LU83117A1 - BETA-LACTAM CORE ANTIBIOTICS, AND PROCESS FOR PREPARING THEM - Google Patents

Description

I 'ΐ_4 D. 51.378 ί _Q τ i \ 7 V% JII / GRAND-DUCHÉ DE LUXEMBOURG Patent N ° ................... ...... .........

du ..., 6 ..... f évrior ..... 1981 Minister for the Economy and the Middle Classes

Title issued ......................................... Intellectual Property Service

SM? LUXEMBOURG

Invention Patent Application I. Application ..... La ..... so.cié.té ..... dlte.; ..... JS. »..... R, .. .... S.QUXBB ...... & ...... SOUNDS, ..... INC. " ., ..... Lawrence.vill.e -......... (i) ... Prince.tQn .... RQa &, „. A ... PRINCETON .., ... ..New ..... Jersey ... ..q85.4q # ... United States ........................

....................... (2) ... acting. ... en ..... qnalit.é ._ „de .... iQandat; air.e ......................... .................................................. .................................................. ......

deposit (s) this .... six .... february ..... ISlqq ..... eighty-one ------------------- ----------------------------- (3) a to ........ 1.5 ....... ........ hours, at the Ministry of the Economy and the Middle Classes, in Luxembourg: 1. this request for obtaining a patent for an invention concerning: ..... "Antibiotics. .... to ..... n.QY.au .... ^ - la.ctame, ...... et.pr.QcMé ..... pQur ..... les .. ............................. (4) ♦ ........ prepare ".. * .... .................................................. .................................................. .................................................. .................................................. ....................

2. the delegation of power, dated PEDKEDCW ^ NeW. Jerseje ........... 3.0 December ..19 8 3. description in French .......... language ............... ....... of the invention in two copies; 4 ....... DD ............. .... drawing boards, in two copies; 5. the receipt of the taxes paid to the Luxembourg Registration Office on ....... 6 ..... February ..... 19.81 ............. .................................................. .................................................. .................................................. .............................................

declares (s) assuming responsibility for this declaration, that the inventor (s) is (are): ..... see on .... back ............. .................................................. .................................................. .................................................. ............................................... (5) claim) for the above patent application the priority of one (or more) application (s) of (6) .................. patent ....... .......................................... registered ki (7) in the United States of America _______________ on ....... 7 ..... February ..... 19.8.0 ........... (No. »..... .119 ..,. 2.7.8.) ..... and. on ..... 2.9 ...... September. ..19.8o ....................... (8) ................ (No .. ....... 18.8, .. 8.93) ..................................... .................................................. .................................................. .................................................. .....................

* in the name of S inventors ............................................ .................................................. .................................................. ......................................... (9) v afiTiiicile T elect (elect ) for him / her and, if designated, for his / her representative, in Luxembourg .................................

a ..... 3.5, bld. Royal................................................. .................................................. .................................................. .................................................. ...... (io) requests (nt) the grant of a patent for the invention for the subject described and represented in the aforementioned appendices ^, - with postponement of this grant to ........ ..... 1.8 ......................................... months. (11)

% e ...... agent), ............ 'v ............ V V

II. Deposit Minutes

The aforementioned patent application has been filed with the Ministry of the Economy and the Middle Classes, Intellectual Property Service in Luxembourg, on:. / '£ "-A Pr. 1 ^' Minister (OfD at ...... 15 .......... hours. / De ^ Économie e ^ JjÎs Classes Moyennes, U4K \ · · / / * fi \ A 68007 __..., ~ _ · _ ^. · _ D. 51.378

CLAIM OF PRIORITY

of the patent application / cfi // JÊ IN THE UNITED STATES OF AMERICA From February 7, 1980

September 29, 198o LAXHisd Brief Description filed in support of a request for

U

PATENT

at

Luxembourg on behalf of: E. R. SQUIBB & SONS / INC.

for: "Antibiotics with β-lactarone nucleus, and process for * preparing them".

1 ------ '-.................-....... I IIHII

Ss-LACTAM.E CORE ANTIBIOTICS,

AND PROCESS FOR PREPARING THEM

The present invention relates to a new family of β-lactam nucleus antibiotics, as well as the use of these compounds as antibacterial agents. It has been discovered that the β-lactam nucleus can be biologically activated by a substituent formed by a sulfonic acid salt, attached to the nitrogen atom of the nucleus.

Ss-lactams having a substituent formed by a sulfonic acid salt (including an "internal salt") in position 1, and an acylamine substituent in position 3 appear to be active against a range of Gram-negative and Gram-negative bacteria positive.

The preferred members of this new family of β-lactam nucleus antibiotics according to the invention are those which are defined by the formula: 1 h h R -NH-C-C-R., A-LsoV.

</

In addition to the β-lactams described above having a sulfonic acid salt substituent in position 1 and an acylamine substituent in position 3, the invention also encompasses ß-lactams having a sulfonic acid salt substituent (including a "internal salt") in position 1, and an amine substituent in position 3. The preferred compounds of. this type have the formula:. __

Ia ^ 2 jl nh2-c-c-r3 C - N-SO®M® r // J. '0

These compounds are useful intermediates for the preparation of the corresponding 3-acylamine compounds.

As used in formulas I and the

AT

2 and throughout the specification, the symbols have the definitions below; R is an acyl radical; R2 is a hydrogen atom or an alkoxy radical of 5 1 to 4 carbon atoms? R ^ and R ^ are the same or different and are each a hydrogen atom or an alkyl, cycloalkyl, phenyl or substituted phenyl radical, or one of R ^ and R4 is a hydrogen atom and the other is an alkoxycarbonyl, alken-1-yl, alkyn-1-yl, 2-phenylethenyl or 2-phenylethynyl radical.

· * © M is a hydrogen ion or a cation. __

The terms "alkyl" and "alkoxy" refer to straight chain or branched chain radicals. Radi-15 waters having 1 to 10 carbon atoms are preferred.

The terms "cycloalkyl" and "cycloalkenyl" refer to cycloalkyl and cycloalkenyl radicals having 3 to 7 carbon atoms.

The term "alkenyl" refers to a straight chain or branched chain radical. Alkenyl radicals having 2 to 10 carbon atoms are preferred.

The term "halogen" denotes a fluorine, chlorine, bromine or iodine atom.

The term "substituted phenyl" denotes a phenyl radical substituted by 1, 2 or 3 amine groups, halogenated atoms, hydroxyl groups, trifluoromethyl radicals and / or lower alkyl or alkoxy radicals.

The term "protected carboxyl" denotes a carboxyl radical which has been esterified by a conventional ester protecting group. These groups are well known in the art; see, for example, U.S. Patent No. 4,144,333. The preferred protected carboxyl radicals are the benzyl, benzhydrylic and t-butyl ester radicals.

The term "acyl" includes all organic radicals derived from an organic acid (i.e., a carboxylic acid) by removal of the hydroxyl group. Certain acyl radicals are of course preferred, but this preference should not be regarded as a limitation of the scope of the invention. Exemplary acyl radicals are those which have been used in the past to acylate β-lactam ring antibiotics, including 6-aminopenicillanic acid and its derivatives and 7-amincephalosporanic acid and its derivatives; see, for example, Cephalosporins and Penicillins, edited by Flynn, Academie Press (1972), the German patent application published under No. 5 2,716,677, Belgian patent No. 867,994, U, S patent.

No. 4,152,432, U.S. Patent No. 3,971,778, U.S. Patent

No. 4,172,199, and British Patent No. 1,348,894. The parts of these references which describe various acyl radicals are incorporated herein by reference. The following list of alkyl radicals is presented to give further examples of the term "acyl"; it should not be considered as limiting this term. Exemplary acyl radicals are: (a) aliphatic radicals having the formula:

O

II

R5-c- in which is an alkyl radical; cvcloalkyle; alkoxy; alkenyl; cycloalkenyl; cyclohexadienyl; or alkyl or alkenyl respectively substituted by one or more halogen atoms, by one or more cyano or nitro groups, or by one or more amine, mercapto, alkylthio or cyanomethylthio radicals.

(b) carbocyclic aromatic radicals having the formula:

Jl R o (Ç -CH-C-,

Rg R? R6 ^ JVR8 »j ^ \ <^ cn2-oc-, 4 R7 R6ħo-CH2X, RÄs.c„ 2-I, or R7 R6 # VR8 ° <^) - ch2-sc- where n is 0 , 1, 2 or 3; and Rg are each independently a hydrogen or halogen atom, a hydroxyl or nitro group, an amine, cyano, trifluoromethyl, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms or aminomethyl group; and Rg is an amine radical, a hydroxyl function, a carboxylic salt, a protected carboxyl radical, a formyloxy radical, a sulfonic salt, a sulfoamine salt, an azide radical, a halogen atom, a hydrazine, alkylhydrazine radical, 10 phenylhydrazine, or [(alkylthio) thioxomethyl] thio.

Preferred carbocyclic aromatic acyl radicals include those having the formula: · ho - (^ - ch2-c-, Q-ch2-Î- \: h2nh2, h 5 ho - \ - ca-c- R9 (Rg is preferably a carboxylic salt or a sulfonic salt) or Q-CJ-R9 (Rg is preferably a carboxylic salt or a sulfonic salt).

5 (c) Heteroaromatic radicals having the formula: 0 0

RloÇh-Ï-, R9 0 R10_O “CH2_C" '0

II

R ^ g-S-CI ^ -C-, or

0 0 II II

Rjq "~ C C-, where n is 0, 1, 2 or 3; 'Rg has the same definition as above; and is a heterocyclic pentagonal, hexagonal or heptagonal ring, substituted or unsubstituted, containing 10 having 1, 2, 3 or 4 (preferably 1 or 2) nitrogen, oxygen or sulfur atoms. Exemplary heterocyclic rings are thienyl, furyl, pyrrolyl, pyridinyl, pyrazinyl, thiazolyl, morpholinyl , pyrimidinyl and tetrazolyl, exemplary substituents are halogen atoms, hydroxyl and nitro groups, amine, cyano, trifluoromethyl, alkyl of 1 to 4 carbon atoms, or alkoxy of 4 carbon atoms.

Preferred heteroaromatic acyl radicals include radicals having the above formulas wherein R g is a '2-amino-4-thiazolyl, 2-amino-6 5-halo-4-thiazolyl, 4-aminopyrimidin- radical 2-yl, 5-amino-1,2,4,4-thiadiazol-5-yl, 2-thienyl or 2-furanyl.

(d) The radicals [[(4-substituted-2,3-dioxo-1-piperazinyl) carbcnyl] amino] arylacetyl having as formula: 0 <? / \

It II '

-C-CH-NH-C-N

U

0 or 5 in which is an aromatic radical (including carbocyclic aromatic radicals such as those having the formula: -------- 'as well as heteroaromatic radicals as they are included in the definition of R10); and R12 is an alkyl, substituted alkyl (or the alkyl radical is substituted by one or more halogen atoms, or by one or more cyano, nitro, amine or mercapto groups), arylmethylene-amine (i.e. -to say: -N = CH-R · ^ or R ^ has the same definition

O

II

As above), arylcarbonylamine (i.e.; -NH-C-R ^ where Ri; l has the same definition as above), or alkyl -carbonylamine.

Preferred [[(4-substituted-2,3-dioxo-1-piperazinyl) -carbonyl] amino] arylacetyl radicals include those in which R ^ 2 is an ethyl, phenylmethyleneamine or 2-furylmethyleneamine radical.

(e) Arylacetyl (substituted oxyimino) radicals having the formula:

O

-c-ç = n-o-r13

Rn in which R ^^ has the same definition as above and R ^ 25 is a hydrogen atom, an alkyl or cycloalkyl radical,

L

7 alkylaminocarbonyl, arylaminocarbonyl (i.e. 0 I! -C-NH-R where Ri; the same definition as above) or a substituted alkyl radical (in which the alkyl radical is substituted by one or several halogen atoms or groups cyano, nitro, amine, mercapto, alkylthio, or aromatic (according to the definition of R ^), carboxyl (including its salts), amide, alkoxycarbonyl, phenylmethoxycarbonyl, diphenylmethoxycarbonyl, hydroxyalkoxyphosphinyl, dihydroxyphosphinyl, hydroxy (phenylmethoxy) phosphinyl, or dialkoxyphosphinyl.

Preferred arvlacetyl (substituted oxyimino) radicals include those in which R ^ is a 2-amino-4-thiazolyl radical. Also preferred are radicals in which R 2 is methyl, ethyl, carboxymethyl, or 2-carboxyisopropyl.

(f) (acylamino) arylacetyl radicals having the formula:

O O

He II

-C-CH-NH-OR,, I 14 R11 in which R has the same definition as above and Rl4 20 is a radical R7 8 (CII ^). -0-, amine, alkylamine, (cyanoalkyl) amine, amide, alkylamide, (cyanoalkyl) amide, "

NH, _ NH „O

Il / ΓΛ I 2 i, -CH2-NH-C- ('NN, -CïI-CH2-C-NH-CH3, - - HCk_ ~ K Vf VS02-N (CH2-CH2- ° H) 2' - fVcH,, γΤΝ \ == / \ = / 3

OH

AT

8

OH QH

fY *! °

The preferred (acylamino) arylacetyl radicals according to the above formula include the radicals in which R 4 is an amine or amide radical. Also preferred are radicals in which is a phenyl or 2-thienyl radical.

(g) The radicals [[[3-substituted-2-oxo-1-imidazoli-dinyl] carbonyl] amino] arylacetyl having the formula:

O

tl O 0

Il II / \ -C-CH-NH-CN NR, c AII 15 ^ L1 CH 2-CH2 in which R has the same definition as above and R ^ is a hydrogen atom or an alkylsulfonvl, arylmethyleneamine radical (i.e.: -N = CH-R ^ where R ^ has the

O

II

same definition as above), -CR ^ g (where R ^ g is a hydrogen atom or an alkyl or alkyl radical substituted by a halogen atom), an aromatic radical (according to the definition of R 15 above), alkyl 'or substituted alkyl (in which the alkyl radical is substituted by one or more halogen atoms or cyano, nitro, amine or mercapto groups).

Preferred [[[3-substituted-2-oxo-1-imidazolidinyl] - · carbonyl] amino] arylacetyl radicals according to the above formula include those in which R ^ is a phenyl or 2-thienyl radical, Also preferred the radicals in which R ^ is a hydrogen atom or a methylsulfonyl, phenylmethyleneamine or 2-furylmethylene-amine radical.

The term "cation", as used throughout this specification, means any atom or group of atoms carrying a positive charge.

AT

0 0 9

The substituent "SOpd" carried by the nitrogen atom of the β-lactams according to the invention includes all the salts of sulfonic acid. Pharmaceutically acceptable salts are of course preferred, although other salts are also usable for purifying the products of the invention or as intermediates for the preparation of pharmaceutically acceptable salts. The cationic part of the sulfonic acid salts according to the invention can be obtained from organic or inorganic bases. Such a cationic part includes, but is not limited to the following ions: ammonium; substituted ammonium, for example * alkylammonium (for example tetra-n-butylammonium, hereinafter called tetrabutylammonium); alkali metals, such as lithium, sodium and potassium; alkaline earth metals, such as calcium and magnesium; pyridinium; dicyclohexylammonium; hydrabaminium; benzathinium; N-methyl-D-glucaminium.

As indicated in formula I and in the definitions according to formula I, M can be a hydrogen ion.

As will be described below, the β-lactams according to the invention can be synthesized. The ß-lactams of formula I which are unsubstituted in position 4 and which are not alkoxylated, that is to say the compounds of formula I in which and R4 are hydrogen atoms, can be prepared by using as starting material 1 6-aminopenicillinic acid or 6-acylaminopenicillanic acid. The β-lactams of formula I in which R2 is an alkoxy radical can be prepared from the corresponding non-alkoxylated β-lactam. Some of the compounds according to the invention can be crystallized or recrystallized from solvents containing water. In these cases, hydration water may form. The present invention does not exclude stoichiometric hydrates as well as compounds containing varying proportions of water which can be obtained by methods such as lyophilization.

AT

f 10. Some of the ß-lactams of formula I have also been prepared by biological means. The cultivation of a strain of the microorganism Chromobacterium yiolaceum SC 11378 gives an acid salt (R) -3 * - · (acetylamino) -3 -methoxy-2-oxo-l-5 azetidinesulfonic. The culture of various bacteria of acetic acid, for example of Gluconobacter species SC 11435 gives an acid salt (R) -3 - [[N- (Dy-glutamyl) -D-alanyl] amino] -3 ~ methoxy -2 ~ oxo-l-azetidinesulfonic.

Ss-lactams bearing a sulfonic acid salt substituent in position 1 and an amine or acyl-amino substituent in position 3 contain at least one chiral center - the carbon atom (in position 3 of the ß-lactam nucleus) to which is attached the substituent amine or acylamine. The subject of the invention is the β-lactams which have been described above, the stereochemistry of the chiral center occupying position 3 of the β-lactam nucleus is the same as the configuration on the carbon atom. position 6 of penicillins of natural origin (penicillin G for example), and the configuration on the carbon atom 20 occupying position 7 of cephamycins of natural origin (cephamycin C for example).

As regards the preferred β-lactams of formulas I and 1a, the formulas developed have been drawn so as to show the stereochemistry on the chiral center occupying position 3. For reasons of nomenclature convention, the compounds of formulas I and la in which is a hydrogen atom have the configuration S, and the compounds of formulas I and la in which R „is an alkoxy radical have the configuration R.

"Z" 30 Also included within the scope of the invention are racemic mixtures which contain the β-lactams described above.

Ss-lactams having a sulfonic acid salt substituent in position 1 of the ß-lactam nucleus and an amine or acylamine substituent in position 3 of the ß-lactam nucleus are active against a range of Gram-negative and Gram-positive organisms . The salt substituent sulfonic acid is essential for the activity of the compounds according to the invention. The A compounds in which R ^ and / where R ^ are a hydrogen atom 11 or an alkyl radical, in particular the methyl radical, show a particularly useful activity.

The compounds according to the invention can be used as agents for combating bacterial infections (including urinary tract infections and respiratory infections) in mammalian species such as domestic animals (eg dogs, dogs, cats, cows, horses, etc.) and humans.

To combat bacterial infections in mammals, a compound according to the invention can be administered to a mammal to be treated, at a rate of approximately 1.4 mg / kg / day to approximately 350 mg / kg / day, preferably at from about 14 mg / kg / day to about 100 mg / kg / day. All the modes of administration which have been used in the past to reach the site of infection by penicillins and cephalosporins are also envisaged with the new family of β-lactams according to the invention. These modes of administration include oral, intravenous, intramuscular, and suppository forms.

The β-lactams are generally prepared according to the invention by introducing a sulfonic acid substituent (a sulfonic group -SO 4 -) on the nitrogen atom occupying the 1 position of the β-lactam nucleus. This sulfonation reaction is easily accomplished by treating the β-lactam with a sulfur trioxide complex or with an equivalent sulfonation reagent such as chlorosulfonate.

The most commonly used sulfur trioxide complexes are the pyridine-sulfur trioxide, lutidine-sulfur trioxide, dimethylformamide- sulfur trioxide, and picoline-sulfur trioxide complexes. Instead of using a complex formed in advance, the complex can be formed in situ, for example using as reagents the chlorosulfonyltrimethylsilyl ester and pyridine. Alternatively, the sulfonation can be carried out by means of an intermediate compound, for example by silylating first the nitrogen atom of the β-lactam nucleus, then by subjecting the silylated compound to an exchange reaction. of the silyl radical with trimethylsilylchlorosulfonate or a similar reagent. Exemplary silylating agents are monosilyl-

AT

12 trifluoracetamide, the combination of trimethyl-silyl chloride / triethylamine and his-trimethylsilyltrifluor- · acetamide.

In general, the sulfonation reaction is carried out in the presence of an organic solvent such as pyridine, or a mixture of organic solvents, preferably a mixture of a polar solvent such as dimethylformamide and a halogenated hydrocarbon. such as dichloromethane.

The product initially formed in the sulfonation reaction is a salt of the sulfonated β-lactam. When the sulfonation complex is the pyridine-sulfur trioxide complex, the product initially formed is the i, -f pyridinium salt of the sulfonated ß-lactam, M being the pyridinium ion in the formula below: yN-S03 " m +

O

These complexes can be transformed into other sulfonic acid salts using standard techniques (eg ion exchange resins, crystallization or extraction of ion pairs). These processing techniques can also be used to purify products. Particularly useful are the conversion of the pyridine salt to the potassium salt using potassium phosphate or potassium ethyl hexanoate; transformation into tetrabutylammonium salt using tetrabutyl ammonium hydrogen sulfate; or conversion to a zwitterion (M + = hydrogen) using formic acid; "·

It should be appreciated that the sulfonation reaction which introduces the sulfonic group on the nitrogen atom of the β-lactam nucleus can take place at various stages of the synthesis, including introduction before the formation of a β-lactam nucleus. lactam, in cases where such a procedure is followed as summarized below. The sulfonation reaction is carried out in the presence of the solvents already described and usually at room temperature. In cases where the amine function is present, it is preferably carried out after protecting the amine function.

fL ~ 13

When, for example, a benzyloxycarbonyl protecting group is used, the sulfonation reaction can be represented as follows: ________, * 2 R4 R2R-4 c6h5ch2oco-nh -: ”R3 ___ ^ H2N R3 1) sulfonation J__N_S0" M + 2) elimination q 3

protection dd III

Other protective groups can be used to protect the amine function, for example a t-butyloxy-carbonyl group, a simple acyl radical such as the acetyl or benzoyl or phenylacetyl radical, a triphenyl-methyl radical, or alternatively the amine function in the form of an azide function. The desired acyl radical (R ^) can then be attached by a conventional acylation reaction.

Exemplary acylation techniques for transforming a compound of formula III into a product of formula I include reaction with a carboxylic acid (R ^ OH), or with a carboxylic acid haloenide or a corresponding carboxylic acid anhydride . When R2 is an alkoxy radical, acylation is best carried out using an acid chloride or an acid bromide. The reaction with a carboxylic acid is most easily carried out in the presence of a carbodiimide such as dicyclohexylcarbodiimide, and a substance capable of forming an active ester in situ, such as N-hydroxybenzotriazole. In these cases, when the acyl radical (R ^ contains reactive functions (for example amine or carboxyl radicals), it may be necessary to protect these functional radicals first, then to carry out the reaction d acylation, and finally to remove the protection of the resulting product. Alternatively, the sulfonation reaction can be carried out with the acyl radical already in place, that is to say carry out the following reaction:

AT

14

• Ro R. RR

2 4 λ2 _4 r1 “NH - t-Rj. χ R] -NH - i-R ^ JL sulfonation '' - +

y) -NH ^ -N-S.O-.M

0 0 ’3

IV V

In the case where R2 is a lower alkoxy radical, there may be an additional variation in that one can introduce the radical R2 ~ lower alkoxy after the sulfonation as well as before the sulfonation using the mode conventional procedure consisting of chlorinating the acylated nitrogen atom which occupies position 3, then reacting the resulting product with a lower alcoholate, according to the reaction: ^ H (R2) alcoholate R2 R4 acyl-N - jr ^ - -acyl-NH - T “" R3 __N-SO ~ M + JN-SO, "M + 0 ^ 3 Ό 3

VI V

In the above reaction, the acyl groups also include easy-to-remove groups which act as protective groups and which can be removed afterwards. the reaction to obtain the product "deacylated" (-NH2).

Furthermore, the β-lactam nucleus can be formed by a cyclization reaction, and the sulfonation reaction can be carried out both before cyclization and after cyclization, that is to say carry out the reaction:>. ... R4.

acyl-NH - Cyclization acyl-NH- -r ^

. ; -NH-SO, ”m + i-N-S03 M

<J 3 0

VII

In this reaction, the acyl radical can also be an easy-to-remove group which acts as a protective group and which, after elimination, makes it possible to obtain ή // 20 the product having substituent -NH0.

15

The starting azetidinones in which R2 is a hydrogen atom and at least one of the substituents R R and is a hydrogen atom can also be prepared from amino acids having the formula: NH „° j * y / R4 XII CH - C.

| R3

.C-OH

0 '5 (at least one of R ^ and R ^ is a hydrogen atom). The amine radical is first of all protected using a conventional protective group, for example the t-butoxy-carbonyl radical (hereinafter designated by the abbreviation "Boc"). The carboxyl function of the protected amino acid is then reacted with an amine salt having the formula: XIII YO-NH ^ Cl "in which Y is an alkyl or benzyl radical, in the presence of a carbodiimide, to obtaining a compound having the formula: ÎV “4 XIV boc-nh-çh-c- ^

C ---: NH-0-Y

O * (at least one of R3 ’and R4 is a hydrogen atom). The hydroxyl group of a compound of formula XIV is transformed into a labile group (V) using a conventional reagent, for example methanesulfonyl chloride (the methanesulfonyl radical is designated below by the abbreviation " Ms "). Other labile substituents (V) which can be used are the benzenesulfonyl and toluene-tallow radicals and the chlorine, bromine and iodine atoms.

The fully protected compound is cyclized, having the formula: l / \ 1

V

16 BOONH-ÇH- XV ^ r3 *.

C-NH-O-Y

O '' (at least one of and R ^ is a hydrogen atom) by treating it with a base, potassium carbonate, for example. The reaction is preferably carried out in an organic solvent such as acetone, under reflux conditions, and a compound is obtained having the formula: * 4 BOC-NH-i-y-R3

XVI I

£ -N-O-Y

(at least one of R3 and R ^ is a hydrogen atom).

Alternatively, it is possible to cyclize a compound of formula XIV without first transforming the hydroxyl group into a labile group. Treatment of a compound of formula XIV with triphenylphosphine and diethylazodicarboxylate ,. gives a compound of formula XVI in which at least one of R3 and R ^ is a hydrogen atom.

The elimination of the protecting group from position 1 of an azetidinone of formula XVI can be achieved by using a reduction by sodium when Y is an alkyl radical, and this gives an intermediate of formula: BOC-NH -i -— R3

XVII

0 ^ -HH

(at least one of R3 and R4 is a hydrogen atom). If Y is the benzyl radical, the catalytic hydrogenation (using palladium on carbon for example) will initially give the corresponding N-hydroxylated compound which, by / J treatment with titanium trichloride, will give a / h 17 intermediate of formula XVII in which at least one of and R4 is a hydrogen atom.

The synthesis involving ring closure of the type described above leads to an inversion of the stereochemical configuration of the substituents R3 and R4.

As already mentioned, the above azetidinone can be sulfonated to form a compound of formula: Ê4 XVIII BOC-NH-i-ψ- R3 Jn-so3 ”m + (at least one of R3 and R ^ is an atom of hydrogen).

Another procedure for preparing a compound of formula I in which R2 is a hydrogen atom and. at least one of R3 and R ^ is a hydrogen atom, uses as starting material an amino acid amide having the formula:

OH

I / r4 NH2-CH- XIX R3 d> c-nh2 (at least one of R3 and R ^ is a hydrogen atom).

The protection of the amine group by a conventional protective group, the benzyloxycarbonyl radical for example (hereinafter designated by the letter Z) or by Boc, and the transformation of the hydroxyl group into a labile group (V) such as Ms, gives a compound of formula: OMs XX ANH-CH-4

I ^ R

0 * C-NH2 3 20 (at least one of R3 and R ^ is a hydrogen atom), A being a protective group.

The sulfonation of a compound of formula XX gives an 18 compound of formula: OMs f ^ -R4

ANH-CH - C

XXI I R3 +

Jn - NHSO "M

(at least one of R3 and R4 is a hydrogen atom).

The cyclization of a compound of formula XXI is carried out using a base, potassium carbonate for example.

The reaction is preferably carried out in a mixture of water and an organic solvent (for example a halogenated hydrocarbon such as 1,2-dichloroethane), under reflux conditions, and a compound of formula is obtained: - ---------------- 'R4 ANH —- T-R3

XXII

„__- N-SO ~ M + CK 3 (at least one of R3 and R4 is a hydrogen atom).

The removal of the protective group from a sulfonated azetidinone of formula XXII, in which A is a protective group, as well as equivalent compounds already described comprising an I-alkoxy radical, by catalytic hydrogenation, gives a compound of formula : U2 R4 h2n - f — R3 • 'XXI11 J_N-SO' Μ * cr 3 15 (at least one of R3 and R4 is a hydrogen atom), formula in which R2 is a hydrogen atom or a radical alkoxy, and this compound can be transformed into the corresponding zwitterion, having the formula: R2 * 4 XXIV NH3 - T “R3 'oJ-ν-303' 19 (at least one of R ^ and is a hydrogen atom) , by treatment with an acid such as formic acid.

Removal of the protective group from a sulfonated azetidi-none of formula XXII in which A is a protective group Boc using acid conditions (using for example formic acid) gives the corresponding zwitterion of formula XIV.

An excellent source of the starting ß-lactams are 6-aminopenicillanic acids and 7’-amino-10 cephalosporanic acids which may carry an optional 6-alkoxy or 7-alkoxy substituent respectively. These compounds have the formulas respectively: R9 2 s / CH_ R ..- NH-- XXVa I | ch3

-N-C-COOH

and JÎ3 s

XXVb r ^ ~ NH- X

J-CH3

COOH

where R3 is a hydrogen atom or an alkoxy radical and R ^ is a hydrogen atom or an acyl radical. By adapting the procedures described in the literature, 3-amino-2-azetidinones can be prepared; see for example Chem. Soc. Special Publication, No. 2.8, page 288 (1977);

The Chemistry of Penicillins, Princeton Univ. Press, page 257, and Synthesis, 494 (1977).

The first step is to desulfurize 6-amino penicillanic acid or 7-amino cephalosporanic acid, by reduction using Raney nickel. The reaction can be carried out in water under reflux conditions; the resulting compound has the following structural formula: l · 20 R2 R.-NH-- XXVI 1 J-N-CH-CH (CH-) - O | 32

COOH

Replacement of the carboxyl group of the compound of formula XXVI by an acetate group, followed by a hydrolysis, gives a 3-amino-3-alkoxy-2-azetidinone of formula I in which is a hydrogen atom or an acyl radical , R ^ is a hydrogen atom or a lower alkoxy radical, and R3 and are hydrogen atoms.,. The treatment of a compound of formula XXVI with cupric acetate and lead tetraacetate in a organic solvent (acetonitrile for example) makes it possible to replace the carboxyl group with an acetate group. The resulting compound can be hydrolyzed using potassium carbonate in the presence of sodium borohydride.

The introduction of a sulphonic group in position 1 of the above 3-amino-3-alkoxy-2-azetidinone can be carried out by reacting the intermediate with a complex of dimethylformamide and trioxide of sulfur.

A starting 3-azido-2-aetidinone can be prepared by first reacting an olefin of formula: R.

i4 XXVII ch2 = c — r3 with a halosulfonylisocyanate (preferably chloro-sulfonylisocyanate) having the formula: XXVIII 0 = C — N — S02-halogen to obtain an azetidinone of formula: R4 Z / -T R3 JN-SO- - halogen θ '2 h

XXIX

21 The reductive hydrolysis of an azetidinone of formula XXIX gives an unsubstituted ß-lactam on nitrogen, having the formula: R4 -f-R3

XXX

J-

For a more detailed description of the reaction sequence described above, reference can be made to the literature; see, for example, Chem. Soc. Rev., 5 ^, 181; (1976) and J. Org. Chem., 35, 2043 (1970).

An azide group can be introduced in position 3; an azetidinone of formula XXX (or its sulfonated equivalent) by reacting the compound with an arylsulfonylazide (such as toluenesulfonylazide) to obtain a starting azetidinone having the formula: R4 N3 R3

XXXI

ck-NH

The reaction works best if the nitrogen of the azetidinone is first protected by a silyl residue (for example t-butyldimethylsilyl, or t-butyldiphenylsilyl), then by giving rise to the anion in position 3 of the nucleus using a strong organic base (lithium diisopropylamine for example) at low temperature, then treating this anion with toluenesulfonylazide. The resulting intermediate 20 is stabilized with trimethylsilyl chloride, and then acid hydrolysis or solvolysis with a fluoride of the nitrogen protecting group gives the compound of formula XXXI.

Alternatively, the compound of formula XXXI can be obtained by first reacting a primary amine of formula: H2N “CH2" ^ y ^ 0 "alkyl or H2N - ^^ - 0-alky] e O-alkyl 22 with - an aldehyde of formula R ^ CH ^ O, to obtain the corresponding Schiff base. A cycloaddition [2 + 2] with an activated form of 3 * ~ azido-acetic acid gives a 3- ^ azido-2-azetidinone having the formula: r4 N3- | -Ï ~ R3 J-— ° 5 in which Q is a radical of formula: O-alkyl - ^ ~ ^ —0-alkyl O-alkyl Elimination of the substituent Q by oxidation gives the compound of formula XXXI.

The 3-acylamino-2-azetidinones can be obtained by first reducing a 3-azido-2-azetidinone of formula XXXI 10 to obtain the corresponding 3-amino-2-azetidinone, then by acylating the 3-amino ~ 2- azetidinone.

As already mentioned in the case where is a lower alkoxy radical, the product can be prepared from the equivalent product in which is a hydrogen atom. The chlorination of the nitrogen amide of a non-alkoxylated compound. gives an intermediate of formula:

Cl H Ra l - R. -N - j — R-, XXXII 1 3 J-N-SC> 3 ~ M +

Reagents and procedures for chlorinating amides on the nitrogen atom are known in the art. Exemplary reagents are tertiary butyl hypochlorite, sodium hypochlorite, and chlorine. The reaction can be carried out in an organic solvent (for example a lower alkanol such as methanol) or else ls \ 23 in · a two-phase mixed solvent (for example a mixture of water and methylene chloride) in the presence a base such as sodium borate decahydrate. The reaction is preferably carried out at reduced temperature.

The reaction of an intermediate of formula XXXI with an alkoxylating agent, for example with an alkali metal alcoholate, gives a product of formula I in which 1 * 2 is an alkoxy radical, in combination with its enantiomer. The reaction can be carried out in an organic solvent, for example in a polar organic solvent such as dimethylformamide, at a reduced temperature.

Another mode of synthesis for preparing the compounds of formula I in which is an alkoxy radical consists in first alkoxylating an intermediate of formula VI in which R ^ NH is a carbamate (R ^ is, for example, the benzyloxycarbonyl radical ) and R2 is a hydrogen atom, then to introduce a sulfonic group in position 1 of the resulting compound. The chlorination of a compound of formula VI using the procedure described above (for the chlorination of a non-alkoxylated compound of formula I in order to obtain a compound of formula XXXII) gives an intermediate of formula:

Cl R.

1 = 4 C H -CH.-O-CO-Νη- = f — Rn XXXIII Ö ^ 2 3 'J-N-Cl 0

Using the alkoxylation procedure described above (allowing a compound of formula XXXII to be transformed into a product of formula I), and then adding a reducing agent such as trimethylphosphite, the compound of formula XXXIII can be transformed in an intermediate of formula: O-alkyl R4

CcH-0-C0-NH --—— f-R,

XXXIV b 5 J

I-NH

/ Λ 24 in combination with its enantiomer.

The above procedures give the products of formula I in which is an alkoxy radical, in the form of a racemic mixture. If necessary, the R configuration enantiomer can be isolated from this racemic mixture, using conventional techniques such as fractional crystallization of an appropriate salt with an optically active organic amine, or by pairwise chromatography. ions, using an optically active cation.

Biological production of the antibiotic EM5117; The salts of (R) -3- (acetylamino) -3-methoxy-2-oxo-1-azetidinesulfonic acid can also be prepared (formula I, R 1 is an acetyl radical and R 2 is a methoxy radical; below by the code EM5117) by culture of a strain of the microorganism Chromobacterium violaceum SC 11.378 which has been deposited in the American Type Culture Collection (ATCC) under No. 31.532.

The microorganism The microorganism used for the production of EM5117 is a strain of Chromobacterium violaceum SC 11.378. A subculture of this microorganism can be obtained from the A.T.C.C.'s permanent collection in Rockville,

Maryland, United States of America. His access number in the 25 collection is A.T.C.C. N ° 31.532. In addition to the specific microorganism described and characterized here, it should be understood that 'mutant species of the microorganism (for example mutant species produced using X-rays, an,' ultraviolet radiation or 'mustard' nitrogen ") can also be grown to produce EM5117.

We can isolate Chromobacterium violaceum SC 11.378, A.T.C.C. N ° 31.532, of a wet soil sample containing the micro-organism, by first applying the soil sample to a medium containing: 35 Earth extract .... 400 ml

Distilled water ....... 600 ml

Yeast extract ... 5.0g

Glucose ............. 10.0 g Agar ............. 17.5 g i / ~ \ 25

This medium is adjusted to pH 6.0 and sterilized in an autoclave at 121 ° C for 20 minutes. After 24 to 72 hours of incubation at 25 ° C., soil applied to the slide is isolated from the colonies of Chromobacterium violaceUm SC "11.378.

5 These isolated colonies are then grown on a medium containing:

Yeast extract ... lg Beef extract .... 1 g NZ amine A ......... 2g 10 Glucose ............ 10 g Agar .... ......... 15 g

Distilled water to make 1 liter.

This medium is adjusted to pH 7.3 and treated in an autoclave at 121 ° C for 30 minutes.

15 Chromobacterium violaceum SC 11.378 is a Gram negative rod often showing "bars", bipolar staining and lipid inclusions. It can move with a single polar flagellum, sometimes with side flagella that are smaller.

20 On nutrient agar, Chromobacterium violaceum SC 11.378 produces purple colonies. Pigmentation is increased on media rich in tryptophan and yeast extract. In a nutritious broth, the microorganism produces a purple ring on the wall of the tube 25 but no confluent film. The violet pigment is soluble in ethanol but insoluble in water and * chloroform.

Chromobacterium violaceum SC 11.378 is mesophilic,. ’Growing over an interval of 15 to 37 ° C; no growth occurs at 4 ° C or above 37 ° C. Casein is strongly hydrolyzed by the microorganism, which is positive for oxidase. In the presence of Chromobacterium violaceum SC 11.378, glucose, fructose and trehalose ferment (method of Hugh & Leifson, 1953). L-arabinose is not used by the microorganism, either by fermentation or by oxidation. Hydrocyanic acid is produced by the microorganism, and hydrolysis by esculoside is negative.

J The fundamental characteristics described above, // Λ 26 form the basis for the identification of the micro-organism as being C hr omoh'ac ter ium y io1ac eum and not Chromob ac ter ium lividum, the only other species of the genus Chromobacterium which has been identified in the 8th edition of the "Manual of 5 Determinative Bacteriology" by Bergey.

Additional strains of Chromobacterium violaceum can also be grown to produce EM5117.

L1 antibiotic

The antibiotic EM5117 is obtained by growing '10 Chromobacterium violaceum SC 11.378, A.T.C.C. N ° 31.532, at 25 ° c or around 25 ° C, under submerged aerobic conditions, on an aqueous nutrient medium containing a source of assimilable carbon and nitrogen. Fermentation is carried out until substantial antibiotic activity has been imparted to the medium, i.e. usually for about 18 to 24 hours, preferably for about 20 hours.

Using the following procedure, EM5117 can be separated from the fermentation medium and purified. After fermentation is complete, the broth can be centrifuged to remove the mycelium, or filtration can be used to separate the mycelium from the broth. After separating the mycelium from the broth, EM5117 can be extracted from the broth. The broth is preferably extracted at pH 5 by extraction of ion pairs using cetyldimethylbenzylammonium chloride in methylene chloride, and is reextracted in aqueous sodium iodide (adjusted to pH 5 with l 'acetic acid). After concentrating the sodium iodide extract in vacuo, an aqueous solution of the residue can be washed with butanol. Concentration of the resulting aqueous solution to dryness gives a residue which is dissolved, as far as possible, in methanol. Centrifugation can be used to separate insoluble material, which is washed with methanol and discarded.

The purification of the antibiotic can be carried out by dissolving the methanol concentrate in a 1: 1 mixture of methanol and water and passing it through a column / chromatography, for example in a column comprising

/ rN

27 of "Sephadex G-10" in the same mixture of solvents. After elution with this same mixture, the active fractions are combined and concentrated to dryness. The residue is mixed with methanol and the insoluble material is filtered and then discarded.

A further purification is carried out by passing the material soluble in methanol through a column of DEAE-cellulose. This column can be eluted with a linear gradient prepared from 0.0 M sodium phosphate buffer 10 pH 5 and 0.1 M sodium phosphate buffer

of pH 5. The active fractions are combined, concentrated and separated and discarded, the substances insoluble in methanol.

Yet another purification of EM5117 is carried out by dissolving this substance in water and passing the solution through a column of "Sephadex LH-20", and eluting this column with water. The active fractions are combined and concentrated. A further purification of EM5117 is then carried out by dissolving the substance in water, passing the solution through a column of "Diaion HP-20AG" and eluting with water. The active fractions are combined and concentrated. The concentrate is dissolved in water and passed through a column of "Dowex 50W-X2", in potassium form, washing with water. The effluent can be concentrated to obtain a crystalline substance which is a relatively pure potassium sel of EM5117.

The process described above for isolation and purification therefore gives the potassium salt of EM5117. Other salts can be prepared corresponding to the form of the ion exchange resin used in the final purification step.

Biological production of the antibiotic EM521Q

It is also possible to prepare salts of (R) -3-35 [[N- (DY-glutamyl) -D-alanyl] amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid (formula I, is the radical Dy-glutamyl-D-alanyl and R2 is the methoxy radical, designated below by the code "EM5210") by culture of various / bacteria of acetic acid.

Λ 28

The microorganism

The microorganism Gluconobacter species SC 11.435 can be used for the production of EM52.10, A subculture of this microorganism can be obtained from “· the permanent collection of ATCC, in Rockville, Maryland , United States of America. His access number in the collection is A.T.C.C. N ° 31.581. In addition to the specific microorganism which is described and characterized here, it should be understood that it is also possible to cultivate mutant species of the microorganism (for example the mutant species produced using X-rays, radiation ultraviolet or "nitrogen mustard") to produce EM5210.

Gluconobacter species SC 11.435, A.T.C.C. No. 31.581, crushed foam containing the microorganism, first incubating the crushed foam in a 10% aqueous pectin solution (pH 2.5) for 7 days at 25 ° C. We can then isolate the micro-organism by applying it on a medium containing: * Spartina patens herb extract ..... 400 ml 20 Distilled water ................ .... 600 ml

Yeast extract ................. 5 g

Glucose ........................... 10 g Crude agar in flakes ......... 17.5 g * 0n prepared the Spartina patens herb extract by adding 500 g of chopped dry grass to 3 liters of tap water, bringing to the boil and cooking for 30 minutes.

The pH of the medium is adjusted to 6.0 and the medium is sterilized in an autoclave at 121 ° C for 20 minutes. After approximately 18 hours of incubation at 25 ° C., pectin enrichment solution is isolated on a slide from the colonies of Gluconobacter species SC 11.435. These isolated colonies are then grown on a medium containing:

Yeast extract ................ 1 g 35 Beef extract .................. 1 g NZ amine A .. ............... 2 g

Glucose ............ 10 g Agar .............. 15 g

Distilled water to make ............ 1 liter.

29

The medium is adjusted to pH 7.3 and treated in an autoclave at 121 ° C. for 30 minutes,

Gluconobacter species SC 11.435 is a pleomorphic Gram-negative rod which can move by 1 to 3 5 polar flagella. It must be aerobic, catalase positive and oxidizing. It differs from Pseudomonas in that it is negative for cytochrome oxidase and that it can withstand extremely acidic conditions. These characteristics indicate that the microorganism is more closely related to acetic acid bacteria than to true Pseudomonas.

On BBL trypticase soy agar, Gluconobacter, species SC 11.435, grows as a mixture of rough and smooth type colonies. The rough type is associated with a pale yellow soluble pigment, while the smooth type is mucoid and pigment-free. The dissociation of the two types is influenced by the conditions of environment, temperature and storage. The activity of EM5210 tends to decrease in cultures in which the rough component is predominant.

On Wart BBL agar (pH 4.8), Gluconobacter, species SC 11.435, grows luxuriantly in the form of piled mucoid muddy colonies. A similar growth is obtained on a malt agar-yeast extract 25 (1% each) adjusted to pH 4.5.

On a medium containing 1% yeast extract, 10% glucose, 3% calcium carbonate and 2.5% agar, a sufficient amount is produced from glucose to form a zone of disappearance of calcium carbonate around 30 'from the growth point. This is a characteristic feature of Acetobacter and Gluconobacter.

