DK155187B - ANALOGY PROCEDURE FOR THE PREPARATION OF CEPHALOSPOR COMPOUNDS IN THE SYN-ISOMER FORM OR TOXIC SALTS THEREOF - Google Patents

Description

DK 155187 B

The present invention relates to an analogous process for the preparation of novel cephalosporin compounds in the syn-isomeric form and to the general formula I shown in the preamble of claim 1 wherein R, Ra and Q have the same meanings or non-toxic salts. thereof. The process according to the invention is characterized by the characterizing part of the claim.

The cephalosporin compounds disclosed herein are named from the substance cepham (J. Am. Chem.

Soc. 1962, 84, 3400). The term cephem refers to the basic cepham structure with one double bond.

BACKGROUND OF THE INVENTION As is known, antibiotics of the cephalosporin series are 76-acylamidoceph-3-em-4-carboxylic acids and their various non-toxic derivatives such as salts, esters, lactones (if such can be formed), amides, hydrates or similar sulfoxides. These antibiotics may contain various substituents, especially 20 at the 3-position, including unsubstituted methyl and methyl groups substituted with various substituents as described in the literature.

In 1945 it was discovered that a strain of the Cephalosporium genus isolated from a sewer outlet in Sardinia could form drug-2s with some antimicrobial effect. A substance isolated from it in 1953 was named cephalosporin C, and in 1961 its structure was found to be H3N + CH (CH2) 3CONH SN.

3Q -ooc ^ nxJ_ch2o.co.ch3 V

COOH

and it was also shown that. one could hydrolyze cephalosporin C under mild acidic conditions to remove the aminoadipoyl group at position 7, thereby gaining the so-called core, 7-aminocephalosporanoic acid, into which other 7 substituents could be introduced.

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2

This is of great importance for cephalosporin C has too low antibiotic activity to be therapeutically valuable, although it must be considered a broad-spectrum antibiotic with some stability to lactamases. Lactamases, more specifically penicillinases from staphylococci in particular, are a major problem, especially in hospitals, because bacteria that produce them are resistant to penicillins. Until 1971, four cepha-losporin antibiotics were traded, namely cephalotine and cephalo-ridin, which must be administered by injection; and cephalexin and cephalo 10 glycine, which have a slightly weaker antibiotic effect but have the advantage of being administered orally. These four cepha-losporin compounds have the formulas

cephalotin

CH2CO. NH -t-ζ S VI

Xs / ^ Ln ^ L_ch2o.co.ch3 COONa 20 Cephaloridine

\ / CH2CO .NH -VII

'0Ρ · Τ ^ - * Ο coo "

Cephalexin

-Ch! Co.NH - ^ VIII

COOH

35 3

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cephaloglycin

/ \ - CH.CO.NH - IX

| J-N ^ J-CK2O.CO.CR3

5 NH2 Cf T

COOH

In 1971, there were three additional cephalosporin compounds under serious examination or clinical trial, and they were all for injection. These three compounds 10 are known as cephacetrxl, cefazoline and cephapirin. These compounds have the formulas

Cephacetril NC.CH „.CO.NH -

2 X

J, _CH2O.CO.CH3 C) COONa 20

Cefazolin? = N \ N — N surface

| N - CH- .CO .NH -f ^. in XI

m = c '^ i I I

\ H n ^ h2s -vs ^ -ch3 25 0 COONa

Cephapirin 30 / - \ /% - SCHj.CO.NH - ^ χΙΙ X = 7 J_Nv ^ '- CH2O.CO.CH3

° I

COONa

If you look more closely at the nature of the group in the 7-position for each of the seven compounds, it will be noted that of the residues associated with the common CONH group, two 35

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4 of them a thienylmethyl group, two of them a single optically active form of the α-aminophenylmethyl group and the other three either a simple cyanmethyl group or more complex heterocyclic groups. There is no feature that holds all these 7-page chains together, 5 no common feature that allows one to predict a particularly useful area for further research or any particular area of particularly outstanding interest.

All of the above cephalosporin antibiotics can be combined together under what is now called the "first generation" of cephalosporin antibiotics. It was antibiotics that had different activity than the penicillins, an activity that extended beyond them, but which was still essentially ineffective against β-lactamase-producing gram-negative organisms.

15 Up to 1971, cephalosporin research was particularly concerned with finding new antibiotics with broad antibacterial spectrum and high activity levels, especially against gram-negative organisms, or providing better cephalosporin antibiotics suitable for oral administration. Cephaloridine is more active against gram-negative bacteria than cephalotine. Cefazoline has greater activity against gram-negative organisms than cephaloridine. It has become clear that there is a class of gram-negative organisms producing β-lactamase and that known cephalosporin antibiotics are generally ineffective against gram-25 negative organisms of this class. This is because the β-lactamases produced by the organisms destroy the cephalosporin. Around 1970, cephalosporins resistant to B-lactamases from gram-negative organisms were found, but these cephalosporins have low general antibacterial activity, including, of course, the organisms that produce the & lactamases, so that complete resistance to β-lactamase is obtained. lactamases produced by gram-negative organisms at the expense of the total antibacterial activity of the compound. This problem has been clinically overcome by administering a mixture of antibiotics 35 of which at least one inhibits and at least one has antibacterial activity, but no such product has ever appeared on the market.

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5

Therefore, there is a very urgent need to develop a high activity level cephalosporin compound against gram-positive and gram-negative bacteria (including B-lactamase-producing gram-negative bacteria) and which also have resistance to B-lactamases produced by the gram. -negative bacteria.

Invention It has now been found that a large number of oxyiminosubstituted 7B-acylamido-cephalosporin compounds not only have high antibacterial activity against gram-positive and gram-negative organisms, but also as a group considered to have high resistance to B-lactamases. of gram-negative organisms. These compounds have the option of geometric isomer in (as opposed to optical isomer), an option that has existed only for very few previously known cephalosporins and whose consequence for the antibiotic properties has never been known and is quite surprising. Namely, it was found that the maximum antibacterial activity of the individual pairs, i.e., syn-isomer and anti-isomer, was always with the syn-isomer. Although the anti-isomers had excellent stability to B-lactamases, they exhibited consistently lower antibacterial activity than the corresponding syn-isomers.

Thus, the present invention is based on the observation that oxyimino-substituted 7B-acylamido-cephalosporins have high resistance to 6-lactamases produced by gram-negative organisms, and that, with extraordinary regularity, the syn isomer exhibiting the single pair of compounds is present. maximal antibacterial activity with gra-positive and gram-negative bacteria in combination with the high B-lactamase resistance.

The compounds of this invention are defined as having the syn-isomeric form (cis form) with respect to the configuration of the group -Rα relative to the carboxamido group. In the present 35 "description, the vision configuration is structured as follows:

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6 R.ij.CO.NH-

V

^ ORa and the anti-configuration in this way: 5 R.C.CO.NH-

II

N

These configurations are assumed on the basis of work by 10 Ahmad and Spencer (Can. J. Chem. 1961, 39, 1340).

To further demonstrate the higher activity of the syn isomers than the anti-isomers, paired compound tests have been performed in the well-known tube dilution test 5 for 10 colony forming units, and with some typical gram-negative and gram-negative bacterial strains, namely three strains of Staphylococcus aureus (gram positive). ) and a strain of each of the gram-negative Escherichia coli, Salmonella typhimurium and Proteus mirabilis. The values found are listed in Table I (i) below. In this as well as subsequent tables 20 stands - for not tested.

In the following Table I (ii) a direct comparison of the activity of the syn isomers with respect to the corresponding anti-isomers has been made. By comparison, the anti-isomer is arbitrarily assigned to each tested isomer pair a number value of 0 (zero), and if the syn isomer has a lower MIC (MIC: least inhibitory concentration) than the anti-isomer, then it is more active. , it is assigned a positive number; the higher MIC value, ie lower antibiotic activity, has a negative number. The actual size of the 30 number value assigned is found by comparing the number of tube dilutions separating the respective MIC values. For example, for the isomer pair A regarding Salmonella typhimurium, it can be seen in Table I (i) that the MIC for the anti-isomer is 125, for the syn-isomer 16. Thus, there is a three-fold difference in concentration (dilution by tube dilution is done geometrically progressively) and vision The isomer is therefore assigned the value +3.

In the case of some compounds, β-lactamase stability is also stated in comparison with 8-lactamase stability.

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the density of cephaloridine arbitrarily employed to the value 1.

It is seen that the syn isomers are consistently more biologically, in most cases substantially more active, than the corresponding anti-isomers.

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Closely related prior art

From Danish patent application no. 381/72 (see DK patent no. 143,853) related compounds of the general formula 5 R ° are known. C. CONH__T „Sn1

il in XIII

V <Λ-γ-κ

COOH

wherein R ° represents phenyl, halogen-substituted phenyl, naphthyl or a heterocyclic group having 5 or 6 ring atoms, of which at least one is S, N or 0, Ru is a hydrogen atom, optionally halogenated alkanoyl group having up to 7 carbon atoms, an alkoxycarbonyl group with up to 7 carbon atoms, a benzoyl or nitrobenzoyl group, or an alkylcarbamoyl or haloalkylcarbamoyl group having 1-7 carbon atoms in the alkyl moiety and R is methyl, vinyl or -Cl 2 Y where Y has the same meaning as in formula I.

Thus, as is well known in the definition of R, these known compounds are syn-hydroxy and acyloxyimino derivatives of 7-acylaminocephalosporins, i.e. having an alcohol or ester function in the 7-side chain, whereas the present compounds of formula I are syn-hydroxyimino ethers. Apart from this important structural difference which does not immediately allow predictions of the biological properties of one class from the knowledge of the other, the known esters have the disadvantage of being unstable in human serum and binding to serum while the present ether of formula I bind less to serum and are much more stable in serum. This is shown in Table 2Q below for a number of reasonably comparable compounds.

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From Danish patent applications Nos. 1240/68 (see DK Patent No. 124,265) and 1239/68, a number of 7-glyoxamidocephalosporins are known in particular, but the descriptions also refer to the formation of α-carbonyl derivatives of these compounds. According to Example 34 of Application No. 1240 / 6® (Patent No. 124,265), sodium 3-acetoxymethyl-7β- (2'-methoxyiminophenylacetamido) -ceph-3-em-4-carboxylate and according to Example 35 of Application No. 1239/68 sodium 3-azidomethyl-7β- (2'-methoxyiminophenyl-acetamido) -ceph-3-em-4-carboxylate. In both of the two examples, it is stated that there is likely to be a mixture of vision and anti-forms of these compounds, which are structurally closely related to the present compounds of formula I, but there is no discussion of these geometric in the descriptions. isomeric conditions or the desirability of separating the two forms, there are no indications of how the pure vision or anti-forms can be recovered, and there is no indication of the aforementioned surprising fact that the forms of vision have significantly higher antibiotic activity than the anti-gnats. In fact, in the above Examples 20 for applications Nos. 1239/68 and 1240/68, the compositions contain predominantly the anti-isomer, namely with the syn / anti ratio of said 3-acetoxymethyl compound 47.6: 52.4 and in The 3-azidomethyl compound of 29.8: 70.2.

