DK161645B - 2- (2-CHLORACETAMIDOTHIAZOL-4-YL) -2-OXYIMINO ACETIC ACID DERIVATIVES FOR USING INTERMEDIATES IN THE PREPARATION OF BETA-LACTAMANTIBIOTICS - Google Patents

Description

in

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The present invention relates to novel 2- (2-chloroacetamido-thiazol-4-yl) -2-oxyiminoacetic acid derivatives for use as intermediates in the preparation of β-lactam antibiotics, especially novel 7- [2- (2-aminothiazol-4-yl) ) -2- (syn) -methoxyiminoacetamido] -cephalo-sporine derivatives of the formula NBL S.

Λ I I 3 N i-N Λ-CH R (I)

\ QT Y

OR6 '

COOH

3 wherein R is hydrogen or a residue of a nucleophilic compound and R 1 is hydrogen or methyl, or pharmaceutically acceptable salts or esters thereof.

The compounds of the invention are characterized in that they have the general formula C1CH-CONH σ Y \ I I (MI)

N _.... J 1-C-C00H

II

N

R6 g wherein R is hydrogen or methyl, or is a salt, a low alkyl ester or a reactive derivative thereof.

The development of synthetic cephalospine derivatives has been directed to the conversion of 7-aminocephalosporanoic acid to various acyl derivatives at the 7-position or to derivatives at the 3-position to provide compounds having a broad antibacterial spectrum or specific antibacterial spectrum. However, the known cephalosporin derivatives do not have satisfactory antibacterial activity.

German Patent No. 2,462,736 discloses cephalosporin derivatives which are closely related to the cephalosporin derivatives of formula I. Among these known cephalosporin derivatives, there are mentioned

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the product "CEFOTIAM". It has been found that the present cephalo-sporin derivatives of formula I have a surprisingly good antibacterial effect over the known, closely related cephalo-sporin derivatives, as illustrated in more detail below.

The cephalosporin derivative of formula Γ appears to have a tautomeric form, i.e. a 2-aminothiazole compound and a 2-iminothiazoline compound, as shown below, although described as the thiazole compound in this specification.

»2« - /) / S.

N_Il-C-CONH-_Y \ \ och3 u Y ^

COOH

; hn S

v \

N_! 1 -C-CONH -_ / Y

H N li PR r3

and 3 / -y-c ^ R

COOH

While the carboxyl group at the 4-position of the compound of formula I may be free, it can form a salt, e.g. with a non-toxic cation such as an alkali metal, e.g. sodium or potassium, a basic amino acid, e.g. arginine, ornithine, lysine or histidine, or a polyhydroxyalkylamine, e.g. N-methylglucamine, diethanolamine, triethanolamine or trishydroxymethylaminomethane. Compound I may form an acidic salt with an inorganic acid such as hydrochloric acid and sulfuric acid, or with an organic acid such as toluene sulfonic acid and benzenesulfonic acid. The 4-carboxyl group may also be one of the biologically active ester forms, e.g. contributes to raising blood levels and prolonging efficacy. Such ester groups include lower alkoxymethyl groups, e.g. methoxymethyl, ethoxymethyl, isopropoxymethyl, α-methoxyethyl and α-ethoxyethyl, α-lower alkoxy-α-substituted methyl groups such as α-lower alkoxy- (C ^ ^) ethyl (e.g. methoxyethyl, ethoxyethyl) , propoxyethyl and i-propoxyethyl), lower alkylthiomethyl groups with

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3 to 3 carbon atoms, e.g. methylthiomethyl, ethylthiomethyl and isopropylthiomethyl, acyloxymethyl groups, e.g. pivaloyloxy-methyl and α-acetoxymethyl, ethoxycarbonyloxy-1-methylmethyl, or α-acyloxy-α-substituted methyl groups (eg α-acetoxy-α-methylmethyl).

Thus, Compounds I are novel compounds which show excellent activity against a wide range of bacteria including gram negative ones such as Escherichia coli, Serratia marcescens, Proteus retgeri, Enterobacter cloacae and Citrobacter freundii and which are resistant to β-lactamase. Compound I may e.g. used as a disinfectant to remove the aforementioned microorganisms from surgical instruments or as an anti-infectious agent, e.g. for the treatment of intraperitoneal infections, respiratory tract infections, urinary tract infections and other infectious diseases caused by the aforementioned microorganisms.

The compounds of the present invention can be converted into β-lactam antibiotics by a method known per se.

(1) Thus, the cephalosporin derivative of formula I is prepared by acylating the 7-amino group in a 7-aminocephalosporin compound of formula V-I-A (II) N J-CH2r3

° T

COOH

wherein the meaning hereinabove has a 2- (2-aminothiazol-4-yl) -2- (syn) -methoxyiminoacetic acid of the formula C1CELCONH-

I I

N U-C-COOH (III) r ^ Or6 followed by removal of the protecting group from the amino group (process I).

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In this process, compound III is used either as a free compound or in the form of a reactive derivative as an acylating agent for the acylation of the amino group at the 7-position of compound II. Thus, the free acid III, an alkali or alkaline earth metal salt of the free acid III (e.g., a sodium, potassium or calcium salt), is subjected to an organic amine salt of the free acid III (e.g. a trimethylamine salt or pyridine salt) or a reactive derivative thereof, such as an acid halide (e.g., acid chloride or acid bromide), acid anhydride, intermediate | the acid anhydride, active amide or active ester the aforementioned acylation reaction. Examples of this active ester include the p-nitrophenyl ester, 2,4-dinitrophenyl ester, pentachlorophenyl ester, N-hydroxysuccinimide ester and N-hydroxyphthalimide ester.

Examples of the mixed acid anhydride may be mentioned a mixed acid anhydride with a carbonic acid monoester {e.g. carbonic monomethyl ester or carbonic acid monoisobutyl ester) and a mixed acid anhydride with a lower alkanoic acid which may be substituted with halogen (e.g. pivalivic acid or trichloroacetic acid). When the carboxylic acid III is used as. the free acid or in the form of a salt, a suitable condensing agent is used. Examples of this condensing agent include N, Nf-disubstituted carbodiimides. e.g., N, Nt-dicyclohexyl arbodiimide, azolides, e.g. N, N1-carbonylimidazole and N, NT-thionyl diimidazole, dehydrating agents, e.g. N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline, phosphorus oxychloride and alkoxyacetylene and 2-halogenopyridinium salts (eg 2-chloropyridinium methyl iodide 'and 2-fluoropyridinium methyl iodide). When such a condensing agent is used, it is assumed that the reaction proceeds via the reactive derivative of the carboxylic acid III. The reaction is usually carried out in a suitably inert solvent.

Examples of such a solvent are halogenated. hydrocarbons, e.g. chloroform and methylene dichloride, ethers, e.g. tetrahydrofuran and dioxane, dimethylformamide, dimethylacetamide, acetone, water and mixtures of such solvents. The amount of said acylating agent is usually within the range of about 1 to 5, preferably 1 to 2 molar equivalents, based on compound II. This reaction is usually carried out at a temperature in the range of -50 ° C to + 40 ° C. The reaction time is from 1 to 10 hours, preferably 1 to 3 hours. After the acylation reaction, the amino group protecting group can be removed if necessary. The removal of the protecting group for the amino group can generally be carried out by methods known per se (e.g., the method described in the

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Japanese Publication No. 520 ^ 3/1975 and in Pure duck

Applied Chemistry, 7, 335 (1963), or an analogous process therefor. It is to be understood that when R of formula I is monohaloacetyl (e.g. monochloroacetyl) and R? a carbamoyloxy group whose amino group is protected, such as N-monohaloacetylcarbamoyloxy (e.g. N-monochloroacetylcarbamoyloxy), these two monohaloacetyl groups (e.g. monochloroacetyl groups) can be removed simultaneously. In this sense, 2, the protecting group R 'of the amino group is preferably a monohaloacetyl group. The reaction to remove the monohaloacetyl group from the amino group is carried out by reacting a compound of formula I whose amino group or groups are protected by monohaloacetyl with thiourea and a basic substance. Usually, this reaction is carried out in a solvent at a temperature near room temperature and in many cases it proceeds completely over about 1 to 10 hours. The solvent may be any solvent which does not interfere with this reaction. Thus, there may be mentioned ethers, e.g. ethyl ether, tetrahydrofuran and dioxane, lower alcohols, e.g., methanol and ethanol, halogenated hydrocarbons, e.g. chloroform and methylene dichloride, esters, e.g. ethyl acetate and butyl acetate, ketones, e.g. acetone and methyl ethyl ketone, water and various mixtures of such solvents.

This reaction to remove the N-haloacetyl group from the N-mono-haloacetylcarbamoyloxymethyl group at the 3-position of compound I does not proceed to a significant extent if thiourea is left alone to affect compound I. However, if compound I is reacted with thiourea and a basic substance, the desired reaction to remove the monohaloacetyl group selectively and smoothly to form the 3-carbamoyloxymethyl compound I. As the basic substance for use in this reaction may be mentioned an alkali or alkaline earth metal salt of a low aliphatic carboxylic acid or an inorganic or organic base having a pKa value of not less than 9.5, preferably in the range of pKa 9.3 to 12.0. Examples of this salt of a low-aliphatic carboxylic acid are the salts of low-aliphatic carboxylic acids having 1 to 6 carbon atoms, such as sodium acetate, potassium acetate, calcium acetate, barium acetate, sodium formate, sodium propionate and potassium hexanoate. Examples of the inorganic base may be mentioned the alkali metal salts of carbonic acid such as sodium carbonate and potassium carbonate. The organic base can e.g. be one of the mono-, di- or tr:

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lower alkyl substituted amines whose lower alkyl moiety has 1 to 4 carbon atoms, e.g. trimethylamine, triethylamine, ethylamine, methylamine, diethylamine, dimethylamine, tributylamine, dibutylamine and butylamine, and 5- to 6-membered cyclic amines substituted at the N-position with lower alkyl of 1 to 2 carbon atoms such as N-methylpyrrolidine, N-ethylpyrrolidine , N-methylpiperazine and N-ethylpiperazine. While thiourea is used in this reaction, the reaction can also be successfully carried out with N- or N, N-substituted thiourea, such as methylthiourea, Ν, Ν-diethylthiourea or N, N-hexamethylthiourea.

(2) 7- [2- (2-Aminothiazol-4-yl) -2- (syn) -methoxyiminoacetamidoj-cepha-losporin derivative of the formula

P S

R

li S

N_ϋ-C-CONH __ / \ (V) II}?

'IT \ o J - ^ - 0 «2R

AND- T

* COOH

Wherein R is hydrogen or chloroacetyl, and R is a group from a nucleophilic compound, prepared by reacting a compound of the formula

2 A

r ^ nh-Zx jl I s N_11-C-CONH - / \

N; _N J-ChA

\ o ^ Y 2

0CH3 COOH

wherein R. is the acylloxy, carbamoyloxy or halogen, with a nucleophilic compound, if necessary followed by removal of the protecting group for the amino group (process 2).

As the acyloxy group represented by the formula TV, e.g. mention is made of acyloxy derived from a low aliphatic carboxylic acid having 2 to 4 carbonic acids.

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atoms which may, if desired, be substituted by oxo, carboxy or ethoxycarbamoyl, e.g. acetyloxy, propionyloxy, 3-oxobutyryloxy, 3-carboxypropionyloxy, 3-ethoxycarbamoylpropionyloxy and 4-carboxybutyryloxy, and acyloxy derived from an aromatic carboxylic acid, which may be substituted with hydroxy, carboxy, carboxy, carboxy, carboxy, carboxy, . mandelyloxy, 2-carboxy-benzoyloxy, 2- (carboethoxycarbamoyl) -benzoyloxy, and 2- (carboethoxysulfamoyl) -benzoyloxy. The halogen represented by R 1 - e.g. be chlorine, bromine or iodine. The group from a nucleophilic compound represented by R 'in formula V means a group from a nucleophilic compound corresponding to a group from a nucleophilic compound re's presented by except the acyloxy groups represented by R 1 and carbamoyloxy. However, for the present reaction, it is generally advantageous to use a compound IV with an acyloxy group derived from a low aliphatic carboxylic acid such as acetyloxy. The nucleophilic compound used in this reaction is a compound corresponding to the group of a nucleophilic compound represented by the symbol R in Formula V. Particularly preferred are the heterocyclic thiol compounds, i.e. mercapto compounds which may contain a substituent. Among the nucleophilic compounds corresponding to the group represented by R 2, mercapto compounds can be used in their free form, although it is advantageous to use them in the form of alkali metal salts, e.g. sodium or potassium salts. This reaction is preferably carried out in a solvent. Eg. water, deuterium or an organic solvent which is readily miscible with water and which does not react with the reactants, e.g. dimethylformamide, dimethylacetamide, dioxane, acetone, alcohol, acetonitrile, dimethylsulfoxide and tetrahydrofuran.

While the reaction temperature and time vary with such factors as the starting material and solvent used, they are generally selected in the range of 0 to 100 ° C, preferably 30 to 70 ° C, and 2 to 4 $ hours, preferably 3 to 15 hours. The reaction is preferably carried out in the vicinity of the neutral point and suitably within the range of from ca. pH 2 to 8, preferably pH 5 to 8. The course of this reaction can sometimes be smoothed by the addition of a quaternary ammonium salt with surface activity such as trimethylbenzylammonium bromide or triethylbenzylammonium bromide or triethylbenzylammonium hydroxide. Moreover, more satisfactory is achieved

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results if the reaction is carried out in an inert gas atmosphere, such as in nitrogen, to prevent air oxidation of the mercapto compound. : (3) The cephalosporin derivative of formula I may also be prepared by subjecting a 7- [2- (2-aminothiazol-4-yl) -2- (syn) -hydroxyiminoacetamido] -cephalosporin derivative of formula 9 SR \ | l I s N-1; 00 "-I— (4 (vi) HJ -" \ J-CH 2 R 3

\ 0 T

OH COOH

wherein R R and R are as previously defined, O-methylation.

The O-methylation is carried out by reacting compound VI with a methylating agent (process 3) *

This Q-methylation reaction is usually carried out in a solvent under ice-cooling or at room temperature (0 to 40 ° C, preferably 5 to 30 ° C), and in many cases, proceeds completely over approx. 5 minutes to 5 hours, preferably 5 minutes to 2 hours. The solvent may be any solvent which does not affect the reaction, such as an ether, e.g. tetrahydrofuran and dioxane, a lower alcohol, e.g. methanol and ethanol, a halogenated hydrocarbon, e.g. chloroform and methylene chloride, an ester, e.g. ethyl acetate and butyl acetate, an amide, e.g. Ν, Ν-dimethylformamide and Ν, Ν-dimethylacetamide, water and mixtures of such solvents. The methylating agent may be a methylating agent commonly used in organic chemistry such as a methyl halide (e.g., methyl iodide and methyl bromide), dimethyl sulfate, or diazomethane.