In the presence of Gluconobacter, species SC 11.435: (i) a soluble brown pigment is produced on agar plates of yeast-glycerol extract and yeast-calcium lactate extract, (ii) acid is produced on glucose, fructose, galactose, mannose, xylose, mannitol and arabinose, but no growth or production of acid occurs on rhamnose, lactose, sucrose or maltose, when these sugars are incorporated / (30 to a medium of Hugh & Leifson,

The biological production of EM5210 is not limited to Gluconobacter, species SC 11.435, but is widely distributed among all of the acetic acid bacteria. Each of the following cultures can be used for the production of EM5210:

Acetobacter pasteurianus subsp. pasteurianum A.T.C.C. 6033

Acetobacter aceti subsp. aceti A.T.C.C. 15973.

10 Gluconobacter oxydans subsp. oxidans A.T.C.C. 19357.

Gluconobacter oxydans subsp. suboxydans A.T.C.C. 23773

Gluconobacter oxydans subsp. oxidans A.T.C.C. 15178 Acetobacter aceti subsp. liquefaciens A.T.C.C. 23751 15 Acetobacter peroxydans A.T.C.C. 12874

Gluconobacter oxydans subsp. suboxydans A.T.C.C. 19441

Acetobacter sp. A.T.C.C. 21780 Gluconobacter oxydans subsp. industralis 20 A.T.C.C. 11894 L1 antibiotic

To obtain the antibiotic EM5210, you can grow · Gluconobacter, species SC 11.435, A.T.C.C. No. 31581, or one of the microorganisms listed above, at a temperature equal to or close to 25 ° C., under submerged aerobic conditions, on an aqueous nutritive medium containing an assimilable source of carbon and carbon 'nitrogen. The fermentation is carried out until substantial antibiotic activity has been imparted to the medium, that is to say usually for about 16 to 24 hours, preferably for about 20 hours.

Using the following procedure, EM5210 can be separated from the fermentation medium and purified. Once the fermentation is complete, the broth is centrifuged to remove the bacteria and the antibiotic is separated from the supernatant of the broth, at pH 3.7, by absorption on an anion exchange resin, for example on "Dowex 1 -X2 ".

The antibiotic is eluted from the resin using sodium chloride / about pH 4, and the eluate is concentrated, then passed through a carbon column. EM5210 is then eluted from charcoal with a 1: 1 mixture of methanol and water.

The active fractions are combined and dried, then they are placed on an anion exchange column, for example 5 "Dowex 1-X2", and elution is carried out with a gradient of sodium chloride buffered to pH 4. Concentration the active fractions and they are desalted on a column of macroreticular styrene-divinylbenzene copolymer resin ("Diaion HP20AG").

The active fractions eluted are concentrated with water and freeze-dried, and this substance represents the sodium salt of EM5210.

* The purification method described above therefore gives the sodium salt of EM5210. Other salts can be obtained by changing the salt used to elute EM5210 in the ion exchange chromatography described above, using, for example, potassium chloride to obtain the potassium salt.

The following examples are specific embodiments of the invention.

20 Example 1

(S) -N- (2-oxo-l-sulfo-3-a'zét'idinyl) -2- phenylacetamide potassium salt Method I: A) l - [(IR) -carboxy-2-methyl (propyl )] - 2-oxo- (3S) - [phenyl-25 [acetyl (amino)]] azetidine

Raney nickel is washed with water by decantation * for several hours until the pH of the water (5 to 6 times the volume of Raney nickel) is 7.6.

4.

• To a solution of 9.0 g of penicillin.G (Na) in 30 500 ml of water, 54 g (90 ml) of Raney nickel are added. The tank, fitted with a reflux condenser, is immersed in a 155 ° C. bath. When the reflux begins, it is continued for 15 minutes. The flask is immediately cooled in an ice-water bath, then the Raney 35 nickel is removed by filtration through "Celite". The pH is adjusted to 3 using dilute hydrochloric acid, and the aqueous solution is concentrated to approximately 150 ml, then cooled,

The oily layer crystallizes when the walls of the container are scraped off. After washing with water and drying under vacuum for 32 3 hours at δΟ’Ο, 3.83 g of the title compound are obtained.

B) 1- [acetyloxy-2-methyl (pr opylj] -2-oxo ~ (3S) ~ [phenyl * · [acetyl (amino)]] azetidine

Nitrogen is bubbled for 15 minutes into a stirred suspension of 608 mg (2 mmol) of the above compound in 20 ml of dry acetonitrile. A water bath at 40-45 ° C is used for several minutes to dissolve all of the acid. The water bath is removed, and powdered cupric acetate monohydrate (182 mg, 10 mmol) is added, then, after one minute of stirring, 886 mg (2 mmol) of lead tetraacetate. The mixture is stirred at room temperature for 20 minutes. The acetonitrile solution is decanted from the precipitate, and the solids are washed with ethyl acetate. The combined solution of acetonitrile and ethyl acetate is evaporated to a residue which is taken up in a mixture of ethyl acetate and water. The ethyl acetate layer is washed successively with water (3 times), with aqueous sodium bicarbonate (pH 7), and with water. The ethyl acetate layer was dried over sodium sulfate and concentrated to a residue (515 mg) which was used without further purification in the next reaction.

C) 2-oxo- (3S) - [phenyl [acetyl (amino)] jazetidine To a solution of 911 mg (2.86 mmol) of the above compound in 21 ml of methanol, 3.5 ml of d water, then 383 mg (2.86 mmol) of potassium carbonate. The mixture is stirred under nitrogen for one minute, then 160 mg (4.30 mmol) of sodium borohydride is added. The reaction mixture is stirred at room temperature for 20 minutes. The methanol is removed in vacuo, and the residue is taken up in ethyl acetate and a small amount of water. Then the pH is adjusted to 2.5. The ethyl acetate layer was washed at pH 7.0 with aqueous sodium bicarbonate and then with a small volume of water, and finally, it was dried over sodium sulfate and evaporated to obtain the crude product (493 mg). The addition of a small amount of ethyl acetate gives 250 mg (yield 43%) of the desired crystalline product. Additional quantities of L · product can be obtained by crystallization or chromatography.

Π 33 D) Potassium salt 'dtf (Sj -Kl · · (2-oxo ^ l-sulfo ^: 3-azetidinyl) -2-phenylacetamide

A pyridine ^ -SO ^ complex (215 mg; 1.35 mmol) is added to a stirred solution of 251 mg (123 'mmol) of the above product in 2 ml of dry dimethylformamide and 2 ml of methylene chloride dry, under nitrogen at room temperature. The mixture is stirred for 3 hours. The solvents are removed in vacuo, and the residue is taken up in a mixture of methylene chloride and water. The pH 10 is adjusted to 6.5 using 2N potassium hydroxide. The aqueous layer is washed with methylene chloride (3 times), filtered and concentrated to a residue. This residue is stirred with 20 ml of methanol, and the potassium sulfate is filtered off. The filtrate is concentrated to a residue which is stirred with 10 to 15 ml of methanol. The solids are collected to obtain 49 ml of the title compound, melting point 189 ° C, decomposition.

Analysis for N ^ O ^ SK:

Calculated: C, 40.99; H, 3.44; N, 8.69; S, 9.93 Found: C, 45.96 '; H, 3.83; N, 9.86; S, 8.99.

The compound has spectral characteristics identical to those of the product obtained in method II below. Method II:

A solution of 660 mg of potassium salt of (S) -2-oxo-3 - [[(phenylmethoxy) carbonyl] amino] -l-azetidine- is stirred in a hydrogen atmosphere for 2 hours. sulfonic (see Example 3) in 13 ml of water, with 200 mg of 10% palladium on carbon. The catalyst is filtered and the filtrate is diluted with an equal volume of acetone and then cooled in an ice bath. In 30 minutes, phenylacetyl chloride (eight fractions of 40 μl) and a 10% potassium bicarbonate solution (pH maintained between 5.2 and 5.8) are added. After 40 minutes, the solution is concentrated in vacuo to remove acetone and applied to a 200 ml column of "HP-20". Elution with water and then with a mixture 9: 1 of water and acetone, gives 160 mg of crude product after examination by thin layer chromatography of the positive fractions in the Rydon test, then // by collection and evaporation. Crystallization from a mixture of methanol and d ether gives 101 mg of the title compound, melting point 210 ° C, decomposition.

Analysis for C ^ H ^^ O ^ SK. ^ / 2 1 ^ 0:

Calculated: C, 39.86; H, 3.65; N, 8.45; S, 9.68; K, 11.80. Found: C, 40.01; H, 3.37; N, 8.59; S, 9.59; K, 11.98 NMR (D20) 3.66 (s, 3), 3.67 (d of d,) = 6.4), 3.90 (trj ~ j) r 4.90 (d of d, j = 6.4), 7.36 ppm (m.5).

Method .III

40 mg of dry methylene chloride are added to a solution of tetrabutylammonium salt of 10 (S) -3-amiho-2-oxo-1-azetidinesulfonic acid (121 mg; see Example 6A) in 3 ml of dry methylene chloride phenylacetic acid and 61 mg of dicyclohexylcarbodiimide. The mixture was stirred for 48 hours at room temperature and filtered to remove the dicyclohexylurea. The solvent is removed in vacuo and the residual compound is taken up in acetone, filtered and the title compound is precipitated by the addition of 5 ml of acetone saturated with potassium iodide. The supernatant is decanted, the residue is washed 3 times with acetone 20 and, after drying, 48 mg of product are obtained, the spectral characteristics of which are in agreement with those of the products of Methods I and II above.

Method IV:

A solution of 2.83 g of pyridine salt of (S) -2-oxo-3-C [(phenyl-methoxy) carbonyl] amino] -1-azetidinesulfonic acid is stirred in a hydrogen atmosphere. (see Example 2) in 36 ml of water, with 707.5 mg of 10% palladium on carbon (175 ml of hydrogen are absorbed). After 2 hours, the suspension is filtered and the filtrate is cooled to 0 ° C. and then diluted with 46 ml of acetone (initial pH = 4.25 adjusted to pH 6.7 with a cold bicarbonate solution 10% potassium). A solution of 2.4 ml of phenylacetyl chloride in 10 ml of acetone is added dropwise over 15 minutes. The pH is maintained between 5.2 and 5.8 by simultaneous addition of a cold 10% potassium bicarbonate solution. After 45 minutes, the suspension is diluted with 93 ml of potassium phosphate 0, 5M (pH = 4.2) and concentrated to remove the acetone. The suspension is filtered / filtered and washed with water. The filtrate and the washings are combined and passed through a 450 ml "HP-20" column. Elution with one liter of 0.5M potassium phosphate (pH = 4.2), one liter of water, then 2.5 liters of a 9: 1 mixture of water and acetone, gives 5 1.285 g of the title compound in fractions 14 to 19 (fractions 1 to 15 were 200 ml, fractions 16 to 21 were 100 ml). The spectral data are in agreement with those obtained in the previous methods. Example 2 10 Pyridine salt (1: 1) of (S) -2-oxo-3 - [[(phenyl-methoxy) carbonyl] amino] -1-azetidinesulfonic acid Method I: A) l - [(IR ) -carboxy-2-methyl (propyl) 3-2-oxo- (3S) - [[(phenyl-methoxy) carbonyl] amino] azetidine 15 A suspension of 6-aminopenicillanic acid is added all at once ( 12.98 g; 0.06 mole) in 140 ml of water containing 5.18 g of sodium bicarbonate (stirred for approximately 10 minutes without complete dissolution), to a well stirred suspension (mechanical stirrer) of Raney 20 nickel ( washed with water at pH 8.0, 260 ml of suspension = 130 g) in an oil bath at 70 ° C. After 15 minutes, the suspension is cooled, filtered, and the filtrate is treated with 5.18 g of sodium bicarbonate and with a solution of 11.94 g (0.07 mole) of benzyl chloroformate in 12 ml of acetone. After 30 minutes, the solution is acidified to pH 2.5 and extracted with methylene chloride.

The organic layer is dried, concentrated, and triturated with a mixture of ether and hexane to obtain one. 6.83 g total of the title compound.

B) 1 - [(acetyloxy) -2-methyl (propyl)] - 2-oxo- (3S) - [[(phenyl-methoxy) carbonyl] amino] azetidine

A solution of 6.83 g (0.0213 mole) of the above acid in 213 ml of acetonitrile is treated with 1.95 g (0.0107 mole) of cupric acetate monohydrate and with 9.5 g 35 (0.0213 mole) lead tetraacetate. The sus pension is immersed in an oil bath at 65 ° C and stirred by bubbling a stream of nitrogen through the suspension until the starting material is consumed. The suspension is filtered and the solids are washed with ethyl acetate / 36. The filtrate and the combined washing waters are concentrated in vacuo and the residue is taken up in 100 ml of ethyl acetate and 100 ml of water and the pH is adjusted to 7. _ The ethyl acetate layer is separated , dried and evaporated to give 6.235 g of the title compound.

C) Phenyl methyl ester of (S) - (2-oxo-3-azetidinyl) carbamic acid

A solution of 3.12 g (0.0093 mole) of the above acetate in 70 ml of methanol and 7 ml of water is cooled to -15 ° C., and 1.33 g of carbonate are added. potassium and 349 mg sodium borohydride. The reaction mixture is stirred at a temperature of from -15 ° C to 0 ° C. Once the reaction is complete (about 2 hours), the mixture is neutralized to pH 7 with 2N hydrochloric acid and concentrated in vacuo. The concentrate is adjusted to pH 5.8, saturated with salt and extracted with ethyl acetate (3 times). The organic layer is dried and evaporated in vacuo. The residue was combined with the substance from a similar experiment and triturated with ether to obtain 3.30 g of the title compound.

D) Pyridine salt of (S) -2-oxo-3 - [[(phenyl-methoxy) carbonyl] amino] -1-azetidinesulfonic acid Method I:

A solution of 440 mg (0.002 mole) of the above azetidinone in 2 ml of dry methylene chloride and 2 ml of dry dimethylformamide, * is stirred for 2 hours under nitrogen, with 350 mg (0.0022 mole) of 'a complex of pyridine and sulfur trioxide. Most of the solvent is then removed in vacuo and the residue is triturated with ethyl acetate to obtain 758 mg of solid, which consists mainly of the title compound.

NMR (D20-CD30D) 3.63 (1H, d of d,; = 6.4), 3.90 (1H, t,; = 6), 4.85 (1H, d of d,} = 6, 4), 5.10 (2H, S), 7.27 (5H, S) 8.0-9.0ppm (m1s 5H).

35 Method II:

18.87 g of chlorosulfonyltrimethylsilyl ester are added dropwise at -20 ° C. to 7.9 g of anhydrous pyridine, with stirring under a nitrogen atmosphere. Once the addition / λ is complete, stirring is continued for 30 minutes at room temperature, then the trimethylchloro-silane is removed in vacuo. A solution of 20 g of the above azetidinone (Method I, part C) in 120 ml of dimethylformamide and 120 ml of methylene chloride is added, and stirring is continued at room temperature for three and a half hours. The solvent is removed by vacuum distillation and the oily residue is crystallized by adding ethyl acetate thereto, giving 31 g of the title compound. The NMR data are identical to that of the product 10 obtained by Method I.

Example 3, Potassium salt of (S) -2-oxo-3 acid - [[(phenylmethoxy) - carbonyl] amino] -1-azetidinesulfonic acid - Method I: The acid pyridine salt (S ) -2-oxo ~ 3- [[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic (135 mg; see Example 2) in 2 ml of 0.5 M monobasic potassium phosphate (adjusted to pH 5.5 with 2N sodium hydroxide), and the solution is passed through a 25 ml column of "HP-20AG". This column is eluted with 100 ml of a buffer, 200 ml of water and 100 ml of a 1: 1 mixture of acetone and water. Fractions 14 and 15 (25 ml) are extremely positive on the Rydon test. Evaporation gives 80 mg of substance which consists mainly of the title compound (spectral data identical to that of the compound obtained by the method below).

Method II:

Pyridine salt of (S) -2-oxo-3- acid is dissolved. '[[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic 30 (600 mg; see Example 2) in 2 ml of water and the solution is mixed with 15 ml of a monobasic potassium phosphate buffer at pH 5.5. A solid forms, and the suspension is cooled to 0 ° C, filtered, washed with cold pad, cold 50% ethanol, ethanol and ether to obtain 370 mg of the title compound (containing an excess of potassium ion according to the analysis). A solution of 280 mg of this salt dissolved in 10 ml of water is passed through a 100 ml column of "ΉΡ-20". This column is eluted with 200 ml of water and then with a 9: 1 mixture of water and ΙΛ 38 of acetone. The fractions are collected (50 ml); the evaporation of fraction 7 gives a solid. Trituration with acetone, filtration and drying under vacuum gives 164 mg of the title compound, melting point 193-196 ° C.

5 Analysis for C-j ^ H ^^ OgSK. 1/21 ^ 0:

Calculated: C, 38.02; H, 3.48; N, 8.06; S, 9.23; K, 11.25 Found: C, 38.19; H, 3.24; N, 8.15; S, 9.12; K, 11.53. NMR (D20) 3.69 (1H, d of d, j = 6.4), 3.91 (1H, t, j = 6), 4.76 (1H, m), 5.16 (2H, S ), 7.43 ppm (5H, S).

10 Method III:

Is suspended in 200 ml of acetonitrile, phenylmethyl ester of (S) - (2-oxo-3-azetidinyl) - carbamic acid (20.0 g, see Example 2C), 21 is added. , 6 ml (25.3 g) monotrimethylsilyltrifluoracetamide, and the mixture is heated to 50 ° C with stirring for one hour.

After cooling in an ice bath to 0 ° C, 17.2 g of trimethylsilyl chlorosulfonate are added dropwise and the solution is stirred at room temperature for 6 hours. 24.2 g of potassium ethylhexanoate in 100 ml of butanol are added to the solution and stirring is continued for another hour. The suspension is poured into one liter of dry diethyl ether and the precipitate is separated by filtration and then dried under vacuum. The compound is dissolved in 500 ml of water, the pH is adjusted to 5.0 with potassium carbonate, the insoluble material is filtered off and the mother liquor is subjected to freeze-drying. The yield of the crude compound is 19.4 g. The compound contains small amounts of chloride. potassium which is eliminated by chromatography, the spectral data being identical to that of the product obtained by method II.

Example 4

Tetrabutylammonium salt (1: 1) of (S) -2-oxo-3- [[(phenylmethoxy) carbonyl] amino] -l-azetidinesulfonic acid 35 Method I:

The pyridine salt (1: 1) of (S) -2-oxo-3 - [[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid (34.3 g;) is dissolved in 800 ml of water. see Example 2). The solution is clarified with the carbon, 30.7 g / 39 of tetrabutylammonium hydrogen sulfate in 80 ml of water are added, and the pH is adjusted to 5.5 with IN potash. The solvent is removed in vacuo until the volume is no more than about 200 ml. The precipitated tetrabutylammonium salt is filtered off and dried in vacuo. The compound can be recrystallized from water or dissolved in methylene chloride, then filtered and precipitated by addition of ether. Yield 34.3 g, melting point 108-110 ° C.

10 Method II:

Dissolve the potassium salt (1: 1) of (S) -2-oxo-3 - [[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfoni-que acid (20.2 g; see Example 3) in 500 ml of water, filtered and 20.3 g of tetrabutylammonium hydrogen sulfate in 100 ml of water are added. The pH is brought to 5.5 with IN potash. The volume is reduced in vacuo to about 100 ml and the precipitated tetrabutylammonium salt is filtered off. The compound is dissolved in 30 ml of methylene chloride, filtered and precipitated by addition of ether, which gives 21 g of the title compound, melting point 109-111 ° C.

Example 5

Potassium salt (1: 1) of the phenylmethyl ester of (3S) -a acid - [[((2-oxo-1-sulfo-3-azetidinyl) amino] carbonyl] -25 benzene acetic A) (S) -3-amino-2-azetidinone; The phenylmethyl ester of (S) - (2-oxo-3-azetidinyl) carbamic acid (3 g is hydrogenated (3 g; see Example 2C). in 100 ml of methanol, in the presence of 1 g of palladium on carbon serving as catalyst. Once the theoretical amount of hydrogen has been absorbed, the catalyst is filtered off and the filtrate is concentrated to dryness. At rest, 1.1 g of the title compound crystallizes.

B) (3S) -g - [[(2-oxo-3-35 azetidinyl) amino] carbonyl] benzeneacetic acid phenylmethyl ester

The above azetidinone (3.0 g) is dissolved in 100 ml of dimethylformamide. The solution is cooled to 0 ° C. and 4.5 g of N-methylmorpholine are added, then (dropwise) / 10.8 g of a- (chlorocarbonyl) - Κλ phenylmethyl ester.

t · I '»4 40 benzene acetic acid in 50 ml acetonitrile, with stirring. The mixture is stirred for about 16 hours at 5 ° C. The solvent is removed by vacuum distillation and 100 ml of water are added to the residue. The aqueous suspension is extracted twice with 100 ml fractions of methylene chloride. The organic layers are combined, · they are washed with sodium bicarbonate, 2N phosphoric acid and water, they are dried over sodium sulphate, they are filtered and they are evaporated to dryness . The residue is crystallized from ethyl acetate and petroleum ether to give 8.7 g of the title compound, mp 164-166 ° C.

, C) Potassium salt (1: 1) of the phenylmethyl ester of (3S) -ot acid - [[(2-oxo-1-sulfo-3-azetidinyl) amino] carbonyl] - benzeneacetic 15 On suspends in 150 ml of acetonitrile 6.9 g of the above compound. 5.7 g of monotrimethyl-silyltrifluoracetamide are added and the solution is heated for 30 minutes at 50 ° C. with stirring. The solution is cooled to 0 ° C. and 3.9 g of 2.0 trimethylsilyl chlorosulfonate are added dropwise. Once the addition is complete, the mixture is heated at 50 ° C for 5 hours. After cooling to 20 ° C., 7.6 g of potassium ethylhexanoate in 10 ml of butanol are added and stirring is continued for 30 minutes. By the addition of 300 ml of ether, the title compound precipitates, and is separated by filtration. The crude product is stirred with 100 ml of dry acetonitrile for 30 minutes and separated by filtration, which gives 4.5 g of the title compound, melting point 118-120 ° C. A new purification of the crude product by chromatography on "HP-20", followed by a cryodessication, gives a pure substance including! the melting point is 188-190 ° C.

Example 6

Potassium salt of acid (S) -3 - [[(2-amino-4-thiazolyl) -. Acetyl] amino] -2-oxo-1-azetidinesulf onic 35 A) Tetrabutylammonium salt of acid ( S) -3-amino-2-oxo-1-azetidinesulfonic

Dissolve in 100 ml of dimethylformamide the tetrabutylammonium salt of (S) -2-oxo-3 - [[(phenyl- // methoxy) carbonyl] amino] -1-azetidinesulfonic acid (2 g; see Λ · 41 Example 4), and hydrogenated for about 30 minutes using as catalyst 1 g of palladium on carbon (10%). The catalyst is filtered off and the dimethylformamide is removed, leaving the title compound in the form of an oil.

NMR (CDCI 3) 3, 82 (1H, t, j = 5.5), 4.05 (d. 1H, d of d, j = 5.5, 2.5 cps).

B) Potassium salt of acid (S) -3 - [[(2-amino ~ 4-thiazolyl) -acetyl] amino] -2-oxo-1-azetidinesulfonic 10 Agitation is carried out at 0 ° C. in 100 ml of dry dimethylformamide 2 g of the above compound, 0.5 g of aminothiazole acetic acid and 0.4 g of hydroxybenzotriazole, at the same time as a solution of 0.7 g of dicyclo- is added dropwise hexylcarbodiimide in 10 ml of dimethylformamide. After the addition is complete, stirring is continued for 12 hours at 20 ° C. The insoluble urea is filtered off and the solvent is evaporated in vacuo. The oily residue is treated with a solution of potassium perfluorobutane sulfonate in 20 ml of acetone at room temperature for 15 minutes. After adding 200 ml of dimethyl ether, the title compound precipitates, it is separated by filtration, dried and purified on a chromatography column containing 300 ml of "HP-20", using water as eluent. The yield is 850 mg of the title compound 25, melting point> 300 ° C.

Example 7

Potassium salt of [3S (±)] - 3 - [[(formyloxy) phenyl-acetyl] amino] -2-oxo-1-azetidinesulfonic acid,. The tetrabutylammonium salt of (S) -3-amino-2-oxo-1-azetidinesulfonic acid (1.5 g; see Example 6A) is cooled to 0 ° C in 100 ml of dimethylformamide and 2 ml propylene oxide. A solution of O-formylmandelic acid chloride in 10 ml of acetonitrile is added dropwise, with stirring. The temperature is maintained for 1 hour and then the solvent is removed by vacuum distillation.

The oily residue is treated with a solution of 2 g of potassium perfluorobutane sulfonate in 15 ml of acetone. After adding 200 ml of ether, the title compound crystallizes, is filtered, giving 1.5 g of product. On, v \ 42 purifies the product by chromatography on "HP-20", m.p.

180-185 ° C, with decomposition.

Example 8.

Potassium salt of [3S (+)] - 3 - [[(formyloxy) phenyl-5-acetyl] amino] -2-oxo-1-azetidinesulfonic acid

Following the procedure of Example 7, but replacing the O-formylmandelic acid chloride by D-O-formylmandelic acid chloride, the title compound is obtained, melting point 120-125 ° C after cryodessication.

10 Example 9

Potassium salt of [3S (-)] acid - 3 - [[(formyloxy) phenyl-acetyl] amino] -2-oxo-1 -azetidinesul’f onique

Following the procedure of Example 7 but * replacing O-formylmandelic acid chloride with LO-formylmandelic acid chloride, the title compound is obtained, containing 1 mole of water, melting point 203-205 ° C. After careful drying, the product melts at 228-230 ° C.

Example 10

Potassium salt of (S) -2-oxo-3 - [(1-oxo-octyl) amino] -20 1-azetidinesulfonic acid

The tetrabutylammonium salt of (S) -3-amino-2-oxo-1-azetidinesulfonic acid (1.5 g; see Example 6A) is cooled to 0 ° C. in 100 ml of dimethylformamide and 2 ml of propylene oxide. At this temperature, a solution of 0.8 g of caprylic acid chloride in 20 ml of dry acetone is added dropwise, and stirring is continued * for 30 minutes. The solvent is evaporated in vacuo and the oily residue is treated with 2 g of potassium perfluorobutane • sulfonate in 15. ml of acetone. - Acetone is removed by vacuum distillation. The residue is dissolved in 5 ml of water and subjected to chromatography on 300 ml of "HP-20" resin, using a 9: 1 mixture of water and acetone as eluent, which gives 0.9 g of the title compound, melting point 173-180 ° C, after cryodessication.

35 Example 11

Potassium salt of [3S (Z)] acid - 3 - [[(2-amino-4-thiazolyl) - [[[hydroxy (phenylmethoxy) phosphinyl] methoxy] imino] acetyl ·] -amino] -2- oxo-1-azetidinesulfonic λ Stir for 24 hours at room temperature! r \ 43 the tetrabutylammonium salt of (S) -3-amino-2-oxo-1-azetidinesulfonic acid (0.8 g; see l 'Example 6A) in 30 ml of dimethylformamide, 0.9 g of (Z) -2-amino-a- [[[hydroxy (phenylmethoxy) phosphinyl] methoxy] imino'] .- 4-5 thiazoleacetic acid, 0, 3 g of hydroxybenzotriazole and 0.7 g of dicyclohexylcarbodiimide. The precipitated urea is filtered off and the solvent is removed in vacuo. The remaining oil is treated with an equivalent amount of potassium perfluorobutane sulfonate in 10 ml of acetone. The title compound is filtered off and purified on "HP-20" resin using water as the eluent to give 500 mg of product, melting point 210-215 ° C, with decomposition .

„Example 12 15 Potassium salt of [3S (Z)] - 3 - [[(2-amino-4-thiazolyl) - (ethoxyimino) acetyl] aminoj-2-oxo-1-azetidinesulfonic acid

The tetrabutylammonium salt of (S) -3-amino-2-oxo-1-azetidinesulfonic acid (1.5 g; see Example 6A) in 100 ml of dimethylformamide is stirred at room temperature for 24 hours. , 0.6 g of hydroxybenzotriazole, 1 g of dicyclohexylcarbodiimide and 0.8 g of (Z) -2-amino-α- (ethoxyimino) -4-thiazoleacetic acid. The solvent is distilled off and the residue is dissolved in 30 ml of acetone. The urea is filtered off and the mother liquor is treated with a solution of 2 g of potassium perfluorobutane sulfonate in 20 ml of acetone. After adding 200 ml of ether, the title compound precipitates, it is separated by filtration and dried. We purify. . cation by column chromatography of "HP-20" using water as eluent, to give 1.1 g of the title compound, melting point 180-185 ° C, with decomposition.

Example 13

Potassium salt of [3S (E)] - 3 - [[(2-amino-4-thiazolyl) (ethoxyimino) acetyl] amino] -2-oxo-1-azetidinesulfonic acid

Following the procedure of Example 12, but replacing (Z) -2-amino-a- (ethoxyimino) -4-thiazoleacetic acid with (E) -2-amino-a- (ethoxyimino ) -4-thiazoleacetic, the title compound is obtained, melting point 160-170 ° C after cryodessication.

Example 14

Potassium salt of [3S (Z) -3 - [[(2-amino-4-thiazolyl) - (2,2,2-trifluorethoxy) imino] acetyl] amino] -2-oxo-1-azetidine acid -5 sulfonic

Following the procedure of Example 12, but replacing the (Z) -2-amino-a- (ethoxyimino) -4-thiazoleacetic acid with (Z) -2-amino-a - [( 2,2,2-trifluorethoxy) imino] -4-thiazoleacetic, the title compound is obtained, melting point 160-170 ° C after cryodessication. Example 15

Potassium salt of (S) -2-oxo-3 - [(l ~ oxopropyl) amino] - 1-azetidinesulfonigue Method I: 15 Cooled to 0 ° C with stirring the tetrabutylammonium salt of (S) -3-amino-2-oxo-1-azetidinesulfonic acid (1.5 g; see Example 6A) in 100 ml of dry dimethylformamide and 4 ml of propylene oxide. At this temperature, 0.5 g of propionic acid chloride in 10 ml of acetonitrile is added dropwise and the mixture is stirred for 2 hours. The solvent is removed by vacuum distillation and the oily residue is treated with an equivalent amount of potassium perfluorobutane sulfonate in 5 ml of acetone. By addition of ether, the title compound crystallizes on filter, giving 0.8 g of product, melting point 135-140 ° C after freeze drying.

Method II:

The tetrabutylammonium salt of the acid is hydrogenated. (S) -2-OXO-3- [[(phenylmethoxy) c.arbonyl] amino] -1-azetidine-sulfonic (4 g; see Example 4) in 100 ml of diglyme with 1 g of palladium on coal as a catalyst. The hydrogenation is complete after 2.5 hours. The catalyst is filtered off and 2 ml of propylene oxide are added. After cooling to 0 ° C., 0.5 g of propionic acid chloride in 10 ml of dry diglyme is added with stirring. After 30 minutes, the solvent is removed in vacuo and the oily residue is treated with an equivalent amount of potassium perfluorobutane sulfonate in / 20 ml of acetone. After addition of ether, the compound of

V

/ k * 45 title crystallized, filtered and recrystallized from a mixture of water and acetone, which gives 0.9 g of product, melting point · 156 - 160 ° C (decomposition).

Example 16 5 Potassium salt of [3S (±)] - 3 - [(hydroxyphenylacetyl) -amino] -2-oxo-1-azetidinesulfonic acid

The tetrabutylammonium salt of (S) -3-amino-2-oxo-1-azetidinesulfonic acid (1.5 g; see example 6A) is stirred in 100 ml of dry dimethylforrnamide for approximately 16 hours with 1 , 5 g of dicyclohexylcarbodiimide, 0.5 g of hydroxybenzotriazole and 0.6 g of mandelic acid.

The solvent is removed in vacuo and the residue is dissolved in 20 ml of acetone. The precipitated urea u is filtered off and the mother liquor is treated with an equivalent amount of potassium perfluorobutane sulfonate. After addition of ether, the title compound precipitates, it is separated by filtration, which gives 1.4 g of crude product. After re-installation in water, the product has a melting point of 138-140 ° C.

20 Example 17

Potassium salt of (S) -3 - [[[(cyanomethyl) thio] -acetyl] amino] -2-oxo-1-azetidinesulfonic acid

The tetrabutylammonium salt of (S) -3-amino-2-oxo-1-azetidinesulfonic acid (1.5 g; see Example 6A) and 0.72 are dissolved in 70 ml of acetonitrile g of [(cyanomethyl) thio] acetic acid, and a solution of 1.04 g of dicyclohexylcarbodiimide in acetonitrile is added dropwise. The mixture is stirred for about 16 hours at 0 ° C. and the precipitated 30-tff. Dicyclohexylurea is filtered off, the filtrate is evaporated and the oily residue is dissolved in acetone. By adding a saturated solution of potassium iodide in acetone, the title compound is precipitated, which gives 1.1 g of product, melting point 150 - 155 ° C.

35 Example 18

Potassium salt of (S) -2-oxo-3 - [(1H-tetrazol-1-yl-acetyl) amino] -1-azetidinesulfonic A solution of 0.005.mole of tetrabutylammonium salt (S) -3-amino-2-oxo-1-azetidinesulfonic acid 46 (see Example 6A) in 70 ml of dimethylformamide, 0.77 g of ΙΗ-tetrazole-l-acetic acid and a solution of 1.13 g of dicyclohexylcarbodiimide in ml of dimethylformamide. The mixture is stirred for about 16 hours at room temperature and the solvent is removed in vacuo. The remaining oil is dissolved in 20 ml of acetone and treated with 0.006 mole of a solution of potassium perfluoro-butane sulfonate in acetone to give 1.5 g of the title compound, point of melting 170 - 175 ° C, 10 with decomposition.

Example 19 -i Potassium salt of (S) -2-oxo-3- [(2H-tetrazol-2-yl- * acetyl) amino] -1-azetidinesulfonic acid * Following the procedure of Example 18, but 15 by replacing 1H-tetrazole-1-acetic acid with 2H-tetrazole-2-acetic acid, the title compound is obtained, melting point 175-177 ° C with decomposition.

Example 20

(S) -2-oxo-3 - [(2-thienylacetyl) -20 amino] -1-azetidinesulfonic acid potassium salt

Following the procedure of Example 18, but replacing ΙΗ-tetrazole-l-acetic acid with 2-thiopheneacetic acid, the title compound is obtained, period; 180 - 190 ° C with decomposition.

25 Example 21

Potassium salt of [3S (Z)] - 3— Ç [(2-amino-4-thiazolyl) - * · (methoxyimino) acetyl] amino3 -2-oxo-1-azetidinesulfonic acid * Cool to 0 ° C the tetrabutylammonium salt of (S) -3-amino-2-oxo-1-azetidinesulfonic acid (prepared as described in Example 6A from 7.9 g of tetrabutylammonium acid salt ( S) -2-oxo-3 - [[(phenylmethoxy) -carbonyl] amino] -1-azetidinesulfonic, and 3.53 g of (Z) -2-amino-a- (methoxyimino) -4- acid are added thiazoleacetic, then a solution of 3.27 g of dicyclohexylcarbodiimide 35 in 10 ml of dimethylformamide. The mixture is stirred for 16 hours at 5 ° C., filtered and the solvent is removed in vacuo. The residue is dissolved in 1 ml. acetone and we filter.

By adding 60 ml of a 10% solution of potassium perfluorobutane sulfonate in acetone, 47 g 4.7 g of crude product are crystallized. This crude product is purified by chromatography on "HP-20", with a particle size 74 to 149 μ, to obtain 3.0 g of the title compound, melting point 235 ° C.

5 Example 22

(3S) -a - [[(2-oxo-1-sulfo-3-azetidinyl) amino] carbonyl] benzene acetic acid dipotassium salt

The potassium salt of the phenylmethyl ester of (3S) -a-10 [[(2-oxo-1-sulfo-3-azetidinyl) amino] carbonyl] benzeneacetic acid is dissolved in 20 ml of anhydrous methanol. mg; see Example 5). Palladium on carbon is added (10 mg, 10%), and the mixture is treated with the hydrogen gene for 15 minutes. The catalyst is filtered off, and the methanol is evaporated in vacuo. The residue is dissolved in 5 ml of water and the pH is adjusted to 6 with IN sodium hydroxide. After cryodessication, a crude product is obtained. This raw material is subjected to chromatography on resin "HP-20" (using water as eluent), which gives 60 mg of the title compound, melting point 80 - 85 ° C.

Example 23

Potassium salt of (S) -3- (acetylamino) -2-oxo-1-azetidinesulfonic acid Method I: 25 A) Potassium salt of (S) -3-amino-2-oxo-i acid -azetidinesulfonic

The potassium salt of (S) -3-, (benzyloxycarbonylamino) -2-oxo-1-azetidinesulfonic acid (169 mg; * see Example 3) is dissolved in 4.0 ml of water, and hydrogen on 30 37 mg of 10% palladium on carbon for 1 hour 40 minutes. The catalyst is filtered off and washed with 1 ml of 50% aqueous acetone.

B) Potassium salt of (S) -3-acetylamino) -2-oxo-1-azetidinesulfonic acid The above solution of the free amine is diluted with 3.5 ml of acetone and stir in an ice bath. 320 μl of acetyl chloride are added, in approximately 15 minutes, in small fractions, alternating the addition of 48 6.5 and 1.2. After 30 minutes, thin layer chromatography on silica gel in a mixture (19: 1) of acetone and acetic acid and the Rydon test indicate that the reaction is almost complete. 5 6 ml of 0.5M monobasic potassium phosphate buffer of pH 5.5 are then added, and the solution is acidified to pH 4.8 with 2N hydrochloric acid. The acetone is removed in vacuo and the aqueous solution thus obtained is passed through a column of 50 ml of "HP-20 AG" to obtain 2.197 g of a solid.

This solid is digested with methanol to obtain 282 mg of an extractable substance which still contains a little salt. The product is subjected to a further purification by passage through a "IRC-50" column, then by acidification to pH 3.8 with subsequent concentration to dryness 15 under vacuum. Trituration with acetone gives 64 mg of the desired product containing about 0.5 equivalent of mineral potassium salts. The final passage through a 200 ml column of "HP-20 AG", followed by lyophilization from 0.5 ml of water, gives 22 mg of product in the form of an amorphous powder than the dried under vacuum for 2 hours at 50 ° C, melting point 170-180 ° C after softening at 100 ° C.

Analysis for C5H705N2SK,

Calculated: C, 24.38; H, 2.87; N, 11.37; K, 15.9 Found: C, 26.06; H, 3.14; N, 9.96; K, 18.04.

Method II:

A solution of 2.0 g of pyridine salt ¢ - * of acid (S) -2 ~ oxo-3 - [[(phenylmethoxy) carbonyl] amino] -1-, azetidinesulfonic (see Example 2) is hydrogenated. in 25 ml of water, 30 in 500 mg of 10% palladium on carbon. After 2 hours, the solution is filtered, cooled to 0 ° C and 40 ml of acetone are added. The pH of the solution is maintained between 5.2 and 5.8 by simultaneous addition of acetyl chloride and a cold solution of 10% potassium bicarbonate. The pH of the solution is adjusted to 4.2 with acetyl chloride and the solution is concentrated in a rotary evaporator to remove the acetone. Chromatography on a 300 ml column of "HP-20 AG" (using water as eluent and taking 25 ml h 49 fractions) gives 900 mg of the title compound in fractions 13 and 14 , contaminated with a little potassium acetate. A new chromatography on "HP-20 AG" gives an analytical sample, melting point 205-210 ° C.

5 Analysis for Ct-H ^^ O ^ SK:

Calculated: C, 24.38; H, 2.86; N, 11.38; S, 13.02; K, 15.88 Found: C, 24.23; H, 2.81; N, 11.25; S, 12.86; K, 15.74 Example 24

(S) -2-oxo-3- [(phenoxyacetyl) -10 amino] -1-azetidinesulfonic acid potassium salt

The tetrabutylammonium salt of (S) -3-amino-2-oxo-1-azetidinesulfonic acid (1.5 g; ^ ’see Example 6A) in 100 ml of dry dimethylformamide is cooled to 0 ° C. We add 2 ml of propylene oxide, then, drop by drop, 1 g

V

Phenoxyacetyl chloride, with stirring. The reaction is finished after one hour. The solvent is removed in vacuo and the residue is treated with an equivalent amount of potassium perfluorobutane sulfonate in 20 ml of acetone. By addition of ether, the title compound (1 g) is crystallized, which is filtered off and dried. After recrystallization from boiling water, the title compound has a melting point of 176-180 ° C.

Example 25

Potassium salt of [3S (R *)] - 3 - [[[[[3 - [(2-furanyl-25-methylene) aminoj-2-oxo-1-imidazolidinyl] carbonyl] amino] -phenylacetyl] acid amino] -2-oxo-1-azetidinesulfonic

The tetrabutylammonium salt of the acid * (S) -2 ^ oxo-3 - [[(phenylmethoxy) carbonyl] amino] -1-azetidine- is hydrogenated

P

* sulfonic (3 g; see Example 4) in 100 ml of dimethyl-30 formamide with 1.5 g of palladium on charcoal as catalyst. After half an hour, the catalyst is filtered off and 1.8 g of dicyclohexylcarbodiimide, 2 g of (R) -a- [[[3 - [(2-furanylmethylene) amino] -] acid are added. 2-oxo-1-imidazolidinyl] carbonyl] amino] benzeneacetic and 0.9 g of hydroxybenzotriale. After 3 hours, the reaction mixture is concentrated to dryness, it is dissolved in 50 ml of dry acetone and the precipitated urea is filtered off. An equivalent amount of potassium perfluorobutane sulfonate is added to 20 ml of acetone, causing the title compound to precipitate. The crystallization is completed by adding 200 ml of ether. After filtration, the title compound is recrystallized from water, which gives a yield of 2 g, melting point 220-225 ° C, with decomposition.

Example 26

Potassium salt of [3S (R *)] acid - 2-oxo-3 - [[[[((2-oxo-l-imidazolidiny1) carbonyl3 amino 3 phenylacety1] amino] -1 -azetidinesulfonic 10 The salt is hydrogenated (S) -2-OXO-3 - [[(phenylmethoxy) carbonyl] amino] -1-azetidine sulfonic acid tetrabutylammonium (3 g; see Example 4) in 100 ml of dry dimethylformamide with 1.5 g of palladium on carbon as catalyst; the catalyst is filtered off after 30 minutes. 1.8 g of (R) -a - [[(2-oxo-1-imidazolidinyl) carbonyl] aminoIbenzene acetic acid, 1.3 g of dicyclohexylcarbodiimide and 0.9 g of hydroxybenzotriazole are added, and the solution is stirred for 2.5 hours. The solvent is removed in vacuo and the residue is dissolved in 50 ml of acetone. The precipitated urea is filtered off and the mother liquor is treated with an equivalent quantity of potassium perfluorobutane sulfonate in 20 ml of acetone. The title compound crystallizes and is filtered off after adding 200 ml of ether to give 1.8 g of product, melting point 210-215 ° C after recrystallization from a mixture of water and acetone.

Example 27

Potassium salt of [3S (Z)] acid - 3 - [[(methoxyimino) - • phenylacetyl] amino j-2-oxo-l-azetidinesulfonic 30 A) [3S (Z)] - 3 - [[( methoxyimino) phenylacetyl] amino] -2-azetidinone

3.58 g of (Z) -a- (methoxyimino) -benzene acetic acid are dissolved in methylene chloride, cooled to 5 ° C. and treated with a solution of 4.53 g of dicylo-35 hexylcarbodiimide in 50 ml of methylene chloride. The mixture is stirred for 30 minutes at 5 ° C and a solution of 1.72 g of 3-amino-2-azetidinone (see Example 5A) in 100 ml of methylene chloride is added. The reaction mixture is kept at 5 ° C for one hour and for two hours at room temperature. After removing the dicyclohexylurea by filtration, the filtrate is evaporated, which gives 6.6 g of crude product. This substance is purified by chromatography on 750 g of silica gel, 5 using as eluent a mixture (7: 3) of methylene chloride and ethyl acetate, which gives 2.9 e of product.

B) Potassium salt of [3S (Z)] acid - 3 - [[(methoxyimino) -phenylacetyl] amino] -2-oxo-1-azetidinesulfonic 10 0.5 ml of pyridine is dissolved in 5 ml of chloride of absolute methylene, the mixture is cooled to -30 ° C. and a solution of 0.93 ml of trimethylsilyl chlorosulfonate in 5 ml of methylene chloride is added. The mixture is stirred at room temperature for 30 minutes and is concentrated in vacuo to dryness. The residue is dissolved in 10 ml of dimethylformamide and treated at room temperature with a solution of 1.23 g of the above azetidinone in 10 ml of dimethylformamide. After stirring for 2 hours at room temperature, the solution is evaporated to dryness to obtain 2.1 g of crude pyridine salt of [3S (Z)] - 3 - [[(methoxyimino) acid. phenyl-acetyl] amino] -2-oxo-1-azetidinesulfonic.