Other biological documentation The following Table III lists the biological activity, determined by the tube dilution method of 10 µ colony forming units and expressed in Y / ml, for a number of the compounds of the invention, including many of the compounds of the present invention. examples provided. In addition, the table lists values for the stability to islet lactamases from three organisms that form such; hereby, the islet lactamase stability of cephaloridine is arbitrarily employed to the value 1.

^ The organisms to which an x is listed are S-lactamase-forming.

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DK 155187 B

19 3 t) t-3

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to ^ oo oo co h oooo oo cn HOP

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DK 155187 B

20 3 a f-a

t £ -> c ^^ oo ου oo oo oo oo HO C

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DK 155187 B

21

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22

Detailed description of the invention

The group Ra of formula I must be capable of forming the desired ether group with the adjacent oxygen atom. Examples of Ra include alkyl groups such as methyl, ethyl, n-propyl, isopropyl / n-butyl, isobutyl, s-butyl and t-butyl; alkenyl groups such as vinyl, allyl, isopropenyl and dimethylallyl; cycloalkyl groups such as cyclopentyl and cyclohexyl. Ra may be substituted as set out in the preamble.

Examples of the heterocyclic group R are thien-2-yl, thien-3-yl, furyl such as fur-2-yl, pyridyl such as pyrid-3-yl, pyrrolyl, N-substituted pyrrolyl such as N-methylpyrrolyl, isothiazole -lyl and 3- or 4-isoxazolyl.

If Q is a substituted methyl group of the formula -C 2 Y, Y may be derived from a wide variety of nucleophilic substances which are distinguished by containing a nucleophilic nitrogen, carbon, sulfur or oxygen atom; such are described in previous patents and other literature on cephalosporin chemistry.

As stated in the claim, the compounds can, if desired, be converted into non-toxic salts thereof.

By the term "non-toxic" in the context of derivatives of the subject compounds is meant such derivatives which are physiologically acceptable in the doses in which they are administered. Examples of such salts are: a) inorganic base salts such as alkali metal salts, e.g., sodium and potassium salts, alkaline earth metal salts such as calcium salts, and organic base salts such as procaine, phenylethylbenzylamine and dibenzylethylene diamine salts, and b) acid addition salts, hydrochloric acid, hydrochloric acid, hydrochloric acid, phosphoric acid, toluene-β-sulfonic acid and methanesulfonic acid. The salts may also take the form of resinates, for example, formed with a polystyrene resin containing amino groups, quaternary amino group or sulfonic acid groups, or a resin containing carboxylic groups such as a polyacrylic acid resin. The resin may optionally be cross-linked, for example, a copolymer 35 of styrene and divinylbenzene containing appropriate groups. In addition, the derivatives may take the form of chelates with heavy metals such as iron or copper.

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23

If, according to the invention, an acylating agent in condensed form and with the general formula III is condensed with an amino-cephalosporan compound of formula II, a carboxyl blocking group may be, for example, the residue of an ester-forming alcohol 5 (aliphatic or araliphatic), phenol, silanol, stannanol or acid; the condensation is optionally carried out in the presence of a condensing agent and if necessary it is followed by removal of the group R.

For example, as a reactive derivative of compound III for use as an acylating agent, an acid halide, in particular an acid chloride or acid bromide, may be used. The acylation can be carried out at temperatures between -50 and + 50 ° C, preferably between -20 and + 30 ° C. The acylating agent can be prepared by reacting the acid III or a salt thereof with a halogenating agent, e.g., phosphorus pentachloride, thionyl chloride or oxalyl chloride. Use of oxalyl chloride with the sodium or potassium salt of the acid III is preferred because the isomerization in these circumstances becomes minimal. The acylation can be carried out in aqueous or non-aqueous media, and examples of suitable media are aqueous ketones such as aqueous acetone, esters such as ethyl acetate, amides such as dimethylacetamide or nitriles such as acetonitrile, or mixtures thereof.

Acylation with an acid halide of the acid III can be carried out in the presence of an acid-binding agent, for example a tertiary amine such as triethylamine or dimethylaniline, an inorganic base such as calcium carbonate or sodium bicarbonate or an oxirane which binds hydrogen halide which is released during the acylation reaction. The oxirane is preferably a lower 1,2-alkylene oxide such as ethylene oxide or propylene oxide.

When using the free acid as a compound of the general formula III, examples of suitable condensing agents may be mentioned carbodiimides, e.g., N, Ν'-diethyl, dipropyl or diisopropylcarbodiimide, N, N'-dicyclohexylcarbodiimide or N-ethyl-N '. -Y-dimetylaminopropylkarbodiimid; a carbonyl compound such as carbonyl diimidazole; or an isoxazolinium salt such as N-ethyl-5-phenylisoxazolinium-3'-sulfonate or N-t-butyl-5-methylisoxazolinium perchlorate. It is desirable to carry out the condensation reaction in an anhydrous reaction medium, for example methylene

DK 155187 B

24 chloride, dimethylformamide or acetonitrile, since it is possible to more precisely control reaction parameters such as temperature.

It is also possible to carry out the acylation with other amide-forming derivatives of the free acid such as a symmetrical anhydride or mixed anhydride, for example with pivalic acid or formed with a halogen formate such as a lower alkyl halide formate. The mixed or symmetrical anhydrides can be formed in situ. Thus, a mixed anhydride can be formed using N-ethoxy-carbonyl-2-ethoxy-1,2-dihydroquinoline. Mixed anhydrides can also be formed with phosphorous acids, eg phosphoric acid or phosphoric acids, or with sulfuric acid or aliphatic or aromatic sulfonic acids such as p-toluenesulfonic acid. A further convenient reactive derivative of the acid III is an activated ester such as a compound of formula 15

R.C.CO.L

L, XIV

ORa wherein L is, for example, an azide, oxysuccinimide, oxybenztriazole, penachlorophenoxy or p-nitrophenoxy group.

A starting material of formula II wherein Q is vinyl can be prepared as described in BE Patent No. 761,897.

If Q of Formula II is methyl, the compound can be prepared by the method described in GB Patent No. 957,569 or from a penicillin compound by the method described in US Patent Nos. 3,275,626 and BE Patents Nos. 747,119 and 747,120. .

Cephalosporin Compounds II having an acyloxymethyl group as substituent at the 3-position can be prepared from a cephalo-sporin with an S-CH 2 Y group, where Y ° represents OH or the residue of an acid HY ° having a pKa value of not above 4 and preferably not more than 3.5 (measured in water at 25 ° C).

Y ° may be chlorine, bromine or iodine, and formyloxy or acetoxy with at least one electron-attracting substituent on the α-carbon atom or a nucleus-substituted benzoyloxy group wherein the nucleus substituent is of the electron-withdrawing type as disclosed in GB Patent No. 1,241,657, and the nucleophilic replacement reaction to introduce the desired substituent

DK 155187 B

25 in the 3 position can be performed as described herein.

If Y ° is a hydroxy group, the desired 3-acyloxymethylcephalosporin may also be obtained by an acylation analogous to that described in GB Patent No. 1,141,293, wherein a process for the preparation of Δ-cephalosporins with a 3-acyloxymethyl substituent from a corresponding 3-hydroxymethyl analog consisting of aralkylating the 4-carboxy group, acylating the 3-hydroxymethyl group of the protected compound and then removing the aralkyl group.

The acylation may be carried out in any convenient manner, for example, using acid chloride, acid anhydride or mixed acid anhydride as an acylating agent, preferably in the presence of acid binding agent, e.g., an organic base such as pyridine or a lower alkylene oxide such as ethylene oxide or the like. l) propylene oxide, carrying out the reaction in solution in an inert anhydrous solvent such as methylene chloride. The acylation can also be carried out in aqueous acetone / sodium bicarbonate solution. The preferred acylating agent is acid chloride.

The acylation reaction should be carried out as soon as possible, since under the acylation conditions, rearrangement to the Δ derivative may occur.

Compounds of general formula III can be used in N-sylated amino form or as an acid addition salt. Useful acid addition salts are, for example, with hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, toluene-p-sulfonic acid or methane sulfonic acid.

Any protecting group on a compound of formula II is preferably formed with an alcohol (aliphatic or araliphatic), phenol, silanol, stannanol or acid, which can be readily decomposed at a later stage in the reaction sequence.

Examples of suitable protected carboxyl groups are those which, as an ester group, contain a group selected from the list below, which, however, cannot be considered an exhaustive list of possible ester groups.

(I) -COOCR ^ R ^ R * 1 wherein at least one of the groups R ^, R ^ and R * 1 is an electron donor such as, for example, p-methoxyphenyl, 2,4,6-trimethylphenyl, 9-anthryl, methoxy, acetoxy or fur-2-yl. The residue (s) of symbols R 1, R 2 and R 1 may be hydrogen or

DK 155187 B

26 organic substituted groups. Suitable ester groups of this type include p-methoxybenzyloxycarbonyl and 2,4,6-trimethylbenzyloxycarbonyl.

(ii) -COOCR 2 R 2 R 1, wherein at least one of the groups R 1, R 9 and R 1 is an electron-withdrawing group such as, for example, benzoyl, p-nitrophenyl, 4-pyridyl, trichloromethyl, tribromomethyl, iodomethyl, cyanomethyl, ethoxycarbonylmethyl, arylsulfonylmethyl, 2-dimethylsulfoniumethyl, o-nitrophenyl or cyan. The residue (s) of symbols R, R ° and Rn may denote hydrogen or organic substituent groups. Examples of suitable ester groups of this type are benzoylmethoxycarbonyl, p-nitrobenzyloxycarbonyl, 4-pyridylmethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl and 2,2,2-tribromoethoxycarbonyl.

We are (iii) -COOCR ^ R ^ R11 wherein at least two of the symbols R, R9 and Rn 15 are hydrocarbon groups such as alkyl groups such as methyl or ethyl or aryl groups such as phenyl, while the remaining symbols-f cr h.

ne R, R9 and R, if there is a residue, denotes hydrogen. Suitable ester groups of this type include t-butyloxycarbonyl, t-amyloxycarbonyl, diphenylmethoxycarbonyl and triphenylmethoxycarbonyl.

(iv) -COOR1 wherein R1 represents adamantyl, 2-benzyloxyphenyl, 4-methylthiophenyl, tetrahydrofur-2-yl or tetrahydropyran-2-yl.