This reaction can proceed smoothly in the presence of a suitable base, except for diazomethane. As such, an inorganic base such as an alkali metal salt of carbonic acid (e.g., sodium carbonate, potassium carbonate) or an alkali metal hydroxide (e.g., sodium hydroxide or potassium hydroxide) is normally used. However, where the stability of Compound VI is to be considered, sodium carbonate, potassium carbonate or the like is preferably used. The reaction can

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y is also carried in a buffer at ca. pH 7.5 to S.5.

The cephalosporin compounds I prepared by the numerous manufacturing processes described above can each be purified by methods known per se, such as column chromatography, extraction, precipitation and recrystallization. If necessary, each of these compounds can be treated by methods known per se to obtain the desired salts, esters and more.

The compound of the invention, i.e. The 2- (2-chloroacetamidothiazol-4-yl) -2-oxoiminoacetic acid derivative III, e.g. are prepared by the numerous different methods which are described in detail below.

(I) First, a 4-halogen-3-oxo-2-oxyimino butyric acid derivative of the formula XCHPCOC-COOR7 N (VII) OR6 6 wherein X is halogen, e.g. chlorine or bromine, R ° is hydrogen or 7 methyl, and R 'is a lower alkyl of 1 to 3 carbon atoms, e.g. methyl, ethyl or propyl, with thiourea to give a 2- (2-amino-thiazol-4-yl) -2-oxyiminoacetic acid derivative of the formula S.

H "N- / 2 II I 7 N._1-c-coor" (VIII)

II

N

OR6 6 * 7 wherein R and R 'have the meaning given above. In the two cases where R is hydrogen and methyl respectively, compound VIII is usually obtained as a mixture of syn and anti-isomers thereof. This reaction is usually carried out by reacting a compound of formula VII with thiourea in an organic solvent such as ethanol, methanol or tetrahydrofuran at ambient or elevated temperature (0-100 ° C, preferably 10-50 ° C). The reaction time is from 1 to

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30 hours, preferably 1 to 5 hours. To isolate the desired syn isomer from the resulting mixture of the syn and anti forms of compound VIII, one of the following methods can be successfully followed. Thus, these processes include fractional crystallization which utilizes the difference in crystallizability or solubility of the isomers of compound VIII as such, a salt of compound VIII of a hydrogen halide (HBr or HCl salt), or a derivative of compound VIII with a protecting group on its 2-amino group, wherein the protecting group (e.g., monochloroacetyl or dichloroacetyl) is introduced by a method known per se, isolation by chromatography and such a method that when the compound VIII or compound VIII with a protecting group on its 2-amino group is hydrolyzed i the ester moiety, in a per se known method for a carboxylic acid derivative of formula III, the syn isomer alone is selectively isolated by utilizing the difference in rate of hydrolysis between the syn and anti isomers.

In the latter process, because of the higher rate of hydrolysis of the anti-isomer than of the syn isomer, the anti-isomer can be selectively hydrolyzed and removed. The reaction by hydrolysis of the ester linkage in Compound VIII with or without a substituent of the 2-amino group of the pendant is usually carried out in the presence of one to several molar equivalents of an alkali metal hydroxide, e.g. potassium hydroxide or sodium hydroxide at a temperature of from 0 ° C to room temperature and in water or a mixture of water and a water-miscible organic solvent, e.g. methanol, ethanol, acetone, tetrahydrofuran, dioxane, Ν ', N-dimethylformamide or N, N-dimethylacetamide. If R 1 of compound VIII is hydrogen, the isolated syn isomer may be converted to a syn isomer of compound VIII, which is methyl, by subjecting the former to compound VIII methylation. This methylation reaction is usually carried out in a solvent under ice-cooling or at temperatures near room temperature and, in many cases, proceeds completely over a few minutes to several hours. The solvent for this purpose can be any type of solvent provided it does not affect the reaction.

Thus, e.g. mention is made of tetrahydrofuran, dioxane, methanol, ethanol, chloroform, methylene dichloride, ethyl acetate, butyl acetate, N, N-dimethylformamide, Ν, Ν-dimethylacetamide and water as well as mixtures of such solvents. As methylating agent may be mentioned methyl

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halides, e.g. methyl iodide and methyl bromide, dimethyl sulfate and diazomethane, just to name a few. In all cases, except where diazomethane is used, compound VIII wherein R 1 is hydrogen is reacted with this methylating agent in the presence of a base such as an alkali metal carbonate (e.g., sodium carbonate or potassium carbonate) or an alkali metal hydroxide (e.g., sodium hydroxide or potassium hydroxide). Some of the physical constants of the thus-obtained isomers of compounds III and VIII are shown below in comparison with the physical constants of the corresponding anti-isomers (see Table 1).

Table 1.

NMR Spectrum Melting Point

Structure (ppm) (° C)

A

syn- H2N "f] 1 d6 ~ DMS0 isomer jj_ [-C-GOOC ^ R. 6, SOs (5-H) 1 $ 5.5 4> 11.6s (OH)

X0H

S

anti-H2N - / \ I d6 ~ DMSO isomer N-1 - C-COOCA 7.50s (5-H) 145.3 4 ^ 12.5s (OH) HO

S

syn-H2N- / 'v \ I CDCl3 163 car isomer J_J-c-COOC-fL · 6.74s (5-H) 164 H ^^ 4.02s (OCH3) xoch3

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Table 1 cont.

NMR Spectrum Melting Point

Structure (ppm) (° C)

S

anti- H2N - / \ I CDC13 114 to isomer N _! - C-COOC-7ς 7} 43s (5-H) 115 |

II ^ 5 4.07s (0CH) I

/ 3 CH30 s syn- H0N - / \ I CDC1, isomer -C-COOCH ^> 74s (5-H) 104 ^ 9 I 3 4.02s (OCH3) XOCH3s anti-H2N- / \ I CDC13 isomer N_ [ -C-C00CH-. 7 µ4 s (5-H) J 3 4.06s (OCH 3) · CH 3 q /

S

syn- ClCH2 CONH - / \ I CDC13 111 to isomer | _ [-C-COOC-,ς 7.15s (5-H) 112 I 3 4, east (och3) XOCH0 '5 S' 'anti-C1CH2 CONH- / N. In CDC13 Si to Somer 1_11-0-0000, ¾ 7.94s (5-H) 82 J, 2 5 4.10s (OCH3)

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Table 1 cont.

NMR Spectrum Melting Point Structure (ppm) (° C)

S

syn- CICEpCONH- / \ I d6 ~ DMS0 170 to isomer J_ [I 7.57s (5-H) 171 ^ C-COOH 3.95s (OCH3) to ^ OCt ^

S

anti-C1CH2C0NH- / \ I d6-DMSO1S2 to isomer | _1-C-C00H #, 00s (5-H) 1 & 3 4, OOs (OCH3) 0Η30κ

S

syn- C1CH2CQNH- / X I CDC13

isomer _ [-C-COOCH 7j24s (5-H) 130, S

J 3 4.02s (OCH3) xoch3 β anti-C1CH2 CONH-A I CDC13 isomer I_ll-O-COOCH, 8.02s (5-H) J, 3 4.12s (OCH3) CH30

Notes: s: singlet.

The methoxyimino (hydroxyimino) group in the nsyn "isomer is cis to the carboxyl group and, the fanti" isomer is trans to the carboxyl group.

(II) The method of selectively preparing compound III

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(syn-isomer) is described below. While the aforementioned reaction of compound VII with thiourea yields a mixture of syn and anti-isomers of compound VIII, the anti-isomer of compound VIII is often predominant. Studies on the conditions of this cyclization reaction shed light on the conditions leading to a selective formation of the desired syn isomer. Thus, if the reaction of compound VII with thiourea to form compound VIII is carried out under the conditions described above, and anti-isomers usually in a ratio of 2: 9 ^ to 50:50.

However, it has been found that if this cyclization reaction is carried out in water or a mixture of water and a water miscible solvent such as methanol, ethanol, acetone, tetrahydrofuran, dioxane, Ν, Ν-dimethylformamide, N, N-dimethylacetamyl or N-methyl-piperidone and in the presence of a basic substance, the syn-isomer of compound VIII is selectively formed (usually at a ratio of ca.

$ 5: 15 to 100: 0). As the basic substance useful for this reaction, mention may be made of alkali or alkaline earth salts of lavariphatic carboxylic acids and inorganic or organic bases having pKa values of not less than 9.5, preferably in the range - from 9, S to 12 , 0th Examples of these low-aliphatic carboxylic acid salts include the salts of low-aliphatic carboxylic acids having 1 to 6 carbon atoms, such as sodium acetate, potassium acetate, calcium acetate, barium acetate, sodium formate, sodium propionate and potassium hexanoate, while the above-mentioned inorganic alkali potassium carbonate. As the organic bases may be mentioned trilavalkyl-substituted amines whose lower alkyl has 1 to 4 carbon atoms. such as trimethylamine, triethylamine and tributylamine, and 5- to 6-membered cydic amines substituted at the N-position by lower alkyl of 1 to 2 carbon atoms, such as N-methylpyrrolidine, N-ethylpyrrolidine, N-methylpiperazine and N-ethylpiperazine. If said Ν, Ν-dimethylformamide, N, N-dimethylacetamide or N-methylpyrrolidone are used as the solvent, it is not always necessary to add the aforementioned basic substance. The reaction temperature and time are generally selected in the range of 0 to 50 ° C, preferably 0 to 30 ° C, ° g in the range of 1 to 30 hours, preferably 1 to 5 hours.

(III) Compound VIII (syn isomer) can also be selectively prepared

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by the following procedure. Thus, in the further search for a process for selectively preparing the syn isomer, it has been discovered that by reacting a 2-aminothiazol-4-ylglyoxylic acid derivative of formula IX with O-methylhydroxylamine, the syn isomer of the methoxyimino compound can be selectively obtained.

s \ C1CH CONH- / \ i 7 (ix) n_U-c-coor '

II

o 7 wherein R has the meaning given above.

Normally, this reaction can be carried out smoothly in a suitable solvent at a pH of approx. 4.0 to 9.0. Said solvent can be any type of solvent unless it affects the reaction. Thus, e.g. ethers such as ethyl ether, tetrahydrofuran and dioxane, lower alcohols such as methanol and ethanol, halogenated hydrocarbons such as chloroform and methylene dichloride, esters such as ethyl acetate and butyl acetate, water and mixtures of such solvents are mentioned. While this reaction proceeds around room temperature, it can be accelerated by heating. The reaction temperature and time are generally selected in the range of 0 to 100 ° C, preferably 0 to 50 ° C, respectively, ranging from 1 to 10 hours, preferably 1 to 5 hours.

The starting material IX for this reaction can be prepared by the reaction described below. Thus, the hydrolysis of a nitron compound of the formula gives

ClCH2 ONH-j / 8 ^ N_i-C-COOR7 (X) o-n-ch3 + η wherein R has the meaning given above, compound IX.

This hydrolysis reaction proceeds smoothly in the presence of a mineral acid and is usually carried out in a solvent. As examples 16

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Hydrochloric acid, sulfuric acid and phosphoric acid are mentioned on this mineral acid. The solvent may be of any desired type which does not affect the reaction. Thus, there may be mentioned ethers, e.g. tetrahydrofuran and dioxane, alcohols, e.g. methanol and ethanol, ketones, e.g. acetone and methyl ethyl ketone, water and mixtures of such solvents. Usually this reaction is carried out under ice-cooling or at room temperature. The starting material X can be obtained by subjecting a compound of formula VIII wherein R 1 is hydrogen and if the amino group at the 2-position is protected, methylation.

The conditions for this methylation reaction are essentially the same as the conditions under which the aforementioned compound VIII wherein R 1 is hydrogen is methylated (cf. the aforementioned process I).

Under the methylation conditions described, the methylation of the syn isomer of compound VIU wherein R 1 is hydrogen does not give a substantial amount of this nitron compound X but the methylation of the anti-isomer of compound VIII wherein R 1 is hydrogen gives the nitron compound. X as a dominant product.

The compound of formula VII may e.g. are prepared by the methods described in the Journal of Medicinal Chemistry, 16-, $ 97 (1973), Helvetica Chimica Acta, _49, 26 (1966), Journal of the American Chemical Society, 60, $ 132 (193) $)> and German Publication No. 2,556,736, or by methods similar thereto. The compound of formula II used in the present invention may e.g. is prepared by a suitable method selected from the methods disclosed in U.S. Patent Nos. 3, $ 75,151 and 3 * 697,515, German Publication No. 2,461.47, German Publication No. 2,607,064 (Dutch Patent Application No. 7,601. 902), German Publication Publication No. 2,619,243, Japanese Publication Publication No. 52.OS3 / I975, German Publication Publication No. 2.46O.33I and No.

2.46Ο.332, or by an analogous method.

Compound IV may e.g. is prepared by Belgian patent specification 719.710 or by an analogous method. Alternatively, it can be prepared using the aforementioned process 1) on compound III and compound II, wherein -CHOR4 is -C2R obtainable by one of the above processes for the preparation of compound II or analogous methods. The compound VI may e.g. is produced by a method analogous to the electric disclosure described in German Publication No. 2,556,736.

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for example, by reacting compound II with the syn isomer of compound VIII wherein Ru is hydrogen.

The present invention is illustrated in more detail below with reference to; Examples 5-7 and 22-23 illustrate the preparation of the intermediates of the invention, Examples 1-4 and 8-21 illustrate the preparation of starting compounds, and Examples 24-43 illustrate the use of the subject intermediates for the preparation of β-lactam antibiotics. and their biological effect. In this specification, "DMSO" is an abbreviation for dimethyl sulfoxide. Resins called "Amberlite" are products manufactured by Rohm & Haas Co. in USA. All temperatures are uncorrected and percentages are by weight unless otherwise stated. The NMR spectra were measured with a Varian Model HA 100 (100 MHz) or T60 (60 MHz) spectrometer with tetramethylsilane as internal or external reference and all δ values are in ppm. The symbol s stands for singlet, d for doublet, t for triplet, q for quartet, m for multiplet, and J is a coupling constant.

