum with absorption bands at 3700-2100 (wide), 1760, 1720, 1660, 1555, 1520, 1350, 1300, 1290, 1240, 1215, 1170, 1155, 1120, 1030, 1000, 920, 860 and 755 cm 1. Crystalline hydrated compound 35 according to claim 5, characterized by the following X-ray diffraction 8702205 i-32 * pattern (given as d-distances in Angstrom units and percent intensities): 12.5 (21); 9.97 (37); 9.51 (22); 6.79 (11); 6.33 (35); 5.29 (10); 4.76 (30); 4.63 (100); 4.39 (54); 4.32 (5); 4.18 (15); 4.09 (31); 3.91 (24); 3.78 (69); 3.69 (17); 3.61 (9); 3.52 (12); 3.48 (21); 3.42 (6); 3.33 (37); 3.15 (36); 3.07 (22); 3.02 (4); 2.96 (36); 2.85 (9); 2.78 (3); 2.75 (3); 2.65 (11); 2.59 (49); 2.50 (6); 2.44 (9); 2.38 (6); 2.31 (3); 2.27 (4); 2.20 (5); 2.16 (9); 2.10 (10), 2.05 (4); 2.01 (9). 8. Process for preparing cephalosporin derivatives, characterized in that compounds according to claim 1 are prepared by acylation of a compound of formula 2, wherein, Z and the dotted line have the meaning defined in claim 1, or a salt or a 7 -N-silyl derivative thereof, with a compound of formula 3, wherein R 2, R 2> R 2 and R 1. have the meaning defined in claim 1, or a salt thereof, or an acylating agent corresponding thereto, 20 ... after which, whenever it is necessary and / or desirable, each of the following reactions is carried out, in any suitable order : 2 i) conversion of a Δ-isomer into a desired Δ ^ -isomer, ii) reduction of a compound in which 25. .... Z is -SO- to a compound in which Z is -S-, iii) conversion of a carboxyl group into a non-toxic, metabolically labile ester function, iv) formation of a non-toxic salt or solvate, 30. v) removal of any carboxyl group blocking and / or N-protecting groups present, and vi) removal of any hydroxy group blocking groups present. 35 The process for preparing the crystalline hydrate according to claim 5, wherein an aqueous solution of (6R, 7R, 2'Z, S) -7-Z2- (2-aminothiazol-4-yl) -2- / is formed. / carboxy) (3,4-dihydroxyphenyl) methoxyimino / acetamid-7-cep-3-em-4-carboxylic acid or an acid addition salt or base salt thereof or a 5702295 τ-33 solvate of the salt, and the hydrate crystallizes therefrom. The process for preparing the crystalline hydrate according to claim 5, wherein an aqueous solution of an acid addition salt or base salt of 5 (6R, 7R, 2 * Z, S) -7- / - (2-aminothiazol-4) is formed -y1) carboxy) - (3,4-dihydroxyphenyl) methoxyimino / acetamido / -cef-3-em-4-carboxylic acid or a solvate thereof and adjust the pH of the solution between 1.0 and 4.0. 11. Pharmaceutical composition comprising as active ingredient at least one compound of formula 1a according to claim 2. A pharmaceutical composition according to claim 11, wherein the active ingredient is a compound according to claim 4. A pharmaceutical composition according to claim 12, wherein the active ingredient is the crystalline hydrated compound according to claim 5. -o-o-o-o- 870. m P NHRi Η H As Η H HiN —f-V ^ Sl SN Ξ ί J,) I I IJZ. J> -KA I - 1-C - CONH-f f> 0 γ H 11. J \ rT T ^ H C00Rl 2 OCHCOORj COOR, j 3 ifS kAn 1 L = J — C-COORs] R ^ i II d NHt N. Rf I ^ OH / NHR, * S ^ N Η HS 1-1 —C— CONH - ^ - I''5 '', - = - C-COOH N. J-NV # \ h II \ O] HN OCHCOOH COOH üCHCOORi 3 ile 1 ° • LA IT XR |, Rra Rf T- CHCOORj NHR HiN-O-CHCOORi Φ. rV & I, ς -.LcO-COORg RJ, 5 ^ 6 ^ 7 y-o-ch-coor2 I Hal —CH COORi - O CH COOR * Φ * & fr Rf T I Xr4 8 Rr 9 Rf 10 870 2 29 5 y .. GLAXO GROUP LIMITED, London, Great Britain