(v) Silyloxycarbonyl groups formed by reaction of a carboxyl group with a derivative of a silanol. The derivative of a silanol is suitably a halogen silane or a silazane of the formula R1: L-.SiD; R11 SiD R1 * Si .NR11 «; R 11 Si.NH.SiR11-; 11 J 11 llz. 0 1 ± 11J 11 T 11

R 2 Si.NH.COR; R% Si.NH.CO.NH.SiR R NH.CO.NR .SiR

J 11 11 J 11 0 J or R C (OSiR n): NSiR where D represents a halogen atom and

J 11J

The various groups R, which may be the same or different, represent hydrogen atoms, alkyl groups such as methyl, ethyl, n-propyl or isopropyl * aryl groups such as phenyl or aralkyl groups such as benzyl. Preferred derivatives of silanols are silic chlorides such as, for example, trimethylchlorosilane and dimethyl dichlorosilane.

The carboxyl group can be regenerated from an ester in any of the usual ways, and, for example, acid and base catalyzed hydrolysis is generally applicable as well as enzymatically catalyzed hydrolysis. However, aqueous mixtures can

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27 can be poor solvents for these compounds and they can cause isomerizations, rearrangements, side reactions and general destruction so that special methods may be desirable.

5 Five suitable methods for deesterification are: 1) Reaction with Lewis acids.

Suitable Lewis acids for reaction with the esters include trifluoroacetic acid, formic acid, hydrochloric acid in acetic acid, zinc bromide in benzene and aqueous solutions or suspensions of mercury compounds. The reaction with Lewis acid can be facilitated by the addition of a nucleophile such as anisole.

2) Reduction.

Suitable reduction reduction systems are zinc / acetic acid, zinc / formic acid, zinc / low boiling alcohol, zinc / pyridine, palladized charcoal and hydrogen as well as sodium and liquid ammonia.

3) Attack of nucleophiles.

Suitable nucleophiles are those containing a nucleophilic oxygen atom or sulfur atom, for example, alcohols, mercaptans and water.

4) Oxidative methods, such as those involving the use of hydrogen peroxide and acetic acid.

5) Radiation.

Compounds of general formula I wherein Q is a group C 1 Y can also be prepared by reacting a compound IV wherein Y 1 is chlorine, bromine or iodine, with a nucleophile corresponding to group Y

The starting material of Formula IV can be prepared as described in BE Patents Nos. 719,711, 719,710, 734,532 and 734,533. The reaction of the 3-halomethylcephalosporin with nucleophiles within the framework specified in the claim can then be carried out as described in Belgian Patent No. 719,711. If Y is a hydroxy group, the compound can be prepared by the methods described in GB Patent No. 1,121,308.

The ceph-3-em starting compounds IV can be prepared by halogenating a 7β-acylamido-3-methylceph-3-em-4-carboxylic acid ester 1-oxide, followed by reduction of the 1

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28 in the reaction sequence as described in BE patent no.

755,256.

The corresponding ceph-2-em compounds can be prepared by the method disclosed in published NL Patent Application No. 6902013 by reacting a ceph-2-em-3-methyl compound with N-bromosuccinimide to form ceph -2-em-3-brorametylforbindelsen.

Visual and anti-isomers can be distinguished by an appropriate technique such as differences in their ultraviolet 10 spectra, thin-layer chromatography or paper chromatography, or by their nuclear magnetic resonance spectra. For example, in solution in DMSO-d6, compounds of general formula I (syn isomers) exhibit the doublet of amide-NH in a lower field than the corresponding anti-isomers. These factors can be exploited by monitoring the reactions.

The compounds can be worked up into pharmaceutical compositions which may contain from 0.1% of the active substance upwards, preferably 10-60% of the active material, depending on the method of administration. If the compositions are in dosage form, each unit preferably contains 50-500 mg of the active substance. The dose used for adult humans will generally be in the range of 100-3000 mg, e.g. 1500 mg per day, depending on the route of administration and frequency.

Some examples serve to further elucidate the method of the invention. Prior to them, there are a number of examples of making starting materials.

Starting material 1 2-Methoxyimino- (thien-2-yl) -acetic acid, syn- and anti-isomer

A solution of 5.85 g of methoxyamine hydrochloride in 60 ml of dry methanol was neutralized to phenolphthalein with a solution of sodium methoxide in methanol (from 2.5 g of sodium and 50 ml of dry methanol). The precipitated sodium chloride was removed by filtration and the filtrate was added to a solution of 10 g of thien-2-ylglyoxylic acid in 60 ml of dry methanol. The resulting solution was refluxed for one hour, cooled and evaporated to an oil. 100 ml of ether were added, the mixture filtered and the filtrate evaporated to 13.06 g of an oil.

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29 12.5 g of the oil was dissolved in 50 ml ether and an ethereal solution of diazomethane was added until a permanent yellow color remained. Excess diazomethane was destroyed by leaving the solution in sunlight for one hour. Evaporation of this solution produced 13.2 g of an oil.

10.33 g of the oil was purified by preparative plate chromatography (silica gel PF254 + 366 ° C, three times eluting with 75% petroleum ether (boiling point 60-80 ° C) in benzene to give 10 a) 3.44 g (27% ) methyl 2-methoxyimino-2- (thien-2-yl) acetate, syn isomer, Xmax (ethanol) 290 nm (e = 11250), xinfl 271 nm (e = 5400), vmax (CHBr +) 1738 and 1230 cm 2 (COOCH 2 in / x values (CDCl 3) include 6.06 (s, CO 2 CH 3) and 5.78 (s, OCH 2); and b) 1.21 g (9.5%) of methyl 2-methoxyimino-2- (thien-2-yl) acetate, anti-isomer, xmax (ethanol) 221 and 288 nm (e = 5020 and 11000), vmax (CHBr 2) 1732 and 1212 cm (COOCH values (CDCl 3) include 6.06 (s, COOCH 2) and 6.00 (s, OCH 2), as well as additional fractions which are isomer mixtures.

8.27 ml of 2N sodium hydroxide were added to a solution of 20.28 g of the syn isomer of methyl 2-methoxyimino-2- (thien-2-yl) acetate in 50 ml of methanol and the solution was stirred at room temperature for 18 hours. hours. 20 ml of water was added and the solution evaporated to remove methanol, then washed with ethyl acetate. The pH of the solution under ethyl acetate (50 ml) was changed to 2 with 2N hydrochloric acid. The layers were separated and the aqueous phase was extracted with ethyl acetate. The organic extracts were combined, dried and evaporated to 2.58 of a white solid. This was crystallized from cyclohexane to give 2.23 g (73%) of the syn isomer of the title compound, m.p. 105.5 ° C, xmax (ethanol) 289 nm (ε = 10100), Xinfl 262 and 271 nm (e = 7750 and 8150), τ values (CDCl ^) include 0.32 (OH) and 5.92 (OCH j).

Corresponding hydrolysis of the anti-methyl ester gave 0.85 g of the anti-isomer of 2-methoxyimino- (thien-2-yl) -acetic acid with 35 xmax (ethanol) 286-287 nm (ε = 10200), τ values (CDCl ) includes 1.31 (OH) and 5.73 (OCH +).

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30

Starting material 2

Syn-t-butoxyiminotien-2-ylacetic acid

A solution of 6.2 g of thien-2-ylglyoxylic acid and 3.36 g of sodium bicarbonate in 100 ml of water was added dropwise to a solution of 5,65 g of t-butoxyamine hydrochloride and 3.78 g of sodium bicarbonate in 100 ml. water at 0-5 ° C and the mixture was stirred at room temperature for 18 hours. The mixture is acidified with 2N hydrochloric acid to pH 2.0 and extracted with ethyl acetate. The combined extracts were washed with water, dried and concentrated to 9.75 g of a solid. Recrystallization from petroleum ether with boiling range 60-80 ° C gave 4.0 g (44% of the title compound, mp 106-107 ° C, xmax (ethanol) 290 nm (ε = 11600), τ values (CDCl ) include 2.46, 2.66, 298 (d doublets, thienyl protons), 8.60 (CiCH 2 ig).

15

Starting material 3-10 2-Alkoxyiminoaryl acetic acids, general procedure

A mixture of the substituted glyoxylic acid and excess 2Q shots (10 to 15%) of the alkoxyamine hydrochloride was suspended in water or aqueous ethanol and stirred and the pH of the mixture was adjusted to between 4 and 5 with sodium hydroxide solution (N to 10N). . A clear solution of pH 4 to 5 was maintained during the reaction by further additions of sodium hydroxide solution and ethanol as needed. The reaction mixture was kept at room temperature until all the keto acid was consumed (it may be necessary to add an additional portion of the more volatile alkoxyamines). The course of the reaction was followed by acidification of a portion, extraction with ethyl acetate and thin layer chromatography of the extract on silica plates (developed with a mixture of chloroform / methanol / acetic acid 18: 2: 1). The alkoxyimino acetic acids were less polar than the keto acids used as starting materials. The reaction times were 2 hours to 2 days. The reactions were carried out at pH 7.8 and when the reaction was complete, the pH of the mixture was adjusted to between 7 and 8 and the ethanol - if present - removed by evaporation. The aqueous mixture was extracted with ether and the extract discarded and the aqueous phase acidified to a pH

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di less than 2 with dilute hydrochloric acid. The mixture was extracted with ethyl acetate and the extract dried and evaporated to a crude product which was purified in one of the following ways: a) Crystallization and recrystallization (if necessary) from a suitable solvent.

b) The crude product dissolved in ether was treated with a small excess of a solution of diazomethane in ether. Excess reagent was destroyed with acetic acid and the solution washed with sodium bicarbonate solution and evaporated to the crude methyl esters.

The esters were separated by preparative thick-layer chromatography or column chromatography on silica and then conventionally hydrolyzed with alkali to give the pure synthetic acid.

c) The mixture of the methyl esters was prepared as under (b) and the isomers separated by crystallization from a suitable solvent and hydrolyzed in a similar manner.

These methods were used to produce the intermediates listed in Table 1 (syn isomers) page 32.

Starting material 11 n-Butoxyimino-thien-2-ylacetic acid, syn isomer 0.6 ml of 1-bromobutane was added to a solution of 1.0 g of methyl syn-hydroxyiminothien-2-yl acetate sodium salt (prepared (continued page 33) 25 30 35

DK 155187 B

32

H 3! -I 3 cQ G B

o co co ό σι ^^ ju 'S' H (D iQ Φ

CD I m H

_ _—— ---- H

/ = 1 / = 1 / = 1 / = 1 / = 1 r = \ r = \ w co o OO co ω cn y = ly = ly = J y »ν = * y = * J3 m ° gss nP s ^ «UP uP S“ “n uP“ ^ Μ * / \ "8 8 8

thaw

P) N.