Example 1 In a solution of 13.3 g of sodium carbonate in 120 ml of water, 10 g of ethyl 3-oxo-2-hydroxyiminobutyrate is dissolved and 30 ml of methanol are added. The mixture is cooled with ice and 15.3 g of dimethyl sulfate is added dropwise with stirring over 3 minutes. After the addition is complete, the ice bath is removed and the mixture is stirred at room temperature for 40 minutes. The reaction mixture (pH 6 or higher) is extracted twice with ethyl acetate and the extracts are combined, washed with water and dried. The solvent is then evaporated under reduced pressure and the residue is subjected to distillation under reduced pressure. In the above process, 9 g of ethyl 3-oxo-2-methoxyiminobutyrate is obtained as a pale yellow oil boiling at 56-61 ° C / 0.3 to 0.4 mm Hg.

<

Elemental Analysis of Cy ΗχιΝΟ:

Calculated: C 46.54 H 6.40 N 6.06

Found: C 46.41 H 6.51 N 7.96 NMR Spectrum (60 MHz, in CDCl3): 2.40 ppm (3H, s, CH 2 CO), 4.10 ppm (3H, s, = N0CH3).

Example 2

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(1) In 120 ml of chloroform 27.3 g of ethyl 3-oxo-2-methoxyimino-butyrate are dissolved and the solution is heated to 40 ° C. Then a solution of 25.3 g of bromine in 30 ml of chloroform is added dropwise over 30 minutes. The mixture is stirred and the reaction proceeds at room temperature for one hour. The reaction mixture is washed with a 5 $ aqueous solution of sodium bicarbonate and then with water and the organic layer is dried. The solvent is then distilled off under reduced pressure to give 36.2 g of ethyl 4-bromo-3-oxo-2-methoxy-iminobutyrate as an oily product.

.Delta. (60 MHz, in CDCl 3): 4.16 ppm (3H, s, OCH 2), 4.36 ppm (2H, s, BrCH 2 CO).

(2) In 20 ml of ethanol, 5 g of the above product is dissolved and then 1.3 g of thiourea is added. The mixture is heated under reflux for 3 hours. After cooling, the precipitate is collected by filtration and dissolved in 20 ml of water, to which is added sodium bicarbonate.

The separated oil is extracted with ethyl acetate. The ethyl acetate layer is washed and dried. The ethyl acetate is then evaporated to give white crystals. Recrystallization from ethanol gives 2.6 g (57.2 $) of ethyl 2- (2-aminothiazol-4-yl) -2- (anti) methoxyiminoacetate as white crystals, mp 114-115 ° C.

Elemental analysis of CgH--N ^O₂S:

Calculated: G 41.91 H 4, $ 4 N 13.33

Found: C 41.71 H 4.75 N 13.07 NMR Spectrum (60 MHz, in CDG1): 4.07 ppm (3H, s, OCH 3), 5.30 ppm (2H, br s, NH 2 ), 7.43 ppm (1H, s, th thiazole 5H).

(3) The filtrate obtained by collecting the first precipitate is concentrated under reduced pressure and sodium bicarbonate is added to the residue. The mixture is extracted with ethyl acetate and the oil obtained from the ethyl acetate layer is purified by column chromatography on silica gel.

In the above process, 59 mg (1.3%) of ethyl 2- (2-aminothiazol-4-yl) -2- (syn) methoxyiminoacetate is obtained as white crystals, m.p.

19 DK 161645 B

section I63-I640C.

Elementary analysis of:

Calculated: C, 41.91; H, 4.34; N, 13.33

Found: C 41.57 H 4.76 N 13.07 NMR Spectrum (60 MHz, in CDCl3): 4.02 ppm (3H, s, AND 4), 5.30 ppm (2H, br s, NH2), 6.74 ppm (1H, s, thiazole 5H).

Example 3

To 600 ml of ethanol is added 121 g of ethyl 4-chloro-3-oxo-2-hydroxyiminoacetate with 47.6 g of thiourea and the mixture is stirred at room temperature for 3 hours. The ethanol is then evaporated under reduced pressure and 350 ml of water is added. The aqueous layer is washed with ether, neutralized with sodium bicarbonate (to pH 7.5) and extracted with ethyl acetate-tetrahydrofuran (1: 1). The organic layer is washed with water and dried. The solvent is then distilled off and 45 g of crystalline product is obtained.

1 g of the above product is purified by column chromatography on silica gel (eluent: ethyl acetate-n-hexane). The first fraction gives 65Ο mg of the anti-isomer of ethyl 2- (2-aminothiazol-4-yl) -2-hydroxy-iminoacetate, and 150 mg of the syn isomer is obtained from the second fraction.

Anti-isomer: white crystals, mp 145.3 ° C. Syn isomer: pale yellowish-white crystals, mp 135.5 ° C.

Elemental Analysis of CyH

Calculated: C 39.06 H 4.21 N 19. 52

Found: (Anti-) C 33.31 H 4.20 N 19.62 (Syn-) C 39.23 H 4.10 N 19.63 NMR Spectrum (60 MHz, id 1 -DMSO): Anti-isomer : 7.10 ppm (2H, br s., NH 2), 7.50 ppm (1H, s., Thiazole 5H), 12.5 ppm (1H, s., OH).

20

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Syn isomer: 6, BO ppm (1H, s., Thiazole 5H), 7.12 ppm (2H, br. S, NH 2), 11.6 ppm (1H, s., OH).

Example 4 In 150 ml of water, dissolve 10.6 g of sodium carbonate and then add a solution of 10.7 g of ethyl 2- (2-aminothiazol-4-yl) -2- (syn) -hydroxy-iminoacetate in a mixture of 150 ml of tetrahydrofuran and 50 ml of methanol. During ice cooling, 12.6 g of dimethyl sulfate is added dropwise over 5 minutes. After the addition is complete, the ice bath is removed and the mixture is stirred at room temperature. During stirring, the separation of white crystals begins. After 3 hours, the majority of the organic solvent is distilled off under reduced pressure and the residue is cooled with ice. The resulting precipitate is collected by filtration, washed with water and dried. By the above process, 5 g of ethyl 2- (2-aminothiazol-4-yl) -2- (syn) methoxyiminoacetate are obtained as white crystals.

By NMR spectrum and other properties, this product is identified as ethyl 2- (2-aminothiazol-4-yl) -2- (syn) methoxyiminoacetate.

Example 5 In 10 ml of Ν, Ν-dimethylacetamide is dissolved 2.15 g of ethyl 2- (2-aminothiazol-4-yl) -2- (syn) methoxyiminoacetate (m.p. 63-64 ° C) and added dropwise under ice-cooling 1.27 g of chloroacetyl chloride. The mixture is stirred under ice-cooling for 30 minutes and then at room temperature for 30 minutes. The reaction mixture is diluted with 50 ml of water and extracted twice, each time with 100 ml of ethyl acetate. The extracts are combined, washed with a 5% aqueous solution of sodium hydrogen carbonate and a saturated aqueous solution of sodium chloride in the order mentioned, and finally dried. The solvent is then evaporated to give 2.04 g of ethyl 2- (2-chloroacetamidothiazol-4-yl) -2- (syn) methoxyiminoacetate as a crystalline product, mp 111-112 ° C.

Elemental Analysis for C- qHHN ^O₂SCI -

Calculated: C, 39.29; H, 3.96; N, 13.74

Found: C 39.15 H 3.91 N 13.69 NMR Spectrum (60 MHz, in CDCl3): 4.00 ppm (3H, s., = NOCH3), 4.24 ppm

21 DK 161645 B

(2H, s., C1CH2 CO), 7.15 ppm (1H, s-, thiazole 5H).

Example 6

To a solution of 9 g of potassium hydroxide in a mixture of 85 ml of water and 452 ml of ethanol is added 9.62 g of ethyl 2- (2-chloroacetamidothiazol-4-yl) -2- (syn) methoxyiminoacetate and the mixture is stirred at room temperature. for 2 hours. The ethanol is distilled off under reduced pressure and after the addition of 85 ml of water, the residue is washed with 100 ml of ethyl acetate. The aqueous layer is adjusted to pH 2 with 10% Ts hydrochloric acid solution and extracted twice with 200 ml portions of ethyl acetate. The extracts are collected, washed with a saturated aqueous solution of sodium chloride and dried. The solvent is then distilled off and 7.63 g of 2- (2-chloroacetamidothiazol-4-yl) -2- (syn) -methoxyiminoacetic acid are obtained as crystals melting at 170-171 ° C.

Elemental Analysis for CgHgN2 O2 SC1:

Calculated: G 34.60 H 2.90 N 15.13

Found: C 34.97 H 3.03 N 14.74 NMR Spectrum (60 MHz, id 1 -DMSO): 3.95 ppm (3H, s. == NOCH 3), 4.40 ppm (2H, -s., C1CH2 CO), 7.57 ppm (1H, s., thiazole 5H).

Example 7 2.3 µg of a 7: $ mixture of the syn and anti-isomers of ethyl 2- (2-aminothiazol-4-yl) -2-methoxyiminoacetate is chloroacetylated with chloroacetyl chloride as in Reference Example 5, and 30 ml of ether are added to the resulting mixture of the syn and anti forms of ethyl 2- (2- * chloroacetamidothiazol-4-yl) -2-methoxyiminoacetate. The precipitated crystals are collected by filtration (product A) - By NMR spectrum and other properties, this product is identified with the ethyl 2- (2-chloroacetamidothiazol-4-yl) -2- (syn) methoxyiminoacetate obtained in Example 5. Yield 600 mg.

The oil obtained by concentration of the filtrate (2.1 + 2 g, a mixture of syn- and anti-isomers) is added to a solution of $ 79 mg of potassium hydroxide in a mixture of 5 ml of water and $ 0 ml of ethanol under ice-cooling,

22 DK 161645 B

and the whole mixture is stirred at this temperature for 15 minutes. The ethanol is distilled off under reduced pressure and the residue is diluted with 50 ml of water and extracted twice with 100 ml portions of ethyl acetate. The ethyl acetate layer is washed with water and dried. The ethyl acetate is then distilled off and 577 mg of ethyl 2- (2-chloroacetamidothiazol-4-yl) -2- (syn) methoxyiminoacetate (product B) is obtained. By NMR spectrum and other properties, the product is identified by the syn isomer of Example 5. Products A and B provide a total yield of 1076 g or a recovery rate of 96.8%.

Example 8 Dissolve 67.8 g of ethyl 4-chloro-3-oxo-2-hydroxyiminoacetate in 600 ml of 50% aqueous tetrahydrofuran solution, adding 155 g of sodium acetate trihydrate and 53.2 g of thiourea. The mixture is stirred at room temperature for 4 hours. The reaction mixture is adjusted to pH 7.0 with sodium bicarbonate and extracted twice with sodium chloride twice with 300 ml of tetrahydrofuran. The extracts are collected, washed with water and dried. The tetrahydrofuran is then distilled off and 27.5 g of ethyl 2- (2-aminothiazol-4-yl) -2-hydroxy-iminoacetate are obtained as crystals. Based on NMR and other data, this product appears to be an 82: 18 mixture of syn- and anti-isomers.

A similar reaction is carried out without the use of sodium acetate. Based on the same criteria, the resulting product is seen to be a 25: 75 mixture of syn- and anti-isomers.

Example 9

The reaction of Example 8 is repeated except that a 50% aqueous ethanol solution is used in place of 50% T aqueous tetrahydro uranium solution. In this case, if sodium acetate is used, an 83:17 mixture of the syn and anti-isomers of ethyl 2- (2-aminothiazol-4-yl) -2-hydroxyiminoacetate is also obtained. In contrast, if sodium acetate is not used, the above reaction gives a 50:50 mixture of syn- and anti-isomers. The ratio of the syn and anti-isomers is determined by NMR spectra and other methods.

Example 10

23 DK 161645 B

The reaction of Example 9 is repeated except that N, N-dimethylacetamide is used in place of 50% 1s aqueous tetrahydrofuran sodium acetate. This process gives a # 5: 15 mixture of syn and anti-isomers of ethyl 2- (2-aminothiazol-4-yl) -2-hydroxyiminoacetate.

Example 11 In 10 ml of 50% aqueous ethanol solution, 200 mg of ethyl 2-amino-thiazol-4-yl glyoxylate are dissolved, followed by the addition of 166 mg of 0-methylhydroxylamine hydrochloride and then 16B mg of sodium bicarbonate. The mixture is stirred in a sealed container at 70 ° C for 5 hours. The reaction mixture is concentrated under reduced pressure and the residue is diluted with 10 ml of water and extracted with ethyl acetate. The ethyl acetate layer is washed with water and dried. The ethyl acetate is then distilled off, and ethyl 2- (2-aminothiazol-4-yl) ~ 2-methoxyiminoacetate is obtained as crystals. Based on NMR and other data, this product appears to be a S3: 17 mixture of syn- and anti-isomers.

Example 12 In 70 ml of ethanol containing 10% HCl, 2.44 g of methyl nitrone of ethyl 2- (2-aminothiazol-4-yl) -2- (anti) hydroxyiminoacetate which is N- (2-aminothiazole ~ 4 (1-yl-ethoxycarbonyl) -methylene methylamine-N-oxide, m.p. The mixture is stirred at room temperature for 16 hours. The reaction mixture is concentrated under reduced pressure and after addition of 10 ml of water, the residue is adjusted to pH 7.5 with a 5% aqueous solution of sodium bicarbonate and extracted with ethyl acetate. The ethyl acetate layer is washed with water and dried. The ethyl acetate is then distilled off and the residue recrystallized from ethanol. By the above process 1.54 g of ethyl 2-aminothiazol-4-yl glyoxylate are obtained as yellow crystals melting at 143.3 ° B.

Elemental Analysis for CyHgN2 O2 S:

Calculated: C 41.9 $ H 4.02 N 13.99

Found: C 41, # 3 H 4.14 N 13.9 °

Example 13

24 DK 161645 B

In 50 ml of 1 N hydrochloric acid, 1 g of the same N- (2-aminothiazol-4-yl-ethoxycarbonyl) -methylene methylamine-N-oxide used in Reference Example 12 is dissolved and the solution is stirred at room temperature for 5 hours. The reaction mixture is neutralized with sodium carbonate and extracted with ethyl acetate. Then, the procedure of Reference Example 12 is repeated and 0.5 g of ethyl 2-aminothiazol-4-yl glyoxylate is obtained. Based on NMR and other data, this product is identified with the product obtained in Example 12.

Example 14 In 20 ml of etharole containing 10% HCl, 1.2 g of methyl nitrone of 2- (2-aminothiazol-4-yl) -2- (syn) hydroxyiminoacetate, i.e. N- (2-Aminothiazol-4-yl-ethoxycarbonyl) -methylene methylamine-N-oxide, m.p. 111.6 °, and the suspension is stirred at room temperature for 16 hours. Then, the procedure of Reference Example 12 is repeated and 0.7 g of ethyl 2-aminothiazol-4-yl glyoxylate are obtained as yellow crystals.

In terms of NMR and other properties, this product is identical to the product of Example 12.