Treatment of this pyridine salt with tetrabutylammonium hydrogen sulfate gives the corresponding tetrabutylammonium salt, which is extracted with methylene chloride and which remains in the form of an oil after evaporation.

The treatment of tetrabutylammonium salt with a. equimolar amount of potassium perfluorobutane sulfonate in acetone, evaporation, and treatment of the residue with ether gives 1.6 g of the title potassium salt, which is purified by chromatography on "HP-20 ".

Elution is carried out with a 90:10 mixture of water and acetone, which gives a product with a melting point of 220 ° C., with decomposition.

AT

52

Example 28

Potassium salt (1: 1) of the acid [3S (Zj] -3 ~ [[(2-amino - 4-thiazolyl) [[2- (diphenylmethoxy) -1, l-dimethyl-2 ~ oxo- ethoxy] -irnlno] acetyl] amino] -2-oxo'-l-azétidlnesulfonique 5 A solution of 0.005 mole of tetra-butylammonium salt of (S) -3-amino-2-oxo-l-azétidinonesulfo acid is treated. -nique (see Example 6A) and 0.006 mole of (Z) -2-amino-α acid - [[2- (diphenylmethoxy) -1, l-dimethyl-2-oxo-ethoxy] -imino] - 4-thiazoleacetic in 60 ml of dimethylformamide, per 0.7 g of hydroxybenzotriazole and 1.13 g of dicyclohexyl-carbodiimide The mixture is stirred for about 16 hours at room temperature, filtered and the filtrate is evaporated. The residue is dissolved in 30 ml of acetone, filtered and treated with 20 ml of a solution of potassium per-15 fluorobutane sulfonate 10% in acetone. After addition of petroleum ether, the title compound precipitates and is treated with ether and then filtered to obtain 3.8 g of product, melting point 190 ° C, with decomposit ion.

20 Example 29

[3S (Z)] - 3 - [[(2-amino-4-thiazolyl) [- 1-carboxy-1-methylethoxy) imino] acetyl] amino] -2-oxo-1-azetidinesulfonic acid dipotassium salt

The potassium salt of the acid [3S (Z)] - 3 - [[(2-amino-4-thiazolyl) [[2- (diphenylmethoxy) -1, 1 is suspended in 5 ml of anisole -dimethyl-2-oxo-ethoxy] imino] acetyl] -amino] -2-oxo-1-azetidinesulfonic (2g; see Example 28), and 25 ml of trifluoroacetic acid are added at -10 ° C. The mixture is stirred for 10 minutes at -10 ° C. . 100 ml of ether are added slowly at -10 ° C., then 50 ml of petroleum ether. The precipitate is filtered to obtain 1.6 g of the trifluoroacetic acid salt. This salt is suspended in 20 ml of water at 0 ° C., the pH is adjusted to 5.5 with diluted potassium hydroxide and purified on a column of "HP-20". The title compound is eluted with water, and it has a melting point of 225 ° C, with decomposition.

h 53

Example 30

Potassium salt of [3 S (Z)] acid - 3 - [[(2-amino-4-thiazolyl) [[2- (Æiphenylmethoxy) -2-oxo-ethoxy] imino] acetyl] -amino] - 2-oxo-1-azetidinesulfonic 5 Following the procedure described in Example 28, but replacing the acid (Z) -2-amino-a- [[2- (diphenyl-methoxy) -1, 1 -dimethyl-2-oxo-ethoxy] imino] -4-thiazoleacéti-que by acid (Z) -2-amino-a - [[2- (diphenylmethoxy) -2-oxo-ethoxy] imino] -4- thiazoleacetic, the title compound is obtained, melting point 180 ° C, with decomposition.

Example 31

Potassium salt of [3S (i)] - 3 - [(azidophenylacetyl) -amino] -2-oxo-1-azetidinesulfonic acid Method I: 15 'A) (±, S) -α-azido-N- (2-oxo-3-azetidinyl) benzeneacetamide

Dissolve (S) -3-amino-2-azetidinone (2.15 g; see Example 5A) and 2.1 g of sodium bicarbonate in 50 ml of a mixture (2: 1) of acetone and water. 5 g of (±) -a-azidobenzene-20 acetyl chloride dissolved in 10 ml of acetone are added dropwise, while maintaining the temperature between 0 and 5 ° C. and the pH at 6.8 with bicarbonate of sodium. After stirring for one hour, the acetone is distilled off and the remaining aqueous solution is adjusted to pH 8 with sodium carbonate, then extracted with three 50 ml portions of methylene chloride. Evaporation of the dried organic layers over sodium sulfate gives 3.6 g of the title compound, in the form of an oil which crystallizes after trituration with ether. Recrystallization from a mixture of methylene chloride and ether gives the title compound, melting point 97-100 ° C.

B) Potassium salt of [3S (±)] - 3 - [(azidiophenyl-acetyl) amino] -2-oxo-1-azetidinesulfonic acid

A solution of 2.45 g of the above azetidinone and 3 g of monosilyl trifluoroacetamide in 20 ml of acetonitrile is kept for one hour at 40 ° C. After cooling to 0 ° C., the solution is treated with 1.88 g of trimethylsilyl chlorosulfonate and the mixture is stirred for 5 hours / under argon. Finally, 6.12 ml of a 2N solution of potassium 2 54 2-ethylhexanoate in n-butanol is added and the stirring is continued for 45 minutes. The solution is poured into 300 ml of ether and the precipitate is separated by filtration. A filtered solution of 1.2 g of precipitate in a phosphate buffer (pH 5.5) is subjected to chromatography on 100 ml of "HP-20". The elution is carried out with: 1) 20 ml i of buffer; 2) 200 ml of water; 3) 200 ml of a mixture (9: 1) of water and acetone; 4) 200 ml of a mixture (3: 1) of water and acetone. The elution is checked by thin layer chromatography (Rydon silica test). 25 ml fractions are taken and from fractions 15 and 16 280 mg of the title compound are obtained. A second column chromatography of this substance gives 120 ml of the title compound, melting point 148 ° C, with decomposition.

15 Method IX:

Dissolve in 30 ml of acetonitrile the tetra-butylammonium salt of (S) -3-amino-2-oxo-1-azetidine-sulfonic acid (2.03 g; see Example 6A) and 0, 9 g of (±) -α-azidobenzene acetic acid, and a solution of 1.03 g of dicyclohexylcarbodiimide in 10 ml of acetonitrile is added.

The temperature is maintained at 0 ° C for one hour and at room temperature for 10 hours. After the resulting precipitate has been filtered off, the solvent is distilled off in vacuo and the oily residue is dissolved in 20 ml of acetone and then treated with 1.70 g of potassium perfluorobutane sulfonate in acetone . The addition of 10 ml of ether crystallizes the title compound melting point 149 ° C, with decomposition.

, "Method III: 2.5 2.5 g of o (-azidiophenylacetyl chloride) are added to a solution of 4.06 g of tetrabutylammonium salt of 3-amino-2 ~ oxo-1-azetidinesulfonic acid (see Example 6A) and 5 g of propylene oxide in 30 ml of acetonitrile After stirring for two hours, the solvent is removed by vacuum distillation and the oil residue is treated with an equivalent amount of potassium perfluorobutane sulfonate in acetone After addition of ether, the title compound crystallizes and is filtered, mp 148-149 ° C, with decomposition.

/ Λ 55

Example 32

Potassium salt of [3S (D)] -3 - [[[[[[(4-methoxy-phenyl) methoxy] carbonyl] amino] phenylacetyl] amino] -2-oxo-1-azetidinesulfonic 5 To a solution 2.03 g of tetrabutylammonium salt of 3-amino-2-oxo-1-azetidinesulfonic acid (see Example 6A) and 1.58 g of D-α acid - [[[(4 -methoxy-phenyl) methoxy] carbonyl] amino] benzeneacetic in 50 ml of acetonitrile, a solution of 1.03 g of dicyclohexylcarbodiimide in 10 ml of acetonitrile is added dropwise. The temperature is maintained at 5 ° C for one hour and at room temperature for 6 hours. Separation of the dicyclohexylurea formed and distillation of the solvent allows the title compound to be obtained in the form of an oily residue. Treatment of this oil in acetone with potassium perfluorobutane sulfonate and ether gives 2.4 g of product, melting point 108 - 111 ° C, with decomposition.

Examples 33 to 37 Following the procedure of Example 32, but replacing (D) -a - [[[(4-methoxyphenyl) methoxy] -carbonyl] amino] benzeneacetic acid with the compound indicated in column I below, the compound indicated in column II below is obtained.

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/ Λ 57

Example 38

Tetrabutylammonium salt of (3S) -3 - [- [[[(methoxy-thio) thioxomethyl] thio] phenylacetyl] amino] - 2-oxo-1-azetidinesulfonic acid 5 A solution of 1.03 g of (±) -ot- [[(methylthio) thioxomethyl] thio] benzeneacetic acid and 1.63 g of tetrabutylammonium salt of 3-amino-2-oxo-1-azetidinesulfonigue acid (see Example 6A) in 30 ml acetonitrile, with 0.8 g of dicylcohexylcarbodiimide dissolved in 10 ml of acetonitrile at -5 ° C. After having stirred for approximately 16 hours, the dicyclohexylurea formed is filtered off and the mother liquor is evaporated. The resulting oily residue is subjected to chromatography on a column of 400 g of silica using as eluent a mixture (8.5: 1: 0.5) of ethyl acetate, methanol and water, which gives 1.3 g of the title compound. Example 39

Potassium salt of 3 (S) -3 acid - [[[[(methylthio) thioxo-methyl] thio] phenylacetyl] amino] -2'-oxo-1-azetidinesulf onic The acetone is dissolved in acetone tetrabutyl ammonium salt of (3S) -3 - [- [[[(methylthio) thioxomethyl] -thio] phenylacetyl] amino-2-oxo-1-azetidinesulfonic acid (1.3 g; see Example 38), and the solution is treated with an equivalent amount of potassium perfluorobutane sulfonate. The addition of ether crystallizes the title compound, which is separated by filtration, to give 0.18 g of product, melting point 157 ° C, with decomposition.

Example 40 • "Potassium salt of [3S (D)] - 3 - [[[[(4-ethyl-2,3-dioxo-1-piperazinyl) carbonyl] amino] -2-thienylacetyl] amino acid ] - '2-oxo-l-azetidinesulfonique

3.25 g of (D) -ct- [[(4-ethyl-2,3-dioxo-1-piperazinyl) carbonyl] amino] -2-thiopheneacetic acid, 4.20 are dissolved in 25 ml of acetonitrile g of the tetrabutylammonium salt of 3-amino-2-oxo-1-azetidinesulfonic acid (see Example 6A) and 1.3 g of N-hydroxybenzotriazole. The dropwise addition of 2.06 g of dicyclohexylcarbodiimide dissolved in 10 ml of acetonitrile at -10 ° C takes 10 minutes and is continued. naked agitation for about 16 hours. The urea formed is separated by filtration, and the solvent is evaporated in vacuo. The remaining oil is dissolved in 50 ml of acetone and, after treatment with an equivalent amount of potassium perfluorobutane sulfate, the title compound 5 crystallizes, melting point 185-187 ° C, with decomposition. Example 41.

Potassium salt of [3S (±)] - 3 - [. (Bromophenylacetyl) -amino] -2-oxo-1-azetidinesulfonic acid

1.4 g of bromophenylacetyl chloride in 10 ml of acetonitrile are added dropwise to a solution of 5 mmol of tetrabutylammonium salt of 3-amino-2-oxo-1-azetidinesulfonic acid, and 3 g of propylene oxide in acetonitrile at 0 ° C. After stirring for three hours, the solvent is distilled off and the remaining oily residue is dissolved in 30 ml of acetone. Add the equivalent amount of potassium perfluorobutane sulfonate in acetone. The addition of ether crystallizes the title compound, melting point 135-137 ° C, with decomposition.

20 Example 42

Potassium salt of [3S (±)] acid - 3 - [[[(aminocarbonyl) -amino] -2-thienylacetyl] amino] -2-oxo-i-azetidinesulfonic Method I: To a solution of 2 g of tetrabutylammonium salt of (S) -3-amino-2-oxo-1-azetidinesulfonic acid and 1.5 g of propylene oxide in acetonitrile, 1.1 g of hydrochloride of (± ) -2-amino-4- (2-thienyl) -5 (4H) -oxazolone. After stirring for three hours at 0 ° C. and for one hour at room temperature, the solvent is distilled off and the oily residue is dissolved in 30 ml of acetone. By adding the equivalent amount of potassium perfluorobutane sulfonate in acetone, the title compound is crystallized. Purification by column chromatography of "HP-20", using water as the eluent, gives the title compound, mp 218-222 ° C, with decomposition.

Method II: To a suspension of 2 g of (±) -a-t (aminocarbonyl) amino] -2-thiopheneacetic acid, 10 mmol of E ^ 59 tetrabutylammonium salt of (S) -3-amino acid -2-oxo-1-azetidinesulfonic acid and 100 ircnoles of N-hydroxybenzotriazole in 50 ml of acetonitrile at 0 ° C., 10 mraoles of dicyclohexylcarbodiimide dissolved in 15 ml of 5-acetonitrile are added (with stirring). Stirring is continued for one hour at -15 ° C and for about 16 hours at room temperature.

After the dicyclohexylurea has been filtered off, the solvent is evaporated off and the oily residue is dissolved in 50 ml of acetone. Addition of the slow equivalent amount of potassium perfluorobutane sulfonate in acetone gives a crystalline product. Purification by column chromatography on "HP-20", using water as the eluent, gives the title compound, mp 220-223 ° C.

15 Example 43

Potassium salt of [3S (±)] - 3 - [[[[(methylamino) -carbonyl] amino j-2-thienylacetyl] amino] -2-0x0-1-azetidine sulfonic acid

0.54 g of 20 (±) -a - [[(methylamino) carbonyl] amino] -2-thiopheneacetic acid and 1.00 g of tetrabutylammonium salt of (S) acid are dissolved in 20 ml of acetonitrile -3-amino- 2-oxo-1-azetidinesulfonic (see Example 6A), and 0.5 g of dicyclohexylcarbodiimide is added at a temperature of 0 ° C. After stirring for 8 hours, the dicyclohexylurea is filtered off and, after removing the solvent by distillation, the oily residue is dissolved in 50 ml of acetone and the equivalent amount of potassium perfluorobutane sulfonate is added. We separate it. crystalline product by filtration and purification is carried out on "HP-20" using water as eluent, melting point 205 ° C, with decomposition.

Example 44

Potassium salt of [3S (±) 3 —3 - [[[(amino-oxoacetyl) -amino] -2-thienylacetyl] amino] -2-oxo-1-azetidinesulfonic acid 35 Following the procedure of 'Example 42, method II, but replacing (±) -a - [(amino-carbonyl) amino] -2-thiopheneacetic acid with (±) -a - [(amino-oxoacetyl) amino] - 2-thiopheneacetic, the * the title compound is obtained, melting point 218-222 ° C.

//AT

60

Example 45

Potassium salt of the acid [3S (R *)] -3- [[[[((4-ethyl-2,3-dloxo-1-pipëraz iny1) carbony1] amino] phenylacéty1] amino] -2-oxo- l-azetidinesulfonique 5 Following the procedure of Example 40, but replacing the acid (D) -a - [[(4-ethyl-2,3-dioxo-1-piperazinyl) carbonyl] amino] -2 -thiophene acetic acid (R) —a— [[(4-ethyl-2,3-dioxo-1-piperazinyl) carbonyl] amino] -benzene acetic, the title compound is obtained, melting point 155-157 ° C, with decomposition.

Example 46

Potassium salt of 3- (acetylamino) -3-methoxy-2- oxo-1-azetidinesulfonic acid Method I: 15 A) Mixed sodium and potassium salt of 3 - [(N-acetyl-N -chloro) amino] -2 ~ oxo-l-azetidinesulfonic A solution of potassium salt of 3- (acetylamino) acid -2-oxo-l-azetidinesulfonic (172 mg, see Example 23) in 17 ml of methanol containing 4% of sodium borate decahydrate cooled between -15 ° and -10 ° C, 110 μl of tert-butyl hypochlorite are added. The mixture is stirred cold for one hour and 45 minutes, poured into 50 ml of a 0.5M monobasic potassium phosphate solution, and the pH is lowered to 5.5. The solvent is removed in vacuo and the residue is dissolved in a minimum quantity of water, then it is subjected to chromatography on "HP-20 AG", with a particle size 74 to 149 μ (140 ml). Elution with water gives an oil (63 mg) which crackles. tallise at rest. Trituration with a mixture of 30 methanol and ether, then with ether alone gives a solid (53 mg), melting point 124 ° C, slow decomposition. B) Potassium salt of 3- (acetylamino) -3-methoxy-2-oxo-1-azetidinesulfonic acid

Dissolve in 1.5 ml of dry dimethylformamide 35 37 mg of the mixed salt of 3 - [(N-acetyl-N-chloro) amino] - 2-oxo-1-azetidinesulfonic acid and the solution is added to a solution. 50 mg of lithium methylate in 1 ml of methanol at -78 ° C. After having stirred for 15 minutes at -78 ° C., 10 ml of a 0.5 M solution of monobasic potassium phosphate 61 is added, then the solution is acidified to pH 4 with IN hydrochloric acid. To this solution is added 70 mg of tetrabutylammonium hydrogen sulfate and the solution is extracted four times with methylene chloride. The combined extracts are dried over sodium sulfate and the solvent is removed in vacuo to obtain 55 mg of an oil. Chromatography on 5.5 g of silica gel gives the oily product (41 mg) in the form of the tetrabutylammonium salt (by elution with a mixture of 8% methanol and 92% methylene chloride. oil (31 mg) in water and passed through an 0 ion exchange column (5 ml of "AG 50W-X2", form K, particle size 37 to 74 y, 0.6 meq / ml) Removal of the water under vacuum gives an oil which is crystallized from a mixture of methanol and ether. By triturating twice with ether, the product is obtained in the form colorless powder (11 mg); melting point 182-183 ° C, decomposition.

Method II: 20 A) Tetrabutylammonium salt of 3-amino-3-methoxy-2-oxo-1-azetidinesulfonic acid

A 4% solution of sodium borate decahydrate in 100 µl methanol is added to a suspension of 30 mg of 10% palladium on charcoal in 2 ml of methanol, and the mixture is stirred under an atmosphere of hydrogen for 15 minutes. Tetrabutylammonium salt of 3-methoxy-2-oxo-3 - [[(phenylmethoxy) carbonyl] amino] - 1-azetidinesulfonic acid (60 mg; see Example 49) is added in 2 ml. of methanol, and the mixture is stirred vigorously for 30 minutes under a hydrogen atmosphere. The catalytic converter is separated by filtration on "Celite" on a "Millipore" filter (0.5 μm), and the solvent is removed from the filtrate under vacuum, then the residue is extracted with methylene chloride. Removal of the solvent under reduced pressure gives 35 mg of the title compound as an oil. B) Potassium salt of 3- (acetylamino) -3-methoxy- 2- oxo-1-azetidinesulfonic acid To a solution of 35 mg of tetrabutylammonium salt ^ / "j of 3-amino-3-methoxy acid -2-oxo-1-azetidinesulfonic acid in 62 10 rtil of methylene chloride at 0 ° C., 2 ml of propylene oxide and 74 μl of acetyl chloride are added. After 2 hours, the solvent is removed under reduced pressure and the residual oil is subjected to chromatography on 5 g of silica gel. Elution with 6 to 8% of methanol in methylene chloride gives 18 mg of an oil which is dissolved in water and passed through an ion exchange resin (3 ml of "AG 50W-X2", form K, 0.6 meq / ml). eluate water under vacuum gives the desired product (10 mg).

Example 47

N- (3-methoxy-2-oxo-1-sulfo-3-azetidinyl) -2-phenylacetamide tetrabutylammonium salt A) N-chloro-N- (2-oxo-1-sulfo-3-) potassium salt Azetidinyl) -2-phenylacetamide To a solution of potassium salt of (S) -N- (2-oxo-1-sulfo-3-azetidinyl) -2-phenylacetamide (50 mg? See Example 1) in methanol containing 4% sodium borate decahydrate (5 ml) cooled in a bath to -5 ° C, 30 µl of tert-butyl hypochlorite is added. After stirring for 32 minutes, the mixture is poured into a 0.5 M potassium phosphate buffer at pH 5.5 at 0 ° C.

The pH of the resulting solution (5.9) is adjusted to 4.5, concentrated in vacuo to remove methanol, and subjected to chromatography on "HP-20 AG", 74-149%. (100 ml). After washing the column with 100 ml of 0.5 M pH 5.5 buffer and water, the desired product is eluted with a 9: 1 mixture of water and acetone. A concentration. vacuum tration gives 50 mg. of the title compound.

B) Tetrabutylammonium salt of N- (3-methoxy-2-oxo-1-sulfo-3-azetidinyl) -2-phenylacetamide To a stirred solution of 160 mg of lithium methylate in 5 ml of methanol cooled to -78 ° C, a solution of 149 mg of potassium salt of N-chloro-N-35 (2-oxo-1-sulfo-3-azetidinyl) -2-phenylacetamide in 10 ml of dry dimethylformamide is added. When the addition is complete, the mixture is stirred for 15 minutes at -78 ° C, poured into 100 ml of a 0.5 M mono-basic potassium phosphate solution, and washed three times with methylene chloride 63. 213 mg of tetrabutylammonium bisulfate are added to the aqueous layer, which is then extracted three times with methylene chloride. The combined extracts are dried over sodium sulfate, and the solvent is removed in vacuo to give 271 mg of an oil. Chromatography of this oil on 25 g of silica gel and elution with a mixture of 4% methanol and 96% methylene chloride gives 149 mg of the product in the form of an oil.

10 Example 48

N- (3-methoxy-2-oxo-1-sulfo-3-azetidinyl) -2-phenylacetamide potassium salt

The tetrabutylammonium salt of N- (3-methoxy-2-oxo-1-sulfo-3-azetidinyl) -2-phenylacetamide (91 mg; see Example 47) is dissolved in water. the solution is passed through an ion exchange column (10 ml of $ "AG 50W-X2", form K). The eluate is concentrated in vacuo and, scraping the walls of the container, the residual oil in a mixture of methanol, acetone 20 and ether After having been triturated twice with ether, the product is obtained in the form of a solid (53 mg): V 1762, 1665 cm " 1; NMR (CD, OD) <5 3.41 (S, 3H, OCH,), ΓΠ.3.Χ Jj 3 3.59 (S, 2H, CH2), 3.84 (ABq, J = 6, 3 Hz, 2H, H4), 7.30 (m, 5H, aromatic).

25 Analysis for ci2Hi3N206'5H2 °:

Calculated: C, 39.88; H, 3.62; N, 7.75 Found: C, 39.62; H, 3.65; N, 7.60.

Example 49 'Tetrabutylammonium salt of 3-methoxy-2-oxo-3- [[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid Method I: A) Tetrabutylammonium salt of 2-oxo acid 3- [N-chloro-N - [(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic Add the tetrabutylammonium salt of 35 (S) -2-0X0-3 acid - [[(phenylmethoxy) carbonyl] amino ] -l-azetidine-sulfonic (0.9 g; see Example 4) dissolved in 80 ml of methylene chloride, to a mixture (cooled to 0 - 5 ° C) of 3.17 g of sodium borate decahydrate and 11.8 ml of a 5.25% sodium hypochlorite solution in 70 ml K \ 64 of water. The reaction mixture is stirred vigorously for 55 minutes while cooling in an ice bath. After diluting the mixture with a solution of 0.5 M monobasic potassium phosphate, the product is extracted with three fractions of 150 ml of methylene chloride. The grouping of the extracts, drying over sodium sulfate and elimination solvent in vacuo give the title compound as an oil (0.94 g).

10 B) Tetrabutylammonium salt of 3-methoxy-2-oxo-3 - [[(phenylmethoxy) carbonyl3amino] -l-azetidinesulfonic A to a stirred solution of 667 mg of lithium methylate in 10 ml of anhydrous methanol to - At 78 ° C., a solution of 0.94 g of tetrabutylammonium salt of 2-ΟΧΟ-3- [N-chloro-N - [(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid in 10 ml is added. dry dimethylformamide. After the mixture was stirred at -78 ° C for one hour, it was poured into a 0.5M monobasic potassium phosphate solution and extracted with three 150 ml fractions of methylene chloride. The combined extracts are dried over sodium sulfate and the solvent is removed in vacuo to obtain 0.83 g of an oil. The desired product is obtained by subjecting this oil to chromatography on 100 g of silica gel and eluting with 4-5% methanol in methylene chloride to obtain 513 mg of an oil: V (net) 1767, 1720 cm “1; BMN (CDC1.J δ 3.40 (S, 0CH ~), ΙΠ3.Χ ό ό 3.03 (ABq, J = 6.5 Hz, H ^), 5.08 (S, CH2), 6.00 (S, NH), 7.27 (S, aromatic).

Method II; To a solution of 400 mg of potassium salt of 3-benzyloxycarbonylamino-3-methoxy-2-oxo-1-azetidinesulfoni-that in water, a solution of 0.1 M tetrabutylammonium bisulfate (10 , 9 ml, adjusted to pH 4.3 with potassium hydroxide). The mixture is extracted three times with methylene chloride, the extracts are combined, they are dried over sodium sulfate and the solvent is removed in vacuo to obtain 625 mg of a foam whose fsPectral characteristics are roughly those of the product obtained by 65 method I.

Example 50

Potassium salt of 3-methoxy-2-oxo-3-tQphenyl-methoxy) carbonyllamlnoTl-1-azetidinesulfonlque 5 Method I: A) Potassium salt of 2-oxo-3- [N-chloro- N - [(phenylmethoxy) carbonyl ·] amino] -1-azetidinesulfonic A solution of the potassium salt of (S) —2— oxo-3 - [[(phenÿlmethoxy) carbonyl] amino] -l-azetidinesulfonic 10 (1.00 g; see Example 3) in 90 ml of inethanol containing 4% sodium borate decahydrate at -10 ° C, 420 μΐ of tert-butyl hypochlorite are added. After stirring for 2 hours at -10 ° C., 100 ml of a 0.5M monobasic potassium phosphate solution are added and the pH is adjusted to 6 with 1N hydrochloric acid. After concentrating the solution under vacuum at 30 ml, the aqueous solution is subjected to chromatography on "HP-20 AG", 74 to 149 μ (200 ml). After passing a solution of 50 g of monobasic potassium phosphate in 1000 ml of water, then 20 000 ml of water, the product is eluted with a mixture of 10% acetone and 90% water . The solvent is removed in vacuo and the title compound is crystallized from water to obtain 530 mg of a solid, melting point 173-175 ° C.

B) Potassium salt of 3-methoxy-2-oxo-3 - [[(phenyl-methoxy) carbonyl] amino] -1-azetidinesulfonic To a solution of 874 mg of lithium methylate in 10 ml of methanol dry at -78 ° C, 857 mg of potassium salt of 2-oxo-3- [N-chloro-N - [(phenylmethoxy) -, carbonyl] amino] -1-azetidinesulfonic acid are added in 13. ml of 30 dimethylformamide dry. After 15 minutes at -78 ° C, the mixture is poured into 200 ml of a 0.5M monobasic potassium phosphate solution and the pH is adjusted to 5.5 with IN hydrochloric acid. The aqueous mixture was washed with three 100 ml portions of methylene chloride, and 1.169 g of tetrabutylammonium bisulfate was added. The product is extracted with three 200 ml fractions of methylene chloride, dried over sodium sulfate, filtered and concentrated in vacuo. The oil / residual is subjected to chromatography on 150 g of silica gel Λ 66 and the product is eluted with 2 to 4% methanol in methylene chloride / which gives the tetrabutylammonium salt of the product under the form of an oil (701 mg).

Part of this oil (51 mg) is dissolved in water 5 and this solution is passed through an ion exchange column (3 ml of "AG 50W-X2", 37 to 74 y, form K ®, 0.6 meq / ml). The concentration of the eluate under vacuum gives 30 mg of an oil which is crystallized by scraping the walls of the container with acetone: 10 V (KBr) 1760, 1725 cm "1; NMR (D00) δ 3.48 (S, 3H, OCH.), ΙΓ13.Χ 3 3.92 (S, 2H, H4); 5.20 (S, 2H, CH2), 7.42 (S, 5H, aromatic; melting 196-198 ° C.

Method II: A) 1-chloro-3- [N-chloro-N - [(phenylmethoxy) carbonyl] -15 2-azetidinone

A solution of 3 - [[(phenyl-methoxy) carbonyl] amino] -2-azetidinone (440 mg; see Example 2C) in 40 ml of 4% methanolic borax is cooled to 0 ° C., and 0 is added. , 5 ml of t-butyl hypochlorite. After 20 minutes at 0 ° C, the solution is poured into 200 ml of cold water and extracted with two fractions of 100 ml of ethyl acetate. The combined ethyl acetate layer is washed with water, dried, and concentrated in vacuo to give 546 mg of the title compound as an oil.

B) 3-methoxy-3 - [[(phenylmethoxy) carbonyl] amino] -2-azetidinone

Cooled to -78 ° C a solution of 730 mg (0.0025 mole) of 1-chloro-3- [N-chloro-N - [(phenylmethoxy) carbonyl] -30 amino] -2-azetidinone in 5 ml of tetrahydrofuran, and 4 ml of methanol containing 285 mg of lithium methylate are added. After 20 minutes at -78 ° C, 0.6 ml of acetic acid and 0.6 ml of trimethylphosphite are added. The solution is stirred for 5 minutes at -78 ° C, allowed to warm to room temperature and stirred for 30 minutes. The resulting solution is diluted with ethyl acetate, washed with 5 I sodium bicarbonate, water, 5% potassium bisulfate, water, saturated saline, and we dry it.

The removal of the solvent gives an oil which is applied to four plates of silica gel 20 × 20 cm × 1 mm.

Development with a (1: 1) mixture of benzene and ethyl acetate and isolation of the main active band 5 in the ultraviolet, of Rf = 0.25, gives 91 mg of an oil that l 'We crystallize in ether to obtain a solid. Recrystallization from ether gives the title compound, mp 112-114 ° C.

C) Potassium salt of 3-methoxy-2-oxo-3 - [[(10-phenyl-methoxy) carbonyl3amino] -1-azetidinesulfonic

A solution of 25 mg of 3-methoxy-3 - [[(phenylmethoxy) carbonyl] amino] -2-azetidinone in 0.175 ml of dichloromethane and 0.175 ml of dimethylformamide with 55.4 mg of is stirred for 24 hours. a complex of pyridine and sulfur trioxide. The resulting suspension is diluted with 5 ml of cold 0.5 M monobasic potassium phosphate (adjusted to pH 4.5) and extracted with ethyl acetate. The aqueous layer is passed through a 40 ml column of "HP-20 AG". Elution with additional buffer, water and a (9: 1) mixture of water and acetone gives 32 mg of the title compound as an oil which slowly solidifies. Crystallization from acetone gives the title compound, melting point 196 - 198 ° C, with decomposition.

Example 51 potassium salt of 3 - [[1,3-dioxo-2-phenyl-3- (phenylmethoxy) propyl] amino] -3-methoxy-2-oxo-1-azetidine sulfonic acid A) Salt 3 - [[1,3-dioxo-2-30 phenyl-3- (phenylmethoxy) propyl] amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid tetrabutylammonium acid

The tetrabutylammonium salt of crude 3-amino-3-methoxy-2-oxo-1-azetidine sulfonic acid (431 mg, see Example 74), containing borax, is dissolved in 30 ml of dry acetonitrile. 317 µl of dry pyridine is added and the solution is carefully stirred at -10 ° C under dry nitrogen.

568 mg of a- (benzyloxycarbonyl) phenylacetyl chloride in 3 ml of dry acetonitrile are added dropwise. The . thin layer chromatography indicates that the reaction is / λ 68 completed after 15 minutes. A potassium phosphate buffer (0.5 M; pH 5.5; 17 ml) is added and most of the acetonitrile is removed in vacuo.

The residue is diluted with water and extracted "three times 5 with equal volumes of methylene chloride. The extract is dried over anhydrous sodium sulfate and concentrated in vacuo to give 1.032 g of product crude, which is dissolved in 3 ml of methylene chloride and which is subjected to chromatography on a column of silica 10 using as eluent a mixture of methylene chloride and methanol, to obtain 470 mg of the title compound.

B) Potassium salt of 3 - [[1,3-dioxo-2-phenyl-3- (phenylmethoxy) propyl] amino] -3-methoxy-2-oxo-1 “azetidine-15 sulfonic acid

Dissolve in 15 ml of a mixture of 30% acetone and water 470 mg of tetrabutylammonium salt of 3 - [[1,3-dioxo-2-phenyl-3- (phenylmethoxy) propyl] amino acid ] -3-methoxy-2-oxo-1-azetidinesulfonic, and the solution is passed through a column of "Dowex 50WX2" (form K), using the same solvent as eluent. The whole of the eluate is evaporated under vacuum to obtain 345 mg of an amorphous solid which is lyophilized into an amorphous powder, melting point 100-120 ° C.

25 Analysis for C20H19 ° 8N2SK:

Calculated: C, 49.37; H, 3.94? N, 5.76; S, 6.59 Found: C, 49.08; H, 4.00; N, 5.58; S, 6.29

Example 52. Potassium salt of (+) - 3 - [[[(cyanomethyl) thio] - acetyl] amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid To a solution of tetrabutylammonium acid salt 3-amino-3-methoxy-2-oxo-1-azetidinesulfonic acid (414 mg, see Example 74) in 50 ml of dry acetonitrile at -20 ° C, 210 μl of diethylaniline and 169 mg of chloride are added 35 cyanomethylthioacetyl. After 10 minutes, the solvent is removed under reduced pressure and a 0.1 M solution (22.8 ml) of tetrabutylammonium sulphate adjusted to pH 473 is added with potassium hydroxide, then the product is extracted / with three fractions of 50 ml of methylene chloride, Λ 69 is dried, filtered and concentrated under reduced pressure. The oily residue is purified on 60 g of silica gel and the product (294 mg) is eluted with a mixture of 4% methanol in methylene chloride. The purified product 5 is passed through an ion exchange resin (16 ml of "AG 50W-X2" (form K +), 0.6 mequiv. / Ml, 37-74 y. water gives 164 mg of a partially purified product. This product is subjected to a further purification on 100 ml of resin "Diaion AG HP 20" using water as eluent. The solvent is removed under reduced pressure to obtain 126 mg of a product which is triturated with ether to obtain 76 mg of the title compound, melting point 110-120 ° C.

Analysis for CgH ^ ¢ 1 ^ 520 ^ 1 (: 15 Calculated: C, 27.66; H, 2.88; N, 12.10; S, 18.44

Found: C, 27.25; H, 3.00; N, 10.84; S, 17.53.

Examples 53 to 56

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Example 57

Potassium salt of [3S (±) 3 —3— Ç [(2-methylthio) -1-oxopropyl] amino] -2-oxo-1-azetidinesulfonic acid

Following the procedure of Example 31.5, Method II, but replacing (±) -a-azido-benzene-acetic acid with (±) -2- (methylthio) propanoic acid, the compound of the title, melting point 173 ° C, with decomposition.

Example 58 10 Potassium salt of [3S (R *) J -3 - [[[[[2,3-dioxo-4 - [(phenylmethylene) amino] -1-pipërazinyl] carbonyl] amino] -phenylacetyl acid ] amino] -2-oxo-l-azetidinesulfonic

Dissolve in 20 ml of acetonitrile 0.7 g of the acid (R) -û (r [[[2,3-dioxo-4- [(phenylmethylene) amino] -1-piperazinyl] -15 carbonyl] amino ] benzeneacetic acid and the tetrabutylammonium salt of (S) -3-amino-2-oxo-1-azetidinesulionic acid (0.8 g; see Example 6A), while stirring, added dropwise 0.4 g of dicylohexylcarbodiimide at 0 ° C. Stirring is continued for 18 hours and, after filtration, the solvent is distilled off, which leaves an oily residue. This residue is dissolved in acetone and one treated with potassium perfluorobutane sulfonate to precipitate the title compound Purification by column chromatography of "HP-20" using water as eluent. gives the title compound, mp 193-194 ° vs.

Example 59

Potassium salt of [3S (Z)] acid - 3 - [[(2-amino-4-thiazolyl) • [(2-methoxy-2-oxo-ethoxy) imino] acetyl] amino] - '2- oxo-1-30 azetidinesulfonic

1.3 g of (Z) -2-amino-a - [(2-methoxy-2-oxo-ethoxy) imino] -4-thiazole- are dissolved in 50 ml of acetonitrile. acetic acid and the tetrabutylammonium salt of (S) -3-amino-2-oxo-1-azetidinesulfonic acid (2.03 g; see Example 35 6A), and "dropwise added at 0 ° C. 1.03 g of dicyclo-hexylcarbodiimide dissolved in 5 ml of acetonitrile After having been stirred for 15 hours and separating the dicyclohexylurea by filtration, the solvent is distilled off, the remaining oily residue is dissolved in acetone and treated with the equivalent amount of potassium perfluorobutane sulfonate. The title compound is isolated and purified by column chromatography on "HP-20" using water as the eluent, to give the compound of 5 titer, melting point 195 - 198 ° C.

Example 60

Potassium salt of [3S (R *)] acid - 3 - [[[[[3 - [[(4-chloro-phenyl) methylene] amino] -2-oxo-1-imidazolidinyl] carbonyl3 -amino] phenylacetyl] amino] -1-azetidinesulfonic 10 The tetra-butylammonium salt of (S) -3-amino-2-oxo-1-azetidine-sulfonic acid is stirred together for 12 hours (1.5 g; see l 'Example 6A) in 100 ml of absolute diglyme (diethylene glycol dimethyl ether), 1.5 g of (R) -a - [[[3 - [[(4-chorophenyl) methylene] amino] amino acid) -15 2- oxo-1-imidazolidinyl] carbonyl] amino] benzeneacetic, an equivalent amount of dicyclohexylcarbodiimide and 0.5 g of hydroxybenzotriazole. The solvent is removed in vacuo and the residue is dissolved in 50 ml of acetone, then filtered.

The acetone is distilled off and the residue is dissolved in 200 ml of methylene chloride. The solution is washed with aqueous sodium bicarbonate and then with an aqueous solution of sodium chloride. The methylene chloride layer is dried over sodium sulfate, evaporated to dryness and crystallized by addition of ether.

The compound is recrystallized from a mixture of acetone and ether. The resulting white crystalline powder is dissolved in acetone and the solution is treated with an equivalent amount of potassium perfluorobutane sulfonate in acetone. The title compound precipitates and is separated by filtration to give 1.4 g of product, mp 217 - 222 ° C.

Example 61

Potassium salt of [3S (R *)] acid - 3 - [[[[[[2-oxo - [(phenyl-methylene) amino] -1-imidazolidinyl] carbonyl] amino] phenyl-35 acetyl] amino] -1-azetidinesulfonic

The tetrabutylammonium salt of (S) -3-amino-2-oxo-1-azetidinesulfonic acid (2.25 g; see Example / '6A) in 100 ml is stirred together at room temperature for 12 hours. dry dimethylformamide (DMF), an amount.

73 dicyclohexylcarbodiimide equivalent, 2.5 g (R) -a - [[[2-oxo-3 - [(phenylmethylene) amino] -1-imidazolidinyl] -carbonyl] amino] benzeneacetic acid and 0.85 g d 'hydroxybenzo-triazole. After 12 hours, the DMF is removed in vacuo and the residue is dissolved in 50 ml of acetone. The precipitated urea is filtered off and the mother liquor is treated with an equivalent quantity of potassium perfluoro-butane sulfonate in 20 ml of acetone. After adding 100 ml of ether, the title compound precipitates out and is separated by filtration. The purification is carried out by dissolving the compound in a mixture of DMF and acetone and precipitating it with water, which gives 1.5 g of product, melting point 224-226 ° C, with decomposition.

15 Example 62

Potassium salt of [35 (R *)] acid - 3 - [[[[[[3-methyl-sulfonyl) -2-oxo-1-imidazolidinyl] carbonyl] amino3phenyl-acetyl] amino] -2-oxo- 1-azetidinesulfonic

The tetrabutylammonium salt of (S) -3-amino-2-oxo-1-azetidinesulfonic acid (2.25 g; see Example 6A) is stirred for 12 hours in 60 ml of dimethylformamide, with 1, 9 g of (R) -a- [[[3- (methylsulfonyl) -2-oxo-1-imidazolidinyl] carbonyl] amino] benzeneacetic acid, 0.75 g of hydroxybenzotriazole and 2.3 g of dicyclohexylcarbodiimide. The solvent is removed in vacuo and the residue is dissolved in 20 ml of acetone, then filtered. The mother liquor is treated with 1.87 g of potassium perfluorobutane sulfonate in 20 ml of acetone. After addition of ether, the title compound precipitates, giving 2.0 g of substance. The compound is purified by recrystallization from water, and it then has a melting point of 240 -245 ° C, with decomposition.

Example 63

Potassium salt of [3S (R *)] acid - 3 - [(hydroxyphenyl-acetyl) amino] -2-oxo-1-azetidinesulfonic 35 The tetrabutylammonium salt of (S) - 2-amino acid is stirred -oxo-1-azetidinesulfonic (1.5 g; see Example 6A); d & ns 100 ml of dry dimethylformamide, for about 16 hours with 1.5 g of dicyclohexylcarbodiimide, 0.5 g / of hydroxybenzotriazole and 0.6 g of Ra-hydroxybenzene 74 acetic acid. The solvent is removed in vacuo and the residue is dissolved in 20 ml of acetone. The precipitated urea is filtered off and the mother liquor is treated with an equivalent quantity of potassium perfluorobutane sulfonate. Adding ether, the title compound is precipitated, and separated by filtration, giving 1.3 g of crude product. This product is purified by column chromatography on "HP-20" using as eluent a mixture (9: 1) of water and acetone, and the product 10 then has a melting point of 145-150 ° C., with decomposition.

Example 64

Potassium salt of [3S (S *)] - 3 - [(hydroxyphenyl-acetyl) amino] -2 “Oxo-1-azetidinesulfonic 15. Following the procedure of Example 63, but replacing (R) -α-hydroxybenzeneacetic acid with (S) -α-hydroxybenzeneacetic acid, the title compound is obtained, melting point 195-197 ° vs.

EXAMPLE 65 Potassium salt (1: 2.) Of [3S (±)] - 2-oxo-3 - [(phenyl-sulfoacetyl) amino] -1-azetidinesulfonic acid

The tetrabutylammonium salt of (S) -3-amino-2-oxo-1-azetidinesulfonic acid (2.25 g; see Example 6A) is cooled to 0 ° C. in 100 ml of dry diglyme, 2, 4 g of triethylamine and 0.3 g of dimethylaminopyridine. 1.8 g (R) -a- (chlrocarbonyl) benzene-methanesulfonic acid in 20 ml of diglyme are added dropwise. The temperature is maintained for 2 hours, the solvent is removed by vacuum distillation and the residue is dissolved in acetone. The insoluble precipitate is filtered off and the mother liquor is treated with an equivalent quantity of potassium perfluorobutane sulfonate. By adding ether, the title compound is precipitated, which is separated by filtration, which gives 1.4 g of crude product; 35 melting point 240 - 245 ° C with decomposition, after recrystallization from a mixture of water and acetone.

h 75

Example 66

Potassium salt of [3S (Z)] acid - 3 - [[(2-amino-4-thiazolyl) [[(diethoxyphosphinyl) methoxÿ] imino] acetyl] -amino] -2-oxo-1-azetidinesulfonic 5 The tetrabutylammonium salt of 11 (S) - 3-amino-2-oxo-1-azetidinesulfonic acid (2.25 g; see Example 6A) is treated in 100 ml of dry dimethylformamide with 1.87 g of (Z) -2-amino-a - [[(diethoxyphosphinyl) methoxy] imino] -4-thiazoleacetic acid, 0.75 g of hydroxybenzotriazole and 2.29 g of dicyclohexylcarbodiimide, for 12 hours with stirring. The precipitated urea is filtered off and the solvent is removed in vacuo. The remaining oil is treated with an equivalent amount of potassium perfluorobutane sulfonate in 20 ml of acetone. By adding ether, the title compound is precipitated, which is filtered off, giving 2.77 g of crude product. Purification of this crude product by column chromatography on "HP-20" using as eluent a mixture (9: 1) of water and acetone, gives the title compound, melting point 155-160 ° C , with decomposition.