__——-----——-- K

- - - 3 W

P) p) pJ P) CT P) PJ H'ffi> ✓ '<- <· N -' * '' '-µ · s— ^ 13 3 cq tn

-O CD Η H 00 CD 00 H W

ifc ·· Η Η H UI tO CO O, 3 - I H o I -CX3 Ό o co - - oo σ ui i · to Ui Ui —J IH - I co o

H «3 O

- o

UI

tow WN UW WM CON --- 10 (0 -. 10 H

0 - Π- V

CTiCOUJHWHUlHCOtO SOOtO 0 (31 S

UllUlOUltOOJOWU) ΚΟΙΌ OH h w to - - - - O - Η I π - T ω co fi> fc. OJ OJ U> tO Qi to - - H ·

^^> <J \ · * KD CD

cd oo η σι · »H Ί S" «ui ui oo σ ui] - oo ui ui 3 0>> ^" »O Η * Ό

00 -J -J O m S ~ J ~ J CO .. Ø H

t-i eo o σ> Ji »cd to co to toQiQi> - O pi CD t n T - H 3 oo CO cn o · Π Q ^ H · - - - σι ffl fi 3

-o to to · - Co to cQ

ui ui oo in to to to to toto ffiB 3 ut-OO ^ Oo oooo rrm

00 ι £ * UI UI CD CO CD PjX

S> ^^ p ui ui ui U1 0

Η Η H to H HH

co ui σι H to to o

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cQ

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33 by treating methyl syn-hydroxyiminothien-2-yl acetate with 1 equivalent of sodium methoxide) in 15 ml of benzene / dimethylformamide 2: 1, and this mixture was stirred for 17 hours at room temperature and then poured into water. The aqueous solution was extracted with ethyl acetate, washed with water, dried and evaporated to give 0.88 g of the syn methyl ester as a pale yellow oil.

4.0 ml of 2N sodium hydroxide was added to a solution of 0.85 g of the syn-methyl ester in 10 ml of methanol and the mixture was allowed to stand at room temperature for 18 hours. The methanol was removed by evaporation; the aqueous residue was diluted with water, washed with ether and acidified to pH 2.0 with 2N hydrochloric acid. The mixture was extracted with ethyl acetate; the combined extracts were washed with water, dried and evaporated to give 0.74 g (81%) of the title compound as a pale yellow oil; 15 τ values (DMSO-d6) include 2.30, 2.7-3.0 (thien-2-yl protons), 5.84 (OCH2) and 9.10 (CH3).

Starting material 12 2-Methoxymethoxyimino- (thien-2-yl) -acetic acid, syn isomer

A solution of sodium methoxide in methanol (about 0.2 M) was added to 0.5 g of the syn isomer of methyl 2-hydroxyimino-2- (thien-2-yl) acetate and the resulting solution evaporated to a yellow oil which, by azeotropic treatment with petroleum ether (boiling point 40-60 ° C) gave 0.49 g (88%) of the sodium salt of the syn isomer 2 of methyl 2-hydroxyimino-2- (thien-2-yl) ) -acetate. To a stirred solution of 0.49 g of this sodium salt in 5 ml of benzene / dimethylformamide 2: 1 was added 0.22 ml of chloro dimethyl ether.

After ten minutes, the reaction mixture was poured into saturated sodium bicarbonate solution and extracted with benzene. The combined extracts were washed with water, dried over sodium sulfate and evaporated to a yellow oil in an amount of 0.62 g (100%), the syn isomer of methyl 2-methoxymethoxyimino (thien-2-yl) acetate,

Xmax (ethanol) 288 nm (ε = 10300), vmax (CHBrr) 1730 (C00CH3),

1660 cm 2 (-C = N-), τ values (CDCl 3) include 2.61-2.83 (mul-35 J

tipped, thien-2-yl), 4.83 (singlet, CH2), 6.06 (CC> 2CH3), 6.56 (singlet, CH2OCH3).

A solution of 4 ml of 2N sodium hydroxide and methanol was added to 0.4 g of the methyl ester. After 30 minutes poured

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The reaction mixture was stirred in water and washed with ethyl acetate. The aqueous layer was acidified to pH 1 with 2N hydrochloric acid and extracted with ethyl acetate. The ethyl acetate phase was dried and evaporated to a colorless oil which was treated azeotropically with 5 petroleum ether (bp 40-60 ° C) to give 0.21 (55%) of a white solid, syn-2-methoxymethoxyimino- (thien-2-yl) -acetic acid, m.p. 61.2 ° C, λmax (ethanol) 286 nm (ε = 10400), νmax (nujol) 1732, 2600 cm 1 CO (COOH), τ (DMSO-d6) 2.26 (multiplet; thienyl H5), 2.7 -2.9 (multiplet, thienyl and H4), 4.88 (singlet, OC2 -), 6.6 (singlet, OCH2).

Starting material '13

Syn-2-cyclopentyloxyimino-2- (fur-2-yl) -acetic acid 2.80 g of fur-2-yl glyoxylic acid and 3.3 g of cyclopentyloxamine hydrochloride were dissolved in a mixture of 100 ml of water and 50 ml of ethanol and the pH of the solution value was set to 5.0. The solution was stirred for 19 hours, the alcohol was evaporated and the solution was acidified to pH 1.5 under ethyl acetate. The acidic mixture 2Q was extracted into ethyl acetate and the combined extracts washed, dried and evaporated to give 4.38 g of crude acid. This acid was treated with charcoal in benzene for 15 minutes and filtered and the filtrate evaporated to give a solid which was recrystallized twice from cyclohexane to give 2.28 g (51%) of the title acid, m.p. 96.6-25 q 97.7 C, xmax (ethanol) 277.5 nm (ε = 15600).

Starting materials 14-16 2-Alkoxyimino-2-arylacetic acids 30 General Methods

A mixture of the substituted glyoxylic acid and an excess (10 to 15%) of the alkoxyamine hydrochloride was suspended in water or aqueous ethanol and stirred and the pH of the mixture was adjusted to between 4 and 5 with sodium hydroxide solution 35 (n to 10N). A clear solution having a pH of 4-5 during the reaction was maintained by further additions of sodium hydroxide solution and ethanol if necessary.

35

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The reaction mixture was kept at room temperature until all keto acid was consumed (it may be necessary to add an additional portion of the more volatile alkoxyamines). The progress of the reaction was followed by acidification of a sample, extraction with ethyl acetate and thin layer chromatography of the extract on silica plates (developed with a mixture of chloroformimethanol: acetic acid; 18: 2: 1). The alkoxyimino acetic acids were less polar than the keto acids used as starting material. The reaction times were two hours to two days. When the reaction was complete, the pH of the mixture was adjusted to between 7 and 8 and if ethanol was present it was removed by evaporation.

The aqueous mixture was extracted with ether, the extract was discarded and the aqueous phase acidified to pH <2 with dilute hydrochloric acid. The mixture was extracted with ethyl acetate or ether, the extract dried and evaporated to give the crude product which was purified by one of the following methods: a) Crystallization and recrystallization (if necessary) from a suitable solvent.

b) The crude product dissolved in ether was treated with a small excess of a solution of diazomethane in ether. The excess reagent was destroyed with acetic acid and the solution washed with sodium bicarbonate solution and evaporated to give the crude methyl esters. The esters were isolated by preparative thick-layer chromatography or column chromatography on silica and then hydrolyzed in conventional manner with alkali to give the acids in visual form and purified by crystallization from a suitable solvent.

These methods were used to prepare the intermediates (syn isomers) shown in Table 2.

30 35

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Starting material 17

Propoxyiminothien-2-ylacetic acid, syn-isomer

A mixture of 3.12 g of thien-2-yl glyoxyl acid, 2.8 g of n-propoxyamine hydrochloride, 75 ml of ethanol and 75 ml of water was adjusted to pH 4.5-5 with 2N sodium hydroxide solution and stirred at room temperature. A clear solution of pH 4.5-5 was maintained by further additions of base and ethanol as needed. After four hours, a further portion of 1.4 g of n-propoxyamine hydrochloride was added and the mixture was stirred for a further three 10 hours (pH kept at 4.5-5) and then allowed to stand overnight. The ethanol was evaporated and the remaining solution was diluted with water and acidified and extracted with ethyl acetate. Evaporation of the dried (MgSO 4) ethyl acetate solution gave a mixture of the syn and anti-form of the title acid as an oil in an amount of 4.8 g.

The acid mixture was esterified in conventional manner with di-azomethane to give 3.175 g of a mixture of the syn and anti-methyl ester.

The ester mixture in 50 ml of methanol was treated with 14 ml of 20N sodium hydroxide solution for ten minutes at room temperature. The methanol was rapidly removed by evaporation and the residue in water was extracted with ethyl acetate. Evaporation of the dried (MgSO4) ethyl acetate solution gave 0.416 g of the syn methyl ester.

This ester in 10 ml of methanol was treated with 1.7 ml of 2N sodium-25 hydroxide solution and kept at room temperature for 26 hours. Conventional acidic insulation gave the title compound as 0.235 g of an oil. τ values (DMSO-d6) include 2.28, 2.7-2.9 (thienyl), 5.90 (O-CH 2).

Starting material 18

General method of converting a 2-alkoxyiminoaryl acetic acid to its acid chloride without isomerization

A solution of an equivalent of the pure syn- (or anti-) 2-alkoxyiminoaryl acetic acid in methanol (about 2-4 ml / mmol) was treated with an equivalent of sodium methoxide in methanol at 0-25 ° C and the mixture was evaporated to formation of the sodium salt which can be dried by azeotropic treatment with some portions of bone

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38 zen, and / or by drying in vacuo over phosphorus pentoxide.

An equivalent of the anhydrous sodium salt was suspended for approx. 5 ml / mmol of dry benzene containing a few drops of dry dimethylformamide and treated with 1-2.5 equivalents of freshly distilled oxalyl chloride. The mixture is stirred at room temperature for one hour and then evaporated to remove benzene. The resulting acid chlorides were not characterized but dissolved in acetone or methylene chloride and were immediately used to acylate the nucleus in the antibiotic.

The following acids are converted to their acid chlorides in the following manner: syn-2-methoxyimino- (thien-2-yl) -acetic acid, syn-2-ethoxyimino- (thien-2-yl) -acetic acid, syn-2-n butoxyimino- (thien-2-yl) -acetic acid, syn-2-t-butoxyiminO- (thien-2-yl) -acetic acid, syn-2- (2-bromoethoxy) -imino- (thien-2-yl) -acetic acid, syn-2- (2-t-butoxycarbonylaminoethoxy) -imino- (thien-2-yl) -acetic acid, syn-2-methoxyimino- (fur-2-yl) -acetic acid, syn-2-t- butoxyimino (fur-2-yl) -acetic acid, syn-2-ethoxyimino- (fur-2-yl) -acetic acid, syn-2-propoxyimino- (thien-2-yl) -acetic acid, syn-2-ethoxyimino (thien-3-yl) -acetic acid.