Example 15

To a mixture of 10 ml of tetrahydrofuran and 5 ml of ethyl acetate is added 1 g of ethyl 2- (2-aminothiazol-4-yl) -2- (anti) hydroxyiminoacetate, m.p. 145.3 ° C, followed by the addition of an excess of diazomethane-ether solution. The mixture is left at room temperature for 2 days. After the residual diazomethane is decomposed with acetic acid, the reaction mixture is concentrated under reduced pressure and the residue is recrystallized from ethyl acetate. In the above process, 0.1 µg of methylnitron compound is obtained, i.e. N- (2-aminothiazol-4-yl-ethoxycarbonyl) -methylene methylamine-N-oxide, as yellow crystals melting at 184-185 ° C.

Elemental analysis for CgH-jNNO₂S:

Calculated: C, 41.91; H, 4.64; N, 13.33

Found: C, 41.56; H, 4.75; N, 18.35

DK 161645 B

+ NMR Spectrum (60 MHz, in CDCl 3): 3.32 ppm (3H, s-, N-CH 2), 5.27 ppm (2H, br s, NH 2), 3.49 ppm (1H , s., thiazole 5H).

Example 16

To a solution of 23 mg of sodium in 3 ml of methanol is added 215 mg of ethyl 2- (2-aminothiazol-4-yl) -2- (anti) hydroxyiminoacetate, m.p. 145.3 ° C and at room temperature 230 mg of methyl iodide are added. . The mixture is stirred for 45 minutes, then concentrated under reduced pressure. The residue is diluted with water (pH 7 or higher) and extracted with ethyl acetate. The ethyl acetate layer is washed with water, dried and concentrated. The residue is recrystallized from tetrahydrofuran-ethyl acetate. In the above process, 160 mg of methyl nitrone compound are obtained as yellow crystals. This product is completely identical to the product obtained in Example 15.

Example 17

The filtrate after collecting the precipitated ethyl 2- (2-aminothiazol-4-yl) -2- (syn) methoxyiminoacetate from the concentrated reaction mixture in the procedure of Reference Example 4 is extracted with tetrahydrofuran-ethyl acetate (1: 1) and the extract is washed with water, dried and concentrated. To the residual tan oil is added 20 ml of tetrahydrofuran and the mixture is refrigerated for 16 hours.

The resulting crystals are collected by filtration and recrystallized from ethyl acetate. By the above process, 1.3 S of methyl nitrone of ethyl 2- (2-aminothiazol-4-yl) -2- (syn) hydroxyimino acetate, i.e. N- (2-aminothiazol-4-yl-ethoxycarbonyl) -methylene-methylamine-N-oxide, as yellow crystals melting at 111.6 ° C.

*

Elemental Analysis for CgH- ^N NO₂S;

Calculated: C, 41.91; H, 4.34; N, 13.33

Found: G 41.39 H 4.91 N 13.44 NMR Spectrum (60 MHz, in CDCl3): 4.14 ppm (3H, s., Ft-CH 2), 5.34 ppm (2H, br. s., NH 2), 6.62 ppm (1H, s., thiazole 5H).

Example 18

26 DK 161645 B

In 10 ml of tetrahydrofuran dissolve 1.5 g of ethyl 4-bromo-3-oxo-2-methoxy-iminobutyrate and after adding 7 ml of water, an additional 2.4 g of sodium acetate trihydrate and 0.9 g of thiourea are added. The mixture is stirred at room temperature for 17 hours, then concentrated under reduced pressure. The concentrate is adjusted to pH approx. 1.5 with dilute hydrochloric acid and wash with ethyl acetate. The aqueous layer is neutralized with sodium bicarbonate and extracted with ethyl acetate. The ethyl acetate layer is washed with water, dried and concentrated under reduced pressure to give 0.1 g of yellow crystals. This product is ethyl 2- (2-aminothiazol-4-yl) -2- (syn) methoxyiminoacetate. Based on NMR and other data, this product is identified with the syn isomer obtained in Example 2.

Example 19 Dissolve 2 g of ethyl 4-bromo-3-oxo-2-methoxy-iminobutyrate in 10 ml of dimethylformamide, after which 1.2 g of thiourea is added. The mixture is reacted at room temperature for 5 hours. To the reaction mixture is added 20 ml of a saturated aqueous solution of sodium chloride and the pH of the mixture is then adjusted to approx. 1.5 with dilute hydrochloric acid. Then, the procedure of Reference Example 1 # is followed and 1.1 g of pale yellow crystals is obtained. Based on 'NMR and other data, this product is identified as a & 7:13 mixture of syn and anti-isomers of ethyl 2- (2-aminothiazol-4-yl) -2-methoxy-iminoacetate. Washing the product with a small amount of ether gives the syn isomer substantially free of the anti-isomer.

Example 20

To 200 ml of water are added 3 g of sodium nitrite and 53 g of methyl acetoacetate, and under ice-cooling and stirring are added dropwise over approx. one hour 200 ml of 4 N sulfuric acid. During this dropwise addition, the temperature of the reaction mixture is kept at 5 ° C. The mixture is stirred within this temperature range for another 2 1/2 hours, then extracted twice with 300 ml portions of ethyl acetate. The extracts are combined and washed twice with a saturated aqueous solution of sodium chloride. Then a solution of 96.7 g of sodium carbonate is divided into 1 27

DK 161645 B

liter of water in three equal portions, by which 3-oxo-2-hydroxyimino-butyrate is extracted from the above ethyl acetate layer (three times).

To the aqueous layer (1 liter) is added 200 ml of methanol and, after cooling with ice, 150 g of dimethyl sulfate are added dropwise with stirring over 10 minutes. After drip is completed, the mixture is stirred at room temperature for 1 1/2 hours and extracted twice with 300 ml portions of ethyl acetate. The extracts are collected, washed with water and dried. The ethyl acetate is then distilled off and the residue is cooled with ice and then solidified. The solid residue is collected by filtration and washed with a small amount of water. In the above process, 52.3 g of methyl 3-oxo-2-methoxyiminobutyrate are obtained as white crystals melting at 64.4 ° C.

Elemental Analysis for C ^H ^ NONO:: Calculated: C, 45.23; H, 5.70; N, S0

Found: C 44.93 H 5.6 N 3, .71 NMR Spectrum (60 MHz, in CDCl3): 2.40 ppm (3H, singlet, -C-CH ~), in π 3 o 3, δ 6 ppm (3H, s., C00CH3), 4.10 ppm (3H, s., ^ NOCH ^).

Example 21 In 150 ml of chloroform 40 g of methyl 3-oxo-2-methoxyiminobutyrate are dissolved and the solution is heated to 40 ° C. Then a solution of 50 ml of chloroform is added dropwise over one hour. Then, the reaction is continued under stirring at room temperature for one hour. The reaction mixture is washed with a 5 µl aqueous solution of sodium hydrogen carbonate and water in the above order and the organic layer is dried. The solvent is then distilled off and 52.1 g of methyl 4-bromo-3-oxo-2-methoxyiminobutyrate is obtained as an oil.

NMR Spectrum (60 MHz, in CDCl 3): 3, & 2 ppm (3H, s., COOCH 3), 4.09 ppm (3H, s., = N-OCH 3), 4.27 ppm (2H, s ., BrCHgCO).

Dissolve 52 g of methyl 4-bromo-3-oxo-2-methoxy-iminobutyrate in 350 ml of tetrahydrofuran, then add 250 ml of water and further

DK 161645 B

28 & 9.1 g of sodium acetate trihydrate and 33.2 g of thiourea. The mixture is stirred at room temperature for 1 hour. To the reaction mixture is added 200 ml of a 5 $ Ts aqueous solution of sodium bicarbonate, then extracted with ethyl acetate. The organic layer is washed with water, dried and concentrated under reduced pressure to remove the solvent. To the concentrate is added 200 ml of ether and the resulting precipitate is collected by filtration. In the above process, 24 µg of methyl 2- (2-aminothiazol-4-yl) -2- (syn] methoxyiminoacetate are obtained as crystals melting at 164.9 ° C.

Elemental Analysis for CyH ^N ^O₂S:

Calculated: C, 39.06; H, 4.21; N, 19.52

Found: C 3 ^, 73H, 4.15 N, 19.33 NMR Spectrum (60 MHz, in CDCl:): 3 -, 4 ppm (3H, s, COOCH ^), 4.02 ppm (3H s., = NOHH ^), 5.74 ppm (2H, br s., NH₂), 6.74 ppm (1H, s., thiazole 5H).

Example 22 In 90 ml of Ν, Ν-dimethylacetamide, dissolve 21.5 g of methyl 2- (2-aminothiazol-4-yl) -2- (syn) -methoxyiminoacetate, and 13.6 g of chloroacetyl chloride are added dropwise under ice-cooling. The mixture is stirred under ice-cooling for 30 minutes and then at room temperature for 30 minutes. After adding 500 ml of water, the reaction mixture is extracted twice with ethyl acetate. The extracts are collected, washed with a 5 $ aqueous solution of sodium bicarbonate and water in that order, and dried. The solvent is distilled off and 25 g of methyl 2- (2-chloroacetamidothiazol-4-yl) -2- (syn) methoxyiminoacetate are obtained as crystals melting at 130 ° C.

Elemental analysis for C ^H ^N ^O₂SC1:

Calculated: C, 37.03; H, 3.45; N, 14.40

Found: G 37.30 H 3.40 - N 14.33 NMR Spectrum (60 MHz, in CDCl3): 3.90 ppm (3H, s., COOCH , = NOCH 3), 4.26 ppm (2H, s., ClCl 2 CO), 7.24 ppm (1H, s., Thiazole 5H).

Example 23

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29

To a solution of 19.2 g of potassium hydroxide in a mixture of 170 ml of water and 900 ml of ethanol is added 20 g of methyl 2- (2-chloroacetamidothiazol-4-yl) -2- (syn) methoxyiminoacetate and the solution is stirred at room temperature. for 2 hours. The ethanol is distilled off under reduced pressure and after the addition of 170 ml of water, the residue is washed with 200 ml of ethyl acetate. The aqueous layer is adjusted to pH 2 with 10 2 hydrochloric acid and extracted twice with 300 ml portions of ethyl acetate.

The extracts are collected, washed with a saturated aqueous solution of sodium chloride and dried. The solvent is distilled off and 16 µg of 2- (2-chloroacetamidothiazol-4-yl) -2- (syn) -methoxyiminoacetic acid is obtained as crystals. In terms of NMR spectrum and other properties, this product is found to be identical to the product obtained in Example 6.

Example 24 (1) 6 g of 7- (5-carboxy-5-benzamidovalerylamido) -3- (N-chloroacetyl) -carbamoyloxymethyl-3-cephem-4-carboxylic acid are suspended in 60 ml of methylene dichloride containing 7.6 ml of N, N- dimethylaniline and 2.25 ml of phosphorus trichloride are added under cooling at -50 ° C. The mixture is stirred at -30 ° G for 1/2 hour until a clear solution is obtained. To this solution is added 4.17 g of phosphorus pentachloride and the mixture is stirred at -25 ° C for 2 1/2 hours. It is then cooled to -40 ° C and 37 ml of cold methanol are added quickly. The mixture is stirred at -5 ° C for 25 minutes, then diluted with 22 ml of water and adjusted to pH 3.5 with dilute aqueous ammonia solution. The reaction mixture is left at 5 ° C for one hour and the resulting precipitate is collected by filtration. By the above process 1.76 g of 7-amino-3- (N-chloroacetyl) -carbamoyloxymethyl-3-cephem-4-carboxylic acid is obtained as a colorless crystalline product.

Elemental analysis for 01 ^ Η1261Ν ^ 0 ^ 3:

Calculated: G 37.76 H 3.46 N 12.01

Found: C 36.02 H 3.66 N 11, dl 30

DK 161645B

NMR Spectrum (60 MHz, in CF 3 COOH): 3.76 ppm (2H, broad s. 2-CH 2), 4.35 ppm (2H, s., -NHCOCH 2 Cl), 5.42 ppm (2H, broad s., 6H, 7H), 5.46 ppm (2H, quartet, -CH2COONH).

(2) In Ν, Ν-dimethylacetamide dissolves 1, -05. g of the 7-amino-3- (N-chloro-acetyl) -carbaraoyloxymethyl-3-cephem-4-carboxylic acid obtained above (1) and 996 mg of 2-chloroacetamido-thiazole-4 are added under ice-cooling. yl-a- (anti) -methoxyiminoacetylchlorid hydrochloride. The mixture is stirred under ice-cooling for 15 minutes and then at room temperature for 2 hours. After the addition of 50 ml of water, the reaction mixture is extracted twice with 100 ml portions of ethyl acetate. The organic layers are combined, washed with a saturated aqueous solution of sodium chloride and dried over magnesium sulfate. The ethyl acetate is then distilled off. In the above process, 2.2 g of 7-C (2-chloroacetamidothiazol-4-yl) -α- (anti) -methoxyimino] -acetamido-3- (N-chloroacetyl) -carbamoyloxymethyl-3-cephem-4 are obtained. carboxylic acid as a white powder.

Note: Preparation of 2-chloroacetamidothiazol-4-yl-α- (anti) -methoxy-iminoacetyl chloride.

(i) In 100 ml of dimethylacetamide 10 g of ethyl α- (anti) -methoxy-imino-α- (2-aminothiazol-4-yl) acetate is dissolved and 5.91 g of chloroacetylene hydrochloride are added dropwise with cooling. chloride. B-land is not stirred at room temperature for one hour and then poured into ice-water. The mixture is extracted with ethyl acetate and the organic layer is washed, dried and distilled to remove the solvent. The method is obtained. 12.66 g of ethyl α- (anti) -methoxyimino-α- [2- (chloroacetamido) -thiazol-4-yl] -acetate as crystals, melting point & 1-B2 ° C.

Elemental analysis for c10H12N3 ° 4SC1:

Calculated: G 39.29 H 3.96

Found: C, 36.74; H, 3.56

The nuclear magnetic resonance spectrum (60 MHz, in CDCl 3) of this product gives singlets, one at 4.1G ppm, attributable to methoxy protons, one at 4.24 ppm, which. can be attributed to chloroacetyl protons, and one at 7.94 ppm, which can. is attributed to thiazole-5-hydrogen.

31

DK 161645 B

(ii) 12.66 g of ethyl α- (anti) methoxyimino-α- [2- (2-chloroacetamido) thiazol-4-yl] acetate are added to a solution of 11.74 g of potassium hydroxide in a mixture of 25 ml of water and 500 ml of ethanol. The mixture is stirred at room temperature for 20 minutes and the ethanol is distilled off under reduced pressure and the residue is diluted with water. The mixture is acidified with 1N hydrochloric acid and the resulting precipitate is collected by filtration. By the above process, 10.54 g of α- (anti) -methoxyimino-α- [2- (chloroacetamido) -thiazol-4-yl-acetic acid, m.p. 162-163 ° C, is obtained.