Example 67

Potassium salt of [3S (Z)] - 3 - [[(2-amino-4-thiazolyl) [[2- (1,1-dimethylethoxy) -2-oxo-1-phenylethoxy] -imino] acid acetyl3 amino 3-2-oxo-1-azetidinesulfonic acid The tetra butylammonium salt of (S) -3-amino-2-oxo-1-azetidine sulfonic acid (2.25 g; see Example 6A) in 60 ml of dimethylformamide, with 2.4 g of (Z) -2-amino-a - [[2-, (1,1-dimethylethoxy) -2-oxo-1-phenylethoxy acid ] imino] -4- thiazoleacétique, 1 g of hydroxybenzotriazole and 1.5 g of dicyclohexylcarbodiimide, for 12 hours. The solvent is removed in vacuo and the residue is dissolved in 50 ml of acetone. The precipitated urea is filtered off and the mother liquor is treated with an equivalent amount of potassium perfluorobutane sulfonate. By adding ether, the title compound is crystallized, which is separated by filtration. The purification of this compound is carried out by chromatography on a column of "HP-20" using / as eluent a mixture (7: 3) of water and acetone, 76 which gives 1 g of product, point of melting> 250 ° C, with decomposition.

Example 68

Potassium salt of [3.S (Z)] -3- [[(2-amino-4-5 thiazolyl) [(1H-tetrazol-5-ylmieth.oxy) imino] acetyl3amino] -2-oxo acid -l-azetidinesulfonic

Treating the tetrabutylammonium salt of (S) -3-amino-2-oxo-1-azetidinesulfonic acid (1.9 g; see Example 6A) in 60 ml of dimethylformamide, with 1.4 g 10 d (Z) -2-amino-a- [(1H-tetrazol-5-ylmethoxy) imino] -4-thiazoleacetic, 0.7 g of hydroxybenzotriazole and 1.4 g of dicyclohexylcarbodiimide, with stirring, for 24 hours . After removing the solvent in vacuo, the residue is dissolved in acetone and the precipitated urea is filtered off. The mother liquor is treated with an equivalent quantity of potassium perfluorobutane sulfonate in 10 ml of acetone. The title compound is precipitated by adding 200 ml of ether. The purification is carried out by column chromatography on "HP-20" using water 20 as eluent, and 1.05 g of product is obtained, melting point 250 ° C, with decomposition.

Example 69

Potassium salt of [3S (Z)] - 3 - [[(2-amino-4-thiazolyl) [(phenylmethoxy) imino] acetyl] amino] -2-oxo-l-25 azetidinesulfonic acid

Stir the tetrabutylammonium salt of (S) -3-amino-2-oxo-1-azetidinesulfonic acid (1.5 g; see Example 6A), 1.23 g of (Z) -2 acid -amino-a - [(phenyl-. methoxy) imino] -4-thiazoleacetic, 0.57 g of hydroxybenzo-triaole and 1.14 g of dicyclohexylcarbodiimide, in 60 ml of dimethylformamide at room temperature for 24 hours . The precipitated urea is filtered off, the solvent is removed and the residue is treated with an equivalent amount of potassium perfluorobutane sulfonate in 10 ml of acetone. By adding 200 ml of ether, the title compound is precipitated, which is separated by filtration and which is purified by column chromatography on "HP 20" using a mixture (9: 1) as eluent. of water and / of acetone, which gives 1 g of substance, melting point 77,200 ^ (3, with decomposition.

Example 7 0

Potassium salt (1: 1) of [3S (Zj] -3 - [[((2-amino-4-thiazolyl)] [(carboxymethoxy) iminolacetyl] amino] -2-oxo-l-5 azetidinesulfonic acid

The potassium salt of the acid [3S (Z)] - 3 - [[(2-amino-4-thiazolyl) [[2- (diphenyl-methoxy) -2-oxo- is mixed with 5 ml of anisole ethoxy] imino] acetyl] amino] -2-oxo-1-azetidine-sulfonic (1: 1) (1/3 g; see Example 30). At -15 ° C, 25 ml of trifluoroacetic acid is added and the mixture is stirred for 10 minutes. · 100 ml of ether is added slowly at -10 ° C, then 50 ml of petroleum ether. The precipitate is suspended, while cooling, in 20 ml of water and it is adjusted to pH 5.0 with dilute potassium hydroxide. The product is purified by chromatography on a "HP-20" column to give 3.0 g of the title compound, mp 230-235 ° C, with decomposition.

Example 71

Potassium salt of [3S (Z)] acid - 3 - [[(2-amino-4-20 thiazolyl) [[2-oxo-2- (phenylmethoxy) ethoxy] iminolacetyl] -amino] -2-oxo -l-azetidinesulfonic

Following the procedure of Example 28, but replacing the acid (Z) -2-amino-a - [[2- (diphenylmethoxy) - 1.1- dimethyl-2-oxo-ethoxy] imino] -4- thiazoleacetic with (Z) -2-amino-a - [[2-oxo-2- (phenylmethoxy) ethoxy] - imino] -4-thiazoleacetic acid, the title compound is obtained, melting point 170 ° C approximately, with decomposition.

Example 72. Potassium salt of [3S (Z)] -3- [[[(2-.amino-2-oxoethoxy) imino] (2-amino-4-thiazolyl) acetyl] amino] -2- oxo-l-azetidinesulfonic

Following the procedure of Example 28, but replacing the acid (Z) -2-amino-a - [[2- (diphenylmethoxy) - 1.1- dimethyl-2-oxo-ethoxy] imino] -4- thiazoleacetic with (Z) -2-amino-a - [(2-amino-2-oxo-ethoxy) imino] -4- thiazoleacetic acid, the title compound is obtained, melting point 205-210 ° C , with decomposition.

78

Example 73

Potassium salt of [3S (Z)] acid - 3 - [[(2-amino-4-thiazolyl) (hydroxyimino) acetyl] amino] -2-oxo-i-azetidine-sulfonic 5 Agitation is carried out for one hour , with 10 g of a molecular sieve 4a, a solution of 0.6 g of 90% hydroxybenzo-triazole in 100 ml of dimethylformamide, is filtered, and the filtrate is added to a solution of 0.004 mole of salt (S) -3-amino-2-oxo-1-10 azetidinesulfonic acid tetrabutylammonium (see Example 6A) in dimethylformamide. 0.89 g of (Z) -2-amino-a- (hydroxyimino) -4-thiazoleacetic acid is added, then 0.91 g of dicyclo-hexylcarbodiimide. This mixture is stirred for about 16 hours, evaporated in vacuo and the residue is dissolved in 20 ml of acetone, then filtered. The addition of a potassium perfluorobutane sulfonate solution precipitates the title compound. Chromatography on resin "HP-20" gives 0.44 g of product, melting point> 240 ° C.

20 Example 74

Potassium salt of 3-methoxy-2-oxo-3 - [(2-thienyl-acetyl) aminoj-1-azetidinesulfonic A) Tetrabutylammonium salt of 3-amino-3-methoxy-2-oxo acid -1-azetidinesulfonic acid The tetrabutylammonium salt of (±) -3-methoxy-3 - [[(phenyl-methoxy) carbonyl] amino] -2] oxo-1-azetidinesulfonic acid is dissolved in 15 ml of dry methanol (143 mg, see Example 49). 12 mg (0.1 equiv.) Of Na2B4O7.10 H2O are added, followed by 72 mg of 10% palladium on charcoal.

The mixture is hydrogenated under a pressure of one atmosphere for 15 minutes. The catalyst is filtered off and the filtrate is evaporated in vacuo to give 114 mg of the title compound.

B) Tetrabutylammonium salt of 3-methoxy-2-oxo-3-35 [(2-thienylacetyl) amino] -1-azetidinesulfonic acid

102 mg of the tetrabutylammonium salt of 3-methoxy-3-amxno-2-oxo-1-azetidinesulfonic acid are dissolved in 10 ml of dry acetonitrile. 56.5 µl of pyridine / dry are added and the solution is carefully stirred at -10 ° C under.

79 dry nitrogen. 44 μl of thienylacetyl chloride in 1 ml of dry acetonitrile are added dropwise. The reaction is complete after 15 minutes, as shown by thin layer chromatography. A potassium phosphate buffer (0.5 M; pH 5.5; 4.2 ml) and 8.5 mg of tetrabutylammonium sulphate (0.1 equiv.) Are added, then the greater is removed under vacuum. part of acetonitrile. The residue is diluted with water and extracted with three 20 ml portions of methylene chloride. The extract is dried over anhydrous sodium sulfate and evaporated in vacuo to give 107 mg of a gum. The crude product is purified by chromatography on silica gel using a mixture of methylene chloride and methanol as eluent, to give 66 mg of the title compound.

C) Potassium salt of 3-methoxy-2-oxo-3 - [(2-thienylacetyl) amino] -1-azetidinesulfonic

Dissolve 154 mg of the tetrabutylammonium salt of 3-methoxy-2-oxo-3 - [(2-thienylacetyl) amino] -1-20 azetidinesulfonic acid in 3 ml of aqueous acetone to 30 I, the solution in a column of "Dowex 50W-X2" (form K +) and eluted with the same solvent. The total eluate is evaporated in vacuo to obtain 95 mg of product, which is lyophilized to obtain an amorphous powder, melting point 120-135 ° C.

Analysis for c1oHnN2 ° 6S2K:

Calculated: C, 33.51; H, 3.09? N, 7.82; S, 17.89

Found: C, 33.46; H, 3.08; N, 7.92; S, 17.64.

'Example 75' 1 30, [3S (Z) 1] -3 - potassium potassium salt [[(2-amino-4-thiazolyl) [(carboxymethoxy) imino] acetyl] amino] -2-oxo- i-azetidinesulfonic

Dissolve the potassium salt of the acid [3S (Z)] - 3 - [[(2-amino-4-thiazolyl) [[2-oxo-2- (phenylmethoxy) ethoxy] -35 imino] acetyl] amino ] -2-oxo-1-azetidinesulfonique (0.1 g; see Example 71) in a mixture of 5 ml of ethanol and 5 ml of water, and the solution is hydrogenated at room temperature in the presence of 0.2 g of 10% palladium on / carbon. After 2 hours, we separate it. catalyst by filtration and the remaining solution is subjected to freeze drying, which gives the title compound, m.p.

235 ° C with decomposition.

Example 76 5 Potassium salt of acid 3 - [[(S) - [(aminocarbonyl) amino] -

2- thienylacetyl] amino] -3-methoxy ~ 2-oxo-l-azetidinesulfoni-que, isomer A

To a solution of tetrabutylammonium salt of 3-amino-3-methoxy-2-oxo-1 - azetidinesulfonic acid (277 mg; see Example 46, method II, part A) in 15 ml of dry acetonitrile , at '-20 <5C, 71 μΐ (0.888 mmol) of pyridine and 166 mg of (D) -2-amino-4- (2-thienyl) “5- (4H) -oxazoline hydrochloride are added. This mixture is stirred for 10 minutes and a second fraction of 71 μΐ of pyridine and 166 mg of oxazoline is added. After another 10 minutes, the solvent is removed under reduced pressure and the residue is dissolved in a mixture of water and acetone, then the solution is passed through an ion exchange resin (20 ml of " AG 50W-X2 ", form k" 1.37 - 74 μ). The elimination of water from fractions 2 and 3 gives a mixture of diastereoisomers (248 mg).

This product is purified and the diastereomers are separated on "Diaion AG HP 20" (130 ml), eluting with water. Isomer A is eluted in fractions 23 to 28 (30 mg) and isomer B in fractions 35 to 45 (30 mg) (8 ml fractions). The intermediate fractions (10 mg) are combined with the fractions from other passages and a total of 35 mg of the A isomer and 50 mg of the B isomer are isolated.

30 Analysis for isomer A, m.p. 158-165 ° C for C11H13N407S2.K.1 / 2H20

Calculated: C, 31.05; H, 3.29; N, 13.18

Found: C, 30.95; H, 2.97; N, 12.98

Analysis for isomer B, m.p. 160-170 ° C (decomposition) 35 for CUH 3N407S .K.1 / 2H20

Calculated: C, 31.05; H, 3.29; N, 13.18; S, 15.05

Found: C, 31.17; H, 3.09; N, 13.13; S, 15.09.

/ h 81

Example 77

Dipotassium salt of 3 * -methoxy-2-oxo-3 - [(phenyl-sulfoacetyl) aminol-1-azetidinesulfonigue A stirred solution of tetrabutylammonium salt 5 of 3-amino-3 ~ methoxy-2 -oxo-1-azetidinesulfonic crude (366 mg; see Example 74A). and 38 mg of borax in 35 ml of dry acetonitrile at -10 ° C, under nitrogen, 0.53 ml of dry pyridine is added, then, in two minutes, a solution of α-sulfophenyl chloride monoetherate 10 acetyl (348 mg) in 8 ml acetonitrile. After 20 minutes, the solvent is removed in vacuo and the

residue with 35 ml of 0.5 M potassium phosphate buffer

at pH 5.5. 383 mg of tetra-butylammonium hydrogen sulphate are added and the mixture is extracted three times with methylene chloride. The methylene chloride layer is dried over sodium sulfate and evaporated to obtain 685 mg of crude product.

We. combine this crude product with 115 mg of the crude product from a second test, and the total substance is purified on a column of "SiliCAR CC-4" using methylene chloride, then methanol at 2.4, as eluent. , 6, 8 and 10% in methylene chloride. The product, formed from a mixture (about 6: 1) of racemic diastereomers in the form of the tetrabutylammonium salt, is transformed into the potassium salt by passing it through + a resin "Dowex 50W-X2 ”(Form K), using 20% acetone in water as solvent. The product is lyophilized to give 171-mg of the title compound, mp 205-210 ° C, with decomposition.

30 Analysis for:

Calculated: C, 29.51; H, 2.89; N, 5.74; S, 13.10 Found: C, 29.45; H, 2.74; N, 5.51; S, 12.82. Example 78

3 - [(carboxyphenylacetyl) amino] -35 3-methoxy-2-oxo-1-azetidinesulfonic acid dipotassium salt

The potassium salt of 3 - [[1,3-dioxo-2-phenylr-3- (phenylmethoxy) propyl] -amino] -3-methoxy-2-oxo-1- is dissolved in 5 ml of methanol azetidinesulfonic (39 mg; see / Example 51). 3.9 mg of anhydrous potassium carbonate 82 and 19 mg of 10% palladium on carbon are added and the mixture is hydrogenated under atmospheric pressure for 20 minutes. The catalyst is filtered off and the filtrate is evaporated in vacuo to give a glassy residue (34 mg) which is lyophilized to an amorphous powder, melting point 178-190 ° C, with decomposition.

Analysis for C13H12 ° 8N2S K2 ’0.5 H2 °: '

Calculated; C, 35.20; H, 3.18; N, 6.32; S, 7.23

Found: C, 35.51; H, 2.96; N, 6.29? S, 6.92 ·.

10 Example 79

Potassium salt of [35 (Z)] acid - 3 - [[(2-amino-4-thiazolyl) [(2- (l / l-dimäthy ^ thoxy) -l - (meth.ylthio) - 2-oxo-ethoxy] iminojacétyljamino -2-oxo-l ~ azétidinesulfonique

Following the procedure of Example 73, but replacing (Z) -2-amino-a- (hydroxyimino) -4-thiazoleacetic acid with (Z) -2-amino-a- [ [2- (1,1-dimethylethoxy) -1- (methylthio) -2-oxo-ethoxy] imino] -4-thiazoleacetic, the title compound is obtained, melting point 130 ° C, with decomposition.

20 Example 80

Potassium salt of (±) -3-butoxy-3 - [[(phenyl-methoxy) carbonyl] amino] -2-oxo-1-azetidinesulfonic acid

A solution of 185 mg of tetrabutylammonium salt of 2-oxo-3- [N-chloro-N - [(25-phenyl-methoxy) carbonyl] amino] -1-azetidinesulfonic acid is cooled to -78 ° C. Example 49A) in 1 ml of dimethylformamide, and 3.8 ml of 0.73 M lithium n-butoxide in n-butanol are added. After 15 minutes, a 0.5 M monobasic potassium phosphate buffer is added and the product is extracted into three 40 ml fractions of dichloromethane, dried over sodium sulfate, filtered and concentrated in vacuo to obtain 179 mg of the corresponding tetrabutylammonium salt of the title compound.

To a solution of 109 mg of this tetrabutyl-35 ammonium salt in acetone, 60 mg of potassium salt of perfluorobutyl sulfonic acid in 11 acetone are added. The solvent is removed in vacuo and ethyl acetate is added, the product crystallizes, and it is collected and dried. To obtain 66 mg of the title compound, melting point 83.5 186.5 - 187.5 ° C, with decomposition.

Example 81

Potassium salt of [3 ± (E)] - 3-methoxy-3 - [[(methoxy-imino) [2 - [[(phenylmethoxy) carbonyl] amino] -4-thiazolyl] -5 acetyl] amino ] -2-oxo-1-azetidinesulfonique

A suspension of tetrabutylammonium salt of 3-amino-3-methoxy-2-oxo-1-azetidinesulfonic acid (Example 46, Method II, Part A) (0.175 mmol) and 0.0175 mmol is treated sodium borate in. 2 ml of 10-dichloromethane at 0 ° C., per 28 μl of pyridine and 0.175 mmol of (E) -a- (methoxyimino) -2 - [[(phenylmethoxy) -carbonyl] amino] -4-thiazolylacetyl chloride. After one hour, the mixture is diluted with dichloromethane and stabilized with water. The organic layer is washed with water / 15 with saturated saline solution, dried, and evaporated in vacuo. The residue is purified on silica gel. Mallinckrodt "SilicAR CC-4" (20 g) to obtain 43 mg of the corresponding tetrabutylammonium salt of the title compound.

This tetrabutylammonium salt (43 mg) is dissolved in 0.5 ml of acetone and 20 mg of potassium perfluorobutane sulfonate are added in 0.5 ml of acetone. After adding 3 ml of ether, the solid is collected and dried under vacuum to obtain 28 mg of the title compound, melting point 144-146 ° C, with decomposition.

Example 8 2

Potassium salt of [3 ± (Z)] - 3-methoxy-3 - [[(methoxy-imino) [2 - [[(phenylmethoxy) carbonyl] amino] -4-thiazolyl] -. acetyl] amino] -2-oxo-1-azetidinesulfonic 30 Following the procedure of Example 81, but replacing the chloride of (E) -a- (methoxyimino) -2— [[(phenylmethoxy) carbonyl] to mino ] -4-thiazolylacetyl.by (Z) -a- (methoxyimino) -2 - [[(phenylmethoxy) -carbonyl] amino] -4-thiazolylacetyl chloride, the title compound 35 is obtained, melting point 168 - 172 ° C, with decomposition. Example 83

Acid potassium salt 3 - [[(R) -g - [[(4-ethyl-2,3-dioxo-1-piperaz inyl) carbonyl] amino] phenylacety1] amino j-3-ή methoxy- 2-oxo-1-azetidinesulfonique // Λ 84 * To a stirred solution of 0.69 mmol of tetrabutylammonium salt of 3-amino-3-methoxy-2-oxo-l-azetidine-sulfonic acid (Example 46, Method II, Part A) in 30 ml of dry acetonitrile at -20 ° C, under nitrogen, 242 μΐ 5 of dry pyridine are added, then a solution of 352 mg of chloride, of (R) -a - [[ (4-ethyl-2,3-dioxo-1-piperazinyl) carbonyl] amino] -phenylacetyl in 4 ml of acetonitrile. After one hour, 84 μl of pyridine are added, followed by another 117 mg of chloride. the acid indicated above in 1 ml of acetonitrile The reaction mixture is stirred for 20 minutes, diluted with 24 ml of 0.5 M monobasic potassium phosphate buffer at pH 5.5, and concentrated under vacuum to remove acetonitrile The aqueous residue is extracted three times with methylene chloride and the combined extracts are dried over sulfate of sodium and then they are evaporated, which leaves 546 mg of residue. Passing this residue through a column of silica gel using methylene chloride as eluent, then 2%, 4% and 6% methanol in methylene chloride, don-na two fractions (285 mg and 173 mg) of the corresponding tetrabutylammonium salt of the title compound.

Passing the 173 mg fraction through 4.5 g of "Dowex 50-X2" resin (K) using a mixture of acetone and water as the eluent gives 119 mg of the title compound. A 104 mg fraction of this substance is passed through a column of "HP 20-AG" resin in water. The successive elution with water, 5% acetone in water and 10% acetone in water gives 60 mg of product in the form of a mixture (1: 1 approximately) of 30 diastereoisomers. Lyophilization of the 60 mg fraction gives a solid, melting point 171-172 ° C with decomposition.

Example 84

N- (3-butoxy-2-oxo-1-sulfo-3-35 azetidinyl) -2-phenylacetamide tetrabutylammonium salt

Following the procedure of Example 80, but replacing the tetrabutylammonium salt of 2-oxo-3- [N-chloro-N - [(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid with the salt of 85 N-chloro-N- (2-oxo-1-sulfo-3-azetidinyl) -2'-phenylacetamide tetrabutylammonium (see Example 47A for the preparation of the corresponding potassium salt), the title compound is obtained in the form of an oil: 5 NMR (CDC13) 3.62 (s, 2H, CgHgCHj) 4.03 (ABq, 2H, V = 7cps, C-4 CH2), 6.98 (s, ΙΗ, ΝΗ ) and 7.30 ppm (S, 5H, CßH5).

Example 85

Potassium salt of (R) -3-methoxy-2-oxo-3 - [(phenyl-acetyl) amino] -1-azetidinesulfonic acid 10 An IM solution of dimethylformamide-sulfur trioxide complex is prepared by slowly adding trimethylsilyl chlorosulfonate to dimethylformamide at 0 ° C, then evaporating under 0.1 mm for 30 minutes at 0-25 ° C. Under an argon atmosphere, 50 mg are dissolved. Of (R) -N- (3-methoxy-2-oxo-1-azetidinyl) phenylacetamide in 0.2 ml of anhydrous dimethylformamide and cooled to 0 ° C.

A cold solution of IM dimethylformamide-sulfur trioxide complex (0.428 ml) is added, the mixture is stirred for 2 hours and poured into 15 ml of 0.5 M monobasic potassium phosphate. The solution is extracted twice with dichloromethane (discarded) and 73 mg of tetrabutylammonium bisulfate are added. Extraction with dichloromethane (three 10 ml fractions) gives a viscous oil, after drying and evaporation under vacuum. Chromatography on Mallinckrodt "CC-4" silica gel (50: 1) using 2% methanol in dichloromethane as eluent gives 34 mg of tetrabutylammonium salt of (R) -3-methoxy- acid. 2-oxo-3 - [(phenylacety) amino] -1-, * azetidinesulfonic. An ion exchange on. "Dowex 50W-X2" 30 (K +, 10 equivalents) gives the title potassium salt after lyophilization of the aqueous eluate: melting point 130 ° C., with decomposition, [a] +52 ° (C = 0.5, in water). Example 86

Potassium salt of (S) -3 - [[[[((1-ethyl-4-hydroxy-35 3-methyl-ΙΗ-pyrazolo [.3,4-b] pyridin-5-yl) carbony 1 ] amino] - phenylacetyl] amino] -2-oxo-l-azetidinesulfonigue

Following the procedure of Example 73, but replacing the (Z) -2-amino- <t- (hydroxyimino) -4-Jj thiazoleacetic acid with a - [[(1-ethyl- 4-hydroxy-3- # \ 86 methyl-1H-pyrazolo [3,4-b] pyridin-5-yl) carbonyl] amino] -benzeneacetic, the title compound is obtained, melting point 233-236 ° C., with decomposition.

Example 87 5 Potassium salt of (R) -3-acetyTamino-3-methoxy-2-oxo-l-azé t idiné suifon ic acid A) 3-acetylamino-l- [1-carboxy-2-methyl ( propyl)] - (3R) -3-methoxy-2-oxoazetidine To a 650 mg solution of (6R-cis) -7-acetyl-10 amino-7-methoxy-3-methyl-8-oxo-5 acid -thia-l-azabicyclo [4.2.0] -oct-2-ene-2-carboxylic acid and 191 mg of sodium bicarbonate in water, 11 ml of a commercial suspension of Raney nickel (0 , 6 g / ml), which has been made neutral by washing with water. The mixture is lowered into an oil bath preheated to 170 ° C and refluxed in 2 to 3 minutes, while maintaining the bath temperature at 150-170 ° C. After 15 minutes of reflux, the reaction is stabilized by cooling in an ice bath. Ori separates the catalyst by filtration on "celite" 20 and the pH of the filtrate (11) is adjusted to pH 2 with 1N hydrochloric acid. After extracting the aqueous solution five times with ethyl acetate, the combined extracts are dried over sodium sulfate and the solvent is removed in vacuo to obtain 487 mg of an oil. Chromatography on silica gel gives the product (eluted in chloroform) as an oil (381 mg).

B) 3-acetylamino-1- [1- (acetyloxy) -2-methyl (propyl)] - (3R) -3-methoxy-2-oxoazetidine

The previous azetidinone (464 mg) is dissolved in 15 ml of dry acetonitrile and the solution is purged with argon for 15 minutes. Add 359 mg of copper acetate, stir for one minute to dissolve the salt, and add 797 mg of lead tetraacetate. While the argon continues to bubble through the mixture, the reaction temperature is raised by lowering the flask into an oil bath, preheated and kept at 55-65 ° C for 15 minutes. The mixture is allowed to cool to room temperature, filtered through "Celite" and the filter pad is washed thoroughly with acetonitrile.

/ 7Λ 87

The solvent is removed in vacuo from the combined filtrate and washing water. The residue is taken up in water and extracted four times with ethyl acetate. The combined extracts are dried over sodium sulfate and the solvent is removed in vacuo to give the desired product as an oil (382 mg).

C) (R) -N- (3-methoxy-2-oxo-1-azetidinyl) acetamide

The above oil is dissolved in a mixture of 10 ml of methanol and 1 ml of water, the mixture is cooled in an ice and methanol bath at -10 to -15 ° C, and 194 mg of carbonate of potassium, then 53 mg of sodium borohydride. After stirring at -15 ° to -8 ° C for 110 minutes, the solvent is removed in vacuo, the residue is taken up in water and the pH of the solution is adjusted to 6 with acid. hydrochloric acid 1 N. An exhaustive extraction with ethyl acetate, drying over sodium sulphate and removal of the solvent under vacuum make it possible to obtain 224 mg of an oil. Chromatography of this oil on silica gel, eluting with a mixture of 5% methanol and 95% methylene chloride, gives 169 mg of an oil. The title compound crystallizes from a mixture of ether and pentane giving 131 mg of substance, melting point 106 - 112 ° C (sintering at 103.5 ° C).

D) Potassium salt of (R) -3-acetylamino-3-methoxy-2-oxo-1-azetidinesulfonic acid

Under an argon atmosphere, place in a flask and cool to 0 ° C 50 mg of (R) -3 ~ acetylamino-3-methoxy-2-oxo-1-azetidine. A 1 M solution of dimethylformamide-sulfur trioxide complex in 0.95 ml of dimethylformamide is then added, and the solution is stirred for 15 minutes. The contents of the flask are then poured into 40 ml of a 0.5 M solution of K ^ HPO ^, and extracted twice with 10 ml of methylene chloride. 1.2 equivalents of tetrabutylammonium sulfate is added to the aqueous solution and the resulting mixture is extracted with four 10 ml fractions of methylene chloride. The extracts are then dried over sodium sulfate and concentrated * to obtain 39 mg of product. The Λ 4 ~ 88 tetrabutylammonium salt * is converted into the title potassium salt by passing it through a column of "Dowex 50-X2" (K). The concentration of the aqueous fraction gives 19 mg of the potassium salt, with an NMR spectrum identical to that of the product prepared in Example 46, Method I, part B, and that obtained by isolation from natural sources (Example 165).

Example 88

Potassium salt of acid (±) -3 - [(azidophenylacetyl) ~ 10 amino] -3-methoxy-2-oxo-l-azetidine5ulfonic A) Tetrabutylammonium salt of acid (±) -3 - [( azido-phenylacetyl) amino] -3-methoxy-2-oxo-l-azëtid-inesulfonigue The tetrabutylammonium salt of 3-amino-3-methoxy-2-oxo-l is dissolved in 20 ml of dry acetonitrile -15 azetidinesulfonic (202 mg; see Example 74a). To the well-stirred solution, at -20 ° C, under dry nitrogen, 167 μl of dry pyridine and 96 μl of a-azidophenylacetyl are added.

After 20 minutes, 12 ml of a 0.5 M monobasic potassium phosphate buffer at pH 5.5 is added, and the acetonitrile is removed in vacuo. The aqueous residue is extracted three times with methylene chloride. The extract is dried over anhydrous sodium sulfate and evaporated in vacuo to give 281 mg of crude product in the form of a gum. This gum is purified by chromatography in a column of silica gel (30 g) using as eluent methylene chloride and mixtures of methylene choride and methanol containing up to 6% methanol, which gives 231 mg of the title compound.

"· 30 B) Potassium salt of (±) -3 - [(azidophenylacetyl) - amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid

Passing 231 mg of tetrabutylammonium salt of (±) -3 - [(azidophenylacetyl) amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid in 15 ml of 30% aqueous acetone, 35 "Dowex 50W-X2" column (form K +; 3 ml), and elute with water. The total eluate is evaporated in vacuo to obtain a colorless glass (168 mg) in the form of a 1: 1 mixture of diastereoisomers. Passing the mixture / through a 60 ml column of "HP 20-AG" using water 89 and 10% aqueous acetone as eluent gives 71 mg of a 1: 1 mixture of diastereomers and 70 mg of a 1: 3 mixture of racemic diastereomers. The mixture is lyophilized 1: 1 and dried under vacuum at 40 ° C to obtain the dry product in the form of a hemihydrate, melting point 130 ° C, with decomposition.

Analysis for Cl2Hl2N50g.K. 0.5 H20:

Calculated: C, 35.90; H, 3.26, * N, 17.45; S, 7.98 Found: C, 35.94; H, 3.07; N, 17.24; S, 8.02 10 Example 89

Potassium salt of 3 - [(azldophenylacetyl) amino] acid -

3-methoxy-2-oxo-1-azetidinesulfonic, isomer A

The 1: 3 mixture of racial diastereomers obtained in Example 88B is left to stand in deuterated water at room temperature, and the A isomer crystallizes. After refrigeration, the mother liquor is separated and the crystals (28 mg) are dried under vacuum at 40 ° C, melting point 130 ° C, with decomposition, in the form of a mono-deuterate.

20 Example 90

Potassium salt of acid [3 ± (R *)] - 3 - [[[[((4-ethyl-2,3-dioxo-1 -piperaziny.l) carbonyl] amino] phenylacetyl] amino] -3- methyloxy-2-oxo-1-azetidinesulfonic acid A stirred solution of tetrabutylammonium salt of 3-amino-3-methoxy-2-oxo-1-azetidinesulfonic acid (0.69 mmol; see Example 74A) in 30 ml of dry acetonitrile at -20 ° C, under nitrogen, 242 μl of dry pyridine are added, then a solution of 352 mg of (R) -a - [[(4-ethyl-. '2,3-) chloride dioxo-1-piperazinyl) carbonyl] amino] phenylacetyl in 4 ml of acetonitrile. After one hour, 84 µl of pyridine is added, followed by another 117 mg of the acid chloride in 1 ml of acetonitrile. The reaction mixture is stirred for 20 minutes, diluted with 24 ml of a 0.5 M monobasic potassium phosphate buffer at pH 5.5, and concentrated in vacuo to remove the acetonitrile. The aqueous residue is extracted three times with methylene chloride and the combined extracts of methylene chloride are dried over sodium sulfate and evaporated to give / 546 mg of a residue. The passage of this substance in a 90 column of "SilicAR CC-4", using as eluent methylene chloride then 2% methanol, 4%, and finally 6% in methylene chloride, gives two fractions (285 mg and 173 mg) of purified product in the form of the tetrabutylammonium salt.

Passing the 173 mg fraction through 4.5 g of "Dowex 50-X2" resin (K +) using a mixture of acetone and water as eluent gives 119 mg of potassium salt. A 104 mg fraction of this substance is passed through a column of "HP 20-AG" resin in water. The successive elution with water, 5% acetone in water and finally 10% acetone in water provides 60 mg of product in the form of a mixture (1 : About 1) of diastereomers and 21 mg of product as a mixture (about 9: 1) of diastereomers. Lyophilization of the 60 mg fraction gives the title compound, melting point 171-172 ° C with decomposition.

Analysis for ci9H22N5 ° 9SK * H2 °:

Calculated: C, 41.23; H, 4.37? N, 12.65; S, 5.78 Found: C, 41.39; H, 4.12; N, 12.58; S, 5.63.

Lyophilization of the 21 mg fraction gives the title compound, melting point 171-172 ° C, with decomposition. Analysis for C1gH22N50gSK-H20:

Calculated: C, 41.23; H, 4.37; N, 12.65 Found: C, 41.43; H, 4.11; N, 12.28

Treatment of the 285 mg fraction of the tetrabutylammonium salt with the resin "Dowex 50-X2" (K) gives 145 mg of potassium salt, which is combined with the remaining 15 mg of the above-mentioned fraction. 119 mg. potassium salt obtained by treatment with "Dowex" resin. Passing this substance through an "HP 20-AG" resin, as already described, provides an additional 31 mg of product in the form of a mixture (approximately 1: 1) of diastereomers and an additional 42 mg of product under the form of a mixture (about 9: 1) of diastereoisomers. The total amount of the mixture (1: 1) of diastereoisomers is 91 mg, and the total amount of the mixture (9: 1) of diastereoisomers is 63 mg.

h 91

Example 91

Potassium salt of [3S (Z)] - 3 - [(methoxyimino) [2 - [[(phenylmethoxy) carbonyl] amino] -4-thiazolyl] acetyl] amino -2-oxo-1-azetidinesulfonic 5 A a solution of tetrabutylammonium salt of (S) -3-amino-2-oxo-1-azetidinesulfonic acid (0.170 mmol; see Example 6A) and 0.170 mmol of sodium borate in 2 ml of methylene chloride to 0 ° C, 62 μΐ of pyridine and 0.51 mmol of (Z) -a- (methoxyimino) -10 2 - [- ([phenylmethoxy) carbonyl] amino] -4-thiazoleacetyl chloride are added. The reaction mixture is diluted, after 40 minutes, with methylene chloride and water, then with 0.1 M tetrabutylammonium sulfate buffered to pH 4 (5.1 ml). The organic layer is separated and washed with water adjusted to pH 2, water adjusted to pH 7, water saturated with sodium chloride, dried over sodium sulfate, filtered. , and concentrated in vacuo. The residue is purified on 10 g of "SilicAR CC-4" silica gel and the product is eluted with a mixture of 10% methanol and methylene chloride.

The tetrabutylammonium salt is passed, after dissolving in a mixture of acetone and water, in an ion exchange resin (8 ml, "AG 50W-X2", form K- * ", 74 to 149 The elimination of the vacuum water from the 25 fractions 1 and 2 gives 40 mg of the title compound, melting point 17 2 - 174 ° C., with decomposition.

Analysis for C17H16N50gS2K.H20:

Calculated: C, 37.84; H, 3.33; N, 12.99; S, 11.87

Found: C, 37.95; H, 3.30; N, 12.73; S, 11.53 30 Example 92

Potassium salt of (±) -3-butoxy-2-oxo-3 - [(phenyl-acetyl) amino} -1-azetidinesulfonic acid A) Tetrabutylammonium salt of 3- [chloro (phenyl-acetyl) acid ) amino] -2-oxo-1-azetidinesulfonic 35 A solution of tetrabutylammonium salt of (S) -2-oxo-3 - [[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid is added (350 mg; see Example 4) in 3 ml. of methylene chloride to a suspension of 1.27 g of sodium borate in a 5.25% solution of 4.72 ml 92 of sodium hypochlorite and 20 ml of water, at 0 ° C. After one hour, 25 ml of 0.5 M monobasic potassium phosphate is added and the mixture is extracted three times with 50 ml fractions of methylene chloride. The organic extracts are dried over sodium sulfate, filtered and concentrated in vacuo to obtain 344 mg of the title compound.

B) Tetrabutylammonium salt of (±) -3-butoxy-2-oxo-3 - [(phenylacetyl) amino] -1-azetidinesulfonic acid 10 A solution of 344 mg of tetra butylammonium acid salt 3 is added - [[chloro (phenylmethoxy) carbonyl] -amino] -1-azetidinesulfonic acid in 5 ml of dimethylformamide with 0.73 M lithium n-butoxide in 6 ml of n-butanol, in 1 ml of dimethylformamide at -78 ° C. under atmos-15 inert sphere. After 10 minutes, the mixture is diluted with 175 ml of a 0.5M monobasic potassium phosphate solution. After triple extraction with methylene chloride, the organic extract is dried over sodium sulfate, filtered. and the solvent is removed in vacuo. The residue is purified on 80 g of silica gel (SilicAR CC-4 "and the title compound (130 mg) is eluted with 4 to 8% methanol in methylene chloride.

C) Potassium salt of (±) -3-butoxy-2 ~ oxo-3 - [(phenylacetyl) amino] -1-azetidinesulfonic acid 43 mg of the tetrabutylammonium salt of the acid (±) - are dissolved 3-butoxy-2-oxo-3 - [(phenylacetyl) amino] -1-azetidinesulfonic in a mixture (9: 1) of water and acetone and the solution is placed in a cation exchange column (" Dowex AGMP 50W-X2 ", 74 to 149 y, 5 q, form K +).

The product is eluted with water and the eluate is concentrated in vacuo to give 20 mg of the title compound, melting point 122-125 ° C.

Analysis for C ^ H ^ g ^ OgSK. 1/2 ^ 0:

Calculated: C, 44.66; H, 4.96; N, 6.95; S, 7.94 35 Found: C, 44.77; H, 4.76; N, 6.76? S, 7.75

Example 93

Potassium salt of (±) -3-ethoxy-2-oxo-3 - [(phenyl-acetyl) amino] -1-azetidinesulfonic acid / The tetrabutylammonium salt of the acid is added

AT

93 3- [ehloro (phenylacetyl) amino] -2-oxo-1-azetidinesulfonic (200 mg; see Example 92A) in 4 ml of dimethylformamide, with 0.5 M lithium ethylate in 12, 20 ml of ethanol, at -78 ° C under an inert atmosphere. After 10 5 minutes, the mixture is diluted with 15 ml of a 0.5M monobasic potassium phosphate solution. After triple extraction with methylene chloride, the organic layer is dried over sodium sulfate, filtered. and the solvent is removed in vacuo. The residue is purified on 20 g of silica gel "SilicAR CC-4" and the tetrabutylammonium salt of the product (40 mg) is eluted with 2% methanol in methylene chloride.

The tetrabutylammonium salt is dissolved in a mixture (9: 1) of water and acetone and passed through a cation exchange column ("Dowex AGMP 50W-X2", 74 + at 149 y, 5 g, form K). This product is eluted with water and the eluate is concentrated in vacuo to obtain 25 mg of the title compound, melting point 94 -96 ° C.

Analysis for cj3H; 15N206SK: 20 Calculated: C, 42.62? H, 4.10; N, 7.65; S, 8.74

Found: C, 40.36; H, 3.66; N, 6.77; S, 8.44

Example 94

Potassium salt of [3i (Z)] - 3 - [[(2-amino-4-thiazolyl) (methoxyimino) acetyl] amino] -3-methoxy-2-oxo-l-25 azetidinesulfonic acid

Dissolve the tetrabutylammonium salt of 3-amino-3-methoxy-2-oxo-1-azetidinesulfonic acid (see Example 46, Method II, Part A) in 20 ml of acetonitrile and add 1 ml of pyridine to a vigorously stirred suspension of (Z) -a- (methoxyimino) -2-amino-4-thiazoleacetyl chloride in 20 ml of acetonitrile cooled to 0-5 ° C. After stirring cold for one hour, the mixture is diluted with 100 ml of a 0.5M monobasic potassium phosphate solution (the pH of the mixture is 4.8) and the solvent is removed in vacuo. The residue is taken up in a minimum quantity of water containing a small quantity of acetone. Chromatography on ion exchange resin ("AG 50W-X2", 74-149 y, form K +, 200 ml) A gives the crude product in the form of the potassium salt,

/ K

94 after elution with water. Further purification on resin "HP-20" (200 ml) using water as eluent, gives 59 mg of the product in the form of a powder after trituration with a mixture of acetoninitrile and ether 5 then twice with ether. The product is an amorphous powder which melts slowly and decomposes above 150 ° C.

Analysis for C ^ H ^ Nj-O ^ SK:

Calculated: C, 28.77; H, 2.90; N, 16.78; S, 15.36; K, 9.37 10 Found: C, 27.77; H, 2.82; N, 15.87; S, 13.63; K, 10.11. Example 95

Potassium salt of acid [3R (R *) and 3S (S *) 1-3-Γ Γ Γ (amino-carbonyl) amino] phenylacetyl] amino] -3-methoxy-2-oxo-l-_ azetidinesulfonigue 15 A) Internal salt of (±) -3 - [(aminophenylacetyl) -amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid

The potassium salt of (±) -3 - [(azidophenylacetyl) amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid (209 mg; 209 mg is dissolved in dry inethanol) pie 88). 0.6 ml of anhydrous trifluoroacetic acid and 105 mg of 10% palladium on carbon are added and the mixture is hydrogenated for one hour. The catalyst is filtered off and the filtrate is evaporated in vacuo to obtain 271 mg of crude product.

B) Potassium salt of the acid [3R (R *) and 3S (S *)] - 3- [[[((aminocarbonyl) amino] phenylacetyl] amino] -3 - methoxy-2-oxo-1 -azetidinesulfonigue

271 mg of the potassium salt of (±) -3 - [(aminophenylacetyl) amino] -Ι-ΙΟ methoxy-2-oxo-1-azetidinesulfonic acid are dissolved in 6.5 ml of water. 87 mg of potassium cyanate are added and the mixture is stirred at room temperature for 3 hours. The solution is concentrated in vacuo to about 2 ml and subjected to chromatography in a 100 ml column of "HP 20-AG" using water as the eluent. Isomer A (29 mg) is isolated after lyophilization, melting point 160 ° C., with decomposition. Analysis for C ^ H ^ N ^ O ^ SK monohydrate:

Calculated; C, 36.44; H, 3.99; N, 13.07; S, 7.48 / i Found: C, 36.35; H, 3.79; N, 12.81; S, 7.32.

/ ^ 95

Example 96

Potassium salt of [3R (S *) and 3S (R *)] - 3 - [[[(amino-carbony1) amlno] phenylacetyl] amino] -3-methoxy-2-oxo-1-azetidinesulfonic 5 With the potassium salt of [3R (R *) and 3S (S *)] - 3 - [[[(aminocarbonyl) amino] phenylacetyl] amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid produced in Example 95, 21 mg of potassium salt of [3R (S *) and 3S (R *)] acid - 3 - [[[(aminocarbonyl) amino] -10 phenylacetyl] amino] is also produced -3-methoxy-2-oxo-1-azetidinesulfonic, in the form of a lyophilisate, melting point 160 ° C, with decomposition.

Analysis for C ^ H ^ N ^ O ^ SK sesquihydrate:

Calculated: C, 35.69; H, 4.14; N, 12.81; S, 7.33.

Found: C, 35.98; H, 3.87; N, 12.50; S, 7.32.

Example 97

Potassium salt of [3t (s *)] - 3-methoxy-3 - [[[[[2-oxo-3 - [(phenylmethylene) amino] -1-imidazolidinyl] carbonyl] -amino] -2 -thienylacetyl] amino] -2-oxo-1-azetidinesulfonic 20 A stirred solution of tetrabutylammonium salt of 3-amino-3-methoxy-2-oxo-azetidinesulfonic acid (306 mg of crude substance presumed to contain 274 mg of substance organic, prepared as described in Example 74) in 20 ml of dry acetonitrile at -20 ° C, under nitrogen, 0.30 ml of dry pyridine (3.72 mmol) is added, 484 mg of chlorine chloride (S) - [[Γ t (2-oxo-3-phenylmethylene) amino] -1-imidazolidinyl] carbonyl] amino] -2-thienylacetyl, partially dissolved and suspended in 10 ml of dry acetonitrile. The reaction mixture is stirred and allowed to warm to 0 ° C over one hour.