Examples 1-2

General Process for Preparation of 7B- (2-Alkoxyimino-2-Substituted Acetamido) -3- (Substituted) -Methyl-Ceph-3-Em-4-Carboxylic Acids Using Dicyclohexylcarbodiimide 3o i) Preparation of the 4- carboxylic acid ester

To a solution of the t-butyl or diphenylmethyl ester of 7B-amino-3- (substituted) methylceph-3-em-4-carboxylic acid (1 equivalent) and 1-1.2 equivalents of dicyclohexylcarbodiimide 35 in dry methylene chloride (which have been used with solvents such as (dimethylformamide or dioxane) added at 0-20 ° C

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39 is a solution of 1 equivalent of the 2-alkoxyimino-substituted acetic acid (syn form) in dry methylene chloride. After stirring for 3/3 hours at room temperature, the mixture was filtered, the filtrate was washed successively with 2N hydrochloric acid, saturated sodium hydrogen carbonate solution, water and brine. The organic phase was dried and evaporated to give the desired ester as an oil or foam.

ii) Removal of the protection from the intermediate ester 10 The t-Butyl or diphenylmethyl ester was dissolved x trifluoroacetic acid (5-10 ml / g ester) and allowed to stand at room temperature for 5-10 minutes, then evaporated under reduced pressure. The crude product was taken up in ether or ethyl acetate, extracted into aqueous sodium hydrogen carbonate solution and washed with ethyl acetate. The aqueous layer was acidified with 2N hydrochloric acid and extracted into ethyl acetate. The organic layer was washed, dried and evaporated to give the desired syn-7β- (2-alkoxy-imino-2-substituted acetamido) -3- (substituted) methylceph-3-em-4-carboxylic acid.

20

Example 1 a) t-Butyl-3-acetoxymethyl-7β- [2-methoxyimino-2- (thien-2-yl) acetamido] -ceph-3-em-4-carboxylate, syn isomer was recovered as a solid substance (85%), Xmax (ethanol.) 262-263 nm (ε = 14900), xinfl 280 nm (e = 13280), vmax (CHBr 2) 3400 (NH), 1780 (β-lactam), 1738, 1120 (COOR) and 1684, 1514 cm -1 (CONH); τ (CDCl3) 2.5-2.7, 2.95 (thienyl protons), 5.97 (OCH3), 7.93 (OAc) and 8.48 (t-butyl) Removal of the protection afforded the free acid: b) 3-Acetoxymethyl-7β- [2-methoxyimino-2- (thien-2-yl) -acetamido] ceph-3-em-4-carboxylic acid , syn isomer, (56%) [α] + = + 60 ° (c = 0.8 in dioxane), xmax 262 nm (ε = 15700), xinfl 290 nm (ε = 10700), vmax 3275 ( NH), 1768 (β-lactam), 1728 (OAc), 1700, 2600 (COOH) and 1648, 1520 cm -1 (CONH). T (DMSO-d6) 2.2-2.9 (thienyl), 6 , 08 (OCH-j), 0.13 (NH), 4.12, 4.76 (β-lactam), 6.28, 6.52 (J 18Hz; CH 2 in ring), 4.94, 5, 27

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40 (J 13Hz; CH 2 OAc) and 7.94 (OAc).

Example 2 a) t-Butyl 3-acetoxymethyl-78- [2-methoxymethoxyimino-2- (thien-2-yl) -acetamido] -ceph-3-em-4-carboxylate, syn isomer,

Xraax (ethanol) 260 nm (ε = 12940), νmax (CHBr)) 1778 (B-lac-tam), 1688 and 1510 cm "1 (CONH), x (CDCl13) 2.74 (doubled? NH), 4 , 76 (-OCH2OCH3), 6.49 (OCH2OCH3), 7.91 (OCOCH3), 8.44 (t-Bu), 10 was prepared by method i) and then treated with trifluoroacetic acid by method ii) to give b) 3-Acetoxymethyl-7β- [2-methoxymethoxyimino-2- (thien-2-yl) -acetamido] -ceph-3-em-4-carboxylic acid, syn-isomer, [ot] D = + 51 ° (c = 0.8, DMSO), xmax (pH 6 phosphate buffer) 262.5 nm (ε = 16400), vmax (nujol) 1778 (S-lactam), 1670, 1530 cm 1 (CONH), τ ( DMSO-d6) 0.11 (doublet; NH), 4.89 (OCH2OCH3), 6.62 (OCH2OCH3), 7.96 (OCOCH3).

Example 3 3-Acetoxymethyl-78- (2-t-butoxyiminothien-21-ylacetamido) -ceph-3-em-4-carboxylic acid, syn isomer 7.5 ml 0.53N methanolic sodium methoxide was added dropwise to a stirred mixture. solution of 0.9 g of syn-t-butoxy-25 g imino-txen-2-ylacetic acid in 10 ml of dry methanol at 0-5 C. The mixture was evaporated to remove methanol and dried by azeotrope treatment with dry benzene. 0.63 ml (0.95 g) of oxalyl chloride was added to a stirred suspension of the sodium salt and 3 drops of dimethylformamide in 40 ml of dry benzene. The mixture was stirred at room temperature for one hour and evaporated to remove benzene. The acid chloride in 10 ml of acetone was added dropwise to a stirred solution of 1.09 g of 7S-aminocephalosporanoic acid and 0.682 g of sodium bicarbonate in 20 ml of acetone and 20 ml of water at 0-5 ° C and the mixture was stirred at 35 ° C.

room temperature for two hours. The resulting solution was evaporated to remove acetone and washed with ethyl acetate. The aqueous solution was acidified in the presence of ethyl acetate with 2N

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41 hydrochloric acid to pH 1.5 and extracted with ethyl acetate. The combined extracts were washed with water, dried and evaporated to give 1.15 g (61%) of the title compound as a light yellow foam, [α] 3 = + 61 ° (c = 1.0 in 5 dioxane Xmax (pH 6 phosphate buffer) 260 nm (ε = 12500), vmax (nujol) 1780 (B-lactam), 1666 and 1620 cm 2 (CONH), τ values (DMSO-d6) include 0.28 (duplicate, J 8Hz; NH), 2.36 (1H) and 2.84 (2H) (thien-2-yl protons), 7.98 (OAc), 8 "68 (t-Bu).

Example 4 3-Acetoxymethyl-7β- (2-t-butoxyiminofur-2'-ylacetamido) -ceph-3-em-4-carboxylic acid, syn-isomer. 2.11 g of syn-t-butoxyiminofur-2-ylacetic acid was treated at room temperature with 19.1 ml of 0.525M sodium methoxide. The methanol was removed by evaporation and the resulting tan powder dried over phosphorus pentoxide overnight. 933 mg of the sodium salt in 10 ml of dry benzene containing one drop of dimethylformamide was treated with 0.8 ml of oxalyl chloride at room temperature for one hour. The solvent was removed by evaporation and the residue in 30 ml of acetone was added dropwise to a solution of 1.09 g of 7β-aminocephalosporanoic acid and 672 mg of sodium bicarbonate in 50 ml of water at 0-5 ° C. The resulting solution was stirred at room temperature for 1 1/2 hours. The acetone was removed by evaporation and the aqueous phase was adjusted to pH 8 and washed with a little ether. The aqueous layer was acidified to pH 2.0 under ether and the organic layer was washed with water and dried and evaporated to a pale yellow foam. This was dried over silica and potassium hydroxide beads to afford the title compound in an amount of 1.6 g (86%) as a light yellow

J U Q

foam, [a] D = +62 (c = 1 in dioxane), xmax (pH 6 phosphate buffer) 272 nm (e = 18600), vmax (CHBr 2) 1780 (S-lactam), 1674 and 1510 cm 2 (CONH ), τ values (DMSO-d6) include 0.34 (doubled; NH), 2.17 and 3.34 (fur-2-yl protons), 8.71 (t-Bu).

35

Examples 5-16 42

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General Process for Preparation of 3-Substituted Methyl-76- (2-Substituted Oxyimino-2-arylacetamido) -ceph-3-em-4-carboxylic Acids in Syn

Method_A

A solution of the syn-2-substituted oxyimino-2-arylacetyl chloride (prepared from 1 equivalent of the corresponding sodium salt with oxalyl chloride) was dissolved in acetone and the solution was added dropwise to a stirring, ice-cold (0-5 ° C). solution of 76-aminocephalosporanoic acid or 3-azidomethyl-76-aminoceph-3-em-4-carboxylic acid (1 equivalent) in water containing 2-2.5 equivalents of sodium bicarbonate. The mixture was stirred for 30 minutes to 2 1/2 hours allowing the temperature to rise to room temperature. Acetone was removed by evaporation under reduced pressure, the pH was adjusted to 1.5-2.0 and the product extracted into ethyl acetate (or occasionally ether). The extracts were washed with water or saturated brine, dried and evaporated to a foam or solid.

Method_B

As Method A, but the product was purified by dissolving in aqueous sodium bicarbonate and re-precipitated by acidification.

Method C 25 --------

A solution of 1 equivalent of the acid chloride in question was dissolved in dry methylene chloride (about 5 ml / mmol) and the solution was added to a suspension or solution of 1 equivalent of 76-aminocephalosporanoic acid or 3-azidomethyl-76-aminoceph-3-em-3q. 4-carboxylic acid and 3 equivalents of triethylamine in methylene chloride at 0-5 ° C. After stirring at room temperature for 1 1/2 hours, the solution was evaporated to dryness, dissolved in water and washed with ethyl acetate, and the aqueous layer was acidified to pH 1.5 under ethyl acetate. The ethyl acetate extract was washed with water and saturated. 35 brine and evaporated to a foam or solid.

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Example 17 45

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a) t-Butyl 3-acetoxymethyl-7β- (2-methoxyiminopyrid-3'-yl-acetamido) -ceph-3-em-4-carboxylate, syn-isomer

A suspension of 1.8 g of 2-methoxyiminopyrid-3'-ylacetic acid in 50 ml of dichloromethane was cooled to ca. -10 ° C and treated with 2.08 g of phosphorus pentachloride. The mixture was stirred and kept at ca. -10 ° C for 1 1/2 hours, during which most of the suspension was dissolved. The cold mixture was treated with 3.0 g of t-butyl-70-amino-3-acetoxymethylceph-3-em-4-carboxylate and 2 ml of propylene oxide and stirred for an additional 30 minutes. The solution was partitioned between ethyl acetate and saturated sodium bicarbonate solution and extracted with ethyl acetate. The combined organic extracts were washed with water and then extracted with 2N hydrochloric acid. Unreacted amine was removed by adding excess sodium nitrite to the cooled acidic extract, followed by washing with ethyl acetate after ca. 5 minutes at 0 ° C. The resulting aqueous phase was neutralized with saturated sodium bicarbonate solution and extracted with ethyl acetate. The organic extract was washed with water, dried over magnesium sulfate and evaporated to a mixture of the syn and anti-isomer of the title compound as a foam in an amount of 1.33 g.