Elemental Analysis for CgHgN2 O ^ SCl: t

Calculated: C 34.60 H 2.90 N 15.13

Found: C, 34.53; H, 3.00; N, 14.60

The nuclear magnetic resonance spectrum (60 MHz, in d 2 -DMSO) of the above product shows singlets attributable to methoxy protons at 400 ppm, chloroacetyl protons at 4.36 ppm and thiazole-5-hydrogen at 6.00 ppm.

(iii) In 5 ml of methylene chloride, 555.4 mg of α- (anti) -methoxyimino-α- [2- (chloroacetamido) -thiazol-4-yl3-acetic acid are suspended and 416.3 mg of phosphorus pentachloride are added under ice-cooling. The mixture is reacted with stirring for 30 minutes. To the reaction mixture is added n-hexane and the resulting precipitate is collected by filtration. By the above process 620 mg of α- (anti) -methoxyimino-α- [2- (chloroacetamido) -thiazol-4-yl-acetyl chloride hydrochloride are obtained.

Elemental analysis for CgHyN ^O₂SC ^ HCl:

Calculated: C, 26.69; H, 2.42; N, 12.63

Found: C 26.35 H 2.6L N 12.00 (3) In 50 ml of tetrahydrofuran dissolve 2.2 g of the 7 - [(2-chloroacetamido-thiazol-4-yl) -α (anti) - methoxyimino] -acetamido-3- (N-chloroacetyl) -carbamoyloxymethyl-3-cephem-4-carboxylic acid obtained above (2), then 913 mg of finely powdered thiourea and 1.63 g of sodium acetate trihydrate are added. The mixture is stirred at room temperature for 17 hours. The precipitate is collected by filtration, washed with ethyl acetate.

DK 161645 B

32 ether and dissolve in 10 ml of water. The solution is adjusted to pH 7 with sodium bicarbonate and passed through a column of Amberlite XA-D-2. By the above procedure, 360 mg of sodium 7-C (2-aminothiazol-4-yl) -a- (anti) methoxyimino] -acetamido-3-carbamoyloxy-methyl-3-cephem-4-carboxylate is obtained as a white powder. .

Elemental analysis for C - ^^H HN ^O₂S₂Na-2.5H₂O:

Calculated: C 34.42 H 3.85 N 16.05

Found: C 34.43 H 3.70 N 15.68 NMR Spectrum (60 MHz, in D 2 O): 3.55 ppm (2H, quartet, 2-CB 2), 4.11 ppm (3H, s., = NOCH 3), 4.61 ppm (2H, quartet, -Cf ^ OCON 2), 5.21 ppm (1H, doublet, 6h), 5.82 ppm (1H, doublet, 7H), 7.55 ppm ( IH, s., HQN and H).

γγ N – 4

The antibacterial activity (MIC (T / ml)) of sodium 7-C (2-aminothiazol-4-yl) -a- (anti) methoxyimino] -acetamido-3-carbamoyloxymethyl-3-cephem-4 the carboxylate of this example is shown below.

33

DK 161645 B

Microorganism MIGfY / ml)

Escherichia coli 0-111 0.73

Klebsiella pneumoniae DT 1.56

Klebsiella pneumoniae GN 3B35 6.25

Serratia marcescens IFO 12643 12.5

Serratia TN0024 6.25

Proteus vulgaris IFO 3933 0.39

Proteus mirabilis GN 4359 0.73

Proteus morganii IFO 3163 0.73

Proteus justice 3 TNO 336 0.2

Proteus justice GN 4733 0.73

Enterobacter cloacae IFO 12937 25

Citrobacter freundii GN 99 1.56

Citrobacter freundii GN 1706 3.13

Example 25 (1) In 20 ml of Ν, Ν-dimethylacetamide dissolve 1.05 g of 7 "amino-3- (N-chloro-acetyl) -carbamoyloxymethyl-3-cephem-4-carboxylic acid and add under ice-cooling 369 mg 2 -chloracetamidothiazol-4-ylacetyl chloride hydrochloride. The mixture is stirred under ice-cooling for 15 minutes and then at room temperature for 2 hours. After this reaction, the mixture is diluted with 50 ml of water and extracted twice with 100 ml portions of ethyl acetate. The organic layers are combined, washed with a saturated aqueous solution of sodium chloride and dried over magnesium sulfate. The ethyl acetate is then distilled off and a white powder of 7- (2-chloroacetamidothiazol-4-yl) -acetamido-3- (N-chloroacetyl) -carbamoyloxymethyl-3-cephem-4-carboxylic acid is obtained. Yield: 1.6 g.

Note: Preparation of 2-chloroacetamidothiazol-4-ylacetyl chloride.

Dissolve 4 g of ethyl 2-aminothiazol-4-yl acetate in 15 ml of dimethyl acetamide and add 3.62 g of chloroacetyl chloride dropwise under ice-cooling. The mixture is stirred under ice-cooling for 30 minutes and then at room temperature for another 30 minutes. Then 50 ml of water is added and the mixture is extracted twice with 100 ml portions of ethyl acetate-tetrahydrofuran. The extract is washed with 100 ml of an aqueous solution of sodium bicarbonate and then with 100 ml of a saturated aqueous solution of sodium chloride, then dried. The solvent is then distilled off. By the above process, 2.95 g of ethyl 2-chloroethamidothiazol-4-ylacetate is obtained as an oily product. The entire amount of this oil is suspended in 100 ml of methanol and 12 ml of water containing 761 mg of sodium hydroxide are added under ice-cooling. The mixture is stirred at room temperature for 30 j minutes, after which a greater amount of the methanol is distilled off.

that reduced pressure. To the residue is added 10 ml of water. The watery I

The layers are washed with 50 ml of ethyl acetate and after addition of 20 ml of ethyl acetate it is adjusted to pH 2 with 10 $ * s of hydrochloric acid. The mixture | Shake well and the organic layer is separated, washed with a saturated aqueous solution of sodium chloride and dried. The solvent is then distilled off. The above method is obtained; 1.51 g of 2-chloroacetamidothiazol-4-yl acetic acid as colorless crystals, m.p .: 202-203 ° C. |

Elemental Analysis for C ^ KyClN ^O₂S:

C, 35.83; H, 3.01; N, 11.94

Found: C 36.01 H 2.96 N 11.6L In 20 ml of methylene dichloride, 93 # rag of the above product is suspended and 1 g of phosphorus pentachloride is added under ice-cooling. The mixture is stirred at room temperature for 30 minutes. After the addition of 50 ml of petroleum ether, the precipitate is collected by filtration and washed with 10 ml of petroleum ether. By the above process 1.06 g of 2-chloroacetamidothiazol-4-ylacetyl chloride hydrochloride are obtained as colorless crystals.

Elemental analysis for 0

Calculated: C, 29.04; H, 2.44; N, 9.67

Found: C 28.96 H 2.24 N 9.61 IR Spectrum (KBr): 1780 cm 2 (-CO1).

(2) In 40 ml of tetrahydrofuran dissolve 1.6 g of the 7- (2-chloroacetamido-

DK 161645 B

thiazol-4-yl) -acetamido-3- (N-chloroacetyl) -carbamoyloxymethyl-3-cephem-4-carboxylic acid, obtained in the above (1). To this solution is added S60 mg of thiourea, then sodium acetate trihydrate is added. The mixture is stirred at room temperature for 16 hours. The precipitate is collected by filtration, washed with ethyl ether and dissolved in 10 ml of water. The solution is adjusted to pH 7 with sodium hydrogen carbonate and purified by column chromatography on Amberlite XAD-2. By the above procedure, 152 mg of sodium 7- (2-amino-thiazol-4-yl) -acetamido-3-carbamoyloxymethyl-3-cephem-4-carboxylate is obtained as a white powder.

Elemental analysis for C ^H ^N NO₂S₂Na * 2H₂O:

Calculated: C 35.67 H 3, 5 N 14, $ 5

Found: C 35.97 H 3.88 N 14.64 NMR Spectrum (00 MHz, in D 2 O): 3.52 ppm {2H, quartet, 2-CH 2), 3.61 ppm (2H, s 13 --- --- 0Η CO- -4.7 # ppm (2H, quartet, .- CH2OCONH-), 5.14 ppm -2 g (1H, doublet, 6H), 5.6 $ ppm (1H, "doublet , 7H), 6.52 ppm (1H, s., N / N_

Example 26 (1) In 6 ml of Ν, Ν-dimethylacetamide, 290 mg of 7-amino-3- (N-chloro-acetyl) -carbamoyloxymethyl-3-cephem-4-carboxylic acid is dissolved and 276 mg of 2- {ice-cooling) is added. 2-chloracetamidothiazol-4-yl) -2- (syn) -methoxyiminoacetylchlorid hydrochloride. The mixture is stirred under ice-cooling for 15 minutes and at room temperature for 2 hours. Then, the reaction mixture is diluted with 30 ml of water and extracted twice with 50 ml portions of ethyl acetate. The extracts are collected, washed with 50 ml of a saturated aqueous solution of sodium chloride and dried over anhydrous magnesium sulfate. The ethyl acetate is distilled off and 402 mg of 7-C 2- (2-chloroacetamidothiazol-4-yl) -2- (syn) methoxyimino] -acetamido-3- (N-chloroacetyl) -carbamoyloxymethyl-3-cephem-4 are obtained. -carboxylic acid as a viscous oil.

NMR Spectrum (60 MHz, in CDCl3): 3.50 ppm (2H, quartet, 2-CH 2), 3.99 ppm (3H, s., NOCH +), 4.04 ppm, 4.30 ppm ( 2Hx2, s.x2, C1CH2C0x2), 5.10 ppm (1H, doublet, 6h), 5.73 ppm (1H, doublet, 7H), 7.32 ppm (1H, s., Thiazole, 5H).

"DK 161645 B

(6) The entire amount of the above product is dissolved in 9 ml of tetrahydrofuran, after which 163 mg of thiourea and 3OO mg of sodium acetate trihydrate are added. The mixture is stirred at room temperature for 4 hours. The precipitate is collected by filtration, washed with ether and dissolved in 5 ml of water. The solution is adjusted to pH about 7 with sodium hydrogen carbonate and purified by column chromatography on Amberlite XAD-2. By the above process 53 mg of sodium 7- [2- (2-amino-thiazol-4-yl) -2- (syn) -methoxyiminoacetamido-3-carbamoyloxymethyl-3-cephem-4-carboxylate is obtained as a white powder. .

Elemental Analysis for C - ^^H H NN ^OSSgNa ^H ^O:

Calculated: C 33, 4H, 3.9, N, 15.7 #

Found: C 33.94 H 3, $ 2 N 15.42 NMR Spectrum (60 MHz, in D 2 O): 3.47 ppm (2H, quartet, 2- € H2), 3.92 ppm (3H, s., = NOCH 3), 4.63 ppm (2H, quartet, -CH2OGONH2), 5.27 ppm (1H, doublet, 6H), 5.72 ppm (1H, doublet, 7H), 6.95 ppm (1H, s , thiazole 5H).

Process for the preparation of 2- (2-chloroacetamidothiazol-4-yl) -2- (syn) -methoxyiminoacetyl chloride.

In 5 ml of methylene chloride, 273 mg of the 2- (2-chloroacetamido-thiazol-4-yl) -2- (syn) -methoxyiminoacetic acid obtained in Reference Example 6 is suspended and 203 mg of phosphorus-pentachloride is added under ice-cooling. The mixture is stirred at room temperature for 30 minutes, then washed with petroleum ether. In the above process, 276 mg of 2- (2-chloroacetamidothiazol-4-yl) -2- (syn) methoxyiminoacetyl chloride is obtained as a powder.

Elemental Analysis for CgHyN2O2 SC1 ^ HCl:

Calculated: C, 23.39; H, 2.42; N, 12.63

Found: C, 23.47; H, 2.73; N, 12.12

37 DK 161645 B

Example 27 (1) In 22 ml of dry tetrahydrofuran, dissolve 500 mg of 2- (2-chloroacetamido-thiazol-4-yl) -2- (syn) -methoxyiminoacetic acid and add, with stirring, 1 $ 2 mg of triethylamine. This mixture is cooled to -10 ° C and there is added -s drop of s 245 mg of isobutyl chloroformate. The mixture is stirred at this temperature for 2 hours. To the resulting solution of mixed anhydride is added 1 & 2 mg of triethylamine with a solution (ice-cooled) of 590 mg of 7-amino-3- (1-methyl-1H-tetrazol-5-yl) -thiomethyl-3-cephem-4-carboxylic acid in 1 & ml of a 50% 1s aqueous tetrahydrofuran solution. The mixture is stirred under ice-cooling for one hour and at room temperature for 2 hours. Then, the majority of the tetrahydrofuran is distilled off under reduced pressure and the residue is diluted with 100 ml of water and with 40 ml of ethyl acetate. With stirring, the aqueous layer is then adjusted to pH 2 with 1 N HCl. The phases are separated and the aqueous layer is extracted with 60 ml of ethyl acetate. The ethyl acetate layers are collected, washed with 50 ml of a saturated aqueous solution of sodium chloride and dried. The ethyl acetate is distilled off, and 700 mg of 7- {2- (2-chloroacetamidothiazol-4-yl) -2- (syn) -methoxyimino-ac etamido] -3- (1-methyl-1H-tetrazole-4-yl) is obtained. yl) -thiomethyl-3-cephem-4-carboxylic acid as a viscous oil.

(2) The whole of the above product is dissolved in 15 ml of tetrahydrofuran, after which 226 mg of thiourea and 406 mg of sodium acetate trihydrate are added. The mixture is stirred at room temperature for 4 hours. After this reaction, the precipitate is collected by filtration, washed with ether and dissolved in 10 ml of water. The solution is adjusted to pH approx. 7.0 with sodium bicarbonate and purified by column chromatography on Amberlite XAD-2. By the above procedure 125 mg of sodium 7- [2- (2-aminothiazol-4-yl) -2- (syn) methoxyiminoacetamido] -3- (1-methyl-1H-tetrazol-4-yl) is obtained -thiomethyl-3-cephem-4-carboxylate as a white powder.

Elemental Analysis for C ^H- ^N ^O ^S ^Na ^^O:

Calculated: C 33.74 H 3.54 N 22.13

Found: C 34, Id H 3.57 N 21.79 NMR Spectrum (60 MHz, in D 2 O): 3.59 ppm (2H, quartet, 2-CH 2), 3.93 ppm

38 DK 161645 B

(3H, s., = NOCH ^), 3.9 $ ppm (3H, s., N-CH 2), 4.0 $ ppm (2H, quartet, 3-0¾), 5.12 ppm (1H, doublet, one), 5.72 ppm (1H, doublet, 7H), 6.93 PPm (1H, s., thiazole 5H).