The reaction mixture is diluted with a large volume of methylene chloride, then treated with 22 ml of a 0.5 M monobasic potassium phosphate buffer at pH 5.5. The aqueous layer is washed with methylene chloride and the methylene chloride extract is washed with water, dried over sodium sulfate and evaporated to a residue (508 mg). This residue is subjected to chromatography on 50 g of "SilicAR CC-4", using as eluent / methylene chloride and then 2% and 4% methanol tΛ 96 in methylene chloride, to obtain 251 mg tetrabutylammonium salt of the title compound.

To 251 mg of this salt in acetone, a solution of 107 mg of potassium salt of 5-perfluoro-butanesulfonic acid in several milliliters of acetone is added, ethyl acetate is added and the mixture is washed. precipitated three times with ethyl acetate by centrifugation, then dried under vacuum at 40 ° C under 1 mm for 2 hours to obtain 95 mg of the desired potassium salt as a mixture (1: 2 approximately) of diastereoisomers having a melting point of 200 ° C, with decomposition.

Analysis for C21H21NgOgS2R:

Calculated: C, 42.85; H, 3.60; N, 14.28; S, 10.87

Found: C, 43.02; H, 3.74; N, 13.94; S, 10.71.

15 Example 98

Potassium salt of (± -cis) -4-methyl -2-oxo-3- [[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid A) N-Benzyloxy-t-boc * -allothreonine amide

A solution of 6.9 g of d, lt-boc-20 allothreonine and free amine of 5.3 g of hydrochloride do-benzylhydroxylamine (approximately 0.033 mole, release of ethyl acetate and bicarbonate) are treated. sodium) in 80 ml of tetrahydrofuran with 4.82 g of N-hydroxybenzotriazole and 6.5 g of dicyclohexylcarbodiimide in 20 ml of tetra-25 hydrofuran. After having stirred for approximately 16 hours at room temperature, the suspension is filtered, concentrated in vacuo and subjected to chromatography on a column of 400 ml of silica gel. Elution with 5-10% ethyl acetate in chloroform gives 6.8 g of the title compound in fractions 7-22 (200 ml each).

B) (ί-cis) -N-benzyloxy-S-t-butoxycarbonylamino - ^ - methyl-azetidinone

A solution of 6.8 g of the amide of N-benzyloxv-t-boc-allothreonine in 200 ml of tetrahydro-furan is stirred for approximately 16 hours with 5.24 g of triphenylphosphine and 3.2 ml of diethyl azodicarboxylate.

The solvents are evaporated under vacuum and the "boc" is an abbreviation Dour for "butoxycarbonyl".

// Λ 97 residue chromatography on a column of 500 ml of silica gel. Elution with methylene chloride, followed by crystallization from ether, gives a total of 2.65 g of azetidinone. A new chromatography of the 5 mother liquors and mixed fractions gives an additional 0.6 g. The crystallization of a fraction twice in ether (-20 ° C) gives the analytical sample of the title compound, melting point 140 - 142 ° C.

C) (± -cis) -3-t-butoxycarbonylamino-1-hydroxy-4-methyl-10 azetidinone

A solution of 3.2 g of cis-N-benzyloxy-3-t-butoxycarbonylamino-4-methylazetidinone in 200 ml of 95% ethanol, under hydrogen atmosphere, is stirred with 0.7 g of 10% palladium. % on coal. After 40 minutes, the suspension is filtered (absorption 249 ml) and the filtrate is evaporated and then triturated with ether to obtain, in two harvests, 2.05 g of a solid melting at 134 - 136 ° C.

D) (± -cis-3-t-butoxycarbonylamino-4-methylazetidinone) A solution of 2.05 g of cis-3-t-butyloxycarbonylamino-1-hydroxy-4-methylazetidinone in 60 ml of methanol is treated with a total of 90 ml of 4.5 M ammonium acetate (fractions of 40, 20 and 30 ml respectively) and per 45 ml of 1.5 M titanium trichloride (respective fractions of 20, 10 and 15 ml), the second and third addition being made after 15 and 120 minutes respectively, after 135 minutes the solution is diluted with an equal volume of 8% sodium chloride and extracted with three fractions 300 ml of ethyl acetate. The combined organic layers are washed with a mixture of 100 ml of 5% sodium bicarbonate and 100 ml of saturated salt solution, dried and evaporated. of ether gives, in two harvests, 1.65 g of solid, part of the first crop is recrystallized from ether to obtain the analytical sample, melting point 176-178.5 ° C.

E) (± -cis) -3-benzyloxycarbonylamino-4-methylazetidinone

A solution of 1.55 g of cis-3- / t-butoxycarbonylamino-4-methylazetidinone in 4 ml of 98 methylene chloride and 4 ml of anisole is cooled to 0 ° C., and 50 ml of cold trifluoroacetic acid are added. . After 90 minutes, the solvents are evaporated in vacuo (benzene is added and evaporated three times). The residue is dissolved in 25 ml of acetone, the initial pH (2.5) is raised to 7 with 5% sodium bicarbonate, and 2 ml of benzyl chloroformate are added. The solution is kept at 0 ° C and the fold at 7 for 4 hours and the acetone is removed in vacuo to obtain a suspension which is filtered. The salt filtrate is saturated and extracted with methylene chloride.

The solid is dissolved in methylene chloride and dried. The organic layers are combined, concentrated, and the residue is subjected to chromatography on a column of 200 ml of silica gel. Elution with a 3: 1 mixture of chloroform and ethyl acetate gives 850 mg of the title compound in fractions 4 to 11 (100 ml each). The crystallization of a small sample in ether gives the analytical sample, melting point 165-166 ° C.

F) Potassium salt of (i-cis) -4-methyl-2-oxo-3 - [[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic A suspension of cis-3-benzyloxycarbonylamino-4 -methylazetidinone (0.75 g) in 7 ml of dimethylformamide (dried with a 4A molecular sieve activated at 320 ° C for 25 hours under an argon stream) and 7 ml of methylene chloride (dried over basic alumina), add 1.66 g of a pyridine and sulfur trioxide complex. After stirring for 3 hours at room temperature under nitrogen, an additional amount of pyridine complex and sulfur trioxide (1.66 g) is added. The reaction mixture is then stirred at room temperature under nitrogen for about 16 hours. Dimethylformamide is removed in vacuo to give 4.6 g of a residue which is dissolved in 300 ml of a 0.5M monobasic potassium phosphate solution (40 ° C for 10 to 15 minutes). The solution is cooled and passed through a "HP 20" resin column (3 cm x 60 cm) with 400 ml of 0.5 mon monobasic potassium phosphate, 1 liter of distilled water and a mixture (14: 1) water and acetone, for

/AT

99 obtain 280 mg of product in fractions 13 to 26 (100 ml each). Crystallization from a mixture of methanol and petroleum ether gives 757.5 mg of an analytical sample, melting point 214-215.5 ° C with decom-5 position.

Analysis for ^^ 2 ^ 13 ^ 2 ^ 6 ^:

Calculated: C, 40.90; H, 3.72; N, 7.95; S, 9.10; K, 11.10

Found: C, 40.43; H, 3.60; N, 7.89; S, 8.69; K, 10.82.

Example 99 10 Potassium salt of (3S-trans) -4-methyl-2-oxo-3- [[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid

Following the procedure of Example 98 but replacing d, 1-t-boc-allothreonine with 1-t-boc-threonine, the title compound is obtained, melting point 15 133-135 ° C.

Analysis for C ^ 2H ^ 3N2 ° gSK:

Calculated: C, 40.90; H, 3.72; N, 7.95; S, 9.10; K, 11.10

Found: C, 40.72; H, 3.60; N, 7.99; S, 8.80; K, 10.82.

Example 100 Potassium salt of (3S-trans) -4-methyl-2-oxo-3 - [(phenylacetyl) aminol-1-azetidinesulfonic acid salt A) Tetrabutylammonium salt of (3S) -3 acid amino-4-methyl-2-oxo-1-azetidinesulfonic

Dissolve the potassium salt of (4S-trans) -25 4-methyl-2-oxo-3 - [[(phenylmethoxy) carbonyl] aminol-1-azetidinesulfonic acid (352.4 mg; see Example 99 ) in 20 ml of distilled water, and the solution is treated with 373.5 mg (1 mmol) of tetrabutylammonium hydrogen sulphate. After stirring for 10 minutes at room temperature, it is extracted. solution three times with 10 ml fractions of methylene chloride, after saturation with sodium chloride. The methylene chloride layer is dried over sodium sulfate and evaporated in vacuo to give 536 mg of the tetrabutylammonium salt, which is hydrogenated with 270 mg of 10% palladium on carbon in 25 ml of dimethylformamide. The mixture is filtered through "Celite", and washed twice with 2.5 ml fractions of dimethylformamide to obtain the title compound in solution.

100 B) Potassium salt of (3S-trans) -4-methyl-2-oxo 3 - [(phenylacetyl) amino] -1-azetidinesulfonic acid

The crude tetrabutylammonium salt of the acid (3S) -3-amino 4-methyl-2 “OXO-1-azetidinesulfonic 5 obtained in part A (the filtrate and the washing waters combined) is treated at 0 °. C, with 206 mg of dicyclohexvlcarbodiimide, 153 mg of N-hydroxybenzotriazole and 138 mg of phenyl acetic acid. The reaction mixture is stirred at 0 ° C for one hour, then at room temperature for 2 hours. The resulting precipitate is filtered, the filtrate is evaporated in vacuo, the residue is dissolved in 10 ml of acetone and filtered. The filtrate is treated with 25 ml of acetone saturated with potassium iodide, then with 200 ml of ether. The resulting solid (752.7 mg) is a mixture of the potassium salt and the tetrabutylammonium salt of the title compound. The solid is dissolved in 50 ml of 0.5 M monobasic potassium phosphate and passed through a column of "HP 20" resin. Elution with water and then with a mixture of water and acetone gives several fractions which are combined and which are evaporated to obtain the purified tetrabutylammonium salt. An aqueous solution of this substance is passed through a column of "Dowex 50W-X2" resin (form K +) to obtain the title potassium salt (121.4 mg). Trituration with a mixture of acetone and hexane gives 104.6 mg of the title compound, mp 211-213 ° C.

Analysis for ê ^ O:

Calculated: C, 41.72; H, 4.09; N, 8.11; S, 9.28, -K, 11.32 Found: C, 41.70; H, 4.01; N, 8.07; S, 9.01; K, 11.02. 30 Example 101

Potassium salt of (cis) -4-methyl-2-oxo-3 - [(phenylacetyl) amino] -1-azetidinesulfonic acid

A solution of 320 mg of potassium salt of (± -cis) -4-methyl-2-oxo-3 - [[(phenylmethoxy) carbonyl] -35 amino] -1-azetidinesulfonic acid is prepared (see Example 98) in 20 ml of water containing 483 mg of tetrabutylammonium hydrogen sulphate, and the solution is adjusted to pH 5.5. The extraction with six fractions of 25 ml of methylene chloride gives / 517.3 mg of an oil. A solution of this 1/10 - substance in 15 ml of dimethylformamide is stirred with 400 mg of 10% palladium on carbon in a hydrogen atmosphere for 90 minutes. The catalyst is filtered and the filtrate is stirred with 150 mg of phenylacetic acid, 5,169 mg of N-hydroxybenzotriazole and 247 mg of dicyclohexylcarbodiimide, for 7 and a half hours. The solvent is removed in vacuo and the residue is dissolved in 20 ml of acetone, then filtered. The filtrate is treated with 25 ml of 0.044 M potassium iodide in acetone. The dilution with an equal volume of ether gives 330 mg of a solid which is passed through a column of 50 ml of "HP 20" resin in 20 ml of 0.05 M monobasic potassium phosphate. Elution with 200 ml of water and then with a 1: 9 mixture of acetone and water gives a positive substance in the Rydon test in fractions 6 to 10 (50 ml). Evaporation of fractions 7 to 9 gives 81 mg of a solid. Recrystallization from a mixture of acetonitrile and water gives 46 mg of the title compound, which decomposes at more than 205 ° C. A second crop (6 mg) is obtained from the filtrate. An additional 5 mg is obtained from fractions 6 and 10, by evaporation and recrystallization. Analysis for C ^ H ^^ O ^ SK;

Calculated: C, 42.84; H, 3.89; N, 8.33; S, 9.53; K, 11.62 Found: C, 42.75; H, 3.82; N, 8.32; S, 9.26; K, 11.63 25 Example 102

Potassium salt of [3S- [3g (Z), 4ß]] - 3 - [[(2-amino-4-thiazolyl) (methoxyimino) acetyl] amino] -4-methyl-2-oxo-1 -azetidinesulfonic A) Tetrabutylammonium salt of (3S-trans) -4-30 methyl-2-oxo-3 - [[(phenylmethoxy) carbonyl] amino] -1-azetidine sulfonic acid

Dissolve the potassium salt of (3S-trans) - 4-methyl-2-oxo-3 - [[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid (352.4 mg; see Example 99 ) in 20 ml of water, and 373.5 mg of tetrabutylammonium hydrogen sulfate are added. The aqueous solution is extracted three times with methylene chloride and the combined extracts are dried over sodium sulfate. After removing the solvent, 534.6 mg of the title compound are obtained.

Λ 102 B) Potassium salt of acid [3S ~ [3g (Z), 4ß]] - 3 - [[(2-axnino-4-thiazolyl) (methoxyimino) acetyl] amino] -4-methyl-2 -oxo-l-azetidinesulfonic

A solution of 534.6 mg of tetrabutylammonium salt of (3S-trans) -4-methyl-2-oxo-3- [[(phenylmethoxy) carbonyl] aitiino] -1-azetidinesulfonic acid is hydrogenated in 20 ml of hydrogenated. dimethylformamide, with 220 mg of 10% palladium on carbon, at atmospheric pressure, for 2 hours 45 minutes; the hydrogen absorption is 26.3ml. The mixture is filtered and washed twice with 2.5 ml of dimethylformamide. The filtrate and the washing water (about 25 ml total) are stirred under nitrogen with 161 mg of (Z) - (fi- (methoxyimino) -2-amino-4-thiazoleacetic acid, 136 mg of N-hydroxybenzotriazole and 164 , 8 mg of dicyclohexylcarbo-15 diimide The mixture is stirred under nitrogen for approximately 16 hours, the dimethylformamide is removed in vacuo and the rubbery residue is dissolved in acetone, then filtered to remove the urea. add a solution containing 272 mg (0.8 mmol) of potassium salt of perfluorobutanesulfonic acid in 0.8 ml of acetone.

The suspension is diluted with an equal volume of ether and filtered to obtain 325.5 mg of a crude product which is purified by chromatography on 75 ml of "HP-20 AG". Elution with 400 ml of water and 400 ml of a 9: 1 mixture of water and acetone (fractions of 50 ml) gives 335 mg in fractions 3 to 10. After trituration with a mixture of acetone and hexane, 97.3 mg of an analytical sample is obtained from fractions 3 to 5. Similarly, the trituration of fractions 6 to 10 gives an additional 90.4 mg of product in the form d 'a solid.

Analysis for cioHi2N5 ° 6S2K:

Calculated: C, 29.92; H, 3.01; N, 17.45; S, 15.97; K, 9.74 Found: C, 30.32; H, 3.49; N, 15.82; S, 13.95; K, 10.45. NMR (D20) 1.57 (3H, d,; = 7), 3.97 (3K, S), 4.30 (1H, d of q, 35d = 7.3), 4.70 (1H, d. j = 7), 6.95 ppm (1H, S).

Example 103

Dipotassium salt of [3S- [3g (Z), 4ß]] - 3 - [[(2-amino-4-thiazolyl) [(1-carboxy-1-methylethoxy) imino] acetyl] amino] - / 4-methyl-2-oxo-1-azetidinesulfonic 1 / ^ 103 A) N-benzyloxy-t-boc-threonine amide

A solution of 8.76 g of t-boc-threonine and the free amine of 6.4 g of O-benzyl-hydroxylamine hydrochloride are treated (release of ethyl acetate and 5-bicarbo-nate sodium), in 100 ml of tetrahydrofuran, with 6.12 g of N-hydroxybenzotriazole and 8.24 g of dicyclohexyl-carbodiimide in 20 ml of tetrahydrofuran. The mixture is stirred under nitrogen for 26 hours, filtered, and evaporated in vacuo. The residue is subjected to chromatography on a column of 300 g of silica gel (elution with chloroform and then with a 3: 1 mixture of chloroform and ethyl acetate, which gives 7.2 g of The crystallization from a mixture of ether and hexane gives 4.18 g of the title compound.

B) (3S-trans) -N-benzyloxy-3-t-butoxycarbonylamino-4-methylazetidinone

A solution of 12.67 g of N-benzyloxy-t-boc-threonine amide, 11.5 g of triphenylphosphine and 6.23 ml of diethyl azodicarboxy late in nitrogen is stirred under nitrogen for approximately 16 hours. 380 ml of tetrahydrofuran. The solution is evaporated and subjected to chromatography on a column of 900 g of silica gel. Elution with a 3: 1 mixture of chloroform and ethyl acetate gives 13.69 g of a compound which is crystallized from a mixture of ether and hexane to obtain 9.18 g of the title compound.

C) (3S-trans) -3 ~ t-butoxycarbonylamino-l-hydroxy-4-methyl-azetidinone

A solution of 9.18 g of (3S-trans) -N-30 benzyloxy-3-t-butoxycarbonylamino-4-methylazetidinone in 300 ml of 95% ethanol is stirred in a hydrogen atmosphere with 1. 85 g of 10% palladium on charcoal. After 141 minutes, the suspension is filtered and evaporated in vacuo. The residue is recrystallized from a mixture of ether and hexane to give 5.12 g of the title compound.

D) (3S-trans) -3-t-butoxycarbonylamino-4-methylazetidinone

A solution of 4.98 g of (3S-trans) -3-t-butoxycarbonylamino-1-hydroxy-4-methylazetidinone in / 200 ml of methanol is treated with 132 ml of 4.5 M ammonium acetate. ./λ 104 then with 66 ml of 1.5 M titanium trichloride, and the mixture is stirred for 4.5 hours. The aqueous solution is diluted with an equal volume of 8% sodium chloride and extracted with ethyl acetate to give 3.48 g of crude product. Recrystallization from a mixture of ether and hexane gives 3.3 g of the title compound.

E) (35-trans) -3-benzyloxycarbonylamino-4-methylazetidinone

A solution of 3.3 g of (3S-trans) -3-t-butoxycarbonylamino-4-methylazetidinone in 10 10 ml of dichloromethane and 10 ml of anisole is cooled to 0 ° C. and 1.12 ml of acid are added. trifluoracetic. The solution is stirred for 90 minutes and evaporated in vacuo (benzene is added and evaporated three times). The residue is dissolved in 70 ml of acetone and the solution is adjusted to pH 7 with a 5% sodium bicarbonate solution. A total of 5.33 g of benzyl chloroformate, pH 6.5-7.5, is added over one hour. The mixture was stirred for 30 minutes at pH 7, diluted with 10.0 ml of saturated salt, and extracted with ethyl acetate (three 400 ml fractions). The residue obtained by evaporation is subjected to chromatography on a column of 1 liter of silica gel. Elution with a 4: 1 mixture of chloroform and ethyl acetate gives 2.19 g of compound. Crystallization from a mixture of ether and hexane gives 1.125 g of the title compound.

F) Tetrabutylammonium salt of (3S-trans) -4-methyl-2-oxo-3 - [[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid

A solution of 600 mg of (3S-30 trans) -3-benzyloxycarbonylamino-4-methylazetidinone in 2 ml of dimethylformamide is cooled to 0 ° C., and 4 ml of 0.8 M sulfur trioxide in dimethylformamide is added. The solution is stirred at room temperature under nitrogen for one hour and poured into 80 ml of cold · 0.5 M potassium phosphate 35 · monobasic (adjusted to pH 5.5). The solution is extracted with three 50 ml fractions of methylene chloride (discarded) and 868 mg of tetrabutylammonium bisulfate are added. The resulting solution is extracted with four 75 ml fractions of methylene chloride.

105

The combined organic layers are washed with 8% aqueous sodium chloride, dried, and evaporated in vacuo to give 1.54 g of the title compound.

G) Potassium salt of acid [3S- [3g (Z), 4g]] - 3 - [[(2-5 amino-4-thiazolyl) - [(1-diphenylmethoxycarbonyl-1-methyl-ethoxy) imino ] acetyl] amino] -4-methyl-2-oxo-1-azetidine-sulfonic

A solution of 1.54 g of tetrabutylammonium salt of (3S-trans) -4-methyl-2-oxo-3 - [[(10-phenyl-methoxy) carbonyl] amino] -1-azetidinesulfonic acid is stirred in 45 ml of dimethylformamide, in a hydrogen atmosphere, with 8.00 mg of 10% palladium on carbon, for 2 hours.

The catalyst is filtered and the filtrate is stirred for approximately 16 hours with 1.24 g of (Z) -2-amino-a - [(1-15 · diphenylmethoxycarbonyl-1-methylethoxy) imino] -4-thiazole- acid. acetic, 0.4 g of N-hydroxybenzotriazole, and 580 mg of dicyclohexylcarbodiimide. The suspension is evaporated in vacuo and the residue is triturated with 20 ml of acetone, then filtered. The filtrate (plus 2 ml of wash water) is treated with 868 mg of potassium perfluorobutane sulfonate in 3 ml of acetone. Dilution with 75 ml of ether gives a solid which is isolated by decanting from the mother liquor, trituration with ether, and filtration, to obtain 0.91 g of the title compound. The mother liquor is diluted with an additional 100 ml of ether to obtain a second crop (0.45 g) of the title compound.

H) Dipotassium salt of [3S- [3ct (Z), 4ß]] -3- [[(2-amino-4-thiazolyl) [(1-carboxy-1-methylethoxy) imino] acetyl] - 1 amino] -4-methyl-2-oxo-1-azetidinesulfonic A 140 mg suspension of potassium salt of acid [3S- [3a (Z), 4 (3]]) is stirred at -12 ° C under nitrogen -3- [[(2-amino-4-thiazolyl) [(1-diphenylmethoxycarbonyl-1-methylethoxy) -imino] acetyl] amino] -4-methyl-2-oxo-1-azetidinesulfonic in 0, 5 ml of anisole, and 2.5 ml of cold trifluoroacetic acid (-10 ° C.) are added. After 10 minutes, 10 ml of ether and 5 ml of hexane are added and the suspension is stirred resulting for 5 minutes at -12 ° C., then allowed to warm to room temperature fij The solid is isolated by centrifugation and washed two f (/ \ 106 times with ether. It is immediately adjusted to pH 5 , 5, with 0.4 N potassium hydroxide, a solution of this solid in 5 ml of cold water, then the solution is passed through an 80 ml column of "HP-20 AG". water gives 72 mg 5 d u title compound in fractions 7 to 11 (10 ml) after evaporation (acetonitrile is added and the mixture is evaporated three times) and trituration with ether, m.p. About 250 ° C with decomposition.

Analysis for ci3Hi5N50g ^ 2K2; Calculated: C, 30.51; H, 2.95; N, 13.69; S, 12.53; K, 15.28

Found: C, 29.63; H, 3.20; N, 12.96; S, 11.94; K, 12.78.

NMR (D20) 1.46 (S, 6H), 1.58 (1H, d,; = 7) 4.28 (1H, d of q, j = 7, 2.5), 4.67 (1H, d, j = 2), 6.95 ppm (S, 1H).

The remaining 1.22 g of potassium salt of [3S- [3a (Z), 4ß]] - 3 - [[((2-amino-4-thiazolyl)] [(1- diphenylmethoxycarbonyl-1-methylethoxy) imino] -acetyl] amino] -4-methyl-2-oxo-1-azetidinesulfonic (crops 1 and 2) (4.2 ml of anisole, 16 ml of trifluoroacetic acid, 13 minutes at -15 ° C). Chromatography on a 300 ml column of "HP-20 AG" gives 694 mg of the title compound in fractions 6 to 9 (60 ml), after treatment as above.

Examples 104 to 133

Following the procedure of Example 11, but replacing (Z) -2-amino-a - [[[hydroxy (phenyl-methoxy) phosphinyl] methoxy] imino] -4-thiazoleacetic acid with acid indicated in column I below, the compound indicated in column II below is obtained.

h \ i 107 i τι- i i i q

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112 * Examples 134 and 135

Following the procedure of Example 70, but replacing the potassium salt of the acid [3S (Z)] - 3 - [[(2-amino-4-thiazolyl) [[2- (diphenylmethoxy) - 2-oxo-ethoxy] -5 imino] acetyl] amino] -2-oxo-1-azetidinesulfonic by the compound indicated in column I below, the compound indicated in column II below is obtained.

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Example 136

Potassium salt of [3g (Z), 4ß] -3 - [[(2-amino-4-thiazolyl) (methoxyimino) acetyl] amino] -4-methyl-2-oxo-1-azetidinesulfonic acid 5 On extracted with methylene chloride (four 10 ml fractions) a solution of 51.8 mg of potassium salt of (cis) -4-methyl-2-oxo-3 - [[(phenylmethoxy) -carbonyl] amino acid ] -l-azetidinesulfonic acid and 51 mg of tetra-n-butylammonium bisulfate in 5 ml of water, to obtain 81 mg of an oil. This oil is stirred in a hydrogen atmosphere for 2 hours with 40 mg of 10% palladium on carbon in 4 ml of dimethylformamide. The catalyst is filtered and washed with 1 ml of dimethylformamide. The filtrate and the washings are combined and stirred for approximately 16 hours with 31 mg of (Z) -2-amino-a- (methoxyimino) -4-thiazoleacetic acid, 27 mg of N-hydroxybenzotriazole and 31, 5 mg dicyclohexylcarbo-diimide. The solution is evaporated in vacuo and the residue is triturated with 3 ml of acetone. The resulting suspension is centrifuged and the liquid is treated with 51 mg of potassium perfluorobutanesulfonate. Dilution with 5 ml of ether then filtration gives a solid. Chromatography on 40 ml of "HP-20 AG" gives a substance positive for the Rydon test in fractions 3 to 5 (20 ml) (elution with water). Evaporation and trituration with ether give 23 mg of product in the form of a hygroscopic solid.

Analysis for CiqH12N5 ° 6S2K:

Calculated: C, 29.91; H, 3.01; N, 17.44 30 Found: C, 29.30; H, 3.31; N, 16.66.

NMR (D20) 1.40 (3H, d,; = 7), 3.97 (3H, S), 4.46 (1H, apparent pentet, j = 7), 5.37 (1H, d,; = 7), 6.97 ppm (1H, S). Example 137

Acid (3S ~ cis) -3-amino-4-methyl-2-oxo-l-azetidinesulfonic 35 A) t-boc-l-allothreonine

A suspension of 6.72 g of 1-allothreonine in 70 ml of 50% aqueous dioxane is treated with 9.45 ml of triethylamine and 18.1 g of t-butyl pyrocarbonate. The resulting mixture is stirred at room temperature for 4

U

115 hours, then diluted with 70 ml of water and 140 ml of ethyl acetate. After careful stirring, the layers are separated and the organic layer is washed with 30 ml of a 2: 1 mixture of water and brine. The combined aqueous layers are then re-extracted with 70 ml of ethyl acetate. The aqueous layer is cooled in an ice bath and a 10% potassium bisulfite solution i is added to pH 2.3. The acidified solution is extracted with four 150 ml fractions of ethyl acetate. The combined organic layers are dried over anhydrous sodium sulfate and freed from the solvent to obtain 9.13 g of the title compound.

B) N-methoxy-t-boc-l-allothreonine amide

9.13 g of t-boc-1-allothreonine are dissolved in 85 ml of water and 41 ml of IN potassium hydroxide solution. 5.22 g of methoxyamine hydrochloride and 8.67 g of 1-ethyl-3,3- (dimethylaminopropyl) carbo-diimide hydrochloride are added. The mixture is stirred at room temperature for 4 hours and then saturated with sodium and potassium tartrate. The resulting mixture is extracted with ethyl acetate (four 150 ml fractions) and the organic layer is dried over anhydrous sodium sulfate and freed from the solvent to obtain 7.38 g of the title compound, under the form of a solid.

C) O-methanesulfonyl-N-methoxy-t-boc-1-allothreonine amide

7.32 g of N-methoxy-t-boc-1-allothreonine amide are dissolved in 40 ml of pyridine and the solution is cooled to -20 ° C. under nitrogen. 3 ml of methanesulfonyl chloride is added dropwise using a syringe over 5 minutes. The resulting mixture is slowly warmed to 0 ° C and stirred at this temperature for 3 hours.

500 ml of ethyl acetate are added and the solution is washed with 250 ml of a 3N hydrochloric acid solution cooled in ice, then with 100 ml of a 5% sodium bicarbonate solution . The ethyl acetate layer is dried over anhydrous sodium sulfate and freed from the solvent to obtain 8.64 g of the title compound. as a white solid.

/ χΛ 116 D) (3S-cis) -3-t-butoxycarbonylamino-l-methoxy-4-methyl-azetidinone

We. 8.64 g of O-methanesulfonyl-N-methoxy-t-boc-1-allothreonine amide is dissolved in 530 ml of acetone and 11 g of solid potassium carbonate are added. The mixture is slowly heated to 65 ° C under nitrogen and stirred at this temperature. The reaction mixture is then filtered through "Celite" and the filter cake is washed with ethyl acetate. The filtrate is concentrated and the residue is taken up in 250 ml of ethyl acetate. The ethyl acetate solution is washed with 100 ml of a 1N hydrochloric acid solution and with 100 ml of a 5% sodium bicarbonate solution. The ethyl acetate layer is dried over anhydrous sodium sulfate and freed from the solvent to give 6.63 g of crude product.

E) (3S-cis) -3-t-butoxycarbonylamino-4-methylazetidinone

1.35 g of sodium are dissolved in approximately 300 ml of liquid ammonia at -50 ° C. and 5.87 g of (3S-cis) -3- are added dropwise using a syringe. t-butoxy-20 carbonylamino-1-methoxy-4-methylazetidinone in 35 ml of tetrahydrofuran. An additional 10 ml of tetrahydrofuran is used for rinsing. A little before the end of the addition, about 100 mg of additional sodium is added. The mixture is stirred for an additional 5 minutes, then stabilized by adding 3.35 g of solid ammonium chloride at one time. The ammonia is purged using a stream of nitrogen and 250 ml of ethyl acetate are added to the residue. After filtration and washing of the solid with ethyl acetate, the combined filtrate is freed from the solvent to obtain 4.82 g of the title compound.

F) Tetrabutylammonium salt of (3S-cis) -3-t-butoxycarbonylamino-4-methyl1-2-oxo-1-azetidinesulfonic acid

4.98 g of [3S, 4R] -3-t-butoxycarbonyl-amino-4-methylazetidinone are dissolved in 30 ml of dimethylformamide.

11.9 g of a pyridine-sulfur trioxide complex are added and the mixture is stirred at room temperature under nitrogen. After stirring for 14 hours, an additional 1.8 g of pyridine-sulfur trioxide complex is added and stirring is continued for 80 hours. The reaction mixture is poured into λ 117 into 700 ml of a 0.5M monobasic potassium phosphate solution and washed with three 300 ml fractions of methylene chloride. 8.45 g of tetra-n-butylammonium bisulfate are added to the aqueous solution and the mixture is extracted with four 300 ml portions of methylene chloride. The combined methylene chloride layers are dried over anhydrous sodium sulfate and freed from the solvent to obtain 10.76 g of the title compound in the form of a gum.

10 G) (3S-cis) -3-amino-4-methyl-2-oxo-1-azetidine-sulfonic acid • Dissolve 10.76 g of tetrabutylammonium salt of (3S-cis) -3-t acid -butoxycarbonylamino-4-methyl-2-oxo-1-azetidinesulfonic acid in 50 ml of 95-97 I formic acid, 15 and stirred for 4 hours under nitrogen. A small amount of product from a previous reaction is added as a primer, and the mixture is stirred for another hour. The mixture is stored in a freezer for about 16 hours and then the frozen mixture is warmed to room temperature and stirred for another hour. The solid formed is filtered and washed with methylene chloride to obtain 982 mg of the title compound. The filtrate is diluted with one liter of methylene chloride and kept at -20 ° C for 4 hours. The precipitate which forms is recrystallized from a mixture of water, methanol and acetone to obtain an additional 167 mg of the title compound.

NMR (D2O) 1.63 (3H, d, j = 6.5 cps), IR (nujol) 1775 cm> Example 138 30 Potassium gel of acid [3S- [3g (Z), 4g]] - 3 - [[(2-amino-4-thiazoly ^ mdthoxviminolacetyllamino ^ -methyl-Z-oxo-l-azetidinesulfonique

A solution of 201 mg of (Z) -2-amino-a- (methoxyimino) -4-thiazoleacetic acid and 153 mg of N-35 hydroxy-benzotriazole monohydrate in 3 ml of dimethylformamide is treated with 206 mg of dicyclohexylcarbodiimide. The mixture is stirred at room temperature for 20 minutes under nitrogen and a solution of 180 mg of (3S-cis) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid and 0.14 is added. 118 ml of triethylamine in 2 ml of dimethylformamide (an additional 1 ml of dimethylformamide is used for rinsing), and the mixture is stirred for approximately 16 hours. The suspension is evaporated in vacuo, triturated with 12 ml of acetone, centrifuged and the liquid is treated with 338 mg of potassium perfluorobutane sulfonate. Dilution with 10 ml of ether and filtration gives a solid product which is subjected to chromatography on 200 ml of "HP-20" resin, eluting with water. The 10 fractions 18 to 30 (20 ml each) are combined and lyophilized to obtain 274 mg of the title compound, in the form of a hygroscopic solid.

Analysis for C ^ 0H12 ^ 5 ^ 6S2 ^:

Calculated: C, 29.91; H, 3.01; N, 17.44 Found: C, 30.03; H, 3.21; N, 17.06.

NMR (D20) 1.40 (3H, d,; = 6.5), 3.98 (3H, S), 4.48 (1H, d of t, j = 6.4, 5.5), 5 , 36 (1H, d,, = 5.5), 6.97 (1H, S).

Example 139

(3S-trans) -3-amino-4-methyl-2-oxo-1-azetidine-20 sulfonic acid A) Threonine methyl ester hydrochloride

Under a nitrogen atmosphere, a flask containing 500 ml of methanol is cooled to -5 ° C. (ice / brine), and 130 ml (excess) of thionyl chloride are added, at a rate suitable for maintaining the temperature of reaction between 0 ° and 10 ° C. After having again cooled to -5 ° C., 59.5 g of 1-threonine are added and the mixture is allowed to reach room temperature, then it is stirred for> 16 hours. The mixture is concentrated and evacuated 5 to 10 1 torr for 2 hours to obtain a viscous oil. We use this substance directly in the next step.

B) Threonine amide

The crude product from part A is dissolved in 2.5 l of methanol and cooled to -5 ° C (ice-brine). The gaseous ammonia solution is saturated, the cooling bath is removed and the container is left to hermetically closed for 3 days. After removing the greater part of the unreacted ammonia by suction, 100 g of sodium bicarbonate and 50 ml of water are added and the mixture is reduced to a viscous oil. . C) Benzyloxycarbonylthreonine amide

The crude product from part B (already containing the required amount of sodium bicarbonate) is diluted to a volume of 1 liter with water. To this rapidly stirred solution, 94 g (88 ml of 90% pure substance) of benzyloxycarbonyl chloride are added, in the form of a solution in 80 ml of tetrahydrofuran, over 1 hour. The reaction mixture was then stirred for an additional 16 hours and extracted with ethyl acetate (one 500 ml fraction and two 250 ml fractions). The combined extracts are dried over magnesium sulfate and concentrated. The crystalline residue is then dissolved in 250 ml of hot ethyl acetate and 300 ml of hexane are added, then boiled until a clear solution is obtained. The cooling and filtration of the crystalline mass gives, after drying, 104 g of the title compound.

D) Benzyloxycarbonylthreonine amide o-mesylate

Under an argon atmosphere, 100 g of benzyloxycarbonylthreonine amide are dissolved in 400 ml of anhydrous pyridine and cooled in an ice and salt bath. To this stirred solution, 36.8 ml (54.5 g) of methanesulfonyl chloride is added over 15 minutes. After 2 hours of stirring, an additional 0.3 equivalent of methanesulfonyl chloride is added. The reaction mixture is then stirred for 1 hour and poured into a mixture of 1.5 1 of ice and 2 1 of water. The resulting suspension is stirred for about 30 minutes, then filtered. Drying the crude product at 60 ° C for about 16 hours in a vacuum oven gives 109 g of the title compound.

E) N-35 sulfonyl benzyloxycarbonylthreonine tetrabutylammonium salt of N-35 amide amide O-mesylate

A solution of 17.8 ml of 2-picoline in 90 ml of methylene chloride is cooled to -5 ° C. (ice-brine), and 5.97 ml of chlorosulfonic acid is added at a rate A suitable for maintaining the internal reaction temperature 120 below 5 ° C. The resulting solution is added, using a cannula, to a suspension of 7.56 g of benzyloxycarbonylthreonine amide 0-mesylate in 120 ml of methylene chloride. The resulting heterogeneous mixture is refluxed for approximately 16 hours, giving a clear solution. The solution is poured into 500 ml of a phosphate buffer at pH 4.5 (0.5 M) and it is further diluted with 120 ml of methylene chloride. The separated organic layer is then washed, once with 100 ml of a buffer solution, and the combined aqueous phases are treated with 10.2 g of tetra-n-butyl ammonium hydrogen sulphate, then extracted with methylene chloride (a fraction of 300 ml and two fractions of 150 ml). After drying the combined organic extracts over sodium sulfate, the solution is concentrated to obtain 12.7 g of a foam.

F) (3S) trans) -3-amino-4-methyl-2-oxo-l - azetidine-sulfonic acid

A mixture composed of 5.52 g of potassium carbonate in 20 ml of water and 160 ml of 1,2-dichloroethane is brought to reflux, and 15.5 mmol of tetrabutylammonium salt of O-mesylate is added. N-sulfonyl benzyloxycarbonylthreonine amide in 20 ml of 1,2-dichloroethane (20 ml of the latter is used for rinsing). After having maintained at reflux for 30 minutes, the mixture is poured into a separatory funnel, diluted with 50 ml of water and 100 ml of methylene chloride and the phases are separated. The resulting organic phase is dried over sodium sulfate and concentrated to obtain the crude tetrabutylammonium salt of (3S-trans) -3-benzyloxy-carbonylamino-4-methyl-2-oxo-1-azetidinesulfonic acid . The crude azetidinone, in 250 ml of ethanol, is treated with 0.8 g of 5% palladium on carbon as catalyst, and hydrogen is bubbled through the solution. After 90 minutes, the mixture is filtered on "Celite" with 50 ml of ethanol used for rinsing. The addition of 1.2 ml of formic acid to this solution causes an immediate precipitation of the title zwitterion, which is filtered after stirring for one hour to obtain, after drying under 10 1 torr for 1 hour, 1 , 1 g of product.

A second crop of product is obtained by concentrating the filtrate and adding a new quantity of formic acid to obtain 1.3 g of the title zwitterion.

5 P.f. > 218 ° C., With decomposition.

[ot] D = -41.1 ° (C = 1, H2O) NMR (D20) 1.58 (3H, d, j = 7), 4.80 (2H, M).

Examples 140 to 143

Following the procedure of Example 138, but replacing (3S-cis) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid with (3S-trans) -3 acid -amino-4-methyl-2-oxo-1-azetidinesulfonic and (Z) -2-amino-ic (- (methoxyimino) -4-thiazoleacetic acid by the acid indicated in column I below, we obtains the compound indicated in column II below.

AT

/ 122

I I

r-. i i

i — l r-i O P

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ßi Ë Ο I -H pi fi I -P N X fi + 14-) ¾ -H

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a - · ρ ι o bfiji oa P 4-> acN + J onj) SO Ô <m -h -> m fi va) η i va) p4 l— 'G (Ü X ë —1 fir O 6> -> 1 N Γ — 1 N Ë

I iQ) i i Ο Ο I I fi Ο I -fi rd I O rd O

W H H I 0 W P -P o W - I W fi I U

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fi Q) m -rl (-1 0) ro 0) P fi fi— 0) P * 4-> O Q) X 4-> lO

Ο ό i d o fi h fi> 4 va) en o- d i "u m d 0" u en

H CN-Hfitr'CN -fi -fi 0) p Ο ε cm I S P

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w 123

Example 144

Potassium salt of acid [3S- [3a (Z), 4 ß]] -3- [[(2 ~ -amino-4-thlazolyl) [Cl, l-dimethyl-2 - [(4-nitrophenyl) methoxy] -2-oxo-ethoxy] iminolacetyllamino] -4-methyl-2-oxo-l-azetidine-5 sulfonlgue A (3S-trans) -3-amino-4-methyl-2-oxo- acid suspension 1-azetidinesulfonic acid (0.36 g; see Example 139) in 30 ml of dry dimethylformamide, under nitrogen at 26 ° C., 309 μl of triethylamine are added. After about 5 minutes, a clear solution is obtained and 0.816 g of (Z) -2-amino-a - [[1,1-dimethyl-2 - [(4-nitro-phenyl) methoxy) acid is added. 'J-2-oxo-ethoxy] imino] -4-thiazoleacetic, then 0.334 g of N-hydroxybenzotriazole and 0.453 g of dicyclohexylcarbodiimide, The mixture is stirred for 12 hours at 26 ° C, after which the solvent is removed in vacuo and the residue is triturated with 30 ml of acetone. After stirring for five minutes, the solids are separated and the filtrate is treated with 3.680 g of potassium perfluorobutane sulfonate in 5 ml of acetone. The addition of about 40 ml of ether gives a precipitate which is collected and which is dried under vacuum (1.073 g; second harvest 0.066 g; total 1.14 g).

Analysis for C2oH21N6 ° 10S2K * 1 K ^ O:

Calculated: C, 38.33; H, 3.70? N, 13.41; S, 10.23; K, 6.24 Found: C, 38.30; H, 3.63; N, 13.41; S, 9.88; K, 5.98. Example 145

Acid potassium salt (1: 2) [3g (Z), 4g] -3 - [[(2-amino- 4-thiazolyl) [(1-carboxy --l-methylethoxy) iminojacetyl ·] -amino ] -4-methyl-2-oxo-1-azetidinesulfonic 30 A) Potassium salt of acid [3g (Z), 4g] -3 - [[2-amino-4-thiazolyl) [(1-diphenÿlmethoxycarbonyl- l-methylethoxy) -imino] acetyl] amino] -4-methyl-2-oxo-l-azetidinesulfonic A solution of tetrabutylammonium salt of (cis) -4-methyl-2-oxo-3 acid is stirred - [[ (phenylmethoxy) carbonyl] -35 amino] -l'-azetidinesulfonique (201 mg; prepared from the corresponding potassium salt of Example 98 as described in Example 136), in 5 ml of dimethylformamide, with 90 mg of 10% palladium on calcium carbonate in a hydrogen atmosphere, for 2 hours. The suspension is filtered and the filtrate is stirred for about 16 hours with 146 mg of (Z) -2-amino-a- [1-diphenylmethoxy-carbonyl-1-methylethoxy) imino] -4-thiazoleacetic acid, 73 mg dicyclohexylcarbodiimide and 51 mg of 5-N-hydroxybenzo-triazole, under nitrogen. The suspension is evaporated in vacuo and triturated with 4 ml of acetone. The suspension is filtered and the solid is washed twice with fractions of 2 ml of acetone. The filtrate and the washings are combined and treated with 113 mg of potassium perfluorobutane-10 sulfonate. Dilution with 24 ml of ether gives a solid which is isolated by centrifugation and which is washed three times with ether, which gives 186 mg of the title compound.

B) Potassium salt (1: 2) of the acid [3g (Z), 4g] -3 - [[(2-15 amino-4-thiazolyl) [(1-carboxy-1-methylethoxy) imino] acetyl ] -amino] ^ -methyl ^ -oxo-l-azetidinesulfonique

A suspension of 186 mg of potassium salt of acid [3a (Z), 4a] -3 - [[2-amino-4-thiazolyl) [(1-diphenylmethoxycarbonyl-1-methylethoxy) is cooled to -12 ° C. -20 imino] acetyl] amino] -4-methyl-2-oxo-1-azetidinesulfonic in 0.6 ml of distilled anisole, and 3.0 ml of distilled trifluoroacetic acid (at -10 ° C) are added. The solution is stirred for 10 minutes and 12 ml of ether are added, then 6 ml of hexane. After 5 minutes at -10 ° C and 15 minutes of stirring at room temperature, the solid is isolated by centrifugation and washed four times with ether to obtain 141 mg of substance. This substance is dried in vacuo, sprayed, dissolved in 5 ml of cold water and immediately adjusted to pH 5.6 with 0.4N potassium hydroxide. The solution is passed through a column. 100 ml of "HP-20 AG" and eluted with water.