Chromatography on silica using chloroform / methanol 99: 1 gave 394 mg of the anti-isomer, followed by 683 mg of the syn-isomer as a foam. The syn isomer was dissolved in ethyl acetate which precipitated by the addition of petroleum ether (bp 40-60 ° C). The amorphous solid was collected, washed with petroleum ether and dried to give 484 mg (10%) of the title ester, [α] D = + 49 ° (c = 0.45, DMSO), 30 Xmax (ethanol) 255 nm (e = 16100), vmax (CHBr0) 3396 (NH), 1786 3 (B-lactam), 1735 (OAc), 1724 (COOR), 1682 and 1518 cm χ (CONH), τ (DMS0-d6) 0.14 (d, J 8Hz; NH), 1.26, 1.30, 2.03 and 2.48 (multiples; aromatic protons), 4.10 (q, J 5 and 8 Hz; C-7H) , 4.74 (d, J 5 Hz; C-6H), 5.03 and 5.36 (2d, J 13 Hz; C-3 CH 2), 6.03 (s; OCH 3), 6.28 and 6.52 (2d, J 18 Hz; O 2 H 2), 7.97 (s; OCOCH 3), 8.52 (s; C (CH 3) 3).

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46 b) 3-Acetoxymethyl-7e- (2-methoxyiminopyrid-3'-ylacetamido) - ceph-3-em-4-carboxylic acid trifluoroacetic acid salt, syn isomer A solution of 440 mg of t-butyl-3-acetoxymethyl-76 (2-methoxyiminopyrid-3-ylacetamido) -ceph-3-em-4-carboxylate syn-isomer in 7 ml of trifluoroacetic acid was left at 20 ° C for 15 minutes. The solvent was removed and the residue evaporated three times from benzene. The resulting rubber was dissolved in a small volume of acetone and slowly added to the excess of petroleum ether under stirring (bp 40-60 ° C). The solid was collected by filtration, washed well with petroleum ether and dried to give the title synthetic acid salt as a solvated pale yellow powder in an amount of 561 mg, [a] + = + 30 ° (c = 0.88 in DMSO), xmax (pH 6 buffer) 254 nm (e = 14300), vmax (nujol) 3250 (NH), 2600 and 1720 (COOH), 1782 (P-15 lactam), 1736 (OAc) , 1670 (CF 3 CO 2 -), 1670, and 1550 cm 1 1 (CONH), τ (d 6 36 (multiplets; aromatic protons), 4.06 (g, J 5 and 8 Hz; C-7H), 4.73 (d, J 5Hz; C-6H), 4.93 and 5.26 (2d, J 13 Hz; C-3 CH 2), 5.98 (s; OCH 3), 6.26 and 6.49 (2d, J 18 Hz; C-2 H2), 7.94 (s; OCOCH 3).

25 30 35

Example 18 47

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3-Acetoxymethyl-7 8- [2-aminoethoxyimino- (thien-2-yl) -acetamino] -ceph-3-em-4-carboxylic acid trifluoroacetate salt, syn isomer

A solution of syn- (2-t-butoxycarboxamido) -ethoxyimino-5-thien-2-ylacetyl chloride (from 314 mg of the acid) in 10 ml of acetone was added dropwise to an ice-cooled and stirred solution of 272 mg of 78-aminocephalosporanoic acid. 20 ml of water containing 168 mg of sodium bicarbonate. The solution was stirred for two hours after which the acetone was evaporated. The aqueous mixture was acidified to pH 2.0 under ethyl acetate and extracted with ethyl acetate. The combined extracts were washed (water, brine), dried and evaporated to dryness to give 540 mg (95%) of 3-acetoxymethyl-78- [2-t-butoxycarboxamido-ethoxyimino-thien-2-yl) -acetamido] - ceph-3-em-4-carboxylic acid, τ values (DMSO-d6) include 0.27 (d; NH), 7.96 (s; OCOCH 3), 8.62 (s; C (CH 3) 3) ).

500 mg of the crude acid was dissolved in 5 ml of trifluoroacetic acid and the solution was allowed to stand at room temperature for five minutes and then evaporated to dryness at room temperature. 25 ml of dry benzene was added to the residue and the mixture was re-evaporated to dryness. The oily residue was triturated with ether to give 346 mg (64%) of the title compound as a colorless solid, [α] + = + 39 ° (c = 0.97 in dioxane),

Λmax (pH 6 phosphate buffer) 283 nm (ε = 16500), vmax (nujol) 1780 cm ', τ (DMSO-d6) values include 0.31 (c'ublet, J 8 Hz; 25 NH), 1, 98 (NH 3), 4.08 (07 proton), 5.66 (-OCH 2), 6.8 (-CH 2 -N <), 7.96 (OCCOCH 2).

Example 19 3q 78- [2-t-Butoxyimino-2- (thien-2-yl) -acetamido] -3- (5-methyl-1,3,4-thiadiazol-2-yl) -thiomethylceph-3-em -4-carboxylic acid, syn-isomer

A solution of 2- (t-butoxyimino) -2- (thien-2-yl) -acetic acid in methylene chloride / dimethylformamide 5: 1 was added to a solution of diphenylmethyl-78-amino-3- (5-methyl-1,3) (4-Thiadiazol-2-yl) -thiomethylceph-3-em-4-carboxylate in methylene chloride. Then set

DK 155187 B

48 a solution of dicyclohexylcarbodiimide in methylene chloride to said solution, stirring at 20 ° C. The mixture was left at 5 ° C for several hours and then filtered. The filtrate was washed with 2N hydrochloric acid, thin sodium bicarbonate solution, water and brine, dried over MgSO4 and concentrated under reduced pressure to give the crude diphenylmethyl ester of the title compound. The crude diphenylmethyl ester was dissolved in anisole and treated with trifluoroacetic acid. After a few minutes at 20 ° C, the solution was concentrated under reduced pressure. The residue was dissolved in ethyl acetate and the solution was concentrated again. The residue was partitioned between ethyl acetate and thin sodium bicarbonate solution and the ethyl acetate layer was extracted with thin sodium bicarbonate solution.

The combined extracts were washed with ethyl acetate, then covered with ethyl acetate and brought to pH 2 with concentrated hydrochloric acid.

The layers were separated and the aqueous layer extracted with ethyl acetate. The combined ethyl acetate extracts were washed with water and brine, dried and concentrated in vacuo to a small volume. Then the solution was added dropwise to stirred petroleum ether 20 (bp 40-60 ° C) and the precipitate was collected and washed with petroleum ether. The title acid was obtained in a yield of 58%, [α] D = -76 ° (c = 1 in DMSO), xmax (pH 6 phosphate buffer) 271 nm (ε = 14400), vmax (nujol) include 1780 ( 6-lactam) and 1670 and 1530 cm 2 (CONH), τ values (DMSO-d6) include 0.26 (d, J 8 Hz; NH), 2.38 + 2.80 (ms; thienyl protons), 4 , 08 (dd, J 8 and 5 Hz; H-7), 4.73 (d, J 5 Hz; H-6), 5.38 + 5.73 (2ds, J 13 Hz; C-3 CH 2) , 6.10 + 5.36 (2ds, J 18 Hz; O 2 CH 2), 7.27 (s; CH 3) and 8.64 (s; C (CH 3) 3).

Example 20 a) t-Butyl-3-acetoxymethyl-7β- (2-ethoxyiminopyrid-4-yl-acetamido) -ceph-3-em-4-carboxylate, syn-isomer

A solution of 1.15 g of the syn isomer of t-butyl-3β-acetoxymethyl-7β- (2-hydroxyiminopyrid-4-ylacetamido) -ceph-3-em-4-carboxylate in 20 ml of tetrahydrofuran was treated with excess diazoethane at 20 ° C for 40 minutes. The mixture was diluted with ether, washed with water and extracted with 2N hydrochloric acid.

The aqueous extract was neutralized with saturated sodium bicarbonate.

DK 155187 B

49 bonate solution and extracted with ethyl acetate. Evaporation of the dried organic extract gave the title ester as a foam in an amount of 0.84 g (69%), [α] D = + 47 ° (c = 0.84 in DMSO), Xmax (ethanol) 259 nm (e = 15450), ν max 5 (nujol) 3300 (NH), 1788 (γ-lactam), 1740 (OAc), 1724 (COOR), 1678 and 1540 cm 2 (CONH), τ values (d6-DMSO ) includes 0.12 (d, J 8 Hz; NH), 1.26 and 2.42 (multiples; aromatic protons), 5.69 (q, J 7 Hz; OCH 2 CH 3), 7.93 (s; OCOCH 3) , 8.50 (s; (CH 3) 3) and 8.69 (t, J 7 Hz; OCH 2 CH 3).

B) 3-Acetoxymethyl-73- (2-ethoxyiminopyrid-4-ylacetamido) -ceph-3-em-4-carboxylic acid trifluoroacetic acid salt, syn isomer 0.75 g of t-butyl-3-acetoxymethyl-73- (2 -ethoxyiminopyrid-4-ylacetamido) -ceph-3-em-4-carboxylate had the protection removed with 10 ml of trifluoroacetic acid in the usual manner to give the title salt as a powder in an amount of 0.61 g (73% ), [α] = + 30 ° (c = 0.4 in DMSO), xmax (pH 6 buffer) 257 nm (ε = 12800), vmax (nujol) 3300 (NH), 2600 and 1720 (COOH), 1782 (3-lactam), 1740 (OAc), 1680 (CF-COO "), 1670 and

-1 J 2052 cm2 (CONH), τ values (d6-DMSO) include 0.06 (d, J

8 Hz; NH), 1.20 and 2.28 (multiples; aromatic protons), 5.65 (q, J 7 Hz; OCH 2 CH 3), 7.92 (s; OCCOCH 3) and 8.64 (t, J 7

Hz; OCH2CH3).

Examples 21-26

General Methods for Preparation of 3-Acetoxymethyl-73- (2-substituted oxyimino-2-arylacetamido) -ceph-3-em-4-carboxylic acids in Syn

Method A --------

A solution of the syn-2-substituted oxyimino-2-arylacetyl chloride (prepared from 1 equivalent of the corresponding sodium salt with oxalyl chloride) was dissolved in acetone and the solution was added dropwise to a stirring cold (0-5 ° C). solution of 1 equivalent of 73-aminocephalosporanic acid in water containing 2-2.5 equivalents of sodium bicarbonate. The mixture was stirred for 1/2 to 2 1/2 hours, allowing the temperature to rise to room temperature. Acetone was removed by evaporation.

DK 155187 B

Under reduced pressure, ethyl acetate was added, the pH was adjusted to 1.5-2.0 and the product extracted into ethyl acetate. The extracts were washed with water and saturated brine, dried and evaporated to a foam or solid.