Example 28 (1) In 15 ml of Ν, Ν-dimethylacetamide, 762 mg of 7-aminocephalosporanoic acid is dissolved and 931 mg of 2- (2-chloroacetamido-thiazol-4-yl) -2-methoxyiminoacetyl chloride hydrochloride is added under ice cooling. of syn-isomer). The mixture is stirred under ice-cooling for 15 minutes and at room temperature for 2 hours. The reaction mixture is diluted with 10 ml of water and extracted with 100 ml portions of ethyl acetate. The extracts are collected, washed with 100 ml of a saturated aqueous solution of sodium chloride and dried. The ethyl acetate is distilled off and 1.4 g of 7- [2- (2-chloroacetamidothiazol-4-yl) -2- (syn) methoxyiminoacetamido] cephalosporanoic acid is obtained as an oil.

(2) In 30 ml of tetrahydrofuran, dissolve the entire amount of the above product, then add 500 mg of thiourea and then $ 95 mg of sodium acetate trihydrate. The mixture is stirred at room temperature for 4 hours. The resulting precipitate is collected by filtration, washed with ether and dissolved in 6 ml of water. The solution is adjusted to pH approx. 7.0 with sodium bicarbonate and purified by column chromatography on Amberlite XAD-2. By the above procedure, 7 $ mg of sodium 7- [2- (2-aminothiazol-4-yl) -2- (syn) methoxyiminoacetamido] cephalosporanate is obtained as a white powder.

Elemental Analysis for C-H HN ^OSSgNa ^, 5H₂O:

Calculated: C 36.7 $ H 4.05 N 13.40 'Found: 036.93 Η 3, & 0 N 12.6 $ NMR Spectrum (60 MHz, in D20): 2.07 ppm (3H, s. , 000¾), 3.53 ppm (2H, quartet, 2-0¾), 3.9 $ ppm (3H, s., = 1100¾), 4.75 ppm (2H, quartet, 3-0¾), 5 , 21 ppm (1H, doublet, 6H), 5, $ 1 ppm (1H, doublet, 7H), 7.01 ppm (1H, s., Thiazole 5H).

39 DK 161645 B

Example 29

To 10 ml of water is added 1 g of the sodium 7- [2- (2-aminothiazol-4-yl) -2- (syn) -methoxyimine acetamido] cephalosporanate obtained in Example 3, 270 mg 1,3,4-oxadiazole-5-thiol potassium salt and 7 mg triethylbenzylammonium bromide. The mixture is stirred under a nitrogen gas stream at 60 ° C for 6 hours. After cooling, the reaction mixture is purified by column chromatography on Amberlite XAD-2. By the above process, 110 mg of sodium 7- [2- (2-aminothiazol-4-yl) -2- (syn) methoxyiminoacetamido] -3- (2-methyl-1,3,4-oxadiazole-5) is obtained. yl) -thiomethyl-3-cephem-4-carboxylate as a white powder.

Elemental analysis for C ^ yH Hl6NN °6S3Na * 2H₂O

Calculated: C 35, β5 H 3.54 N 17.21

Found: C 35.73 H 3.72 N 17.01 NMR Spectrum (60 MHz, in DgO): δ> 4-2 ppm (3H, s., Oxadiazole 2-CH 2), 3.55 ppm (2H , quartet, 2-0 +), 4.02 ppm (3H, s, = NOCH 2), 5.13 ppm (1H, doublet, 6H), 5.73 ppm (1H, doublet, 7H), 6, 97 ppm (1H, s-, thiazole 5H).

Example 3.0 (1) To 10 ml of tetrahydrofuran are added 633 mg of 2- (2-chloroacetamido-thiazol-4-yl) -2- (syn) -methoxyiminoacetic acid, 3 & 0 mg of N-hydroxysuccin imide and 630 mg of dicyclohexylcarbodiimide, and the mixture is stirred at room temperature for 45 minutes. The precipitate is filtered off and the filtrate is cooled to 5 ° C. It is then added to a mixed solution of 650 mg of 7-aminodesacetoxy-cephalosporanoic acid and 2 ml of bis- (trimethylsilyl) -acetamide in methylene chloride, which has been cooled previously. The mixture is stirred at room temperature for 16 hours and then the solvent is distilled off under reduced pressure. To the resulting oil is added 50 ml of water with 50 ml of ethyl acetate and the mixture is adjusted to pH approx. 2.5 with 1N hydrochloric acid. The two layers are separated followed by extractions with two 50 ml portions of ethyl acetate. The ethyl acetate layers are collected, washed with water and dried. The ethyl acetate is then distilled off and 1.1 g of 7- [2- (2-chloroacetamidothiazol-4-yl) -2- (syn) -methoxyiminoacetamido] -desacetoxycephalosporanoic acid is obtained as an oil.

40 DK 16 1645 B

(2) The entire amount of the above product is dissolved in 25 ml of tetrahydrofuran, after which thiourea is added and then 632 mg of sodium acetate trihydrate. The mixture is stirred at room temperature for 4 hours. The precipitate is collected by filtration, washed with ether and dissolved in 10 ml of water. The solution is adjusted to pH approx. 7.0 with sodium bicarbonate and purified by column chromatography on Amberlite XAD-2. By the above procedure 120 mg of sodium 7- [2- (2-aminothiazol-A-yl) -2- (syn) -methoxyiminoacetamidol-desacetoxy-cephalosporanate is obtained as a white powder.

Elemental Analysis for C ^H ^N ^O ^S ^Na ·! 5 ^0:

Calculated: C 37.67 H 3.64 N $ 15.6

Found: C 37.37 H 3.9¾ N 15.3 $ NMR Spectrum (60 MHz, in DgO) δ: 1.94 ppm (3H, s, 3-6H3K 3.46 ppm (2H, quartet, 2 -0.00), 4.00 ppm (3H, s., ^ NOCH ^), 5.17 ppm (1H, doublet, 6H), 5.76 ppm (1H, doublet, 7H), 6.99 ppm - ( 1H, s., Thiazole 5H).

The minimum inhibitory concentrations (g / ml) of some of the compounds of the above examples are as follows:

Microorganisms Compound from Example 1 from Example 3 from Example 2 from Example 5 E. coli NIHJ 0.10 0.20 0.10 0.76 E. coli 0-111 0.024 0.05 0.024 0.39 E. coli T-7 0.39 0.76 0.76 6.25 K. pneumonia DT <0.012 0.024 0.024 0.20 K. pneumonia GN 3635 0.05 0.05 0.20 0.20 'Ps. aeruginosa Pd 1 50 25 12.5> 100

Ps. aeruginosa PM 3 3.13 1.56 0.76 25

Ps. aeruginosa P2 25 50 50> 100

Ps. aeruginosa GN 3407> 100 50 50> 100

Serr. marcescens IFO 12643 1.56 3.13 0.76 12.5

Serratia TN 0024 0.20 0.76 0.20 1.56 P. vulgaris IFO 3966 <0.02 0.024 0.024 0.20

41 DK 161645 B

from ex.l from ex.3 from ex.2 from ex 5 P. vulgaris GN 4413 1.56 0.7 # 0.39 1.56 P. rairabilis GN 4359 = 0.02 0.05 0.10 0 , 10 P. morganii IFO 3165 0.39 0.20 0.05 12.5 P. trial 6 (TN0 336) = 0.012 = 0.012 = 0.012 = 0.012 P. trial 5 GN 4733 0.05 0.20 0.20 0.10

Ent. cloacae TN 1262 6.25 6.25 1.56 50

Cit. freundii GN 99 0.20 0.20 0.10 3.13

Cit. freundii GN 1706 0.39 0.39 0.20 6.25

Acinetobacter anitratus 6.25 25 25 12.5 TN 1140

Note: The following abbreviations are used to denote the microorganisms used: E: Escherichia K: Klebsiella Ps: Pseudomonas

Serr: Serratia P: Proteus Ent: Enterobacter

Cit: Citrobacter

Example 31 (1) In 20 ml of dry tetrahydrofuran, dissolve 500 mg of 2- (2-chloroacetamido-thiazol-4-yl) -2- (syn) -methoxyiminoacetic acid and add 162 mg of triethylamine with stirring. The mixture is cooled to -10 ° C and then 245 mg of isobutyl chloroformate is added dropwise. The mixture is stirred at this temperature for 2 hours. To the resulting mixed anhydride solution is added a solution (ice-cooled) of 160 mg of triethylamine and 492 mg of 7-amino-3-carbamoyloxymethyl-3-cephem-4-carboxylic acid in a 50 µs aqueous solution. tetrahydrofuran. The mixture is stirred under ice-cooling for one hour and then at room temperature for 2 hours. Most of the tetrahydrofuran is distilled off under reduced pressure and 100 ml of water and 40 ml of ethyl acetate are added to the residue. The mixture is adjusted to pH approx. 2 with 1 N hydrochloric acid. The two layers are separated and the aqueous layer is extracted twice with 50 ml portions of ethyl acetate. The ethyl acetate layers are collected, washed with water, dried and concentrated. By the above procedure, 650 mg of 7- [2- (2-chloroacetamidothiazol-4-yl) -2- (syn) -methoxyiminoacetamido-3-carbamoyloxymethyl-3-cephem-4-carboxylic acid is obtained as an oil.

42

DK 161645 B

(2) The entire amount of the above product is dissolved in 15 ml of tetrahydrofuran, after which 226 mg of thiourea and 406 mg of sodium acetate trihydrate are added. The mixture is stirred at room temperature for 4 hours. The precipitate is collected by filtration, dissolved in 10 ml of water, adjusted to. pH value approx. 7 with sodium bicarbonate and purified by column chromatography on Amberlite XAD-2. By the above procedure 120 mg of sodium 7- [2- (2-aminothiazol-4-yl) ~ 2- (syn) methoxy-iminoacetamido] -3-carbamoyloxymethyl-3-cephem-4-carboxylate is obtained as a white powder. . In terms of NMR spectrum and other properties, the product is found to be identical to the product obtained in Example 26.

Example 32 (1) In 45 ml of dry tetrahydrofuran dissolve 1.11 g of 2- (2-chloroacetamido-thiazol-4-yl) -2- (syn) -methoxyiminoacetic acid and add, with stirring, 5 mg of tri-n-butylamine. The mixture is cooled to -10 ° C and 544 mg of isobutyl chloroformate is added dropwise. The mixture is stirred at -10 ° C for 2 hours, then a cold solution of 741 mg of tri-n-butylamine and 1.4 g of 7-amino-3- (N-chloroacetyl) carbamyloxymethyl 3-cephem-4-carboxylic acid in 40 ml of a 50 $ aqueous tetrahydrofuran solution. The mixture is stirred under ice-cooling for one hour and at room temperature for 2 hours. Most of the tetrahydrofuran is distilled off under reduced pressure and the residue is diluted with 25 ml of water and washed with 40 ml of ethyl acetate. The aqueous layer is separated and adjusted after the addition of 50 ml of ethyl acetate to pH approx. 2.5 with 1N hydrochloric acid. The mixture is separated into two layers. The aqueous layer is extracted two more times with 50 ml portions of ethyl acetate. The extracts are collected, washed with 1Ό0 ml of a saturated aqueous solution of sodium chloride, dried and concentrated. By. In the above process, 1 g of 7-C 2- (2-chloroacetamidothiazol-4-yl) -2- (syn) -methoxyiminoacetamido-3- (N-chloroacetyl) -carbamoyloxymethyl-3-cephem-4-carboxylic acid is obtained as an oil.

(2) The entire amount of the above product is dissolved in 22 ml of tetrahydrofuran, after which 499 mg of thiourea and then $ 92 mg of sodium acetate trihydrate are added. The mixture is stirred at room temperature for 4 hours. The precipitate is collected by filtration, washed with ether and dissolved in 10 ml of water. The solution is adjusted to pH approx. 7 with sodium bicarbonate and purified by column chromatography on

"DK 161645 B

43

Amberlite XAD-2. By the above procedure, 153 mg of sodium 7- [2- (2-aminothiazol-4-yl) -2- (syn) methoxyiminoacetamido] -3-carbamoyloxymethyl-3-cephem-4-carboxylate is obtained as a white powder. Based on NMR and other data, this product is identical to the compound obtained in Example 26.

Example 3-3 · (1) In 20 ml of tetrahydrofuran dissolve 277 mg of 2- (2-chloroacetamidothiazol-yl) -2- (syn) -methoxyiminoacetic acid and 270 mg of tert.butyl-7-aminodes-acetoxycephalosporanoic acid, after which 206 mg is added. dicyclohexyl carbodiimide. The mixture is reacted with stirring at room temperature for 6 hours. The precipitated urea derivative is filtered off and the filtrate is poured into 50 ml of water and extracted with ethyl acetate. The ethyl acetate layer is washed with 0.5 N hydrochloric acid, water and a saturated aqueous solution of sodium chloride in the order mentioned, dried and concentrated. By the above procedure, 320 mg of tert.butyl 7- [2- (2-chloro-acetamidothiazol-4-yl) -2- (syn) -methoxyiminoacetamido] -desacetoxy-cephalosporanate is obtained as an oil.

NMR Spectrum (60 MHz, in CDCl 3): 1.53 ppm (9H, s., T C 1 H 3), 2.13 ppm (3H, s., 3-CH 2), 3.39 ppm (2H , quartet, 2-CH 2), 4.06 ppm (3H, s, = NOCH 2), 4.29 ppm (2H, s-, CIC 2 CO), 5.06 ppm (1H, doublet, 6H), 5.66 ppm (1H, doublet of doublet, 7H), 7.20 ppm (1H, s., Thiazole 5H), 6.14 ppm (1H, doublet, 7-CONH).

(2) The whole of the above product is dissolved in 12 ml of tetrahydrofuran, after which 100 mg of thiourea and 200 mg of sodium acetate trihydrate are added. The mixture is stirred at room temperature for 6 hours. The reaction mixture is diluted with 30 ml of water and extracted with ethyl acetate. The ethyl acetate layer is washed with water, dried and concentrated. The resulting oil is purified by chromatography on silica gel. By the above procedure 126 mg of tert.butyl 7- [2- (2-amino-thiazol-4-yl) -2- (syn) -methoxyiminoacetamido] -desacetoxycephalosporate is obtained as a powder.