Fractions 8 to 12 (10 ml) are combined and evaporated in vacuo (acetonitrile is added three times and evaporated). The residue is triturated with ether to obtain 101.7 mg of product as a hygroscopic solid.

Analysis for C ^ 3H1gNgOgS2.2K:

Calculated: C, 30.51; H, 2.95; N, 13.69; S, 12.53; K, 15.28. Found: C, 30.11; H, 3.26; N, 13.35; S, 12.12; K, 15.02.

L125

Example 146

[3S- [3a (Z), 4 3]] -3 - [[(2-amino-4-thiazolyl) - [(1-carboxy-1-methylethoxy) imino] acetyl] amino] -4-methyl- 2-oxo-1-azetidinesulfonigue 5 The dipotassium salt of [3S- [3a (Z), 4ß]] - 3 - [[(2-amino-4-thiazolyl) is dissolved in 1.38 ml of water ) - [(1-carboxy-1-methylethoxy) imino] acetyl] amino] -4-methyl-2-oxo-1-azetidinesulfonic (87.3 mg; see Example 103), cooled to 0 ° C, treated with 0.34 ml of 10 hydrochloric acid as IN and the resulting crystals are separated by centrifugation. The wet solid was dissolved in methanol, filtered, concentrated to about 0.5 ml and mixed with 1 ml of water, giving 55.9 mg of the title compound.

Example 147 [3S- [3g (Z), 4g]] - 3 - [[(2-amino-4-thiazolyl) [(1-carboxy-1-methylethoxy) imino] acetyl] sodium salt aninoj- 4-methyl-2-oxo-1-azetidinesulfonic

A sample of 99.7 mg of [3S- [3α (Z), 4ß]] - 3 - [[(2-amino-4-thiazolyl) [(1-carboxy-1-methyl-20-ethoxy) acid is mixed. ) imino] acetyl] amino] -4-methyl-2-oxo-1-azetidine-sulfonic with 0.207 ml of IN sodium hydroxide, and the resulting mixture is gently warmed to dissolve the remaining solid. The water is removed by azeotropic distillation with acetonitrile and the residue is crystallized from a mixture of 0.5 ml of methanol (to dissolve the residue) and 1 ml of acetonitrile, to give 81.8 mg of solid. A second recrystallization from 0.8 ml of methanol gives 47.9 mg, a third from 0.24 ml of methanol and 0.24 ml of absolute ethanol gives 44.8 mg, and a fourth from 0.225 ml of methanol and 0.225 ml of absolute ethanol gives 38.8 mg. The solid is dried at 20 ° C. and 0.01 mm Hg for 18 hours, then equilibrium with atmospheric humidity for 24 hours, which gives 40.9 mg of the title compound.

35 Example 148

Disodium salt of [3S- [3a (Z), 4ß]] - 3 - [[(2-amino-4-thiazolyl) [(1-carboxy-1-methylethoxy) imino] acetyl] amino] - 4 -methyl-2-oxo-l-azetidinesultonic /! A sample 126 of 3.00 g of acid [3S- [3a (Z), 4 (3]] -3- [[((2-amino-4-thiazolyl)) - is suspended in 30 ml of water. [(1-carboxy-1-methylethoxy) imino] acetyl] amino] -4-methyl-2-oxo-1-azetidinesulfonic (see Example 146), and the suspension is treated with IN sodium hydroxide, and use 12.0 ml to obtain the disodium salt of the title The pH is reduced to 6.5 by adding a little "Dowex 50W-X2" (H +).

The mixture is filtered and the filtrate is diluted to 66.3 g with water. A fraction of 6.63 g is separated for other uses. The remaining filtrate is lyophilized to give 2.38 g of solid. Partial equilibration (24 hours) with atmospheric humidity gives 2.54 g of the title compound. Examples 149 to 151

Following the procedure of Example 138, but replacing (Z) -2-amino-ct- (methoxyimino) -4-15 thiazoleacetic acid with the compound indicated in column I below, the compound indicated in column II below.

Αλ 127

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(U I i — i i — I I · i — i I

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128

Example 152

Potassium (1: 2) salt of [3S- [3α (Z), 4a]] - 3 - [[(2-amino-4-thiazolyl) [(1-carboxy-1-methylethoxy) iminojacetyl] -amino1-4-methyl-2-oxo-1-azetidinesulfonic 5 A) Potassium salt of [3S- [3g (Z), 4g]] acid - 3 - [[(2-amino-4-thiazolyl) [(1-diphenylmethoxycarbonyl-1-methyl-ethoxy) imino] acetyl] amino] -4-methyl] -2-oxo-1-azetidine sulfonic

A solution of 440 mg of (Z) -2-10 amino-a - [(1-carboxy-1-methylethoxy) imino] -4-thiazole-acetic acid and 153 mg of N-hydroxybenzotriazole mono-hydrate is treated. in 3 ml of dimethylformamide, with 206 mg of dicyclohexylcarbodiimide. The mixture is stirred at room temperature for 30 minutes under nitrogen, and a solution of 180 mg of (3S-cis) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid is added (see Example 137) and 0.14 ml of triethylamine in 2 ml of dimethylformamide (an additional 1 ml of dimethylformamide is used for rinsing), then the mixture is stirred for approximately 16 hours. The suspension is evaporated in vacuo and triturated with 12 ml of acetone. The suspension is filtered and the solid is washed with two fractions of 3 ml of acetone. The filtrate and the combined washing waters are treated with 338 mg of potassium perfluorobutanesulfonate. Dilution with 30 ml of ether gives a rubbery solid which solidifies slowly. This solid is filtered and washed with ether to obtain 656 mg of the title compound.

B) Potassium salt (1; 2) of the acid [3S- [3a (Z), 4a]] - 3 - [[(2-amino-4-thiazolyl) [(1-carboxy-1-methylethoxy) imino] -30 acetyl] amino] -4-methyl-2-oxo-1-azetidinesulfonic

A suspension of 656 mg of potassium salt of acid [3S- [3a (Z), 4a]] - 3 - [[((2-amino-4-thiazolvl)] [(1-diphenylmethoxycarbonyl) is cooled to -12 ° C. -1-methylethoxy) -imino] acetyl] amino] -4-methyl-2-oxo-1-azetidinesulionic 35 years old 2.3 ml of distilled anisole, and 11.5 ml of trifluoroacetic acid (cooled) beforehand at -10 ° C).

The solution is stirred for 15 minutes and 45 ml of ether are added, then 23 ml of hexane. After 5 minutes at -10 ° C. and 15 minutes of stirring at room temperature, one u.

129 filters the solid and is washed with ether to obtain 457 mg of a very hygroscopic gum. This gum is dissolved in 6 ml of cold water and the solution is immediately adjusted to pH 5.6 with a 0.4N potassium solution.

5 This solution is passed over 200 ml of "HP-20" resin and eluted with water. Fractions 7 to 11 are combined (50 ml each) and lyophilized to obtain 239 mg of the title compound as a solid.

Analysis for: 10 Calculated: C, 29.99; H, 3.10; N, 13.45; S, 12.32

Found: C, 29.94; H, 3.30; N, 13.30; S, 11.93. NMR (D20) 1.44 (3H, d, j = 75), 1.46 (6H, S), 4.48 (1H, dde tj -7 5.5, 5.5), 5.34 (1H, d ,) = 5.5), 6.96 ppm (1H, S).

Example 153 15 (±) -3-amino-4,4-dimethyl-2-oxo-1-azetidinesulfonic acid A) (±) -4,4-dimethyl-2-oxo-1-azëtidine-tert-butyl-diphenylsilane

A solution of 40.5 ml of t-butylchlorodiphenylsilane in 112 ml of dimethylformamide is cooled to 0 ° C. To this solution is added 22 ml of triethylamine. A solution of 12.87 g of 4,4-dimethyl-2-azetidinone in 25 ml of dimethylformamide is then added dropwise over 10 minutes to the cooled triethylamine solution. The resulting cloudy solution is stirred for 18 hours at 5 ° C under argon. This mixture is poured into 400 ml of ice water and extracted with three 150 ml fractions of a 2: 1 mixture of ether and ethyl acetate. The combined extracts are washed with four 100 ml fractions of 0.5M monobasic potassium phosphate buffer, a 150 ml fraction of sodium bicarbonate solution, two 150 ml fractions of water and a 150 ml fraction of saturated sodium chloride solution. The solution was dried over sodium sulfate and concentrated in vacuo to give 33.03 g of the title compound as a solid.

E) (+) - 3-azido-4,4-dimethyl-2-oxo-1-azetidine-tert-butyl-diphenylsilane

A solution of 4.25 ml of 1.6M n-butyl-lithium (in hexane) and 11 ml of tetrahydro is prepared at -50 ° C. under argon, in a 100 ml three-necked flask. - dry furan. A solution of 0.083 g of triphenylmethane in 1 ml of tetrahydrofuran is added. The resulting solution is cooled to -60 ° C, and 1.0 ml of diisopropylamine is added dropwise using a syringe. The whole is stirred for 15 minutes, then cooled to -78 ° C. A solution of 2.3 g of (±) -4,4-dimethyl-2-oxo-1-azetidine-tert-butyldiphenylsilane in 8 ml of tetrahydrofuran is added slowly, using a syringe.

The resulting solution is stirred for 20 minutes at -78 ° C, during which time heavy precipitation occurs and uniform stirring becomes difficult. A solution of 1.33 g of p-toluenesulfonyl azide in 5 ml of tetrahydrofuran is added dropwise. The resulting mixture is allowed to stir at -78 ° C for 20 minutes, and 2 ml of trimethylsilyl chloride is added dropwise. The reaction mixture is warmed to room temperature and stirred for 1 hour. Then the mixture is cooled to 0 ° C and poured into 150 ml of ethyl acetate at 0 ° C. Sufficient 0.5M monobasic potassium phosphate buffer is added to render the aqueous layer and the organic layer clear. The two layers are separated and the organic layer is washed with three 150 ml fractions of a 0.5M monobasic potassium phosphate solution, a 150 ml fraction of sodium chloride solution, a 150 ml fraction. saturated sodium chloride solution, and dried over sodium sulfate. The solution is concentrated in vacuo to give 2.83 g of an oil which, after trituration with hexane, gives 1.67 g of the title compound as a solid.

C) (±) -3-azido-4,4-dimethyl-2-oxo-1-azetidine

1.52 g of (±) -3-azido-4,4-dimethyl-2-oxo-1-azetidine-tert-35-butyldiohenylsilane are dissolved in 25 ml of acetonitrile in a three-necked flask of 50 ml. To this stirred solution, 0.25 ml of 48% hydrofluoric acid is added. The whole is stirred at room temperature, and fractions of 0.5 ml π of 48% hydrofluoric acid are added every 60 minutes until, after 131 6 and a half hours, a total of 3 , 25 ml of 48% hydrofluoric acid was added. The reaction mixture is then cooled to 0 ° C, neutralized with saturated sodium bicarbonate, and extracted with 120 ml of ethyl acetate. The organic layer is then washed with 100 ml of water, 100 ml of a saturated sodium chloride solution, and dried over sodium sulfate. The dry solution is concentrated under vacuum to obtain 1.34 g of oil. This impure oil is subjected to chromatography on 27 g of silica gel with hexane, then with 33% ethyl acetate in hexane, to obtain 0.358 g of the title compound in the form of a solid.

D) Tetrabutylammonium salt of (±) -3 ~ azido-4,4-dimethyl1-2-oxo-1-azetidinesulfonic acid 15 A 0.100 g of (±) -3-azido-4,4-dimethyl-2 -oxo-1- azetidine at 0 ° C., 2.8 ml of a dimethylformamide-sulfur trioxide complex 0.5 m are added under argon. This mixture is allowed to warm to ambient temperature and stirred. for 45 minutes. The solution is then poured into 20 ml of 0.5 M monobasic potassium phosphate buffer at pH 5.5. The whole is washed with three 20 ml fractions of methylene chloride (discarded) and 0.237 g of tetrabutylammonium hydrogen sulphate is added to the aqueous solution. The resulting mixture is extracted with four 20 ml fractions of methylene chloride and the combined organic extracts are washed with 20 ml of 8% sodium chloride solution. The methylene chloride solution is dried (over sodium sulfate) and concentrated in vacuo to give 0.31 g of an oil which, according to the NMR spectrum, consists of 50% dimethylformamide and 50% of the title compound.

E) (±) -3-amino-4,4-dimethyl-2-oxo-l ~ azetidine-sulfonic acid

A solution of 0.155 g of tetrabutylammonium salt of (±) -3-azido-4,4-dimethyl-2-oxo-1-azetidinesulfonic acid is hydrogenated in 0.6 ml of methanol over 10% palladium. on charcoal, for 20 minutes under an atmosphere. The catalyst is filtered off and rinsed with methylene chloride, and the water is combined.

132 rinsing with methanolic solution. This clear solution is treated with 0.123 ml of 97% formic acid. When the acid is added, the solution immediately becomes cloudy. After standing for 1 hour at 5 ° C, the solid is filtered off to obtain 0.0664 g of the title compound, m.p. 200-2Û2 ° C, with decomposition.

NMR (D-O) 1.64 (3H, S), 1.68 (3H, S), 4.42 (1H, S) Δ -1 IR (KBr) 1765 cm.

Example 154 10 Potassium salt of [3 ± (Z) 1—3 - [[(2-amino-4-thiazolyl) - (methoxyimino) acetyl] amino] -4,4-dimethyl-2-oxo- l-azetidine-sulfonic

A solution of 50 mg of N-hydroxybenzotriazole hydrate and 0.323 mmol of (Z) -2-amino-15a- (methoxyimino) -4-thiazoleacetic acid in 0.5 ml of dimethylformamide is treated with 67 mg of dicyclohexylcarbodiimide, under argon, at room temperature. The resulting mixture is stirred for 1 hour, after which time (±) -3-amino-4,4-dimethyl-2-oxo-1-azetidinesulfonic acid (57 mg is added; see Example 153 ), in the form of a solid, then 0.05 ml of triethylamine is added dropwise. The reaction mixture is stirred at room temperature for 16 hours. The dimethylformamide is removed in a high vacuum at 30 ° C., and the residue is diluted in 4 ml of acetone, then filtered. The filter cake is washed with an additional 4 ml of acetone, and 85 mg of potassium perfluorobutane sulfonate are added to the filtrate, followed by ether. Trituration of the resulting gum with ether gives 40 mg of a buff-colored solid which is subjected to chromatography on a 70 ml column of "HP-20 AG". Elution with water gives 20 mg of the title compound in fractions 16 to 40 (5 ml) after evaporation, trituration with a 1: 1 mixture of acetone and hexane and drying, m.p. 225 ° C, with decomposition.

35 Analysis for · Κ:

Calculated: C, 31.80; H, 3.40; N, 16.86; S, 15.43 Found: C, 29.47; H, 3.48; N, 14.98; S, 13.35.

/ h 133

Example 155

Potassium salt of (i) -4,4-dimethyl-2-oxo-3 - [(phenylacetyl) aminol-1-azetidinesulfonic acid

A solution of 45 mg of N-5 hydroxybenzotriazole hydrate and 40 mg of phenylacetic acid in 0.5 ml of dimethylformamide is treated with 61 mg of dicyclohexylcarbodiimide under argon at room temperature.

The resulting mixture is stirred for 1 hour and (±) -3-amino-4,4-dimethyl-2-oxo-1-azetidine-10 sulfonic acid (52 mg; 52 mg; see Example 153), as a solid, then 0.04 ml of triethylamine is added dropwise. The reaction mixture is stirred at room temperature for 24 hours. The dimethylformamide is removed in a high vacuum at 30 ° C. and the residue is diluted in acetone, then filtered. Potassium perfluorobutane, then ether, is added to the filtrate, and the mixture is cooled. The resulting solid was washed with acetone, hexane and dried, giving the title compound as a powder.

20 Analysis for:

Calculated: C, 44.55; H, 4.32; N, 8.00; S, 9.15; K, 11.16

Found: C, 43.83; H, 4.16; N, 7.96; S, 8.76; K, 11.43.

NMR (D20) 1.3 3 (S, 3H), 1.58 (S, 3H), 3.68 (S, 3H), 4.70 (S, 1H), 7.56 ppm (large S, 5H ).

25 Example 156

Potassium salt of (3S-trans-3 - [[(2-amino-4-thiazolyl) oxo-acetyljamjno] -4-methyl-2-oxo-1-azetidine-sulfonic acid A solution of 1.85 g of diphenyl-phosphinyl chloride in 15 ml of dry dimethylformamide, cooled in an ice and methanol bath (-15 to -20 ° C), 2.14 g of acid triethylamine salt (2- amino) -4-thiazolyl) glyoxylic. After stirring for half an hour, a solution of (3S-trans) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid (1.08) is added to the cold mixed anhydride solution. (see Example 139) and 1.92 ml of triethylamine in 5 ml of dry dimethylformamide, then the reaction mixture is stirred at 5 ° C for 24 hours.

/ The solvent is removed in vacuo, the residual dark oil is dissolved in water / v 134, and chromatography on resin "Dowex 50 X 2-400" (form K®, 200 ml). After elution with water (15 ml fractions), the crude product is collected in fractions 13 to 27 (3.37 g). Chromatography 5 on 200 ml of resin "HP-20", eluting with water (fractions of 15 ml), gives the desired product in fractions 18 to 26. The removal of water under vacuum gives the composed of the title in the form of an amorphous powder. Analysis for CgHgN ^ OgS ^ (372.42): Calculated: C, 29.02; H, 2.44; N, 15.04; S, 17.22; K, 10.50 Found: C, 28.87; H, 2.62; N, 14.85; S, 15.09; K, 10.81. Example 157

Trifluoroacetate (1; 1) of the potassium salt of the acid [3S (R *)] - 3 - [[[(aminoacëtyl) amino] phenylacetyl] amino] -2-oxo-15 l-azetidinesulfonique Elimination of the group protector of the potassium salt of the acid [3 S (R *)] -3- [[[[[f [. (4-methoxyphenyl) -methoxy] carbonyl] amino] acetyl] amino J phenylacety1] amino] -2-oxo-1-azetidinesulfonic (see Example 127), using trifluoroacetic acid and anisole, gives the title compound, melting point 165 ° C, with decomposition.

Example 158

Potassium salt of (3S-trans) -3-methoxy-4-methyl- 2-oxo-3 - [[(phenylmethoxy) carbonyl] amino] -1-azetidine-sulfonic A) (3S-trans) -4-methyl-3-methoxy-2-oxo-4 - [[(phenylmethoxy) -carbonyl1amino1azetidine

A solution of 2.5 g (0.0106 mole) of (3R - trans) -4-methyl-2-oxo-3 - [[(phenylmethoxy) -30 carbonyl] amino] azetidine is cooled to 0 ° C. prepared from d-threo-nine with a yield of 12.6%, essentially as described for the racemic cis isomer in Example 98C), in 112 ml of 4% borax in methanol, and added 3.5 ml of t-butyl hypochlorite. After 20 minutes, the solution is poured into 1 liter of cold etonextra.it water with two 750 ml fractions of cold ethyl acetate. The organic layer is washed with two 750 ml fractions of cold water, with a saturated salt solution, then dried and evaporated to give 3.05 g of crude N, N'- 4Λ 135 dichloramide.

A solution of 426 mg of lithium methylate in 20 ml of dry methanol is cooled to -78 ° C. and diluted with 40 ml of dry tetrahydrofuran. A solution of the above chloramide in 20 ml of tetrahydrofuran (-78 ° C) is added over 5 seconds using a syringe. After 20 minutes at -78 ° C, 2 ml of acetic acid and 2 ml of trimethylphosphite are added. After 40 minutes at room temperature, the solution is poured into 500 ml of water and extracted with two 300 ml portions of ethyl acetate. The organic layer is washed with water, dried, and evaporated to obtain an oil. Chromatography on a 200 ml silica gel column, eluting with a 3: 1 mixture of chloroform and ethyl acetate, gives a total of 1.25 g of the title compound.

B) Potassium salt of (3S-trans) -3-methoxÿ-4-methyl-2-oxo-3- [[(phenylmethoxy) carbonyl] amino] -1-azetidine sulfonic acid 20 Cool to 0 ° C a solution of 800 mg (0.00303 mole) of (3S-trans) -4-methyl-3-methoxy-2-oxo-4 - [[(phenylmethoxy) carbonyl] amino] azetidine in 2 ml of dimethylformamide, and 4 ml of a dimethylformamide-sulfur trioxide complex are added. After one hour at 0 ° C. and 4 hours at room temperature, the solution is poured into 80 ml of 0.5 M monobasic potassium phosphate (adjusted to pH 5.5) and extracted with two fractions of 50 ml of methylene chloride (throw away). The aqueous layer was treated with 1.04 g of tetrabutylammonium sulfate and extracted with dichloromethane to give 1.42 g of oil.

This oil is dissolved in acetone and treated with 1.04 g of potassium perfluorobutane sulfonate in 10 ml of acetone. Dilution with 250 ml of ether and further trituration of the oily solid gives 584 mg of crude product. Chromatography on 200 ml of "HP-20 AG" resin gives 418 mg of purified product in fractions 13 to 16 (100 ml) (elution with 1 liter of water, then with a 9: 1 mixture of water and d 'acetone). Trituration of / 114 mg of this substance with ether gives 104 mg of an analytical sample.

Analysis for C ^ H ^^ O ^ SK.I ^ O · '

Calculated: C, 39.06; H, 4.04; N, 7.01; S, 8.03; K, 9.78 Found: C, 38.91; H, 3.62; N, 6.91; S, 8.06; K, 9.51.

5 NMR (D20) 1.33 (3H, d, j = 7), 3.46 (3H, S), 4.22 (2H, d of d, J = 6), 5.18 (2H, S) , 7.43 ppm (5H, S).

Example 159

Potassium salt of (3S-trans) -3-methoxy-4-methyl "2-oxo-3 - [(phenylacetyl) amino] -1-azetidinesulionic acid The tetrabutylammonium salt of (3S acid) is prepared -trans) -3-amino-3 ~ methoxy-4-methyl-2-oxo-l-azetidine-sulfonic by catalytic hydrogenation of the potassium salt of (3S-trans) -3-methoxy-4-methyl- 2-oxo-3 - [[(phenyl-methoxy) carbonyl] amino] -1-azetidinesulfonic (see Example 158) after conversion to tetrabutylammonium salt, following the procedure of Example 88, but using the tetrabutylammonium salt of (3S-trans) -3-amino-3-methoxy-4-methyl-2-oxo-1-azetidine-sulfonic acid and phenylacetyl chloride, the title compound is obtained.

Analysis for c13Hi5N2 ^ 6SK:

Calculated: C, 42.61; H, 4.31; N, 7.65

Found: C, 39.67; H, 4.09; N, 7.30.

NMR D20) 1.29 (BH, d, j = 7), 3.45 (3H, S), 3.73 (2H, S), 4.36 (2H, d of d,; = 6), 7.38 ppm (5H, S).

Example 160

Potassium salt of [3S- [3g (Z), 4ß]] - 3 - [[(2-amino-4-thiazolyl) [[2- (diphenylmethoxy) -2-oxo-ethoxy] imino] acetyl ] -amino '] - 2-mé thy 1-4-oxo-l-azétidinesul tonique 30 Following the procedure of Example 138, but replacing the acid (Z) -2-amino-a- (methoxyimino ) -4-thiazoleacetic with (Z) -2-amino-a - [[2- (diphenylmethoxy) -2-oxo-ethoxy] imino] -4-thiazoleacetic, and first treating the acid ( 3S-cis) -3-amino-4-methyl-2-oxo-1,3-azetidinesulfonic by triethylamine, the title compound is obtained, melting point 155-160 ° C, with decomposition.

L

137

Example 161

Potassium salt of [3S ^ - [3a (Z), 4g]] -3- [[[(carboxy-methoxy) iroino] (2-aminO "4-thiazolvl) acetyl] amino] -4-methyl- 2-oxo-1-azetidinesulfonic 5 The elimination of the protective group of the potassium salt of the acid [3S- [3α (Z), 4 $]] -3- [L (2-amino-4-thiazolyl) [ [2- (diphenylmethoxy) -2-oxo-ethoxy] imino] acetyl] -amino] -2-methyl-4-oxo-1-azetidinesulfonic (see Example 160), using trifluoroacetic acid and anisole, 10 gives the title compound, which decomposes at> 250 ° C. Example 162

Acid potassium salt (S) -3 - [[[((2,6-dichloro-4-pyridiny1) thio] acetyl] amino] -2-oxo-1-azetidinesulfonic Acylation of the tetrabutylammonium salt of l (S) -3-amino-2-oxo-1-azetidinesulfonic acid (see Example 6A) with the 4-nitrophenyl ester of [(2,6-dichloro-4-pyridinyl) thiolacetic acid , followed by treatment with potassium perfluorobutanesulfonate, gives the title compound, melting point 212-214 ° C.

20 Example 163

Potassium salt of acid [3S (R *) 1--3- [[[[((4-amino-2,3-dioxo-1-piperazinyl) carbonyl] amino] phenylacetyl] amino] -4-methyl- 2-oxo-l-azetidinesulfonic

The potassium salt of the phenylmethyl ester of the acid [3S (R *)] - [4- C [[2- [(4-methyl-2-oxo-1-sulfo-3-azetidinyl)] is hydrogenated. amino] - 2-oxo-1-phenylethyl] amino] carbonyl] -2.3 -dioxo-1 -piperazinyl] -carbamic (see Example 149) using hydrogen gas and 10% palladium on carbon of catalyst to give the title compound, mp 165 ° C, with decomposition.

Example 164

[3S (Z)] -3- [[(2-amino-4-thiazolyl) '1 [(1-carboxy-1-methylethoxy) imino'jacetyl] amino] sodium salt (1: 2) -2-oxo-1-azetidinesulfonic 35 The protecting group of the tetra-butylammonium salt of [3S (Z)] - 3 - [[(2-amino-4-thiazolyl) - - [[2- ( diphenylmethoxy) -1, 1-dimethyl-2-oxo-ethoxy] imino] -acetyl] amino] -2-oxo-l-azetidinesulfonic (see Example ß 28) using trifluoroacetic acid and anisole , for i ^ \ 138 obtain the title compound, melting point 185 ° C, with decomposition, after transformation into disodium salt using aqueous sodium hydroxide, and purification on "HP-20". Examples 165 to 168 Following the procedure of Example 11, but replacing the acid (Z) -2-amino-a - [[[hydroxy (phenyl-methoxy) phosphinyl] methoxy] imino] -4- thiazoleacétique by the acid indicated in Column I below, the compound indicated in Column II below is obtained.

h 139

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140

Example 169

(Trans) -3-amino-4-ethyl-2-oxo-1-azetldinesulfonic acid A) t-boc-N-methoxy- £ -threo-ethylserinamide

1.33 g of threo-D, L-3-ethylserine are dissolved in 50 ml of 2N potassium hydroxide and 5 ml of t-butanol. After adding 2.46 g of di-t-butyl pyrocarbonate, the two-phase mixture is stirred for 4 hours at room temperature. 1.25 g of O-methylhydroxylammonium are added and the pH is adjusted to 4 with IN hydrochloric acid. 1.92 g of 1-ethyl-3- (3-dimethylamino-propyl) carbodiimide hydrochloride are added and the pH is adjusted again to 4. After stirring for one hour, the reaction mixture of sodium chloride is saturated and extracted with four 50 ml portions of ethyl acetate. The ethyl acetate extracts are combined and dried over magnesium sulfate. Removal of the solvent in vacuo gives 1 g of the title compound.

B) t-boc-O-methanesulfonyl-N-methoxy-B-threo-propionamide

10.5 g of t-boc-N-methoxy-8-threo-ethyl-20-serinamide are dissolved in 65 ml of pyridine. 4.65 ml of methanesulfonyl chloride are added dropwise at 0 ° C.

After stirring for 3 hours at room temperature, the reaction mixture is poured into 200 g of ice and 300 ml of IN hydrochloric acid. The pH is adjusted to 4 with concentrated hydrochloric acid. After extracting with three fractions of 85 ml of ethyl acetate, the combined extracts are dried over magnesium sulphate and they are concentrated in vacuo. The residue is treated with carbon tetrachloride and concentrated again. By stirring with ether and then filtering, 6.9 g of the title compound are obtained.

C) (trans) -3-t-butoxycarbonylamino-4-ethyl-1-methoxy-2-azetidinone

4.15 g of anhydrous potassium carbonate and 125 ml of dry acetone are brought to reflux and 3.4 g of t-boc-O-methanesulfonyl-N-methoxy-3-threo-propionamide are added in 25 ml of 'acetone. After one hour, the reaction mixture is cooled and filtered, and the filtrate is concentrated in vacuo. The oily residue is stirred with hexane

AT

141 to obtain 2.2 g of the title compound.

D) (trans) -3-t-butoxycarbonylamino ~ 4-ethyl - 2-azetidinone

3 g of (trans) -3-t-butoxycarbonylamino-4-ethyl-1-methoxy-2-azetidinone are added to 170 ml of liquid ammonia, 5 at -78 ° C under nitrogen, and 1 is added in five batches , 68 g sodium, with stirring, in 5 minutes. The agitation is continued for 30 minutes. Ammonium chloride is then added slowly until the blue color of the reaction mixture disappears. After removing the ammonia under nitrogen, the solid is extracted with two fractions of 100 ml of ethyl acetate. Removal of the solvent, followed by drying under vacuum, gives 2.7 g of the title compound.

E) Tetrabutylammonium salt of (trans) -3-t-butoxycarbonylamino - ^ - ethyl ^ -oxo-1-azetidinesulfonic acid To a solution of 2 ml of absolute pyridine in 20 ml of dry dichloromethane, 3, 7 ml of trimethylsilylsulfonyl chloride in 5 ml of dry dichloromethane. The addition is carried out at -30 ° C, under nitrogen, in 10 minutes. After stirring at room temperature for 30 minutes, the flask is evacuated to obtain a complex of pyridine and sulfur trioxide. 2.67 g of (trans) -3-t-butoxycarbonylamino-4-ethyl-2-azetidinone and 20 ml of dry pyridine are added to the flask, then the flask is placed in an oil bath preheated to 90 ° vs. After 15 minutes, a clear solution is obtained which is poured into 200 ml of an IM solution of dibasic potassium phosphate. After adding 27 g of dibasic potassium phosphate and 100 ml of water, a clear solution is obtained. This solution is extracted with two 60 ml portions of ethyl acetate. Tetrabutylammonium hydrogen sulfate is added to the aqueous layer, and the aqueous solution is extracted with three 100 ml portions of dichloromethane, then the combined organic layers are dried over magnesium sulfate. Concentration in vacuo gives 6.9 g of the title compound.

F) (trans) -3-amino-4-ethyl-2-oxo - l-azetidine-sulfonic acid

Stirred for 3 hours at room temperature Ζ / Λ 142 6.75 g of tetrabutylammonium salt of (trans) -3-t-butoxycarbonylamino-4-ethyl-2-oxo-1-azetidinesulfonic acid in 40 ml of 98% formic acid. 60 ml of dichloromethane are added and the mixture is cooled for approximately 16 hours. The resulting precipitate is separated by filtration and then dried under vacuum to obtain 0.85 g of the title compound, melting point 185 ° C, with decomposition.

Example 170

(Trans, Z) -3 - [[(2-amino-4-10 thiazolyl) [(1-carboxy-1-methylethoxy) imino] acetyl] amino] -4-ethyl-2-oxo dipotassium salt -l-aetidinesulfonic A) Dipotassium salt of (trans, Z) -3 - acid [[(2-amino-4-thiazolyl) [(l-diphenylmethoxycarbonyl-l-methylethoxy) imino] -acetyl] amino] -4 -ethyl-2-oxo-1-azetidinesulfonic 15 0.55 g of (trans) -3-amino-4-ethyl-2-oxo-1-azetidinesulfonic acid and 335 mg of triethylamine are dissolved in 50 ml of dry dimethylformamide . 1.14 g of (Z) -2-amino-a - [(1-diphenylmethoxycarbonyl-1-methylethoxy) -imino] -4-thiazoleacetic acid are added, with stirring at 0 ° C, then 20,450 mg are added d1hydroxybenzotriazole and 0.69 g of dicyclohexylcarbodiimide. After having stirred for approximately 16 hours at 0 ° C., the flask is placed under vacuum. 25 ml of dry acetone are added to the solid, with stirring. The mixture is filtered and 0.94 g of potassium perfluorobutanesulfonate is added to the filtrate, followed by 100 ml of ether. After standing for 1 hour at 0 ° C., the solid is filtered, washed with ether and dried under vacuum, which gives 1.58 g of the title compound.

B) Dipotassium salt of (trans, Z) -3 - [[(2-amino-4-30 thiazolyl) [(1-carboxy-1-methylethoxy) imino1acetynamino] -4- ethyl-2-oxo-1 acid -azetidinesulfonic A solution of 1.31 g of dipotassium salt of (trans, Z) -3 acid - [[(2-amino-4-thiazolyl) [(1-diphenylmethoxy-carbony1-1-methylethoxy) imino] acetyl ] amino] -4-ethyl-2-oxo-35 1-azetidinesulfonic in 10 ml of anisole, 5 ml of trifluoroacetic acid are added over 10 minutes at -15 ° C. After stirring for 2 hours at -10 ° C, a clear solution is obtained. At -30 ° C., 80 ml of dry ether are added and the resulting precipitate is filtered and then treated with 5 ml i / ~ \ 143 of water. The pH is adjusted to 5.5 with IN potassium hydroxide at 0 ° C., and the mixture is filtered to remove the unprocessed starting material. The filtrate is subjected to chromatography on "HP-20" with water as eluent. Lyophilization gives 185 mg of the title compound, melting point 160 ° C, with decomposition.

Example 171

Potassium salt of [3S (Z)] acid - 3 - [[(2-amino-4-thiazolyl) [(4-hydroxy-4-oxobutoxy) imino] acetyl] amino] -2-10 oxo-l -azetidinesulfonic The elimination of the protective group of the potassium salt of the acid [3S (Z)] - 3 - [[(2-amino-4-thiazolyl) [[4- (diphenylmethoxÿ) -4-oxobutoxy] imino] acetyl] amino] -2-oxo-1-azetidinesulfonic (see Example 166), using trifluoroacetic acid and anisole, gives the title compound, melting point> 200 ° C.

Example 172

Internal salt of acid (S) -3 - [[2- (aminomethyl) benzoyl] -amino] -2-oxo-1-azetidinesulfonic The elimination of the protective group of the potassium salt of acid (S) -2-oxo-3 - [[2 - [[[(phenylmethoxy) -carbonyl] amino] methyl] benzoyl] amino] -1-azetidinesulfonic using hydrogen gas, palladium on carbon and hydrochloric acid , gives the title compound, mp 162-165 ° C.

Example 173

Potassium salt of acid (S) -3 - [[[2- (4-formyl-1-piperazinyl) -5-hydroxypyrido [2,3-d] pyrimidin-6-yl] carbonyl] -amino] - 2-oxo-1-azetidinesulfonic acid The tetrabutylammonium salt of (S) -3-amino-2-oxo-1-azetidinesulfonic acid (see Example 6A) is condensed with 2- (4-formyl-1 -piperazinyl) -5-hydroxy-6 - [(4-nitro-phenoxy) carbonyl] pyrido [2,3-d] pyrimidine, and the resulting compound is treated with potassium perfluorobutanesulfonate in acetone, to give the title compound, melting point 290 ° C, with decomposition.

/ h 144

Example 174

(3S-trans) -α-Γ [(4-methyl-2-oxo-1-sulfo-3-azetidinyl) amino1carbonyl] benzeneacetic acid dipotassium salt

The (3S-trans) -3-amino-4-methyl-2-5 oxo-1-azetidinesulfonic acid (see Example 139) is condensed with α- (carboxyl) benzene acetyl chloride, and the resulting compound with triethylamine and potassium perfluorobutane to obtain the title compound, mp 147 ° C, with decomposition.

10 Example 175

[3S-trans] -3-amino-4-cyclohexyl-2-oxo-1-azetidine-sulfonic acid A) g- (t-butoxycarbonylamino) -ß-cyclohexyl-ß-hydroxy-threo-propionic acid 15 suspension 15 g of β-cyclohexyl-06 * amino-8-hydroxy-threo-propionic acid in 150 ml of acetonitrile and 70 ml of water. 17.8 g of triethylamine are added and the mixture is heated, with stirring, to 60 ° C. At this temperature a clear solution is obtained and 21.0 g of di-t-butyl pyrocarbonate is added, and stirring is continued at 60 ° C for one and a half hours. The solvent is removed in vacuo and 50 ml of water are added. The aqueous layer is extracted with ethyl acetate to a pH of 2, which is adjusted by adding 3N hydrochloric acid. The organic layer is separated, dried over sodium sulfate and evaporated to dryness. The remaining crystalline substance is filtered with petroleum ether to give 20.4 g of the title compound, mp 113-115 ° C.

B) a- (t-butoxycarbonylamino) -ß-cyclohexyl-ß-hydroxy-N-methoxy-threo-propionamide

20.2 g of a- (t-butoxycarbonylamino) -ß-cyclohexyl-ß-hydroxy-threo-propionic acid and 7.6 g of O-methylhydroxylamine hydrochloride are suspended in 350 ml of water and 175 ml of t-butanol. The mixture is adjusted to pH 4 with potassium carbonate. 16.4 g of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide are added and the pH is kept at 4, with stirring, for one and a half hours. The t-butanol is removed in vacuo and the remaining aqueous solution is saturated with sodium chloride and then extracted twice with 100 ml portions of ethyl acetate. The organic layers are combined, dried over sodium sulfate and evaporated to dryness.

The remaining crystals are separated by filtration with petroleum ether to give 18.6 g of the title compound, mp 125-127 ° C.

C) a- (t-butoxycarbonylamino) -ß-cyclohexyl-ß- (methane-tallow onyloxy) -N-methoxy-threo-propionamide 10.3 g of a- (t-butoxycarbonylamino) -ß- cyclohexyl are dissolved -p-hydroxy-N-methoxy-threo-propionamide, with stirring, in 100 ml of dry pyridine. The solution is cooled, with stirring, to 0 ° C, and 9.3 g of methanesulfonyl chloride are added dropwise. After one hour at 0 ° C, an additional 3.3 g of methanesulfonyl chloride is added and stirring is continued for another hour. The solution is poured into 300 ml of ice water, 200 ml of ethyl acetate are added and the pH is adjusted to 3 with dilute sulfuric acid. The organic layer is separated, dried with sodium sulfate and the solvent is removed in vacuo. The remaining solid is collected with petroleum ether to give 19.0 g of the title compound, melting point 150-152 ° C.

D) [3S-trans] -3- (t-butoxycarbonylamino) -4-cyclohexyl-l-25 methoxy-2-azetidinone

18.7 g of dV- (t-butoxycarbonylamino) -ß-cvclohexyl-jj- (methanesulfonyloxy) -N-methoxy-threo-propionamide are dissolved in 500 ml of dry acetone. 9.8 g of potassium carbonate are added and the suspension is heated to the temperature of 30 reflux, with stirring, for 5 hours. The insoluble mineral substance is filtered off and the solvent is removed in vacuo, then the remaining oil is dissolved in 30 ml of ethyl acetate. By adding petroleum ether, the title compound is precipitated, which is filtered off (12.9 g), melting point 110-112 ° C.

E) [3S-trans] -3- (t-butoxycarbonylamino) -4-cyclohexyl-2-azetidinone

1 g of [3S-trans] -3- (t-butoxycarbonylamino) -4-cyclohexyl-1-methoxy-2-aetidinone is added to 50 ml of liquid ammonia 146, with stirring. 0.154 g of sodium is added in 5 to 6 fractions over 5 minutes. At the end of this time, an additional 0.025 g of sodium is added and stirring is continued for 5 minutes. 0.89 g of ammonium chloride is added and the ammonia is removed. The residue is extracted with hot ethyl acetate. The organic extract is evaporated to dryness and the remaining crystals of the title compound are filtered with petroleum ether, giving 0.5 g of product, melting point 130-132 ° C.

10 F) Pyridine salt of [3S-trans] -3- (t-butoxy-carbonylamino) -4-cyclohexyl-2-oxo-1-azetidinesulfonic acid

5.3 g of [3S-trans] -3- (t-butoxycarbonyl-amino) -4-cyclohexyl-2-azetidinone are dissolved in 20 ml of methylene chloride and 80 ml of dimethylformamide. After adding 60 mmol of a pyridine-sulfur trioxide complex, the solution is stirred for 6 hours at room temperature. Removal of the solvent in vacuo gives 11.3 g of the title compound as an oil.

G) Tetrabutylammonium salt of [3S-trans ~ [-3- (t- 20 butoxycarbonylamino) -4-cyclohexyl-2-oxo-1-azetidinesulfoni-acid

11.3 g of pyridine salt of [3S-trans] -3- (t-butoxycarbonylamino) -4-cyclo-hexyl-2-oxo-1-azetidinesulfonic acid are dissolved in 250 ml of water. 9.0 g of tetrabutylammonium hydrogen sulphate are added with stirring, and the pH is adjusted to 6.5 with 1N potash. The aqueous solution is extracted twice with 200 ml fractions of methylene chloride. The organic fractions are dried with sodium sulfate, filtered and the sol-vant is distilled off, giving 8 g of the title compound, melting point 135-138 ° C.

H) [3S-trans] -3-amino-4-cyclohexyl-2-oxo-1-azetidine-sulfonic acid

3.8 g of tetrabutylammonium salt of [3S-trans] -3- (t-butoxycarbonylamino) -4-cyclohexyl-2-oxo-1-azetidinesulfonic acid are stirred in 20 ml of formic acid for 3 hours , then 20 ml of methylene chloride are added. The title compound precipitated (1.0 g) is filtered off, mp 217-219 ° C.

AT

147

Example 176

Potassium salt of [3S- [3a (Z), 4ß]] - 3 - [[(2-amino-4-thiazolyl) (methoxyimino) acetyl] amino3-4-cyclohexyl-2-oxo ~ l- 0.25 g of [3S-trans] -3-amino-4-cyclohexyl-2-oxo-1-azetidinesulfonic acid are dissolved in 30 ml of dry dimethylformamide and 0.12 g of triethylamine, shaking. When a clear solution has been obtained, 0.2 g of (Z) -2-amino-a- (methoxyimino) -4-thiazoleacetic acid, 0.16 g of hydroxybenzotriazole and 0.42 g of dicyclohexylcarbodiimi are added. -of. Stirring is continued for 48 hours at room temperature. The precipitated urea is filtered off and the solvent is removed in vacuo. The residue is dissolved in 10 ml of acetone and 0.41 g of potassium perfluorobutane-sulfonate is added. After adding 50 ml of ether, the title compound precipitates, and is filtered. Column chromatography using resin "HP-20" and a 9: 1 mixture of water and acetone as eluent, gives 0.36 g of product, melting point 200-205 ° C (after lyophilization-20 tion ).

Example 177

[3S- [3g (Z), 4ß]] - 3 - [[(2-amino-4-thiazolyl) [(1-carboxy-1-methylethoxy) imino] acetyl] amino] -4 acid dipotassium salt -cyclohexyl-2-oxo-1-azetidinesulfonigue 25 A) Potassium salt of acid [3S- [3g (Z), 4ß]] - 3 - [[(2-amino-4-thiazolyl) [(1- diphenylmethoxycarbonyl-1-methyl-ethoxy) imino3acetyl] amino] -4-cyclohexyl-2-oxo-1-azetidine-sulfonic

[3S-trans] -3-amino-4-cyclohexyl-30 2-oxo-1-azetidinesulfonic acid (0.2 g; see Example 175) is dissolved in 30 ml of dimethylformamide and 0.09 g of triethylamine, with stirring. 0.12 g of hydroxybenzotriazole, 0.30 g of (Z) -2-amino-a - [(1-diphenylmethoxycarbonyl-1-methyl-ethoxy) imino] -4-thiazoleacetic acid and 0.33 g of dicyclohexyl-35 carbodiimide, and stirring is continued at room temperature for 12 hours. The precipitated urea is separated by filtration and the mother liquor is evaporated to dryness.