5

Metode_B

As method A, but the product was extracted into ether. Cephalosporin derivatives prepared in these ways are listed in Table 5 below.

10 15 20 25 30 35

DK 155187 B

51

! tO N) tO to tO S

σι cn ifc * to h CO jr 7 = j 7 = T ~ 7 = / = i ~ β r = T *

ti0 ^ = ιω®) = J y = l y = l y = i y = J

Ω O Hi Hi Q

-, if rN ro ro »O co [I p s S3 I- 'i- ·> 3 1¾

U) Η H

> - * ·

--- 1 J

ω 'to>>>> jf / \

Q O O

i »* g ^ _______ - p ~ j ++ + + + + + c · / ° b“ b “b“ o “o. jfo “Λ- /? 0 S- t --- = - o ^ M, tO tO [Ο tO M tO CON) tOtO „„ _ P \ v / ~~ ι ο σι o σι ό οσι o «ο 3 2? 12, 'SU / - O CO -J »£> to cn O1C0 01 O 3 3 * ® /

U1 U1 ^ f S

_____ to -—-- o H HH ΜΗ H HH HH i? cn tocotoui σι o σι co „u co co o co -o oo -o to co -jn O OO OO O OO O U1 0 OO OO O oo oo Η Η Η Η H1 t — 1 ό -o -o -o - j -j ro o 3 i S gsssg% 1 if II s | s | ro m i_i- <_i. cj. i_i. C? J, Η π 0 0 0 0 1 H · CO CO I—> (—1 {- <(-> _ Il

o OOOOO

* - S '"1 cn to CO Η H w ™ ^

σ> o σι co £ »^« & S

_______ (- π o Qj co σι oo cn 00 cn to S ° n. 0 £ ·

· ->> «^ *« VJ \ U

cn HOtOON) JS- Js- O ^ I cn I 00 I i g <00 - co— to to N x> · »*> .- y σι σι oo oo“ σ _? --------------- Qi .p * ^^^ Kai · * N ^^^>

(—1 Η Η H o O

cn 00 o <n_h_tN ___ - g

Ό -OOOOO

o cn cn 00 σι 00 j ^.

ro

Examples 27-32 52

DK 155187 B

General Procedure for Preparation of 73- (2-Substituted Oxyimino-2-arylacetamido) -3- (Substituted) -ceph-3-em-4-carboxylic Acids in Syn-Form Using Dicyclohexylcarbodi-imide solution of a diphenylmethyl ester of 7B-amino-3- (substituted) -ceph-3-em-4-carboxylic acid (1 equivalent) and 1-1.2 equivalents of dicyclohexylcarbodiimide in dry methylene chloride was added at 0-20 ° C. 1 equivalent of syn-2-alkoxyimino-2-aryledacetic acid in dry methylene chloride. After stirring for 45 minutes to three hours at room temperature, the mixture was filtered and the filtrate was evaporated to dryness. The residue in ethyl acetate was washed successively with sodium bicarbonate solution, water and brine. The organic phase 15 was dried and evaporated to a foam or solid. The resulting ester was purified by crystallization or chromatography on silica and characterized by proton magnetic resonance spectrum or thin layer chromatography.

ii) From the esters thus obtained, the protection 20 was removed by dissolving in a mixture of trifluoroacetic acid (3-10 ml) per ml. ester) and anisole (1-5 ml per g ester) and allowed to stand at room temperature for five to ten minutes, after which the solvents were removed by evaporation under reduced pressure.

The crude acid was shaken with ether or ethyl acetate and excess aqueous sodium bicarbonate, and the aqueous layer was washed with ethyl acetate. The aqueous phase was separated, covered with ethyl acetate and acidified to pH 1-2 with 2N hydrochloric acid. The organic layer was washed, dried and evaporated to the desired syn-7B- (2-alkoxyimino-2-arylacetamido) -3- (substituted) -ceph-3-em-4-carboxylic acid, and the compounds thus obtained. are listed in Table 6 below.

35

DK 155187 B

53 X | to lo lo to to µ 1-3 tO I— 'O Ό 00 Ό gg * G · ft e * ------ r *:% / = 1 ΠΛ r = \ r = \ r = \ r = \ * <* r + cn ω o cn o cn * y = ly = l yj yj y = ly = Å 8 g, __ / * t £ - ** / * \ S- = s I — 1 = sg 'g

U) (o Ϊ0 Q

cn w, - ft g -r?

Q CO CO W X

S ____v / ~ ff G S. ° Is £ to * f In z4 $ 8 S v q- / cn r + W Q-a I tu / S A CO® to

S

S's ι-i to H-J C-- ~ ~ ___ .. i-1

0 QJ B

F o + 111 (1 o

H D ¢ - tfc »00 -J t £> 00 {DO

H- S ιΡ »o oo cn to CT

5 © O OOOOO Q

tQ O 3 cn ft 1 —_______

about

IQ to 3 ^ to two to two 3 σιοΦ ^ -ο-ο ^ σ 3 3 5! ft O (-> CT to CO H 00 0)

Hl - 'ft »·· X σ tf i cn cn tn ——------— 1 1 ~~

X "H (- tO t-1 tO H

K cn -o to oo H cn ft O tO CO I —To CD Π rt o o o o o o

(D O OOOOO

H tn (D --—-——-- cn l-i ^ Η Λ {_ι · ^ · Η '-' Η- ^ Η ^ - (- 1

(D n-J 3-J 30 3> j 5 -i P- O 3 I

ft (i oo tcoøooCoopoo c oo Qi tn ft (- V i_j. oo Øfi uo u.io uo cj. σ Qi3 X! »r + 0 Ρ 0 0 0 0 ft H- g H- {- to i— t— i - * (—1 i- * 3 -o rt 0 - ^ w o-- ^ O ft 3 tn 53 g

ft II

1 -—---

O OOOO o <H

____ * »^ ** (D

Qj to totototo CO X & <! (D to two to o to σ ft! M h

Ml O Qi o - o F- p „ro

ω oo oococnoocn oo oo 2M

5> '-' - '> Ρ3Π3 Ό cn to to cn σι ft n Hi <o σ to cn to to 0

P- M

tn _

ft - S

ft 3

OOtfatOrfs.J ^ tO '-' Cn ^ CJl'Ott '· 0 O' - CQ

c + * ^ * * * »O · * O” O 'O- * O O

H HOOcocn-otdoUSotao ffio i µ. it »it» oocnto tootocntocn Qj p w - ^ σι p.

ft a O cn cn js »cn to cn o \ o σ

iq o φ »σ to cn Ό HK

H · I

H-

Example 33 54

DK 155187 B

(6R, 7R) -7- [2- (Fur-2-yl) -2-methoxyiminoacetamido] -3-pyridinium methylceph-3-em-4-carboxylate (syn isomer)

A solution of 4.89 g of the syn isomer of (6R, 7R) -3-5 acetoxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido] -ceph-3-em-4-carboxylic acid in 50 ml of water and 4.0 ml of pyridine were heated to 80 ° C for 70 min. The cooled reaction mixture was evaporated under reduced pressure to a small volume to remove excess pyridine. The mixture was diluted with water to ca. 70 ml and washed successively with 2 x 30 ml of methylene chloride, a solution of 5 ml of Amberlite w LA2 resin in 50 ml of methylene chloride and 3 x 30 ml of methylene chloride. The washings were washed themselves with 30 ml of water, the aqueous phases were combined and evaporated to a yellow glass in an amount of 2.17 g under reduced pressure at <40 ° C.

The crude product began to crystallize by heating with 10 ml of dimethylacetamide and the process was completed by diluting with 10 ml of acetone and cooling in a refrigerator for 16 hours to give 1.68 g of a solvate of the title compound, (pH Buffer (260 nm (e = 18,800); VαΧ η 20 vmax (nujol) 1770 (3-lactam), 1678 and 1550 (CONH and 1620 cm (CO2); (D20; 100 MHz) 0.98, 1.39, 1.87 (pyridine protons), 2.30 , 3.15, 3.38 (furyl protons, syn isomer), 4.15 (d, J4Hz; 7-Η), 4.72 (d, J4Hz; 6-Η), 6.04 (s; CH 3) , 6.33, 6.80 (dd, J 18Hz; 2-H2), 4.38, 4.63 (dd, J14Hz; CH 2 N +), 6.96, 7.10, 7.93 (dimethylacetamido, 0 , 7 moles) and 7.78 (acetone, 0.15 moles).

Example 34 (6R, 7R) -7- [2-Cyclopentyloxyimino-2- (fur-2-yl) -acetamido] -3-pyridinium methylceph-3-em-4-carboxylate (syn isomer)

Reaction of 8.3 g of the syn isomer of (6R, 7R) -3-acetoxymethyl-7- [2-cyclopentyloxyimino-2- (fur-2-yl) -acetamido] -ceph-3-em-4-carboxylic acid with 5.5 ml of pyridine in 120 ml of water gave 2.71 g of solid. 2.0 g of this in 25 ml of water were treated. 35 with an aqueous solution of perchloric acid (60% by weight) to pH 1.2.

The precipitated salt was collected, washed with a little ice-cold water and dried to 1.7 g of solid. 1.5 g of this solid

DK 155187 B

55 dissolved in the least possible volume of Ν, Ν-dimethylacetamide (ca.

3.0 ml) was treated with 0.45 ml triethylamine, triturated and refrigerated for 16 hours. The crystals were collected, washed with ice-cold Ν, Ν-dimethylacetamide and then with ether to give 1.23 g of the title compound. Recrystallization of a portion of 1.0 g of this material from a mixture of acetone and water (3.0 ml, mixing ratio 85:15) gave 0.68 g of the pure, title compound, λ (pH 6 buffer) 260 and 282 nm (ε = 18,000;

IILclX

18,600); V (d ^-DMSO; 100 MHz) 0.32 (d, J 8Hz; NH), 0.55, 1.32

ISLAND

and 1.74 (pyridine protons), 2.15 and 3.36 (furyl protons, syn isomers), 4.17 (dd, J 5 and 8 Hz; 7-H), 4.78 (d, J 5 Hz; 6-H), 4.26 and 4.60 (dd, J 14 Hz; C CN *), 5.28 and 8.0-8.6 (cyclopentyl protons), 6.34 and 6.76 ( dd, J 18 Hz; 2-H2) · 15

Example 35 (6R, 7R) -7- [2-Methoxyimino-2- (thien-2-yl) -acetamido] -3-pyridinylmethylceph-3-em-4-carboxylate (syn-isomer) - 2Q A solution of 10.0 g of the syn isomer of (6R, 7R) -3-acetoxymethyl-7- [2-methoxyimino-2- (thien-2-yl) -acetamido] -ceph-3-em-4-carboxylic acid 100 ml of water containing 8 ml of pyridine was stirred for 1 hour at 80 ° C. The reaction mixture was cooled and then evaporated to a slight volume to remove excess pyridine. The mixture was diluted with water to ca.