NMR Spectrum (60 MHz, in CDCl 3): 1.52 ppm (9H, s., TC 3 H 2), 2.10 ppm (3H, s., 3-CH 2), 3.40 ppm (2H , quartet, 2-CH 2), 4.00 ppm (3H, s., = NOCH 2), 5.05 ppm (1H, doublet, 6H), 5.96 ppm (1H, doublet of doublet,

«DK 161645 B

7H), 6.66 ppm (1H, s., Thiazole 5H), S, 2 $ ppm (1H, doublet, 7-CONH) - (3) The entire mixture of the above product is dissolved in a mixture of 1 ml of trifluoroacetic acid and 0.1 ml of anisole, and the solution is stirred at room temperature for 1 1/2 hours, then ether is added. The resulting precipitate is collected by filtration and washed with ether. By the above process, 70 mg of 7- [2- (2-aminothiazol-4-yl) -2- (syn) -methoxyiminoacetamido] -desacetoxycephalosporanoic acid trifluoroacetate is obtained as a powder.

According to NMR spectrum (60 MHz, in D 2 O containing NaHCO 3), this product is identical to the product obtained in Example 30.

Example 34

By acylating the 7-amino group in the corresponding cephalosporin compounds in a similar manner as described in Example 2 (Process A) and using sodium 7- [2- (2-aminothiazol-4-yl) -2- (syn) -methoxyiminoacetamido 3-cephalosporanate and heterocyclic thiol compounds similar to those described in Example 4 (Process B) prepare the following compounds: (a) Sodium 7- [2- (2-aminothiazol-4-yl) -2- (syn) - methoxyiminoacetamido] ~ 3- (2-methyl-1,3,4-thiadiazol-5-yl) -thiomethyl-3-cephem-4-carboxylate (process B).

NMR Spectrum (60 MHz, in D 2 O): 2.37 ppm (3H, s., Thiazole 2-GH +), 3.52 ppm (2H, quartet, 2-CH 2), 3.95 ppm (3H, s , = NOCH3), 5.1 # PPm (1H, s., 6H), 5.73 ppm (1H, s., 6H), 5.73 ppm (1H, s-, 7H), 6.95 ppm (1H, s., Thiazole 5H).

(b) Disodium 7- [2- (2-aminothiazol-4-yl) -2- (syn) -methoxyiminoacetamido 3-3- (2-carboxymethyl-1,3,4-thiadiazol-5-yl) - thiomethyl 3-cephem-4-carboxylate (process B).

NMR Spectrum (60 MHz, in D 2 O): 3.56 ppm (2H, quartet, 2-CH 2), 3.96 ppm (3H, s., = NOCH 3), 4.13 ppm (2H, s., CH 2 ClNa), 5.20 ppm (1H, doublet, 6H), 5.74 ppm (1H, doublet, 7H), 6.97 ppm (1H, s., Thiazole 5H).

45 DK 161645 B

(c) Sodium 7- [2- (2-aminothiazol-4-yl) -2- (syn) -methoxyiminoacetamido] -3- (1,2,3-triazol-5-yl) -thiomethyl-3-cephem -4-carboxylate (process B).

NMR Spectrum (60 MHz, in D 1 O): 3.57 ppm (2H, quartet, 2-CH 2), 3.94 ppm (3H, s., = NOCH 3), 5.21 ppm. (1H, doublet, 6H), 5.72 ppm (1H, doublet, 7H), 6.94 ppm (1H, s., thiazole 5H), 7.95 ppm (1H, s., triazole 4H).

(d) Disodium 7- [2- (2-aminothiazol-4-yl) -2- (syn) -methoxyiminoacetamido] -3- (1-carboxymethyl-1,2,3,4-tetrazol-5- yl) -thiomethyl-3-cephem-4-carboxylate (process B).

NMR Spectrum (60 MHz, in D 2 O): 3.55 ppm (2H, quartet, 2-CH 2), 3.96 ppm (3H, s., = NOCH 3), 4.72 ppm (2H, s., - N-OH 2 COONa), 5.16 ppm (1H, doublet, 6H), 5.72 ppm (1H, doublet, 7H), 6.95 ppm (1H, s., Thiazole 5H).

(e) 7- [2- (2-Aminothiazol-4-yl) -2- (syn) -methoxyiminoacetamido] -3- [1- (2-N, N-dimethylaminoethyl) -1,2,3,4- tetrazol-5-yl] -thiomethyl-3-cephem-4-carboxylic acid betaine (process A, B).

CH

NMR Spectrum (60 MHz, in D90); 3.01 ppm (6H, sj-N 2), 3.50 ppm * NCH 3 (2H, quartet, 2-CH 2), 3.96 ppm (3H, s., = NOHH 2)} 5.16 ppm (1H , doublet, 6H), 5.74 ppm (1H, doublet, 7H), 6.96 ppm (1H, s., thiazole 5H).

(f) Sodium 7- [2- (2-aminothiazol-4-yl) -2- (syn) -methoxyiminoacetamido] -3- (6-methyl-1-oxopyridazin-3-yl) -thiomethyl-3-cephem -4-carboxylate (process B).

NMR Spectrum (60 MHz, in D 2 O): 2.60 ppm (3H, s., Pyridazine 6-CH 3), 3.52 ppm (2H, quartet, 2-CH 2), 3.96 ppm (3H, s. , = NOCH3), 5.21 ppm (1H, doublet, 6H), 5.76 ppm (1H, doublet, 7H), 6.95 ppm (1H, s., Thiazole 5H).

The minimum inhibitory concentration (dg / ml) of some of the compounds obtained as mentioned above is as follows:

Microorganism Compound ____ (a) _ (e) E. coli NIHJ 0.20 0.20 E. coli 0-111 0.10 0.024 E. coli T-7 1.56 1.56 K. pneumoniae DT 0.05 0 , 10 K. pneumoniae GN 3635 0.39 0.20

Serr. marcescens IFO 12643 0.73 1.56

Serratia TN 0024 0.73 0.73 P. vulgaris IFO 3933 0.10 0.20! P. vulgaris GN 4413 1.50 1.56 j P. mirabilis GN 4359 0.20 0.39; P. morganii IFO 3163 0.10 0.20 P. justice 3 (TNO 336) = 0.012 0.024 P. justice GN 4733 0.39 0.73 j

Ent. cloacae IFO 12937 3.13 6.25 1

Cit. freundii GN 99 0.20 0.20 I

Cit. freundii GN 1706 0.73 0.73 j

Example 35 Dissolve 230 mg of sodium carbonate in a mixture of 20 ml of water and 10 ml of methanol, then add 477 mg of 7- [2- (2-aminothiazol-4-yl) - 2- (syn) -hydroxyiminoacetamido] - eephalosporansyre. To this mixed solution, 300 mg of dimethyl sulfate is added dropwise under ice-cooling and stirring. After 25 minutes, 300 mg of potassium carbonate and 300 mg of dimethyl sulfate are added. After another 25 minutes, the reaction mixture is concentrated under reduced pressure and subjected to column chromatography * on Amberlite XAD-2, eluting with water. By the above process, sodium 7- [2- (2-aminothiazol-4-yl) -2- (syn) methoxyiminoacetamido] cephalosporanate is obtained. According to NMR spectrum, etc. this product is identical to the compound obtained in Example 28.

Example 36 (1) To a suspension of 5.54 g of 2- (2-chloroacetamidothiazol-4-yl) -2- (syn) -methoxyiminoacetic acid in 70 ml of methylene chloride is added 2.42 g of triethylamine to obtain a solution. Under ice-cooling and stirring, 4, 16 g of phosphorus pentachloride are added in a single dose to the above solution. After 5 minutes, the ice bath is removed and the mixture is stirred at room temperature for 20 minutes, then concentrated under reduced pressure. To the residue is added 150 ml of hexane and then decanted twice. After the addition of 90 ml of anhydrous tetrahydrofuran, the precipitated triethylamine hydrochloride is filtered off to give a solution of 2- (2-chloroacetamidothiazol-4-yl) -2- (syn) methoxyiminoacetyl chloride in tetrahydrofuran.

On the other hand, to a suspension of 4.2 $ g of 7-aminodes-acetoxycephalosporanoic acid (7-ADCA) in a mixture of 50 ml of water and 50 ml of tetrahydrofuran under ice-cooling is added 4.44 g of triethylamine to form a homogeneous solution. During ice cooling, the previously prepared acid chloride solution is added dropwise to the above solution over 15 minutes. The mixture is stirred at room temperature for 2 hours, then a saturated aqueous solution of sodium chloride is added. The mixture is adjusted to pH approx. 2 with dilute hydrochloric acid and extracted with ethyl acetate. The ethyl acetate layer is washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and concentrated to give $ g of a yellowish-white powder. The powder is washed with 50 ml of methanol and the insoluble components are collected by filtration. By the above procedure 4.6 g of 7- [2- (2-chloroacetamidothiazol-4-yl) -2- (syn) -methoxyiminoacetamido] -desacetoxycephalosporanoic acid is obtained as a white powder.

NMR Spectrum (60 MHz, id 1 -DMSO): 2.04 ppm (3H, s, 3- $ H 2), 3.50 ppm (2H, broad singlet, 2-CH 2), 3.92 ppm ( , 4.40 ppm (2H, s-, C1CH2 CO), 5.1 $ ppm (1H, doublet, 6H), 5.7 $ ppm (1H, doubletx2, 7H), 7, 50 ppm (1H, s., Thiazole 5H).

<(2) The above product is reacted and treated in the same manner as in Example 30, (2), and sodium 7-t2- (2-aminothiazol-4-yl) -2- (syn) methoxyiminoacetamido] is obtained. desacetoxycephalosporanate as a white powder. According to NMR spectrum and other properties, this product is identical to the product obtained in Example 20.

Example 37 Sodium 7- [2- (2-aminothiazole-4) is suspended in 25 ml of dimethylformamide.

48 DK 161645 B

yl) -2- (syn) -methoxyiminoacetamido] -desacetoxycephalosporanate, and 3.75 g of iodomethyl pivalate are added under ice-cooling and further 1

3 ml of dimethylformamide. After 17 minutes, 100 ml of ethyl acetate · is added to the reaction mixture and the insoluble components are filtered off. The filtrate is washed with water, a $ 5 aqueous solution of sodium hydrogen carbonate, and a saturated aqueous solution of sodium chloride in the above order and dried over magnesium sulfate. The ethyl acetate is then distilled off and the resulting oil (2.4 g) is purified by chromatography on silica gel. In the above procedure, 1 g of pivaloyloxyraethyl-7- [2- (2-aminothiazol-4-yl) -2- (syn) methoxyiminoacetamido3-desacetoxycephalosporanate is obtained as a white powder.

Elemental analysis for C20H25N5 ° 7S2:

Calculated: C, 46.95; H, 4.92; N, 13.09

Found: G 46.92 H 4.66 N 13.13 NMR Spectrum (60 MHz, in CDCl3): 1.24 ppm (9H, s. -C (CH2) +, 2.16 ppm, 3, 44 ppm (2H, doublet, 2-01 ^), 4.10 ppm OH, s., OCH ^), 5.16 ppm (1H, doublet, 6H), 5.94 ppm (2H, s., -0CH20 ), 6.86 ppm (1H, s., Thiazole 5H).

Example 33 0.7 g of the 7- [2- (2-chloroacetamidothiazol-4-yl) -2- (syn) -methoxy-iminoacetamido] -desacetoxycephalosporanoic acid obtained by the procedure of Example 11 (1) is dissolved in an ice-cooled solution of 149 mg of triethylamine in 7 ml of dime thylph ormamide. After the addition of 715 mg of iodomethyl pivalate, the mixture is stirred for 15 minutes. To this reaction mixture is added 40 ml of ethyl acetate and the mixture is washed with water, a 5 $ T aqueous solution of sodium bicarbonate and a saturated aqueous solution of sodium chloride in the order in which it is dried over magnesium sulfate. The ethyl acetate of these is distilled and 0.8 g of crude pivaloyloxymethyl-7- [2- (2-chloroacetamido-thiazol-4-yl) -2- (syn) -methoxyiminoacetamido] -de sacetoxycephalo sporate is obtained as a brown oil.

This product is dissolved in 3 ml of dimethylacetamide, after which 206 mg of thiourea is added. The mixture is stirred at room temperature for 16 hours. To this is added 40 ml of ethyl acetate and the mixture is washed twice.

«DK 161645 B

with 30 ml portions of a saturated aqueous solution of sodium chloride and dried over magnesium sulfate. The ethyl acetate is distilled off and the resulting tan oil (0.4 g) is purified by chromatography on silica gel. In the above procedure, 0.2 g of pivaloyl-oxymethyl-7- [2- (2-aminothiazol-4-yl) -2- (syn) -methoxyiminoacetamido] -desacetoxycephalosporanate is obtained as a white powder.

According to NMR spectrum and other properties, this product is identical to the product obtained in Example 37.

Example 39

To a suspension of $ 31 mg of 2 ~ (2-chloroacetamidothiazol-4-yl) -2- (syn) methoxyiminoacetic acid in 10 ml of methylene chloride is added 360 mg of triethylamine and 624 mg of phosphorus pentachloride. The mixture is stirred at room temperature for 20 minutes, then 100 ml of hexane is added. The separated oil is recovered by decanting the hexane and dissolved in 1 $ ml of tetrahydrofuran to give a solution of 2- (2-chloro-acetamidothiazol-4-yl) -2- (syn) -methoxyiminoacetyl chloride.

On the other hand, 9 $ 4 mg of 7-amino-3- (1-methyl-1H-tetrazol-5-yl) -thiomethyl-3-cephem-4-carboxylic acid and 660 mg of triethylamine are dissolved in 15 ml of a 50% solution. aqueous tetrahydrofuran solution, and under ice-cooling, the previously prepared acid chloride solution is added dropwise to this solution. The mixture is stirred under ice-cooling for 2 hours, after which the reaction mixture is diluted with water, adjusted to pH approx. 2 with dilute hydrochloric acid and extracted with ethyl acetate. The ethyl acetate layer is washed with a saturated aqueous solution of sodium chloride and dried over magnesium sulfate. The ethyl acetate is distilled off and the residue is treated with ether. The resulting crystals are collected by filtration. By the above process 1.3 g of 7- [2- (2-chloroacetamidothiazol-4-yl) -2- (syn) methoxyiminoacetamido] -3'- (1-methyl-ΙΗ-1e trazole-5 yl) thiomethyl-4-carboxylic acid.