The remaining oil is dissolved in 5 ml of acetone and treated. the solution with 0.3 g of potassium perfluorobutanesulfonate / 7 / \ 148 and poured into 100 ml of ether, with stirring. The potassium salt of [3S- [3α (Z), 4ß]] - 3 - [[(2-amino- 4-thiazolyl) [(1-diphenylmethoxycarbonyl-1-methylethoxy) -imino] acetyl] amino acid ] -4-cyclohexyl-2-oxo-1-azetidinesulfoni-5 which (0.61 g) precipitates and is separated by filtration.

B) [3S- [3α (Z), 4ß]] - 3 - [[(2-amino-4-thiazolyl) [(1-carboxy-1-methylethoxy) iminolacetyl] -amino] acid dipotassium salt 4-cyclohexyl -2-oxo-l · -azetidinesulfonic

0.61 g of potassium salt of [3S- [3a (Z), 4ß]] - 3 - [- ([(2-amino-4-thiazolyl)] [(1-diphenylmethoxycarbonyl-1- methylethoxy) imino] acetyl] amino] - 4-cyclohexyl-2-oxo-1-azetidinesulfonic acid in 6 ml of anisole, cooled to -15 ° C, and 5 ml of trifluoroacetic acid are added dropwise, with stirring . The temperature is maintained for one hour and then lowered to -30 ° C.

About 100 ml of dry ether are added, at a rate such that the temperature does not exceed -10 ° C. The precipitated compound is filtered off and subjected to chromatography on "HP-20" resin, using a 9: 1 mixture of water and acetone as eluent, to give 0.3 g of the compound. titer, melting point 119-120 ° C, with decomposition (after lyophilization).

Example 178

Potassium salt of the acid [3S- [3g, 4ß]] - 4-cyclohexyl-3-25 [[[[[[3 - [(2-furanylmethylene) amino] -2-oxo-l-imidazolidinyl] - carbony1 ] amino 3 phenylacety1] amino] -2-oxo-l-azetidine-sulfonic

[3S-trans] -3-amino-4-cyclohexyl-2-oxo-1-azetidinesulfonic acid (0.1 g; see Example 175) is dissolved in a mixture of 30 ml of dry dimethylformamide and . 0.05 g of triethylamine, with stirring. 0.14 g of [[[(2-furanylmethylene) amino]] -2-oxo-1-imidazoli-dinyl] carbonyl] amino] phenylacetic acid, 0.06 g of hydroxy-benzotriazole and 0.17 g are added. g of dicyclohexylcarbodiimide, and the solution was stirred for 5 days at room temperature. The solvent is removed in vacuo, the residue is dissolved in 10 ml of acetone and the precipitated urea is filtered off. The mother liquor is stirred with 0.15 g of potassium perfluoro- / butanesulfonate and diluted with 50 ml of ether.

k \ 149

The precipitate is filtered and subjected to chromatography with "HP-20" resin using a 9: 1 mixture of water and acetone as eluent, which gives 0.14 g of product, melting point. 195-200 ° C, with decomposition 5 (after lyophilization).

Example 179

Potassium salt of [3S- [3Qt (R *), 43]] -4-cyclohexyl- 3- [[3- (4-ethyl ~ 2,3-dioxo-1-piperazinyl) -1,3 -dioxo-2-phenylpropyl] amino] -2-oxo-1-azetidinesulfonic acid [3S-trans] -3-amino-4-cyclohexyl-2-oxo-1-azetidinesulfonic acid is dissolved (0.1 g; see Example 175) in 30 ml of dimethylformamide and 0.5 g of triethylamine, with stirring. Add (R) -a - [[(4-ethyl-2,3-dioxo-1-piperazinyl) carbonyl] amino] benzeneacetic acid, 0.06 g of hydroxybenzotriazole and 0.17 g of dicyclohexylcarbodiimide and the mixture is stirred for about 16 hours at room temperature. The solvent is removed by vacuum distillation and the remaining oil is dissolved in 10 ml of acetone.

The precipitated urea is filtered off and the mother liquor is stirred with 0.15 g of potassium perfluorobutanesulfonate, then diluted with 50 ml of ether. The precipitate is separated by filtration and subjected to chromatography on resin "HP-20", using as eluent a 9: 1 mixture of water and acetone, which gives 0.15 g of the title compound , melting point 175-180 ° C (after lyophilization).

Example 180

Potassium salt of (trans, Z) -3 - [[(2-amino-4-thiazolyl) (methyloxyimino) acetyljamino] -2,4-ethyl-2-oxo-l-30 azetidinesulfonic acid

Following the procedure of Example 170, part A, but replacing the (Z) -2-amino-a - [(1-diphenylmethoxycarbonyl-1-methylethoxy) iminoI-4-thiazole-acetic acid with (Z) -2-amino-a- (methoxyimino) -4-35 thiazoleacetic acid, the title compound is obtained, melting point 190 ° C, with decomposition.

Example 181

Acid (±) - (trans) -3-amino-2-oxo-4-phenyl ~ l-azetidine-sulfonic 150 A) (±) - (trans) -2-oxo-4-phenyl-l-azetidine-tert -butyl-diphenylsilane

A solution of 20.56 g of tert-butylchlorodiphenylsilane in 45 ml of 5-dimethylformamide is cooled to 0 ° C. under argon, and it is treated with 10.4 ml of triethylamine, then with (±) -2-oxo -4-phenyl-1-azetidine. After several hours at 0 ° C., the resulting mixture is treated with an additional 1 ml of triethylamine and with 2.11 g of tert-butyl-chlorodiphenylsilane and the mixture is left to stir for 65 hours at 5 ° C. The reaction mixture is poured into 300 ml of ice water and extracted with three 125 ml fractions of a 3: 1 mixture of ether and ethyl acetate. The organic extracts are washed with a phosphate buffer at pH 4.5 (three 50 ml fractions), with 50 ml of a saturated sodium bicarbonate solution, with two 50 ml fractions of water, with a solution saturated with sodium chloride, and dried over sodium sulfate. Filtration and concentration in vacuo give a solid which is washed with hexane to obtain, after drying under high vacuum, 15 g of the title compound as a solid.

B) (t) - (trans) -3-azido-2-oxo-4-phenyl-1-azetidine-tert-butyldiphenylsilane "

A 50 ml flask equipped with a stirring bar, a gas inlet and a partition wall is dried with a argon torch, and charged with n-butyl lithium (0.65 ml d '' a 1.6 M solution in hexane) which is cooled to -40 ° C and which is dissolved in 2 ml of tetrahydrofuran. 0.16 ml of diisopropylamine is added dropwise, the resulting mixture is stirred for 30 minutes and cooled to -78 ° C. A solution of (±) - (trans) -2-oxo-4-phenyl-1-azetidine-tert-butyldiphenylsilane (400 mg) in 1.5 ml of tetrahydrofuran is added dropwise over approximately 5 minutes. After stirring for a further 20 minutes, the solution is treated with 204 mg of p-toluenesulfonyl azide in 0.5 ml of tetrahydrofuran. The resulting mixture is stirred for 10 minutes at -78 ° C and treated dropwise with 0.4 ml of chlorotrimethylsilane. After stirring for a further 10 minutes, the cooling bath is removed and the reaction mixture is stirred // λ 151 at room temperature for 2.5 hours. Then, while cooling to 0 ° C, 20 ml of ethyl acetate are added, followed by 8 ml of a phosphate buffer at pH 4.5. The organic layer is washed with two additional 8 ml fractions of the buffer, with three 10 ml fractions of a 5% sodium bicarbonate solution, with 10 ml of a 50% sodium chloride solution, with 10 ml of a saturated solution of sodium chloride, then dried over sodium sulfate. Filtration and concentration in vacuo gives 500 mg of an oil, which is subjected to flash chromatography with a mixture of 5% ethyl acetate and hexane, which gives 253 mg of the title compound .

C) (±) - (trans) -3-azido-2-oxo-4-phenyl-l-azetidine 15 A solution of 17 g of (±) - (trans) -3-azido is dissolved in 240 ml of methanol -2-oxo-4-phenyl-1-azetidine-. crude tert-butyldiphenylsilane, then the solution is treated dropwise at 0 ° C with 35 ml of concentrated hydrochloric acid. The cooling bath is removed, the reaction mixture is stirred at room temperature for one hour, then cooled again to 0 ° C, after which a saturated sodium bicarbonate solution is added until neutral. The resulting mixture is extracted with ethyl acetate (a fraction of 300 ml and four fractions of 100 ml) and the organic extracts are washed with a 1: 1 mixture of 5% sodium bicarbonate and 50% sodium chloride, then with a saturated solution of sodium chloride, and dried over sodium sulfate. Filtration and concentration in vacuo gives 15 g of a thick oil which is subjected to chromatography: on 100 g of silica gel with a mixture of 20% ethyl acetate and hexane, which gives 460 mg of the title compound.

D) tetrabutylammonium acid of (t) - (trans) -3-azido-4-35 phenyl-1-azetidinesulfonic acid

A solution of 300 mg of (±) - (trans) -3-azido-2-oxo-4-phenyl-1-azetidine in 3 ml of dimethylformamide is cooled to 0 ° C. under argon, and treated dropwise. with a complex of dimethylformamide and sulfur trioxide fi / "\ 152 (4.78 ml of a 0.5 M solution in dimethylformamide).

The cooling bath is removed, the reaction mixture is stirred at room temperature for 2 hours and poured into 80 ml of 0.5 M mono-basic potassium phosphate at pH 5.5. The solution is extracted with dichloromethane (to be discarded) and 541 mg of tetrabutylammonium bisulfate is added. The resulting mixture is extracted with dichloromethane and the organic extracts are washed with 10% sodium chloride solution, then dried over sodium sulfate. Filtration and concentration in vacuo gives 800 mg of an oil, comprising * about 40% of the desired product, the remainder being formed by dimethylformamide. This mixture is used without purification in the next step.

E) (±) - (trans) -3-amino-4-phenyl-1-f-azetidine-sulfonic acid

A solution of tetrabutylammonium salt of (+) - (trans) -3-amino-4-phenyl-1-azetidine-sulfonic acid in 4 ml of methanol is hydrogenated over 30 mg of platinum oxide, under 1 atmosphere and at room temperature. After 15 minutes, the apparatus is placed under vacuum and fresh hydrogen is introduced. After another 45 minutes, the reaction is complete, and the apparatus is purged with nitrogen. After several days at room temperature in 200 ml of a 4: 1 mixture of dichloromethane and methanol, the aggregation of the catalyst is complete and filtration is carried out. Then the filtrate is concentrated in vacuo to 18 ml and 0.2 ml of 97% formic acid is added. After cooling to 5 ° C for several hours, the resulting solid is filtered and washed with dichloromethane to obtain, after drying, ~ 150 mg of the title compound as a solid.

Analysis for:

Calculated: C, 44.62; H, 4.17; N, 11.57; S, 13.23 35 Found: C, 43.36; H, 4.31; N, 11.09; S, 13.02.

Example 182

Potassium salt of (i) - (trans) -2-oxo-4-phenyl-3- [(phenylacetyl) amino] -1-azetidinesulfonic acid A) Tetrabutylammonium salt of (±) -trans- acid 2-oxo-153 4-phenyl-3 - [(phenylacetyl) amino] -1-azetidinesulfonic

A solution of 52 mg of N-hydroxybenzo-triazole monohydrate and 46 mg of phenylacetic acid in 0.3 ml of dimethylformamide is treated with 70 mg of solid 5-dicyclohexyl-carbodiimide under argon at 0 ° C. The cooling bath is removed and the resulting mixture is stirred at room temperature for one hour. After diluting with an additional 0.3 ml of dimethylformamide, (±) - (trans) -3-amino-2-oxo-4-phenyl-1-azetidine-10 sulfonic acid is added in the form of a solid (75 mg, see Example 181), then 0.05 ml of triethylamine is added dropwise. The reaction mixture is stirred at room temperature for 23 hours, filtered and the filter cake is washed with dimethylformamide. The filtrate is added to 20 ml of 0.5 M monobasic potassium phosphate (pH 4.5), the mixture is washed with three 8 ml portions of ethyl acetate (discard), and 105 mg is added (0.31 mmol) of tetrabutylammonium bisulfate. The resulting mixture is extracted with three fractions of .15 ml of dichloromethane.

The extracts are washed with two 15 ml portions of 10% sodium chloride solution, thickened with 10 ml of saturated sodium chloride solution, and dried over sodium sulfate. Filtration and concentration in vacuo give (after heating to 32 ° C in high vacuum) 165 mg of an oil, comprising about 40% of the desired compound, and 60% of dimethylformamide.

B) Potassium salt of (±) -trans-2-oxo-4-phenyl-3 - [(phenylacetyl) amino] -1-azetidinesulfonigue acid

A solution of tetrabutylammonium salt of (±) -trans-2-oxo-4-phenyl-3 - [(phenylacetyl) amino] -1-azetidinesulfonic acid salt in 1.5 ml of acetone is treated with 41 mg (0.121 mmol) of potassium perfluorobutanesulfonate. Dilution with 12 ml of ether gives a glass which, after trituration with ether, provides 43 mg of a solid which contains about 20% of an impurity itself containing a tetrabutylammonium part. The solid is dissolved in 50% aqueous acetone and the solution is passed into 1 ml of an "Dowex 50W-X2" (K®) ion exchange resin.

. Removal of the solvent gives a solid which is washed with / Λ 154 acetone, then with hexane, and which is dried at 60 ° C under high vacuum. The yield of the title compound is 15 mg.

Analysis for .K: 5 Calculated: C, 51.23; H, 3.80; N, 7.03; S, 8.05; K, 9.81 Found: C, 50.44; H, 4.20; N, 7.01; S, 7.59; K, 9.40.

Example 183

Acid potassium salt (±) - (trans, Z) -3 - [[(2-amino-4- 'thiazolyl) (methoxyimino) acetyl] amino] -2-oxoT-4-phenyl-1-10 azetidinesultonic

A solution of 52 mg of N-hydroxybenzotriazole hydrate and 69 mg of (Z) -2-amino-a- (methoxyimino) -4-thiazoleacetic acid in 0.3 ml of dimethylformamide is treated, per 70 mg of solid dicyclohexylcarbodiimide, .15 under argon, at room temperature. The resulting mixture is stirred for one hour, (+) - (trans) -3-amino-2-oxo-4-phenyl-1-azetidinesultonic acid (75 mg; see Example 181) is added under form a solid, then 0.05 ml of triethylamine is added dropwise. The reaction mixture is stirred at room temperature for 23 hours. Dimethylformamide is removed under high vacuum at 30 ° C., and the residue is triturated with 2 ml of acetone, then filtered. The filter cake was washed with two additional 3 ml · fractions of acetone and 86 mg of potassium perfluorobutanesulfonate was added to the filtrate. Dilution with 10 ml of ether gives a rubbery solid which is triturated and washed with acetone and hexane to obtain, after drying, 82 mg of the title compound in the form of a solid.

30 Analysis for C25H ^ 4N5 ° gS2 * K:

Calculated: C, 40.26; H, 3.16; N, 15.65; S, 14.33; K, 8.74 Found: C, 38.60; H, 3.19; N, 15.07; S, 13.87; K, 7.5. Example 184

Potassium salt of (cis) "2-oxo-4-phenyl-3 - [(phenyl-35 acetyl) aminol-1-azetidinesulfonic A) N-benzylidene-2,4-dimethoxybenzylamine

12.0 g of 2,4'-dimethoxy-benzylamine hydrochloride are added to 100 ml of a 1N sodium hydroxide solution and the mixture is extracted with 125 ml of ethyl acetate. The organic layer is dried 155 over anhydrous sodium sulfate and freed from the solvent to obtain 10.2 g of 2,4-dimethoxybenzylamine as an oil. This amine is dissolved in 150 ml of benzene; 6.47 g of benzaldehyde and 0.6 g of p-toluenesulfonic acid monohydrate are added. The mixture is heated to reflux, removing the water using a Dean-Stark separator, and within two hours the calculated amount of water (1.1 ml) has separated.

The mixture is cooled to room temperature. When further cooled, the benzene solution deposits a little precipitate. The benzene is removed under reduced pressure and 60 ml of petroleum ether are added to the residue. An oily layer separates with a little more precipitate. 10 ml of benzene are added to make the layers homogeneous and the remaining precipitate is filtered. The filtrate is freed from the solvent to obtain 14.2 g of the title compound in the form of an oil.

B) (±) - (cis) -4-phenyl-1- (2,4-dimethoxybenzyl) -2-oxo-3-azidoazetidine 20.62 g of α-azidoacetic acid are dissolved in 25 ml of chloride methylene, under nitrogen. To this solution, 3.24 g of triethylamine and 1.02 g (4.0 mmol) of the imine of N-benzylidene-2,4-dimethoxybenzylamine dissolved in 10 ml of methylene chloride are added. The mixture is cooled in an ice bath and 3.36 g of trifluoroacetic anhydride is added slowly; the solution takes on a dark color. After stirring for one hour in an ice bath, the mixture is warmed to room temperature and stirred for a further 15 minutes. The solution is then washed with 60 ml of water, with two 50 ml fractions of a 5% sodium bicarbonate solution, and with 60 ml of a 1N hydrochloric acid solution. The organic layer is dried over anhydrous sodium sulfate and freed from the solvent to give 1.72 g of crude product in the form of a dark gum. This gum is treated with charcoal several times and the resulting brown mixture is subjected to chromatography on 40 g of silica gel, eluting with a 1: 1 mixture of petroleum ether and ethyl acetate. The combined fractions 156 give crystals by rapid cooling in a bath of dry ice and acetone. Using these crystals as a primer, the product is recrystallized from a mixture of petroleum ether and ethyl acetate to obtain 817 mg of the title compound in the form of needles which melt on warming to room temperature .

C) (i) - (cis) -4-phenyl-2-oxo-3-azidoazetidine

737 mg of (±) - (cis) -4-phenyl-1- (2,4-10 dimethoxybenzyl) -2-oxo-3-azidoazetidine are dissolved in 25 ml of acetonitrile and the mixture is heated to 80-83 ° C. under nitrogen. To the resulting solution are added, in one hour, 943 mg of potassium persulfate and 570 mg of potassium monoacid phosphate, both dissolved in 25 ml of water. After the addition, the mixture is further heated to 80-83 ° C for 7 hours. The mixture is cooled and the pH is adjusted to 6-7 by adding solid potassium monoacid phosphate. Most of the acetonitrile is removed under reduced pressure and the resulting mixture is extracted with 60 ml of chloroform. The chloroform layer is washed with 60 ml of water, dried over anhydrous sodium sulfate and the solvent is removed to obtain a crude product in the form of an oil. The crude product is subjected to chromatography on 40 g of silica gel, eluting with a 1: 1 mixture of petroleum ether and ethyl acetate. The combined fractions give crystals, and the product is recrystallized from a mixture of petroleum ether and ethyl acetate to obtain 267 mg of the title compound.

D) Tetrabutylammonium salt of (±) - (cis) -4-30 phenyl-2-oxo-3-azido-1-azetidinesulfonic acid i Cooled to 0 ° C under nitrogen 162 mg of (±) - ( cis) -4- phenyl-2-oxo-3-azidoazetidine, and 3.5 ml of an approximately 0.5 M solution of a dimethylformamide complex is added dropwise using a syringe and sulfur trioxide. The resulting clear solution was stirred at 0 ° C for 15 minutes. The mixture is then poured into 50 ml of a 0.5 M monobasic potassium phosphate solution and washed with three 50 ml fractions of methylene chloride. 292 mg of tetra-n-butylammonium bisulfate are added to the aqueous solution and the mixture is extracted with six 50 ml fractions of methylene chloride. The combined layers of methylene chloride are dried over anhydrous sodium sulfate and freed from the solvent to obtain 5,272 mg of the title compound as a gum.

E) Potassium salt of (±) - (cis) -2-oxo-4-phenyl-3 - [(phenylacetyl) amino] -1-azetidinesulfonic acid

293 mg of tetrabutylammonium salt of (±) - (cis) -4-phenyl-2-oxo-3-azido-1-azetidinesulfonic acid are dissolved in 4 ml of ethanol, and the solution is hydrogenated with 80 mg of platinum oxide serving as catalyst, under 1 atmosphere. After having stirred for one hour, the catalyst is filtered on a "Millipore" filter with "Celite"; some catalyst particles pass through the filter giving a black filtrate. The ethanol is removed under reduced pressure and the residue is dissolved in 4 ml of dimethylformamide. 81 mg of N-hydroxybenzo-triazole monohydrate, 78 mg of phenylacetic acid and 117 mg of dicyclohexylcarbodiimide are added, and the mixture is stirred for about 16 hours under nitrogen. The suspension is evaporated in vacuo and triturated with 10 ml of acetone. The resulting suspension is filtered on a "Millipore" filter with an upper part of "Celite" and the brown filtrate is treated with 193 mg of potassium perfluorobutanesulfonate. When 20 ml of ether are added, an eraser separates. The liquid is removed and the gum is washed with ether. The gum is dissolved in 10 ml of methanol. When ether is added, a small amount of precipitate forms. The mixture is filtered and the colored filtrate is treated with a new quantity of ether. The precipitate formed is filtered and recrystallized twice from a mixture of ether and methanol to obtain 26 mg of the title compound.

Analysis for cl7Hl505N2SK.2H20: 35 Calculated: C, 46.99; H, 4.41; N, 6.45.

Found: C, 47.24; H, 4.19; N, 6.34.

Example 185

Potassium salt of (cis, Z) -3 - [[(2-amino-4-a thiazolyl) (methoxyimino) acetyl] amino] -2-oxor-4-phenyl-l- L158 azetidinesulfonic acid

Dissolve the potassium salt of (cis) -2-oxo-4-phenyl-3 - [(phenylacetyl) amino] -1-azetidinesulfonic acid (560 mg; see Example 184, part D) in 5 ml 5 ethanol, and the solution is hydrogenated with 110 mg of platinum oxide as catalyst, under 1 atmosphere.

After having stirred for one hour, the catalyst is filtered on a "Millipore" filter with "Celite"; particles of the catalyst pass through the filter giving a black filtrate. The ethanol is removed under reduced pressure and the residue is dissolved in 4 ml of dimethylformamide. 168 mg of N-hydroxybenzotriazole monohydrate, 221 mg of (Z) -2-amino-a- (methoxyimino) - 4-thiazoleacetic acid and 227 mg of dicyclohexylcarbodiimide are added, and the mixture is stirred for about 16 hours nitrogen.

The suspension is evaporated in vacuo and triturated with 15 ml of acetone. The resulting suspension is filtered on a "Millipore" filter with "Celite", and the filtrate is treated with 372 mg of potassium perfluorobutanesulfonate.

20- When 15 ml of ether are added, an eraser separates. The liquid is removed and the gum is washed with ether. The gum is dissolved in 5 ml of water and passed through 150 ml of "HP-20" resin, eluting with water. Fractions 16 to 34 are combined (30 ml each) and lyophilized to obtain 201 mg of the title compound as a solid.

Analysis for c15H140gN5S2K1 / 5 H2 °:

Calculated: C, 36.73; H, 3.49; N, 14.28; S, 13.07; K, 7.97

Find ; C, 36.65; H, 3.00; N, 13.99; S, 13.48; K, 8.30.

30 Example 186

Acid (cis) -3-amino-2-oxo-4- (2-phen.ylethylen) -1-azetidine sulfonic A) N- (3-phenyl-2-propenylidene) -4-methoxyaniline

12.32 g of p-anisidine are dissolved in 160 ml of methylene chloride and 20 g of anhydrous magnesium sulfate are added. The mixture is cooled in an ice bath and 13.22 g of trans-cinnamaldehyde are added. The mixture is stirred under nitrogen for 2 hours and then filtered. ^ The filtrate is evaporated to obtain a solid.

159

The crude product is recrystallized from a mixture of methylene chloride and petroleum ether to obtain 20.96 g of the title compound in the form of a solid.

B) (±) - (cis) -3-azido-1- (4-methoxyphenyl) -2-oxo-4- (2-5 phenylethenyl) azetidine

24.26 g of 2-azidoacetic acid is dissolved in 100 ml of methylene chloride and the solution is cooled in an ice bath. To the solution, 48.57 g of triethylamine and 14.24 g of N- (3-phenyl-2-10 propenylidene) -4-methoxyaniline dissolved in 250 ml of methylene chloride are added. To the resulting solution, 50.41 g of trifluoroacetic anhydride is added dropwise over one hour. After stirring for one hour in an ice bath, the mixture is warmed to room temperature and stirred for about 16 hours. The mixture is then diluted with 250 ml of methylene chloride and washed with 750 ml of water, two 750 ml fractions of a 5% sodium bicarbonate solution, and 750 ml of a hydrochloric acid solution. -20 than IN. The organic layer is dried over anhydrous sodium sulfate and freed from the solvent to obtain a solid. The crude product is recrystallized from ethyl acetate to obtain 11.39 g of the title compound in the form of a solid.

2 5 C) (±) - (cis) -3-azido-2-oxo-4- (2-phenylethenyl) azetidine To a solution of 10.22 g of ceric ammonium nitrate in 13 ml of water at 0 ° C, 1.99 g of (±) - (cis) -3-azido-l- (4-methoxyphenyl) -2-oxo-4- (2-phenylethenyl) -azetidine dissolved in 65 ml of acetonitrile are added , in 15 minutes 30 (an additional 10 ml of acetonitrile are used for rinsing). The mixture is stirred for a further 15 minutes at 0 ° C., diluted with 750 ml of ethyl acetate, washed with six fractions of 600 ml of water, dried over anhydrous sodium sulfate, and freed from the solvent to obtain an oil. The crude product is subjected to chromatography on 90 g of silica gel, first eluting with 250 ml of a mixture of 30% ethyl acetate and petroleum ether, then a mixture of 50% ethyl acetate and petroleum ether. Fractions 160 11 to 16 are combined (50 ml each) and evaporated to obtain 802 mg of the title compound as an oil.

D) Tetra-n-butylammonium salt of (t) - (cis) -3-azido-2-oxo-4- (2-phenylethenyl) -1-azetidinesulfonic acid 5 334 mg of (±) - is dissolved (cis) -3-azido-2-oxo-4- (2-phenylethenyl) azetidine in 3 ml of dimethylformamide and 868 mg of a pyridine and sulfur trioxide complex are added. The mixture was stirred at room temperature for 40 hours under nitrogen, then poured into 200 ml of a 0.5M monobasic potassium phosphate solution, then washed with 30 ml of methylene chloride. 530 mg of tetra-n-butyl ammonium bisulfate are added to the aqueous solution and the mixture is extracted with four 50 ml portions of methylene chloride. The combined organic layers are washed with two 100 ml fractions of water, dried over anhydrous magnesium sulfate and the solvent is removed to obtain 824 mg of the title compound in the form of a gum.

E) Acid (±) - (cis) -3-azido-2-oxo-4- (2-phenylethenyl) -1-20 azetidinesulfonic

300 mg of the tetra-n-butylammonium salt of (±) - (cis) -3-azido-2-oxo-4- (2-phenylethenyl) -1-azetidinesulfonic acid are dissolved in 4 ml of tetrahydrofuran, and we stir quickly. To this mixture, 600 mg of zinc powder is added, followed by 0.8 ml of an IM monobasic potassium phosphate solution. The mixture is heated to 45 ° C and stirred at this temperature for 3 hours. The mixture is then filtered and taken up in 40 ml of methylene chloride and 10 ml of water. The aqueous layer is further extracted with three 40 ml portions of methylene chloride and the combined layers of methylene chloride are freed from their solvent to obtain 256 mg of a foam. This crude product is dissolved in a small amount of a mixture of about 30% acetone and water and passed through 7.5 ml of "Dowex" resin (K) (0.7 meq / ml) eluting with 40 ml of water. The eluent is evaporated to obtain 151 mg of foam, which is dissolved in 2 ml of water and which is acidified to pH 2 with an acid solution "... - —— · - 161 nitrile to dissolve the precipitate and the resulting solution is passed over 15 ml of "HP-20" resin, first eluting with 150 ml of water, then with a mixture of 10% acetone and water. Fractions 2 to 13 were combined (15 ml each) and evaporated to obtain 101 mg of the title compound in the form of a foam.

Example 187

Acid potassium salt (t) - (cis, Z) -3 - [[(2-amino-4-thiazolyl) (methoxyimino) acetyl] ar.ino] -2-oxo-4- (2-phenyl -10 ethenyl) -1-azetidinesulfonic

A solution of 68 mg of (Z) -2-amino-α- (methoxyimino) -4-thiazoleacetic acid and 51 mg of N-hydroxybenzotriazole monohydrate in 2 ml of dimethylformamide is treated with 69 mg of dicyclohexylcarbodiimide. The mixture is stirred at room temperature for 30 minutes under nitrogen. (Cis) -3-amino-2-oxo-4- (2-phenylethenyl) -1-azetidinesulfonic acid (90 mg; see Example 186) and 34 mg of triethylamine are added and the mixture is stirred for 20 hours under nitrogen. The suspension is evaporated in vacuo and triturated with 10 ml of acetone. The suspension is filtered and the filtrate is treated with 113 mg of potassium perfluorobutanesulfonate. Dilution with 30 ml of ether and filtration gives 169 mg of a solid product, which is dissolved in a small amount of a mixture of about 10% acetonitrile and water and which the passed over 34 ml of "HP-20" resin, first eluting with 150 ml of water, then with a mixture of 10% acetone and water. Fractions 16-19 are combined (15 ml each) and freed from the solvent to obtain 110 mg of the title compound as a solid.

Analysis for C1 ^ H1g0gN2S2K.H20:

Calculated: C, 40.23; H, 3.57; N, 13.80; S, 12.63; K, 7.70

Found: C, 40.03; H, 3.05; N, 13.61; S, 12.31; K, 7.56 35 Example 188

(Cis) -3-amino-4- (methoxycarbonyl) -2-oxo-l-azetidine-sulfonic acid A) [(4-methoxyphenyl) imino] - / acetic acid methyl ester Λ 162

A 1 liter three-necked flask, dry, equipped with a nitrogen inlet and a stirring bar, is loaded with 56.88 g of magnesium sulphate, then with a solution of 19.43 g of anisidine recrystallized from 250 ml of dichloromethane. After having cooled to 0 ° C., a solution of 19.92 g of methyl glyoxylate hemiacetal in 250 ml of dichloromethane is added over an hour and a half. After stirring for a further 20 minutes at 0 ° C, the reaction mixture is filtered under suction, dried over sodium sulfate, filtered and concentrated in vacuo to a quarter of its volume. 300 ml of hexane are added and the solution is concentrated in an oil which solidifies half after standing under high vacuum at 5 ° C.

B) (cis) -3- (1,3-dihydro-1,3-dioxo-2H-iso-indol-2-yl ·) -4-15 methoxycarbonyl-2-oxo-l- (4-methoxyphenyl) azetidine

A 500 ml three-necked flask, dry, equipped with a stirring bar, a tap bulb, a partition and a nitrogen inlet, is loaded with a solution of 21, 09 g of [(4-20. Methoxyphenyl) imino] acetic acid methyl ester in 150 ml of dichloromethane, and the mixture is cooled to 0 ° C. 19.2 ml (0.14 mol) of triethylamine are added dropwise, followed by a solution of 28.4 g of (N-phthalimido) acetyl chloride in 150 ml of dichloromethane, over one hour. The resulting mixture was stirred for 1.5 hours at 0 ° C, then diluted with 2.5 L of dichloromethane. The organic solution is washed with two 500 ml fractions of monobasic potassium phosphate at pH 4.5, two 500 ml fractions of 5% sodium bicarbonate, 500 ml of a saturated sodium chloride solution, then dry over sodium sulfate.

Filtration and concentration in vacuo give a solid which is washed with ethyl acetate, with cold acetone and with hexane to obtain 18.65 g of product.

35 C) (cis) -4- (methoxycarbonyl) -1- (4-methoxyphenyl) -2-oxo-3 - [[(phenylmethoxy) carbonyl] amino] azetidine

A 500 ml dry flask equipped with a nitrogen inlet, a stirring bar and a partition / separation is loaded with 18.65 g of (cis) -3- (1,3- dioxo-2H-iso-163 indol-2-yl) -4-methoxycarbonyl-2-oxo-l- (4-methoxyphenyl) -azetidine and 325 ml of dichloromethane. The resulting suspension is cooled to -30 ° C, and 3.52 ml of methyl hydrazine is added dropwise. The reaction mixture is warmed to 0 ° C and stirred for one hour.

An additional 0.4 ml of methyl hydrazine is added and the mixture is stirred for 10 minutes. This sequence of operations is repeated until a total of 2.9 equivalents (7.7 ml) of methyl hydrazine has been added. The solvent is removed in vacuo; 200 ml of fresh dichloromethane are added, and the mixture is concentrated again. This sequence of operations is repeated two more times, the resulting foam is dried under high vacuum for 20 minutes, redissolved in 225 ml of dichloromethane, and allowed to stand at room temperature for about 16 hours, during which time an amount considerable solid precipitates. The mixture is filtered under nitrogen, the filtrate is cooled to 0 ° C (nitrogen atmosphere) and treated with 17 ml of diisopropylethylamine, then with 7 ml of benzyl chloroformate, drop by drop. The reaction mixture is stirred at 0 ° C for 30 minutes, then at room temperature for 1.5 hours. The mixture is washed with two 300 ml fractions of a monobasic potassium phosphate buffer at pH 4.5, with two 300 ml fractions of 5% sodium bicarbonate with 300 ml of a saturated chloride solution of sodium, then dried over sodium sulfate and filtered. Concentration under vacuum gives a foam which, after trituration with ether, gives 9.9 g of the title compound in the form of a solid. D) (cis) -4- (methoxycarbonyl) -2-oxo-3 - [[(phenylmethoxv) - carbonyl] amino] -1-azetidine

A solution of 8.59 g of ceric ammonium nitrate in 60 ml of a 1: 1 mixture of acetonitrile and water is treated with a suspension of 2 g of (cis) -4- (methoxy-carbonyl). ) -1- (4-methoxyphenyl) -2-oxo-3 - [[(phenylmethoxy) - carbonyl] amino] azetidine in 50 ml of acetonitrile, in 10 minutes. The reaction mixture is stirred for a further 10 minutes at room temperature and diluted with 100 ml ethyl acetate. The separated aqueous / ΙΛ 164 layer is washed with three 40 ml fractions of ethyl acetate and the combined organic extracts are washed with three 70 ml fractions of 50% sodium bicarbonate. The basic washing waters are rewashed with 50 ml of ethyl acetate and the combined organic extracts are washed with aqueous sodium sulfite, 100 ml of 5% aqueous sodium carbonate, two 100 ml fractions of a 5% sodium chloride solution, two 50 ml fractions of a saturated sodium chloride solution, then stirred on charcoal 10 "Darco G-60" for 30 minutes. Sodium sulfate is added, then the mixture is filtered and concentrated in vacuo to give an oil which, after trituration with ether, gives 685 mg of the title compound as a solid.

E) Tetrabutylammonium salt of (cis) -4- (methoxy-carbonyl) -2-OXO-3-Ç [(phenylmethoxy) carbonyl] dmino] -1-azetidinesulfonic acid

Stirred under argon for three hours at 80 ° C. a mixture of 100 mg of (cis) -4- (methoxycarbonyl) -2-oxo-3-20 [[(phenylmethoxy) carbonyllamino] -1-azetidine and 172 mg of '' a complex of pyridine and sulfur trioxide in 1 ml of pyridine. The reaction mixture is poured into 70 ml of 0/5 M monobasic potassium phosphate (pH 5.5) and extracted with four 30 ml fractions of dichloromethane (to be discarded). 122 mg of tetrabutylammonium hydrogen sulphate are added to the aqueous layer, which is then extracted with four 30 ml fractions of dichloromethane. The organic extracts are washed with 8% sodium chloride solution, dried over sodium sulfate and filtered. Concentration in vacuo gives 186 mg of the title compound as a viscous oil.

F) (cis) -3-amino-4- (methoxycarbonyl) -2-oxo-1-azetidinesulfonic acid

A solution of 186 mg of tetra-butylammonium salt of (cis) -4- (methoxycarbonyl) -2-oxo-3 - [[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid in 2 ml of methanol is hydrogenated. , on 95 mg of 10% palladium on charcoal, for an hour and a half under 1 atmosphere. The catalyst is separated by filtration and rinsed with fs \ 165 dichloromethane, then the filtrate is treated with 97% formic acid and cooled to -50 ° C (the presence of a primer crystal at this stage is necessary to induce crystallization). Once crystallization has started, the mixture is allowed to stand for approximately 16 hours at 10 ° C. The resulting solid is washed with dichloromethane, hexane, and dried in vacuo, this yes gives 50 mg of the title compound.

Example 189 10 Potassium salt of (cis) acid -3 - [[2-amino-4-thiazolyl) - [(1-diphenylmethoxycarbonyl) -1-methylethoxy] imino] acetyllamino -4- (methoxycarbonyl) - 2-oxo-1-azetidinesulfonic A solution of 34 mg of N-hydroxybenzotriazole hydrate and 101 mg of 2-amino-cr acid [[1-15 (diphenylmethoxycarbonyl) -1-methylethoxy] imino] -4 is treated -thiazole-acetic in 0.5 ml of dimethylformamide, with 45 mg of solid dicyclohexylcarbodiimide, and the mixture is stirred under argon for 45 minutes at room temperature.

Then (cis) -3-amino-4- (methoxy-carbonyl) -2-oxo-1-azetidinesulfonic acid (45 mg; see Example 188) is added as a solid, followed by 0.03 ml of triethylamine is added dropwise. The reaction mixture is stirred at room temperature for approximately 16 hours. Dimethylformamide is removed in a high vacuum at 30 ° C and the residue is triturated with acetone.

The supernatant is treated with 67 mg of potassium perfluorobutanesulfonate. A dilution with ether gives a solid which is washed with ether and dried under vacuum to obtain 93 mg of the title compound.

30 Example 190

(Cis) acid dipotassium salt -3- [[(2-amino-4-thiazolyl) - [(1-carboxy-1-methylethoxy) imino] acetyl] amino] -4- (methoxy-carbonyl) -2 -oxo-l-azetidinesulfonic

A suspension of 35 · potassium salt of (cis) acid -3 - [[2-amino-4-thiazolyl) - [[1- diphenylmethoxycarbony1) -1-methylethoxy] imino] is stirred at -12 ° C. under argon. acetyl] amino] - 4- (methoxycarbonyl) -2-oxo-1-azetidinesulfonic acid in 0.4 ml of anisole, and 0.9 ml of cold β-trifluoroacetic acid is added. After an hour and a half, 4 ml /// Λ I - 166 of ether and 2 ml of hexane are added and the resulting suspension is stirred for 15 minutes at -10 ° C, then for 15 minutes at ambient temperature. The solid is isolated by centrifugation and washed with ether. The pH of a suspension of this substance in 0.5 ml of cold water is adjusted to 6 with IN potassium hydroxide, then it is passed through a 30 ml column of "HP-20 AG". Elution with water gives 30 mg of the title compound after evaporation (acetonitrile is added and the mixture is evaporated twice).

10 Analysis for: Ci4Hj5K2 ^ 5 ^ 9 ^ 2:

Calculated: C, 31.15; H, 2.81; N, 12.98 Found: C, 29.08; H, 3.03; N, 12.19.

Example 191

(S) - (trans) -3-amino-4-ethynyl-2-oxo-1-azetidine-sulfonic acid A) 2- (Trimethylsilyl) ethynylmagnesium bromide

20 ml of dry tetrahydrofuran, 2.20 ml of trimethylsilylacetylene 20 and 5.05 ml of a 3.06 M solution of bromide are introduced into a 50 ml flask dried with a torch and maintained under a positive nitrogen pressure. methylmagnesium in ether. The mixture is stirred for 140 minutes to give the title compound.

B) (S) - (trans) - 4- [2- (trimethylsilyl) ethynyl] -2-oxo-3 - [(triphenylmethyl) aminojazetidine 25 To a 250 ml three-necked flask dried over whitewash, add 6 .00 g of (S) - (cis) -4- (methylsulfonyl) -2-oxo-3 - [(triphenylmethyl) amino] azetidine. The flask is purged with nitrogen and then maintained under a positive nitrogen pressure. After cooling the reaction mixture in a bath of dry ice and isopropanol, 4.65 ml of a 3.06 M solution of methylmagnesium bromide in ether, stirring quickly. The 2- (trimethylsilyl) ethynylmagnesium bromide solution prepared in part A is added, using a "Teflon" tube, under positive nitrogen pressure (the flask containing the reagent is rinsed with 7 ml tetrahydrofuran). Once the addition is complete, the cooling bath is removed. After 45 minutes, add. a solution of 3.5 g of potassium bisulfate in 20 ml 167 of water. Most of the tetrahydro-furan is removed on a rotary evaporator. The residue is transferred to a separatory funnel with ether and water. The aqueous layer is separated and extracted twice with ether. The combined ethereal layers are washed once with a saturated sodium chloride solution, then dried over sodium sulfate and filtered. Removal of the solvent gives a foam which is subjected to chromatography on a silica column. Elution with 2 10 liters of dichloromethane, 1 liter of a mixture of 1% ether and dichloromethane, 2 liters of a mixture of 2% ether and dichloromethane and 1.5 liters of a mixture of 10% ether and dichloromethane (fraction 1 = 1000 ml; fractions 2 and 3 = 500 ml; 4-terminal fraction = 250 ml) gives 1.30 g of the title compound in fractions 2 to 8 and 1.80 g of the corresponding trans isomer in fractions 12 to 19. Fractions 9 to 11 contain 1.19 g of a mixture of the cis and trans isomers.

C) (S) - (trans) -4-ethynyl-2-oxo-3 - [(triphenylmethyl) amino] - 20 azetidine

2.97 g of (S) - (trans) -4- [2- (trimethyl-silyl) ethynyl] -2-oxo-3 - [(triphenylmethyl) amino] azetidine are dissolved in 30 ml of dichloromethane and 330 are added mg tetrabutylammonium fluoride (containing 20 to 25% water). After 20 minutes, the solvent is removed in vacuo. The residue is taken up in ethyl acetate and water.

The organic layer is separated, washed once with water and once with saturated aqueous sodium chloride solution, dried over sodium sulfate and filtered. Removal of the solvent gives an oil which is stirred for 15 minutes with 60 ml of pentane to obtain 2.35 g of the title compound in the form of a powder (after drying under vacuum).

D) (S) - (trans) -3-amino-4-ethyl-2-oxo-1-azetidine-sulfonic acid

404 mg of (S) - (trans) -4-ethynyl-2-oxo-3 - [(triphenylmethyl) amino] azetidine and 560 mg of a pyridine and sulfur trioxide complex are added to a 25 ml flask. .

/ After purging the flask with nitrogen, 4.0 ml of dry pyridine is added and the mixture is heated to 80-85 ° C for 3 hours. The mixture is added to a rapidly stirred mixture of 4.0 ml of concentrated hydrochloric acid, 50 ml of water and 50 ml of ethyl acetate. The pH is adjusted to 3.15 5 with sodium carbonate. The aqueous layer is separated and extracted once with ethyl acetate. The combined organic layers are washed once with a saturated aqueous solution of sodium chloride, dried over sodium sulfate and filtered. Removal of the sol-10 before vacuum gives a foam which is taken up in 10 ml of dichloromethane. Formic acid (98%, 8 ml) is added, and after 15 minutes the mixture is concentrated to 4 ml and 10 ml of dichloromethane are added to obtain a solid in suspension in the solution. Filtration gives 100 ml of the title compound as a solid (obvious discoloration with melting at> 180 ° C).

Example 192

Acid potassium salt [3S- [3g (Z), 4ß]] - 3-f [(2-amino- 4-thiazolyl) (methoxyimino) acetyl] aminoJ-4-ethynyl-2-oxo-l- 20 azetidinesulfonic

Weighed into a 10 ml flask 100 mg of (Z) - 2-amino-a- (methoxyimino) -4-thiazoleacetic acid, 85 mg of N-hydroxybenzotriazole monohydrate and 113 mg of dicyclohexyl-carbodiimide. The flask is purged with nitrogen and cooled down in an ice water bath. Then 0.6 ml of dimethylformamide is added and the mixture is stirred for 10 minutes, after which an additional 0.6 ml of dimethylformamide is added. (S) - (trans) -3-amino-4-ethyl-2-oxo-1-azetidinesulfonic acid (95 mg; see Example 30 191) is added as a solid with 1, 0 ml of dimethyl formamide and 56 μl of triethylamine. The cooling bath is removed and the mixture is stirred for 22 hours.

3 ml of acetone are added and the solids present are filtered off, followed by washing with an additional 4 ml of acetone. All the solvents are removed in vacuo and the residue is taken up in 5 ml of methanol, then 162 mg of potassium perfluorobutanesulfonate are added and dissolved. After standing, a solid is deposited which is isolated by centrifugation to obtain 68 mg of the title compound,

L

169 melting point> 230 ° C.