200 ml and insoluble material were filtered off. The filtrate was stored in a refrigerator for a few days and more solid was filtered off and discarded. The filtrate was washed successively with (R) 2 x 60 ml of methylene chloride, a solution of 10 ml of "Amberlite" 3q LA2 resin in 100 ml of methylene chloride and 3 x 60 ml of methylene chloride, and the washings were washed themselves with 60 ml of water. The combined aqueous phase was evaporated under reduced pressure at a temperature <40 ° C to give 3.6 g of a yellow glass. The crude product was dissolved in 16 ml of hot dimethylacetamide and treated dropwise with ace-tone until no more solid precipitated. By filtration, the solid was changed to a rubber; this was redissolved in water and evaporated to dryness as above, yielding 2.35 g of dark yellow solid. One was prepared

DK 155187 B

56 column of XAD-2 resin (500 g; 4 x 80 cm) and washed with ca. 1 liter of water. The crude product in 100 ml of water contained some ethanol was introduced into the column and eluted with water and then with water containing increasing amounts of ethanol. Separation progress was monitored by UV spectrometry. The fractions eluted with ethanol / water 1: 1 were combined and evaporated to a slight volume under reduced pressure at a temperature <40 ° C and finally lyophilized to give the title compound as a white foam 10 in an amount of 0.25 g, λ (pH 6 buffer) 259 nm (ε = 16.700); f 3.X-1 vmax (nu7 ° - * -) 1778 (β-lactam), 1670 and 1550 (CONH) and 1620 cm (CO 2 "); * (dg-DMSO; 100 MHz) 0.22 (NH), 0.52, 140, 184 (pyridinium protons), 2.36 and 2.80-3.0 (thienyl protons, syn isomers), 4.27 (7-H), 4.86 (6-H), 4.27 and 4.82 (2d, J 14 Hz; CH 2 N +), 6.14 (s; CH 3) and 6.39 and 6.90 (2d, J18 Hz; 2.H2).

Example 36 (6R, 7R) -3- (3-Carboxypyridinium methyl) -7- [2- (fur-2-yl) -2-metho-2H-iminoacetamido] -ceph-3-em-4-carboxylate sodium salt (syn - isomer) _

Reaction of 2.46 g of nicotinic acid with 4.45 g of the syn isomer of sodium (6R, 7R) -3-acetoxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido] -ceph-3 -em-4-carboxylate gave 1.51 g of the title compound, = = 69 ° (c = 1 H 2 O); max (pH 6 buffer) 270 nm (ε = 17,600) τ (D20; 100 MHz) 0.69, 0.96, 1.11 and 1.88 (pyridine protons), 2.32, 3.16 and 3.39 (furyl protons, syn isomer) 4.12 (d, J4Hz; 7-H), 4.68 (d, J4Hz; 6-H), 4.32 and 4.60 (2d, J14Hz; -CH2N), 6 , 01 (s; 30 CH 3) and 6.29 and 6.76 (2d, J18Hz; 2-H2).

Example 37 (6R, 7R) -3- (3-Carboxymethylpyridinium methyl) -7- [2- (fur-2-yl) -2-methoxyiminoacetamido] -ceph-3-em-4-carboxylate, sodium salt (syn isomer ) _-

Reaction of 2.74 g of pyrid-3-ylacetic acid with 4.45 g of the syn isomer of sodium (6R, 7R) "3-acetoxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido] -ceph-3-em-4-carboxylate 57

DK 1S 518 7 B

gave 0.91 g of the title compound at + = + 76 ° (c = 1 in HoO); λ (pH 6 buffer) 270 nm (ε =

U £ ulciX

20,000); τ (D20; 100 MHz) 1.20, 1.59, and 2.02 (pyridinium protons), 2.37, 3.19, 3.41 (furyl protons, syn isomers), 4.12 (d, J 4Hz; 7-H), 4.76 (d, J 4Hz; 6-H), 4.46 and 4.70 (2d, J 14Hz; CH 2 S), 6.05 (s; CH 3), 6.24 ( s; CH 2 Co 4), 6.38 and 6.82 (2d, J18Hz; 2-H2).

Example 38 (6R, 7R) -3- (2-Carboxypyridinium methyl) - 7- [2- (fur-2-yl) -2-methoxyiminoacetamido] -ceph-3-em-4-carboxylate, sodium salt (syn. isomer) __

Reaction of 2.46 g of 2-carboxypyridine with 4.45 g of sodium (6R, 7R) -3-acetoxymethyl-7- [2- (fur-2-yl) -2-methoxyimino-3-acetamido] -ceph-3 -em-4-carboxylate (syn-isomer) gave the title compound in an amount of 0.21 g, λ (pH 6 buffer) 275 nm (ε = 19,900); τ (Do0; 100 MHz) 1.12, 1.46, 1.96, 2y) 01 (pyridine protons) 2.34, 3.16, 3.40 (furyl protons, syn isomer), 4.16 (d, J 5Hz; 7-H), 4.41 (s; 20 CH 2 S), 4.76 (d, J 5Hz; 6-H), 6.03 (s; CH 3), 6.43 and 6.79 (2d, J18Hz; 2-H2).

Example 39 · (6R, 7R) -7- [2- (Fur-2-yl) -2-methoxyiminoacetamido] -3- (2-methyl-pyridinium methyl) -ceph-3-em-4-carboxylate (syn. isomer) 3.47 g of pyrid-2-ylacetic acid hydrochloride was dissolved in ca. 25 ml of water and treated with sodium hydroxide solution to pH 7. The solution was evaporated to near dryness under reduced pressure and reacted with 4.45 g of sodium (6R, 7R) -3-acetoxymethyl-7- [2- (fur-2 yl) -2-methoxyiminoacetamido] ceph-3-em-4-carboxylate. There was thus obtained 0.68 g of the title compound with λ (pH 6 buffer) 271 nm (ε = 19,400); v (nujol) 1768 (B-lactam), 1658 and 1526 (CONH)

IH 3.X

35 and 1600 (CO 2); τ (D20; 100 MHz) 1.29, 1.63, 2.12 and 2.16 (pyridine protons), 2.33, 3.18 and 3.41 (furyl protons, syn isomer), 4.16 (d, J5Hz; 7-H), 4.76 (d, J5Hz; 6-H), 4.45 and 4.70 (2d, J 15Hz; CH 2), 6.03 (s; OCH 3), 7 , 18 (s; CH 3) and 6.50 and 6.80 (2d, J18Hz; 2-H2).

DK 155187 B

58

Sodium and potassium salts of the compounds of the Examples were prepared in a conventional manner by reaction with sodium and potassium 2-ethyl hexanoate, respectively.

To illustrate such salt formation, preparation of 5 sodium (6R, 7R) -3-acetoxymethyl-7 - [(Z) -2- (fur-2-yl) -2-methoxyiminoacetamido] -ceph-3 is described. em-4-carboxylate.

A solution of approx. 42 g of syn- (6R, 7R) -acetoxymethyl-7- [(Z) -2-fur-2-yl) -2-methoxyiminoacetamido] -ceph-3-em-4-carboxylic acid, produced in 545 ml of wet ethyl acetate (about 1.7% H 2 O), stirred and a solution of 18.3 g of sodium 2-ethyl hexanoate in 180 ml of ethyl acetate was added. The solution was stirred for 30 minutes to produce a thick suspension. The product was isolated by filtration, washed with 4 x 50 ml of ethyl acetate and dried at 40 ° C in vacuo to give the desired sodium salt in an amount of 15 o of 35.4 g and containing 4.4% m / m of water (by Karl Fischer / analysis).

20 25 30 .35

Claims (2)

RCCO.NH-II [Λ -n-Aq ORa I COOH 1q wherein R represents an unsaturated heterocyclic group of five or six ring members, at most two of which ring members are selected from S, N and 0 and optionally substituted by a halogen atom or a hydroxy, C 1-6 alkyl, nitro, amino, C 1-6 alkylamino, di- (C 1-6 alkyl) amino, C 1-6 alkanoyl, (C 1-6 alkanoyl) amido, C 1-6 alkoxy - or C 1-6 alkylthio group, R a alkyl, C 2-6 alkenyl, 1-propynyl, propargyl, C 1-6 cycloalkyl or phenyl or any of these groups substituted with one of the substituents hydroxy, methoxy, ethoxy, bromo, nitro, amino, carboxy and benzoyl, and Q is methyl, vinyl or the group -Cl 2 Y, where Y is (i) the residue of a pyridine base selected from pyridine, carbamoylpyridine and aliphatic substituted pyridines having a maximum of 8 carbon atoms, (ii) azido, 6 (Iii) the group -SR wherein R is alkyl or an unsaturated hetero-cyclic group having a 5- or 6-membered heterocyclic ring wherein at most four of the ring members are selected from and O, N and S and which are unsubstituted or substituted by a methyl group, (iv) hydroxy, (9) the group -O.CO.R, where R is alkyl, vinyl, propylene, phenyl or C C is cycloalkyl, (vi) the group -O.CO.NH (CH2) mX / where X is hydrogen, fluoro, chloro, bromo or iodo and m is an integer of 1-4, (vii) non-toxic salts thereof, characterized in that: a) condensing a compound which may optionally be DK 155187 B in N-silylated amino or acid addition salt form, of the general formula Wherein H2 has the meaning given above and R1 is a hydrogen atom or a carboxyl blocking group, with an acid in the syn-isomeric form and of the general formula RCCOOH 15. ii III N \ a-OR c1 * wherein R has the meaning given above and R has the same meaning as Ra or is a hydrogen atom, or a reactive derivative thereof, or b) translates a compound in the syn-isomeric form and of the general formula RCCO .NH —- ^ 25 || IV ', wherein R, R and R have the meanings given above and Y' is a chlorine, bromine or iodine atom or an acetoxy group, with a nucleophile corresponding to the group Y, where desired and if desired, one or more of the following reactions are carried out: a * (I) ether forms a compound where R is a hydrogen 3 * atom to form a compound of formula I wherein R is R having the meaning given above, DK 155187 B (II) deacylates in a manner known per se, a compound wherein Q is a group -CH 2 Y and Y herein is an acyloxy group to form a compound of formula I wherein Q is the group - CE 2 Y and Y herein are a hydroxy group, 5 (III) acylated in a manner known per se, a compound wherein Q is the group -CH2Y and Y herein a hydroxy group to form a compound of formula I wherein Q is the group -CH2Y and Y herein an acyloxy or carbamoyloxy group, and (IV) removes any carboxyl protecting groups present which are recovered is the desired compound of formula I, optionally after conversion to a non-toxic salt thereof. 15 20 25 30 35