This product is identical to the intermediate obtained by the first part of the process described in Example 27. 5 g of the product prepared as above is dissolved in 20 ml of dimethylacetamide and, under ice-cooling, 1.53 g of thiourea is added. The mixture is stirred at room temperature for 15 hours. To this reaction mixture,

50 DK 161645 B

tea's 200 ml of ice-water and the pH of the mixture is adjusted to 3.5 with sodium bicarbonate. The resulting precipitate is collected by filtration and dissolved in a 10 µl aqueous solution of sodium bicarbonate. The solution is then passed through a column packed with Amberlite XAD-2. By this purification procedure 1.5 µg of sodium 7- [2- (2-aminothiazol-4-yl) -2- (syn) methoxyiminoacetamido] -3- (1-methyl-1H-tetrazol-5-yl) - thiomethyl 3-cephem-4-carboxylate as a white powder.

According to NMR spectrum and other properties, this product is identical to the product obtained in Example 27.

Example 40 In 10 ml of dimethylformamide is dissolved 1 g of sodium 7- (2- (2-aminothiazol-4-yl) -2- {syn) -methoxyiminoacetamido] -3- (1-methyl-1H-tetrazol-5-yl) ) "Thiomethyl 3-cephem-4-carboxylate, and 0.05 g of iodomethyl pivalate are added under ice cooling and stirring. The mixture is stirred for 15 minutes. After the addition of 1 + 0 ml of ethyl acetate, the reaction mixture j is washed with water, a 5 $ Ts aqueous solution of sodium bicarbonate and a saturated aqueous solution of sodium chloride in that order and then dried over magnesium sulfate. The ethyl acetate is distilled off under reduced pressure and the residue is dissolved in a small amount of ethyl acetate and filtered. To the filtrate is added ether and then cooled. The resulting precipitate is collected by filtration. By the above process 0.4 g of pivaloyloxymethyl-7- [2- (2-aminothiazol-4-yl) -2- (syn) -methoxyiminoacetamido] -3- (1-methyl-1H-tetrazol-5-yl) is obtained. -thiomethyl-3-cephem-4-carboxylate as a white powder.

Elemental analysis for C 2, 27, 9, 7, 7:

Calculated: C 42.27 H 4> 34

Found: C 42.29 H 4.40 NMR Spectrum (60 MHz, in CDCl3): 1.22 ppm (9H, s. 3, 20 ppm (2H, broad singlet, 2-CH 2), 3 s94 ppm, 4.06 ppm (3Hx2, s.x2, N-CH2 and OCH2), 5.94 ppm (2H, s., -OCH-pO), 5.12 ppm (1H, doublet, OH), 6.06 ppm (1H, doubletx2, 7H), 4.44 ppm (2H, doublet, 3-CH2), 6, B1 ppm (1H, s., Thiazole 5H).

DK 161645 B

Example 41 Dissolve 2,776 g of 2- (2-chloroacetamidothiazol-4-yl) -2- (syn) methoxyimino acetic acid and 1.2 g of triethylamine in 20 ml of methylene chloride, then add 2.08 g of phosphorus pentachloride. The mixture is stirred at room temperature for 20 minutes, then 150 ml of hexane is added.

The resulting oily precipitate is separated and dissolved in 20 ml of tetrahydrofuran to give a solution of 2- (2-chloroacet-amidothiazol-4-yl) -2- (syn) -methoxyiminoacetyl chloride. On the other hand, 3 »143 g of 7-amino-3-acetylacetoxymethyl-3-cephem-4» Carboxylic acid and 2.20 g of triethylamine are dissolved in 50 ml of a 50 $ Ts aqueous tetrahydrofuran solution. To this is added dropwise under ice-cooling and stirring the previously prepared acid chloride solution. The mixture is stirred under ice-cooling for 2 hours, then water is added. The mixture is adjusted to pH 2.0 with dilute hydrochloric acid and extracted with ethyl acetate. The ethyl acetate layer is washed with a saturated aqueous solution of sodium chloride and dried over magnesium sulfate. The ethyl acetate is then distilled off and ether is added to the residue. The resulting crystalline product is collected by filtration. By the above process, 4.163 g of 7- [2- (2-chloraeetamido-thiazol-4-yl) -2- (syn) -methoxyiminoacetamido] -3-acetylacetoxymethyl-3-cephem-4-carboxylic acid are obtained.

NMR Spectrum (60 MHz, in d / -DMSO): 2.14 ppm (3H, s., -C-CH.J, 3.60 ppm

o r, J

0 (4H, broad s., -C-CH 2 -C- and 2-CH 2), 3.86 ppm (3H, s., 0CHJ, 4.34 ppm χχ CC} oo (2H, s., CICH CO), 4.91 ppm (2H, quartet, 3-CH 2), 5.13 ppm (1H, doublet, 6H), 5.80 ppm (1H, doubletx2, 7H), 7.40 ppm (1H, s , thiazole 5H).

t

Example 42

In 20 ml of dimethylacetamide are dissolved 4.00 g of the 7-[2- (2-chloroacetamidothiazol-4-yl) -2- (syn) methoxyiminoacetamido] -3-acetylacetoxy-methyl-3-cephem-4-carboxylic acid, obtained in Example 41, after which 1.06 g of thiourea is added. The mixture is stirred at room temperature for 17 hours, after which 100 ml of ether is added. The oily precipitate is separated and dissolved in a 5% aqueous solution

"DK 161645 B

dZ

sodium bicarbonate. The solution is lyophilized and the resulting powdered product is added to 50 ml of methanol. The insoluble constituents are filtered off and the filtrate is added to 300 ml of ether. The precipitate is collected by filtration. By the above process, 3> 150 g of sodium 7- [2- (2-aminothiazol-4-yl) -2- (syn) methoxyimino-acetamido] -3-acetylacetoxymethyl-3-cephem-4-carboxylate · I are obtained. 10 ml of water are dissolved 933 mg of the above product, 350 mg of 1- (2-N, N-dimethylaminoethyl) -1H-tetrazole-5-thiol and 168 mg of sodium hydrogen carbonate. The mixture is stirred at 55 ° C for one hour and then the reaction mixture is passed directly through a column packed with Amberlite XAD-2 for purification. By the above procedure, 170 mg of sodium 7- [2- (2-aminothiazol-4-yl) -2- (syn) methoxyiminoacetamido 3-3- [1- (2-N, N-dimethylaminoethyl) tetrazol-5-yl] -thiomethyl-3-cephem-4-carboxylate as a white powder. According to NMR spectrum and other properties, this product is identical to the product obtained in Example 34.

The following table shows the protective effect (ED mg / kg) of the compounds prepared according to the above examples on infected mice.

Table. !

Example No.

for the sample Administration ED ^ q (mg / kg) 26 SC 0.015 (CER: 1.25) 27 SC 0.022 (CER: 1.25) 28 SC 0.018 (CER: 1.25) 30 SC 0.111 (CER: 1.25) 40 Oral 0.11 (CEX: 2.51) 42 Oral 0.27 (CEX: 2.51) * Laboratory Animal: male (ICR / SLC) 5 mice per group per single dose. Infection: intraperitoneal with Escherichia coli 0-111 Observation period: 7 days.

(): control SC = subcutaneous CER = cephaloridine

53 DK 161645 B

lsj-CH2OOBH- | -A + coo "CEX = cephalexin </ -CHCO - j / S \, NH0. -N" -CH0 2 1 / Y 3

COOH

Example .43 (1) To a suspension of 55.6 g of 2- (2-chloroacetamidothiazol-4-yl) -2- (syn) -methoxyiminoacetic acid in 600 ml of methylene chloride is added 24.3 g of triethylamine to obtain a solution. During ice cooling and stirring, 41 µg of phoshorpentachloride is added in two doses to the above solution. After 5 minutes, the ice bath is removed and the mixture is stirred at room temperature for 20 minutes, then concentrated under reduced pressure. To the residue is added 1 liter of hexane and then decanted twice. After the addition of 600 ml of anhydrous tetrahydrofuran, the precipitated triethylamine hydrochloride is filtered off to give a solution of 2- (2-chloroacetamidothiazol-4-yl) -2- (syn) -methoxy-iminoacetyl chloride in tetrahydrofuran.

On the other hand, a suspension of 54.7 g of 7-amino-3-carbamoyloxymethyl-3-cephem-4-carboxylic acid is added to a mixture of 400 ml of water and 400 ml of tetrahydrofuran 6 g of triethylamine under ice-cooling to prepare an homogeneous solution. During ice cooling, the previously prepared acid chloride solution is added dropwise to the above solution over 30 minutes. The mixture is stirred at room temperature for 2 hours, then a saturated aqueous solution of sodium chloride is added. The mixture is adjusted to pH 2 with dilute hydrochloric acid and extracted with ethyl acetate. The ethyl acetate layer is washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and concentrated to give 97.3 g of 7- [2- (2-chloroacetate)

54 DK 161645B

! amidothiazol-4-yl) -2- (syn) -methoxyiminoacetamido] -3-carbamoyloxy-methyl-3-cephem-4-carboxylic acid. According to NMR spectrum and other properties, this product is identical to the product obtained in Example 31 (1).

NMR Spectrum (60 MHz, 1 D -DMSO): 3.56 ppm (2H, broad singlet, 2-CH 2), 3.95 ppm (3H, s, OCH 3), 4.35 ppm (2H, s., C1CH2 CO), 4.76 ppm (2H, quartet, 3-CH2), 5.19 ppm (1H, doublet, 6H), 5.64 ppm (1H, doubletx2, 7H), 6.56 ppm (2H , s., OCONHg), 7.46 ppm (1H, s., thiazole 5H).

(2) Dissolve 97,3 g of the product prepared as above in (1) in 500 ml of Ν, Ν-dimethylacetamide and add 31.2 g of thiourea to the solution under ice-cooling. The mixture is stirred at room temperature for 15 hours. To this reaction mixture is added 2 liters of ether and then the oily product is separated. A suspension of this oily product in 300 ml of water is adjusted to pH 7.0 with sodium hydrogen carbonate. The solution thus obtained is passed through a column packed with Amberlite XAD-2. This purification procedure yields 20.2 g of sodium 7- [2- (2-aminothiazol-4-yl) -2- (syn) methoxyiminoacetamido] -3-carbamoyloxymethyl-3-cephem-4-carboxylate as a white powder. According to NMR spectrum and other properties, this product is identical to the product obtained in Example 26 or 31.

The structures and properties (IR spectrum) of compounds 1 to 33 obtained by the above methods are listed in the following table. In this table, IR spectrum (cm- \ KBr) means characteristic absorption bands due to the β-lactam moiety.

Table.

R2NH q V \ II s

N_l-C-CONH - / X

H J ^ -k J-CH 2 R 3

OCH3 cqqM

Connect 9 µM IR

Rank R E (about_1, KBr)

DK 161645 B

55 1 H -s \ J After 1760 2 H We? After 1765

-so V

H _ 3 H We? After 1? 58 -sAV ch5 CH-, 4- 'H j | fj After 1760

C IL

_-_: _ 5____

CH

5 Η ίι [Γ 5 After 1763 -sAsAch, 6 H V? After 1765 -sAsAnh2 7 H V r? After 1760 -s 4C> HHCOOCH, b 7 /

56 DK 161645 B

- Connect to TT? difference ΈΓ R ^ Μ _Ί no .__ (cm, KBr) 8 Η? ~ ΐί After 1765 -sAs> ch2co m2 9 H !? "π After 1768

-sArH

CH2CO] ®i2 I

10 H? “? After 1768 -sAn> nh2 ch3 11 H -OH Na 1? 60 12 H? "j? CH, Na 1765 -δΛςΛθΗοΝ <3 S CBj 13 H 0 1765 14 H -r y — CQNHp 0 1765 15 · H ΐ ~ f! Na 1768 -S A g jJ- SOØCQONa

TT - TT

16 Η I, cf. After 1765 CH2C00Na

Improvement R2 R3 M _i no (cm, KBr)

"DK 161645 B

17 Η? “Π Na 1765 -SArii 'CHgSO ^ Na Οϊζ 18 H -OCOCH ^ -CH2OCOC-CH5 1760 CH, 7 CH3 19 H-OCNOH -CHoOCOOC-CH, 1765 C. -d. I 7 ___ CH3__ CHj 20 H J “{? -CH 2 OCOC-CH 5 1765 _S_ ^ N "N CH, _; __ H__3___ CH3 21 H f __; __ CH ^ ___ 22 Η H -CHOCOOCgH ^ 1760 CH, 7 23 H -OCOCH ^ -CHOCOOCp 1763 CH, 7 CH ^ 24 H -OCONH9 '-CHOCOC-CH, 1763 2 ii 7 CH5 CHj

N - N

25 H _ Jt N -CHOCOOC ^ 1765 S T CH, CH, 5 7

Connected? µ m IR

division R R (cm -1 KBr)

58 DK 161645 B

26 H 11? -CHOC000, -, ¾. 1768; -s-Il N »^ 2 0 ^ 'K'L' CH, 1 H 3! N - N CH5 27 H -sA ^ N -CHOCOC-CH, 1765 0H2CH2N <CH5 ° H, iH? ! CH3! 28 Η H <JjQ 1760

It o 29 H -OCOCH ^ <^ 0 1763 tt

ISLAND

50 h -ocom2 '1763 tt o J1 * AJ ¢ 0' "0¾__O__ M B -.- O ° Φθ 1765

H O

«\ 55 * AJ '° 00" r H "

's nx O.

gh0ch0nC 5 6¾ 3

Claims (7)

59 DK 161645 B 1. 2- (2-Chloroacetamidothiazol-4-yl) -2-oxyiminoacetic acid derivatives for use as intermediates in the preparation of (5-lactamanti-biotics, characterized in that they have the general formula C1CH-CONH SY) (III) N_Li -C-COOH II NV fi nor6 g wherein R is hydrogen or methyl, or is a salt, a lower alkyl ester or a reactive derivative thereof. A compound according to claim 1, characterized in that it is used to prepare a cephemic compound of the formula S NH? - / -! is NUC-CONH - / \ (I) Il I -¾ Nn c> -N., - CHOR or nor 2 COOH 3 wherein R is hydrogen or a residue of a nucleophilic compound, pharmaceutically acceptable salt or a pharmaceutically acceptable ester thereof. Compound according to claim 1, characterized in that it is 2- (2-chloroacetamidothiazol-4-yl) -2- (syn) -methoxyiminoacetic acid. A compound according to claim 1, characterized in that it is 2- (2-chloroacetamidothiazol-4-yl) -2- (syn) -methoxyiminoacetyl chloride. Compound according to claim 1, characterized in that it is 2- (2-chloroacetamidothiazol-4-yl) -2- (syn) -methoxyiminoacetyl chloride hydrochloride. 60 DK 161645 B A process for preparing a compound according to claim 1 of the formula! Wherein R is hydrogen or methyl and R is lower alkyl, characterized in that a compound of formula H2Y 7 II N L · 6 r n wherein R ° and R have the above meanings are reacted with chloroacetyl chloride before or after isolation of the syn isomer. i i j