Example 193

Potassium salt of acid (S) -3 - [[[(2,5-dichlorophenyl) -thio] acetyl] amino] -2-oxo-1-azetidinesulfonic 5 100 mg of 3-amino acid is dissolved -2-oxo-1-azetidinesulfonique in 2 ml of dry dimethylformamide with 0.083 ml of triethylamine. 123 mg (0.602 mmol) of 2,5-dichlorophenylthioacetic acid, 81 mg of N-hydroxybenzotriazole hydrate and 124 mg of dicyclo-10 hexylcarbodiimide are added, the mixture is stirred for 2 hours at room temperature, then for 2 hours. days at 5 ° C. The solvent is removed in vacuo, the residue is taken up in water, filtered through "Celite", and the filtrate is washed with ethyl acetate. The aqueous layer is combined with dichloromethane, 612 mg of tetrabutylammonium bisulfate are added, the pH is raised to 3 with IN potassium hydroxide solution, and after having extracted a total of three times with dichloromethane, dried over sulfate of sodium from the combined extracts and the solvent is removed in vacuo to give an oil. A solution of this oil in acetone is added to a solution of 612 mg of potassium perfluorobutanesulfonate in acetone, which causes the product to precipitate. After adding a small amount of ether, the solid was collected by filtration, washed several times with acetone and dried to obtain 206 mg of a powder.

Analysis for C ^ H ^ ls ^ OgC ^ K:

Calculated: C, 31.21; H, 2.14; N, 6.62-; Cl, 16.75 Found: C, 27.90; H, 2.11; N, 5.84; Cl, 18.04.

30 Example 194

Potassium salt of (3S-trans) -3 - [[[(2,5-dichloro-phenyl) thio] acetyl] amino] -4-methyl-2-oxo-1-azetidine-sulfonic acid

(3S-trans) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid (250 mg; see Example 139) is dissolved in 2 ml of dimethylformamide with 193 µl of triethylamine. 285 mg of 2,5-dichlorophenylthioacetic acid, 213 mg of N-hydroxybenzotriazole hydrate and 287 mg of dicyclo-dj hexylcarbodiimide are added. After stirring for 16 hours / 7 / Λ 170 approximately at room temperature, the mixture is filtered and the solvent is removed in vacuo. The residue is taken up in water and filtered. The filtrate is washed with ethyl acetate, decanted with dichloromethane and 4.2 mmol of tetrabutylammonium bisulfate is added.

After a total of three extractions with dichloromethane, the combined extracts are dried over sodium sulfate and the solvent is removed in vacuo, which gives 920 mg of an oil. To a solution of this oil in acetone is added 946 mg of potassium perfluorobutanesulfonate dissolved in acetone. A solid precipitates slowly, it is collected, washed twice with ether, and dried to obtain 306 mg of a powder. Chromatography on a column of 100 ml of "HP-20" resin, eluting with a mixture of 20% acetonitrile and 80% water, gives the desired product, which crystallizes on evaporation from a mixture of water and methanol. Trituration of the residue with acetone gives 233 mg of a powder, melting point 212-213 ° (with decomposition).

20 Analysis for ci2HllN2 ° 5C ^ 2S2K:

Calculated: C, 32.95; H, 2.54; N, 6.41; Cl, 16.21; S, 14.66 Found: C, 32.91; H, 2.60; N, 6.42; Cl, 16.50; S, 13.77. Examples 195 and 196

Following the procedure of Example 138, but replacing (3S-cis) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid with (3S-trans) -3 acid -amino-4-methyl-2-oxo-1-azetidinesulfonic and (Z) -2-amino-a- (methoxyimino) -4-thiazoleacetic acid by the acid indicated in column I below, we obtain the compound shown in column II below.

• Al 171

I - I

I — 1 1 — I I — 1 J — i [> ί i — ι (1) cû fl m | 3 ·

^ Md I 'ii — ι D1 C

** Pd d) * · ι — I Ή O

- Λ G> rG Ή

*> 1 · Η * -P O -P

Pi X Ό Pi Md P -H

'- ο Ή ^ ÜH Hl

ö (h -P ö id S O

η Ό \ (ü η h ω ft

1—1> 1 N> -> i (U E

I Pd P I G P! O

σι ι i w «ü · Η ü rn i h m pd τΐ 'Θ 1-1-1 i— (VH Td I — I O 1 — I 4-) 0) O X (d O Ul) ü

Td G O Ό G N (d -ιH Ή I -rl Ή id i> ‘ü Ë CN O Ë! (D

(Ü (Tj I CÖ iTj * —I

- ^ H - - I *

H H ί> ι H H O Q

> i £ l> Ί X ° (ü -P -U (U -P O P *

Td Md «U Td Md I 00

U Ë ISO H

ë p ι E td ι P 0 "i 0 0 H 0

H X I Ή X> i O

Kl O 1—1 in o Pd -h in ι o in ι -p in td ο a id o “ai p -P C -H (D -P G E Ή

0 -H E P Ο Ή I

ae d t? cdE ^ a)

td ι — ι · Η id I TJ

<d - ι — I £ φ - I — I

Td -> iO Pd 'o -P

-p m '—'PG

ι — l ^ \ (D rH ι — 1 * H * H

(D I ü P Q) 1 E 0 in ro P in in m p & d)

1 P

- td1 I

ι I · Η '—-

-P iH

-P MO> i

\ <1) O -P

0 id vu G - ü

1 H P

O> t O

X G X (D

yes op ι Pd ι o1

0 ft O -H

G G -P

-H X - -H \ CL)

Ë O E O

PP PP

- id - H

1—1> 1> —1> 1

1 Pd IG

- I .— \ ÎU

Pi 'i (G Pd - - - Ot m ι — ι

ω where had o

Ό G Ό G

• H · Η · Η · Η

ü E O E

<i P (<P

ü)

iH

P- LD KO

E cr \ σ \

Cl) ι — I ι — 1

X

H

// Λ 172

Example 197

Potassium salt of [3S (R *)] acid - 3 - [[[(amino-oxoacetyl) -amino] (4-hydroxypheny1) acetyl] amino] -2-oxo-1-azetidine-sulfonic 5 Next the procedure described in Example 28, but replacing the acid (Z) -2-amino-a - [[2- (diphenyl-methoxy) -1, l-dimethyl-2-oxo-ethoxy] imino] -4-thiazole-acetic with (R) - [(amino-oxoacetyl) amino] (4-hydroxyphenyl) acetic acid, the title compound is obtained, melting point 128 ° C, with decomposition.

Example 198

Potassium salt of [3S (R *)] acid - 3 - [[(2-amino-4-thiazolyl) [[[3 - [(2-furanylmethylene) amino] -2-oxo-l-imidazolidinyl] carbonyl] amino] acetyl] amino] -2-oxo-1-15 azetidinesulfonic

Following the procedure of Example 6, but replacing the aminothiazoleacetic acid with (R) -2-amino-a - [[[3 - [(2-furanylmethylene) aminoJ-2-oxo-1 -imidazolidiny1] carbonyl] amino] -4-thiazoleacetic, the title compound is obtained, melting point> 250 ° C;

Example 195

Organic production of EM5117 Fermentation of 9 liters

Chromobacterium violaceum SC 11.378, 25 A.T.C.C. No. 31532 on the following sterilized agar medium (A):

Grams

Yeast extract 1

Beef extract 1 30 NZ Amine A 2

Glucose 10 Agar 15

Distilled water to make 1 liter.

The pH is adjusted to 7.3 before sterilization at 121 ° C for 30 minutes.

The contents of a surface growth "louoe" of the microorganism are used to inoculate each of three 500 ml Erlenmeyer flasks, each containing 100 ml of the. following sterilized medium (B): // - 173

Grams

Oatmeal 20

Tomato cream 20

Tap water to make 1 liter.

5 The pH is adjusted to 7, Q before sterilizing at 121 ° C for 15 minutes.

The flasks are then incubated at 25 ° C on a rotating shaker (300 rpm; 5 cm stroke) for approximately 24 hours.

After appropriate incubation as described above, transfers of 1% (volume / volume) are carried out from the growth culture flasks into one hundred 500 ml Erlenmeyer flasks each containing 100 ml of the following sterilized medium (C): 15 Grams

Oatmeal 20

Tomato cream 20

Glucose 30

Tap water to make 1 liter.

The pH is adjusted to 7.0 before sterilization at 121 ° C for 15 minutes.

After inoculation, the flasks are incubated at 25 ° C on a rotating shaker (300 rpm, 5 cm stroke) for approximately 18 to 24 hours. At the end of this time, the contents of the flasks are collected and the broth is centrifuged, which gives about 9 liters of supernatant broth.

250 liter fermentation

A surface growth "magnifying glass" of an inclined agar culture (medium A) of Chromobacterium violaceum SC 11.378, A.T.C.C. No. 31532, for inoculating each of five 500 ml Erlenmeyer flasks each containing 100 ml of sterilized medium (B). These flasks are then incubated at 25 ° C on a rotating agitation device (300 rpm; 5 cm stroke) for approximately 24 hours. After the appropriate incubation as described above, 1% (volume / volume) transfers are carried out from the culture flasks grown in five 4-liter Erlenmeyer flasks

/AT

174 each containing 1.5 liters of sterilized medium (B). After inoculation, the flasks are incubated at 25 ° C on a rotating shaker (300 rpm; 5 cm stroke) for approximately 24 hours. After the appropriate incubation as described above, a transfer of 1% (volume / volume) is carried out into a fermentation tank equipped with an agitator containing 250 liters of the sterilized medium (C). After inoculation, the fermentation is continued under the following conditions: temperature 25 ° C; 3 10 pressure 0.7 manometric atmosphere; aeration 0.28 m / min; stirring 155 rpm. Uconn is added as necessary as an anti-foaming agent. After approximately 18 to 24 hours, the fermentation is complete. The fermentation broth is then adjusted to pH 5.0 using 15 l hydrochloric acid and the contents of the broth of the tank are centrifuged, which gives about 230 liters of supernatant broth.

Isolation and purification

The 250 liter fermentation supernatant broth is adjusted to pH 5 using sulfuric acid and filtered using 3-5% diatomaceous earth ("Celite"). The filtrate is extracted from the broth with two 30 liter fractions of 0.005 M cetyldimethylbenzylammonium chloride in methylene chloride.

The combined lower phase is extracted with 6 liters of 0.05 M sodium iodide which has been adjusted beforehand to pH 5 with acetic acid. The lower phase is discarded and the upper phase is concentrated in vacuo to 500 ml.

The concentrated substance is extracted with 400 ml of n-butanol. The upper phase is discarded and the lower phase is concentrated to dryness under vacuum. The residue is dissolved; (if possible) in 150 ml of methanol. The insoluble material is discarded and the methanolic solution is concentrated to dryness in vacuo to give 38.6 g of crude antibiotic.

The crude product is dissolved in 10 ml of a mixture (1: 1) of methanol and water and subjected to chromatography on a column of 500 ml of dextran gel

L

175 crosslinked ("Sephadex G-10") in the same mixture of solvents, eluting at 2 ml / minute and collecting 20 ml fractions. The active fractions (19 to 26) are combined and concentrated in vacuo. The residue (5.23 g) is mixed with 50 ml of methanol. The insoluble matter is filtered and discarded. The filtrate is concentrated in vacuo.

The residue, comprising 5.0 g of material, is dissolved in 10 ml of 0.01 M sodium phosphate buffer at pH 5 and passed through a column of DEAE-10 cellulose ("Whatman DE52" cellulose ) packed and balanced in the same pad. The column is eluted at 5 ml / minute with a linear gradient prepared from 4 liters of a 0.01 M sodium phosphate buffer at pH 5 and 4 liters of a 0.1 M sodium phosphate buffer at pH 5.15, collecting 20 ml fractions. The active fractions (192 to 222) are combined and concentrated in vacuo, and the material insoluble in methanol is removed, washing thoroughly with methanol. Removal of the solvent leaves 576 mg of substance.

These 576 mg of residue are dissolved in 4 ml of water and the pH is adjusted to 5 with approximately 1 ml of 0.1 N sodium hydroxide solution. The solution is subjected to chromatography on a column of crosslinked and alkylated dextran gel. ("Sephadex LH-20") in water, eluting at 1 ml / minute and collecting 25 ml fractions. The active fractions are combined (38 to 44) and concentrated, which gives 459 mg of residue.

348 mg of this residue is dissolved in water and the solution is passed through a column of macroreticular styrene-divinylbenzene copolymer resin ("Diaion HP 20 AG"); the column was prepared beforehand by washing it with methanolic potassium hydroxide, methanol, methanolic hydrochloric acid, methanol and water, then packing it in water. The column is eluted with water at 1 ml per minute, collecting 10 ml fractions. The active fractions are combined (36 to 43) and concentrated, giving 186.4 mg of substance. Chromatography (in the same way) of another 100 mg of the, 459 mg of residue from the previous operation gives

U

176 51.5 mg of substance. At this stage, the 186.4 mg of substance and the 51.5 mg of substance consist of almost pure EM5117, as shown by the thin layer chromatography and the nuclear magnetic resonance spectra.

5 The 186.4 mg fraction of EM5117 from the previous operation is dissolved in water and passed through a column of ion exchange resin ("Dowex 50W-X2", 74-149 μ), potassium form), washing with a volume of water equal to twice the volume of the resin. The concentration of the effluent gives 189.0 mg of a crystalline solid. This substance is recrystallized by dissolving it in 0.38 ml of water and adding 3.42 ml of methanol to it. The resulting mixture is cooled on ice and filtered, giving 14545 mg of crystals. Two other recrystallizations carried out in this way in a 1: 9 mixture of water and methanol give 95.9 mg of potassium salt of EM5117, m.p. 194 ° C with decomposition.

Optical rotation in water at 21 ° C (c = l) 2 0 λ (nm) [and] 589 + 94.3 ° 579 + 98.6 546 +113.1 436 +203 25 365 +348

The following fermentation media are effective for the production of EM5117, and can be substituted for media (B) and (C) of the previous example.

MIDDLE D

30 Grams

"Nutrisoy" flour 30 (nutritious soy)

Glucose 50 "Yeastamine" (yeast) 2.5 35 CaC03 3

Distilled water to make one liter.

h i

MIDDLE E

177

Grams

Yeast extract 4

Malt extract 10 5 Glucose 30

Glycerol 2

Distilled water to make 1 liter. Adjust the pH to 7.3 before sterilization. MIDDLE F

10 Grams

Glycerol 10 L-asparagine 5 kh2po4 1

Na2HP04 2 15 Glucose 50

MgS04.7H20 0.2

Yeast extract 2.5

Tap water to make 1 liter.

MIDDLE G

20 Grams (NH4) 2S04 2 L-asparagine 5

Glucose 50

Glycerol 10 25 KH0PO. 3 φθ \ 7

MgS04.7H20 0.2

Yeast extract 2.5

Distilled water to make 1 liter.

30 Adjust the pH to 7.0.

MIDDLE H

Grams K2HP04 7 KH2P04 3 35 Na citrate 0.5

MgS04 0.1 (nh4) 2so4 1

Glucose 30 ^ Yeast extract 2.5 ft / A Distilled water to make 1 liter.

(y l

MIDDLE I

178

Grams

"Nutrisoy" flour 10 (NH4) 2S04 5 5 Glucose 50

Yeast extract 2.5

CaCO ^ * 5

Distilled water to make 1 liter.

MIDDLE J

10 Grams

Oatmeal for babies "Gerber" 20

"Contadina" tomato cream 5

Glucose 20 15 Tap water to make 1 liter.

Adjust pH to 7.0

MIDDLE K

Grams

Oatmeal for babies 20 "Gerber" 5

Cream of tomage "Contadina" 20

Glucose 20

Tap water to make 1 liter.

Adjust pH to 7.0

25 MIDDLE L

Grams

Yeast extract 4

Malt extract 10

Glucose 34

30 MIDDLE M

"Amberex" grams (1003) 5

Glucose 30

Tap water to make 1 liter.

35 MIDDLE N

Grams "Amberex" 5 "Cerelose" 33

Tap water to make 1 liter.

h 179

Example 196

Biological production of EM5210 Gluconobacter, species SC 11.435, A.T.C.C. No. 31581 on the following sterilized agar medium 5 (A):

Grams

Yeast extract 1

Beef extract 1 NZ amine A 2 10 Glucose 10 Agar 15

Distilled water to make 1 liter.

The pH is adjusted to 7.3 before sterilization at 12 ° C for 30 minutes.

A surface growth agar of the inclined agar culture (medium A) of Gluconobacter, species SC 11.435, is used to inoculate each of three 500 ml Erlenmeyer flasks each containing 100 ml of the following sterilized medium (B ); 20 Grams

Yeast extract 4

Maltese extract 10

Dextrose 4

Distilled water to make 1 liter.

The pH is adjusted to 7.3 before sterilization at 121 ° C for 15 minutes.

After inoculation, the flasks are incubated at 25 ° C on a rotating shaker (300 rpm; 5 cm stroke) for approximately 24 hours. After appropriate incubation as described above, 1% (volume / volume) transfers are made from the culture flasks grown in one hundred 500 ml Erlenmeyer flasks each containing 100 ml of the following sterilized medium (C): 35 grams

Yeast extract 5

Glucose 10

Distilled water to make 1 liter.

This medium is sterilized at 121 ° C for 15 minutes.

AT

180

After inoculation, the flasks are incubated at 25 ° C on a rotating shaker (300 rpm; 5 cm stroke) for 18 hours. After which the contents of the flasks are collected and the broth is centrifuged to give "about 9 liters of supernatant broth. Isolation and purification (small scale)

The active substance of the supernatant broth (10 liters) is made to absorb by a column of 500 g of a strongly basic anion exchange resin comprising 10 quaternary ammonium groups attached to a network of styrene-divinylbenzene copolymer ["Dowex AG1-X2 "(Cl)], wash with water and. eluted with 5% sodium chloride in 0.01 M aqueous NaH 4 PO 4. The eluate is concentrated in vacuo.

The residue is adsorbed on 250 g of charcoal which is washed with water. The EM5210 is eluted with a 1: 1 mixture of methanol and water. The active fractions are combined and concentrated under vacuum, which gives crude EM5210.

1ΈΜ5210 crude is subjected to chromatography on a 280 ml column of a strongly basic anion exchange resin ["Bio. Rad AG 1-X2" (Cl)] using a linear gradient prepared from '' one liter of water and one liter of 2M pyridinium acetate (pH 4.5). The active fractions are combined and concentrated in vacuo.

The partially purified EM5210 is further concentrated by filtration of the residue on a column of 500 ml of crosslinked dextran gel ("Sephadex G-10"), eluting with water. The active fractions are combined and concentrated, giving 26 mg of EM5210. The potassium salt of EM5210 is prepared by passing EM5210 through a column of cation exchange resin ("Dowex 50-X2") in the potassium form.

Isolation and purification (large scale)

The broth filtrate of a 250 liter fermentation (pH 3.7) of Gluconobacter, species SC 11.435, is absorbed with 10.8 kg of a strongly basic anion exchange resin ["Dowex 1-X8 "(Cl)]. This resin is washed with water and eluted with sodium chloride / 5% in 0.01 M sodium diacid phosphate. Actives 181 the active fractions are combined and concentrated to a small volume. The precipitated salt is eliminated and the filtrate is desalted by passing it through a column of charcoal (1.1 kg of "Darco" from 420 to 840 y) in water. The column is washed with water and the EM 5210 is eluted with a 1: 1 mixture of methanol and water. The active fractions are combined and concentrated. The residue (16 g) is dissolved in water and subjected to column chromatography (600 ml) of a strongly basic ion exchange resin ["Bio. Rad AG 1-X2" (Cl), 37 -74 y], eluting with a linear gradient prepared from one liter of water and one liter of 10% sodium chloride in 0.01 M sodium diacid phosphate.

The active fractions are combined, concentrated in vacuo to a small volume, and the precipitated salts are filtered off. The filtrate is passed through a column of macroreticular styrene-divinylbenzene copolymer resin. The column is eluted with water. The active fractions are combined, concentrated to a small volume, and freeze-dried to give 120 mg of the sodium salt of EM5210.

To transform this sodium salt into a lithium salt, an ion exchange resin column ("Dowex 50 W-X2", lithium form) is used. The sodium salt (100 mg) is dissolved in 0.5 ml of water, passed through the column and eluted with water. The active fractions are combined and subjected to freeze-drying directly, which gives 95 mg of the lithium salt of EM5210 in the form of an amorphous solid.

The free acid (internal salt) of EM5210 is prepared by: passing a base salt of EM5210 through a column of weakly acidic ion exchange resin in the H + form. One can, for example, pass about 2.5 mg of the lithium salt through a column of "Bio.Rad Bio - Rex 7 0" (H) 35 and elute with water to obtain 1.45 mg of l 'free acid (internal salt).

Chemical properties of EM5210 1) Positive "Ninhydrin" test (Ninhydrin is the analytical reagent for amino acids).

/ Ιλ 182 2) Acid hydrolysis (6N hydrochloric acid at 115 ° C for 16 hours) gives two major positive spots for Ninhydrin by chromatography on paper ("Whatman" No. 1, mixture (5: 1: 4) butanol, acetic acid and water), and a weak Ninhydrin-positive stain which quenches fluorescence excited by ultraviolet rays. The two major positive spots for Ninhydrin are D-glutamic acid and D-alamine.

Physical characteristics of EM5210 10 1) UV spectrum of sodium salt in water: final absorption.

2) IR - Major peaks of the lithium salt in potassium bromide: 1770, 1640, 1530, 1384, 1242 and 1051 cm "1.

3) NMR - Chemical shifts of the lithium salt in deuterated water, ppm of falling field of the TSP: 1.40 (d, J = 7Hz), ca. 2.14 (m), ca. 2.44 (m), 3.49 (s), 3.73 (t, J = 6Hz), 3.94 (s), 4.28 (m).

Optical rotation of the free acid (internal salt) in water at 24 ° C (C = 0.15%) (pH 2.7); X (nm) [a] 589 + 73 ° 578 + 79 ° 546 + 91 ° 25 436 + 159 ° 365 + 263 °

The following fermentation media have been shown to be effective for the production of EM5210 and may be substituted for media (B) and (C) in the text.

30 MIDDLE D

Grams

Oatmeal 20

Tomato cream 20

Tap water to make 1 liter.

35. Adjust the pH to 7.0 before sterilization at 12 ° C

for 15 minutes.

/ λ

Lv · "

MIDDLE E

183

Grams "Yeastamine" 5 "Cerelose" 11 5 Tap water to make 1 liter.

Sterilization at 121 ° C for 15 minutes.

MIDDLE F

Grams

Glucose 5 10 'Tartaric acid 2

Yeast extract 0.5 (nh4) 2po4 1 (NH4) 2S04 2 K2HP04 0.5 15 NaH2P04 0.5

MgS04.7H20 0.2

CaC03 1

Distilled water to make 1 liter.

Adjust the pH to 6.0 before sterilization at 121 ° C for 15 minutes.

MIDDLE G

Grams

"Nutrisoy" flour 30

Glucose 50 25 "Yeastamine" 2.5

CaC03 7

Distilled water to make 1 liter. Sterilization at 121 ° C for 30 minutes.

MIDDLE H

30 Grams NZ Amine A 10 "Cerelose" 33 "Yeastamine" 2.5

Tap water to make 1 liter.

35 Sterilization at 121 ° C for 15 minutes.

h

MIDDLE I

184

Grams

"Nutrisoy" flour 15

Soluble starch 15 5 Glucose 50

CoC12.6H20 0.005

CaCo3 10

Distilled water to make 1 liter.

Sterilization at 121 ° C for 30 minutes.

10 Biological activity

The following methodology is used to determine the minimum inhibitory concentration (hereinafter designated by the initials CIM) of the β-lactams according to the invention.

Test microorganisms are grown in approximately 15 to 20 ml of antibiotic test broth ("Difco") by inoculating (in tubes) the broth with a "magnifying glass" of the microorganism from of an inclined culture of BHI agar ("Difco"). These inoculated tubes are incubated at 35 ° C for 18-20 hours. It is assumed that these cultures contain 10 colony forming units (hereinafter referred to as the initials CFU) per milliliter. The cultures are diluted 1: 100 to obtain a final level of inoculum of 10 CFU; dilutions are made with K-10 * broth.

The compounds are dissolved in the appropriate diluent at a concentration of 1000 µg / ml. Double dilutions are made in K-10 broth, which gives a range of 1000 yg / ml to 0.5 yg / ml. 1.5 ml of each dilution are placed in individual square Petri dishes to which 13.5 ml of K-10 ** agar are added.

The final concentration of the antibiotic in the agar ranges from 100 yg / ml to 0.05 yg / ml. Microorganism growth plates containing only agar are prepared and inoculated before and after the test plates. The microorganisms are applied to the agar surface of each plate with the Denley Multipoint inoculator (which delivers approximately 0.001 ml of each microorganism), which makes it possible to obtain a level 4 / (of final inoculum 10 CFU on the agar surface.

Λ 185

The plates are incubated at 37PC for 18 hours and the MICs are determined. The ICD is the minimum concentration of compound that inhibits the growth of the microorganism.

5 * K-10 broth is a beef and yeast broth containing:

Beef extract 1.5 g

Yeast extract 3.0 g

Peptone 6.0 g 10 Dextrose 1.0 g

Distilled water in q.s. to make 1 liter ** K-10 Agar

Beef extract 1.5 g

Yeast extract 3.0 g 15 Peptone 6.0 g

Dextrose 1.0 g Agar 15.0 g

Distilled water in q.s. to make 1 liter.

The tables which follow are the results obtained when the efficacy of the β-lactams according to the invention is tested against various microorganisms. The number following each microorganism indicates the number following each microorganism indicates the number of the microorganism in the collection of E.R. Squibb & Sons, Inc., Princeton, New Jersey, United States of America. A dash (-) in the tables means that the compound tested did not show any activity vis-à-vis the particular microorganism at 100 yg / ml. The initials "N.E." mean that the compound has not been tried.

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Claims (28)

205 1. ß-lactam, characterized in that it carries a Θ Φ 0 substituent sulfonic acid salt -SO ^ M, in which M is a hydrogen atom or a cation, in position 1, and 5 a substituent amine -NH2 or a protected form thereof in position 3. 2. ß-lactam, characterized in that it carries a 0 0 0 substituent sulfonic acid salt -SO ^ M, in which M is a d atom hydrogen or a cation, in position 1, and an azide-N ^ substituent in position 3. 3. ß-lactam according to claim 1, characterized in that it carries a salt-sulfonic acid substituent 0 0 0 - SO ^ M, in which M is a hydrogen atom or a cation, in position 1, an amino substituent -NH2 in position 3, and an alkyl substituent in position 4. 4. ß-lactam according to claim 1, characterized in that it carries a sulfonic acid salt substituent Θ ffi 0 -SO ^ M, in which M is a hydrogen atom or a cation, in position 1, an amine substituent -NH2 in position 20 3, and a substituting cycloal kyle, phenyl or substituted phenyl, in position 4. 5. ß-lactam according to claim 1, characterized in that it carries a substituent salt of sulfonic acid 0 0 0 -SO ^ M, in which M is a d atom hydrogen or a cation at position 1, an amine substituent -NH2 at position 3, and an alkoxycarbonyl, alken-1-yl, alkyn-1-yl, 2-phenylethenyl or 2-phenylethynyl substituent at position 4. 6. ß-lactam according to claim 1, characterized in that it carries a substituent salt of sulfonic acid 0 0 0 30 -SO ^ M, in which M is a hydrogen atom or a cation, in position 1, an azide-N ^ substituent in position 3, and an alkyl substituent in position 4. 7. ß-lactam according to claim 1 , characterized in that it carries a sulfonic acid salt substituent 0 0 0 35 -SO ^ M, in which M is a hydrogen atom or a cation, in position 1, an azide-N ^ substituent in position 3, and a cycloalkyl, phenyl or substituted phenyl substituent, in position 4. A 206 8 p> -lactarne according to claim 1, characterized in that it carries a substituent salt of sulfonic acid -SOpi, in which M is a hydrogen atom or a cation, in position 1, an azide-N ^ substituent in position 3, and a substituent alkoxycarbonyl, alken-1-yl, alkyn-1-yl, 2-phenylethenyl or 2-phenylethynyl, in position 4. 9. ß-lactam according to claim 3, characterized in that carries a sulfonic acid salt substituent 0 0 $ -SOgM, in which M is a hydrogen atom or a cation, in position 1, an amine substituent -NH2 in position 3, and a methyl substituent in position 4. 10 · Ss-lactam according to claim 1, having the formula: = R2 h 15 - 1 NH -C-C-R 1-Α-κφ, · o * 3 wherein R2 is a hydrogen atom or an alkoxy radical of 1 to 4 carbon atoms; and R ^ are the same or different and are each a hydrogen atom or an alkyl, cycloalkyl, phenyl or substituted phenyl radical, or one of and R ^ is a hydrogen atom and the other is an alkoxycarbonyl radical , alken-1-yl, alkyn-1-yl, 2-phenylethenyl or 2-phenylethynyl; and M is a hydrogen atom or a cation. 11. ß-lactam according to claim 1, having the formula: f \ R = 2 = 4 N "C-C-R- I 1 Θ 9 30 J - N-SO M 0 in which R2 is a hydrogen atom or an alkoxy radical of 1 to 4 carbon atoms R ^ and R ^ are the same or different and are each a hydrogen atom or an alkyl radical , cycloalkyl, phenyl or substituted phenyl, or one of R ^ and R ^ is a hydrogen atom and the other is an alkoxycarbonyl, alken-1-yl, alkyn-1-yl radical, 2 -phenylethenyl or 2-phenylethynyl; and M, is a hydrogen atom or a cation. h 0 207 12, ß-lactam according to claim 10 wherein llf wherein 1 * 2 is a hydrogen atom. 13. ß-lactam according to claim 10 or 11, in which and J * 4 are methyl radicals. 5 14. ß-lactam according to claim 10 or 11, wherein 1 * 2 and R4 are hydrogen atoms. 15. ß-lactam according to claim 10, or 11, wherein 1 * 2 is a hydrogen atom and R4 is a methyl radical. 16. ß-lactam according to claim 10 or 11, wherein 10 is a methyl radical and R4 is a hydrogen atom. 17. ß-lactam according to one of claims 10 to 16, in which the amine substituent (NH2 ~) in position 3 is protected. 18. ß-lactam according to claim 17, wherein the amine substituent is in the form of an azide group. 19. ß-lactam according to claim 18, having the formula: R 4 N, -CH-C Rq II ♦ cN-SO- M 0 ^ 3 20 wherein R2 and R4 are the same or different and are each an atom of hydrogen or an alkyl radical, and + M is a hydrogen atom or a cation. 20. ß-lactam according to claim 19, wherein R ^ and R4 are the same or different and are each a hydrogen atom or a methyl radical. 21. ß-lactam, characterized in that it carries a 0 0 0 substituent sulfonic acid salt -SC ^ M, in which M is a hydrogen atom or a cation, in position 1, and an acylamine substituent in position 3. 22. ß-lactam according to claim 21, characterized in that it carries a substituent salt of sulfonic acid 0 0 0 -SOpYI, in which M is a hydrogen atom or a cation, in position 1, a substituent acylamine at position 35 3, and an alkyl substituent at position 4. 23. ß-lactam according to claim 21, characterized in that it carries a substituent salt of sulfonic acid -SCUM, in which M is a atom hydrogen or a cation, in position l, an acylamine substituent in position 3, and 208 24 · ß-lactam according to claim 21, having the formula: s2? 4 R, -NH-CC — R-3 1. I Se Λ-N-so ~ îr gold in which R1 is an acyl radical; is a hydrogen atom or an alkoxy radical of 1 to 4 carbon atoms; R3 and R4 are the same or different and are each a hydrogen atom or an alkyl, cycloalkyl or phenyl radical; and, in addition, when one of R 1 and is a hydrogen atom, the other may be an alkenyl, styryl, alkynyl, alkoxy, alkylthio, carboxyl radical or an alkyl ester thereof, hydroxymethyl, alkylsulfonyl - lower methyl, phenylsulfonylmethyl, in which the phenyl radical may be substituted by a methyl radical or a halogen, halomethyl, mercaptomethyl or a benzyl or triphenylmethyl-thio derivative thereof, azidomethyl or aminomethyl; and M is a hydrogen atom or a cation. 25. A ß-lactam according to claim 24, wherein R3 is a hydrogen atom. 26. ß-lactam according to claim 25, wherein R ^ and R ^ are methyl radicals. 27. ß-lactam according to claim '25, wherein R ^ and R ^ are hydrogen atoms. 28. The β-lactam according to claim 25, wherein r3 is a hydrogen atom and R ^ is a methyl radical. 29. β-lactam according to claim 25, wherein R ^ is a methyl radical and R ^ is a hydrogen atom. 30 . ß-lactam according to one of claims 24 to 29, in which R ^ is an aliphatic group having the formula: F1 R.-C- 5 in which R5 is an alkyl radical; cycloalkyl; A 209 alkoxy; alkenyl; cycloalkenyl; cyclohexadienyl; or alkyl or alkenyl substituted by one or more halogen atoms or cyano, nitro, amine, mercapto, alkylthio or cyanomethylthio groups. 5 31. ß-lactarne according to one of claims 24 to 29, in which is an aromatic carbocyclic group having the formula: R7 R9 R7 / - wRo ο * 7 R6 # V8 ° - o-ch2-c- R7 •, - -. R0 0 v3 ~ s‘Ch2 ^ ~ '* R7 Γ' 'y — CH -S-C- Λ 210 where n is 0, 1, 2 or 3; Rg and Rg are each, independently, a hydrogen or halogen atom or a hydroxyl, nitro, amine, cyano, trifluoro-methyl group, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 5 4 atoms of carbon, or aminomethyl; and Rg is an amine, hydroxyl group, a carboxylic salt, a radical, protected carboxyl, formyloxy, a sulfonic salt, a sulfoamine salt, an azide group, a halogen atom, a hydrazine, alkylhydrazine, phenylhydrazine radical or 10 [(alkylthio) thioxomethyllthio. 32. 3-lactam according to one of claims 24 to 29, in which is a heteroaromatic group having the formula: 0 O II RlO-ÇH-c-, * 9 O R10 “° -CH2 ^ - 'O II R ^ q“ S-CH2-C-, or 0 0 II II R10-CG- where n is Q, 1, 2 or 3; Rg is an amine, hydroxyl group, a carboxylic salt, a protected carboxyl group, formyloxy, a sulfonic salt, a sulfoamine salt, an azide group, a halogen atom, or a hydrazine, alkylhydrazine, phenylhydrazine or [( alkylthio) -thioxomethyl] thio; and R ^ is a substituted or unsubstituted pentagonal, hexagonal or heptagonal hetero-cyclic aromatic ring containing 1, 2, 3 or 4 nitrogen, oxygen or sulfur atoms. 211 33. ß-tarne lake according to one of claims 24 to 29, in which is a group of formula: 0 0 Il II Γλ -C-CH-NH-CN N-Rn, i W jLI o ^ o in which is (1) a group of formula: in which Rg, R ^ and Rg are each, independently, a hydrogen or halogen atom, a hydroxyl, nitro, amine, or cyano group, a trifluoromethyl radical, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms, or aminomethyl; or (2) a substituted or unsubstituted pentagonal, hexagonal or heptagonal heterocyclic ring containing 1, 2, 3 or 4 nitrogen, oxygen, or sulfur atoms; and R ^ is a group of formula O 11 -N = CH-R ^ or —NH-CR ^ ^, an alkylcarbonylamine, alkyl or alkyl radical substituted by one or more halogen atoms or cyano, nitro, amine or mercapto. 34. (3-lactam according to one of claims 24 to 29, in which is a group of formula: O. Il -CC = N-0-R, o R11 in which is (1) a group of formula: in which Rg , R ^ and Rg are each, independently, a hydrogen or halogen atom, a hydroxyl, nitro, amine or cyano group, or a trifluoromethyl radical, / J alkyl of 1 to 4 carbon atoms, alkoxy of 1 with 4 carbon atoms 212 or aminomethyl, or (2) a heterocyclic pentagonal, hexagonal or heptagonal ring, substituted or unsubstituted, containing 1, 2, 3 or 4 atoms of nitrogen, oxygen or sulfur; and is a hydrogen atom, an alkyl, cycloalkyl, alkylaminocarbonyl, -ONH-R ^ radical, or an alkyl substituted by one or more halogen atoms or cyano, nitro, amine, mercapto, alkylthio, R. ^, carboxyl, carboxylic salt, amide, alkoxycarbonyl, phenylmethoxycarbonyl, diphenylmethoxycarbonyl, hydroxy-alkoxyphosphinyl, dihydroxyphosphinyl, hydroxy (phenyl-me thoxy) phosphinyl or dialkoxyphosphinyl. 35. ß-lactam. according to one of claims 24 to 29, -in which is a group of formula: fï î -C-CH-NH-C-R ,. 1 14 R11 - 15 in which R ^ is (1) a group of formula: ·? 7 in which Rg, R-, and Rg are each, independently, a hydrogen or halogen atom, a hydroxy, nitro group , amine or cyano, or a trifluoromethyl radical, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms, or aminomethyl, or else (2) a substituted or unsubstituted pentagonal, hexagonal or heptagonal heterocyclic ring containing 1, 2, 3 or 4 atoms of nitrogen, oxygen or sulfur; and R ^ is a group of formula: R6-TO ^ L (CH2) n-0-25 in which n is equal to 0, 1, 2 or 3, or an amine, alkylamine, (cyanoalyl) amine, amide radical, alkylamide, y (cyanoalkyl) amide, or a group of formula: 213 NH NH0 O | l / —Λ I 1 II -CRj-NH-C-ΟN, -CH-CH2-C-NH-CH3, H0 \ —S00 -N (CH9-CH ~ -0H) 9, \ X> “CH3 \ = N \ = / 2 22 N OH 0H OH 36. ß-lactarne according to one of claims 24 to 29, in which R is a group of formula: 0 II 0 0 -C-CH-NH-C-N N-R R .. I I 11 CH 2-CH2 in which R ^ is (1) a group of formula: R? in which Rg, R ^ and Rg are each, independently, a 5 'hydrogen or halogen atom, a hydroxyl, nitro, amine or cyano group, or a trifluoromethyl radical, alkyl of 1 to 4 carbon atoms, alkoxy from 1 to 4 carbon atoms, or aminomethyl, or alternatively (2) a heterocyclic pentagonal, hexagonal or heptagonal, substituted or unsubstituted, containing 1, 2, 3 or 4 atoms of nitrogen, oxygen or sulfur ; and R ^ g is a hydrogen atom or 0 an alkylsulfonyl radical, -N = CH-R ^, -CR ^ g ^ in which R ^ g is a hydrogen atom, or an alkyl radical, or 214 substituted alkyl by a halogen atom, alkyl, or alkyl substituted by one or more halogen atoms or cyano, nitro, amine or mercapto groups. 37. β-lactam according to claim 34, wherein 5 is the 2-amino-4-thiazolyl radical. 38. ß-lactam according to claim 37, which is a salt of [3S (Z)] - 3 - [[(2-amino-4-thiazolyl) (methoxy-imino) acetyl] amino] -2- oxo-l-azetidinesulfonio, ue. 39. ß-lactam according to claim 37 which is a salt of [3S (Z)] - 3 - [[(2-amino-4-thiazolyl) [(carboxy-thoxy) imino] acetyl acid ] amino] -2-oxo-1-azetidinesulfonic. 40. ß-lactam according to claim 37, which is a salt of [3S (Z)] - 3 - [[(2-amino-4-thiazolyl) [(1-carboxy-1-methylethoxy] imino acid) ] acetyl] amino-2-oxo-1-azetidinesulfonic 41. ß-lactam according to claim 37, which · is a salt of [3S— [3a (Z), 4ß]] - 3 - [[ (2-amino-4-thiazolyl) -methoxyimino) acetyl] amino] -4-methyl-2-oxo-1-azetidine-sulfonic. 42 ß-lactam according to claim 37 which is a salt of [3S- [3a (Z), 48]] - 3 - [[[(carboxymethoxy) imino] - - (2-amino-4- thiazolyl) acetyl] amino] -4-methyl-2-oxo-1-azetinesulfonic. 43 ·· ß-lactam according to claim 37, which is a salt of [3S- [3a (Z), 4ß]] - 3 - [[(2-amino-4-thiazolyl) - [(1 -carboxy-1-methylethoxy) imino] acetyl] amino] -4-methyl1-2-oxo-1-azetidinesulfonic. 44 - ß-lactam according to claim 37, which is the dipotassium salt of [3S- [3a (Z), 4ß]] - 3 - [[(2-amino-4-thiazolyl) [(1- carboxy-1-methylethoxy) imino] acetyl] amino] - 4-methyl-2-oxo-1-azetidinesulfonic. 45. ß-lactam according to claim 37, which is a salt of [3S- [3a (Z), 4a]] - 3 - [[2-amino-4-thiazolyl) -methoxyimino) acetyl] amino] -4 acid -methyl-2-oxo-1-azetidine-sulfonic. 46 - ß-lactam according to claim 37; which is a salt of [3S— [3a (Z), 4a]] - 3 - [[(2-amino-4-thiazolyl) - [(1-carboxy-1-methoxyethoxy) imino] acetyl] amino ] -4-methyl-2-oxo-1-azetidinesulfonic. 215 47 · ß-lactarne according to claim 36, in which is a group of formula: 0 II OO II II -C-CH-NH-CN N_r15 5 0,1 '48. ß-lactam according to claim 47, in which R1 is a group of formula: 0 0. c II Il--C-CH-NH-CN N — N = CH — 177 Λ 1 1 TJ Q CH2 — CH2 5 49. ß-lactam according to claim 47, which is a salt of [3S (R *)] - 3 - [[[[[3 - [(2-furanylmethylene) -amino) -2-oxo-1-imidazolidinyl] carbony1] amino] phenylacetyl] - amino] -2-oxo-1-azetidinesulfonic. 50. ß-lactam according to claim 47, which is a salt of the acid [3S- [3a (R *), 4ß]] - 3 - [[[[[[3 - [(2-furanylmethylene) amino]] -2-oxo-1-imidazolidinyl] carbony1] amino] -phenylacetyl] amino] -4-methyl-2-oxo-1-azetidinesulfonic. 51. ß-lactam according to claim 47, which is a salt of the acid [3S- [3a (R *), 4a]] - 3 - [[[[[[3 - [(2-furanyl-methylene) amino] -2-oxo-1-imidazolidinyl] carbonyl] amino] -phenylacetyl] amino] -4-methyl-2-oxo-1-azetidinesulfonic, 52. ß-lactam according to claim 33, wherein R1 is a group of formula: OO II II / \ -C-CH-NH-C — NN — CH0CH. . A 0 216 53. (3-lactam according to claim 52, which is a salt of [3S (R *)] - 3 - [[[[(4-ethyl-2,3-dioxo-1-piperaz) acid inyl) carbonyl] amino] phenylacetyl] amino] -2-oxo-1-azetidinesulfonic 5 54. ß-lactam according to claim 52 which is a salt of [3S— [3a (R *), 4ß] acid ] -3 - [[[[(4-ethyl-2,3-dioxo-1-piperaz iny1) carbonyl] amino] phenylacetyl] amino] -4-methyl1- 2-oxo-1-azetidinesulfonic 55. ß-lactam according to claim 52 which is a salt of the acid [3S- [3a (R *), 4a]] - 3 - [[[[((4-ethyl-2,3-dioxo-1- piperaz inyl)) carbonyl] amino] phenylacetyl] amino] -4-methyl-2-oxo-1-azetidinesulfonic. 56. A composition comprising an effective amount of a β-lactam having the formula: = 2 = 4 R -NH-C-C-R, J: -i-sc® o * 3 in which is an acyl radical; R2 is a hydrogen atom or an alkoxy radical of 1 to 4 carbon atoms; R ^ and R ^ are the same or different and are each a hydrogen atom or an alkyl, cycloalkyl or phenyl radical; and, in addition, in the case where one of R ^ or R ^ is a hydrogen atom, the other may be an alkenyl, styryl, alkynyl, alkoxy, alkylthio, carboxyl radical or an alkyl ester of this. last, hydroxymethyl, lower alkylsulfonyl-methyl, phenylsulfonylmethyl in which the phenyl radical may be substituted by a methyl radical or a halogen, halomethyl, mercaptomethyl or a benzyl or triphenylmethylthio derivative thereof, azidomethyl or aminomethyl; and M is a hydrogen atom or a cation, as well as a pharmaceutically acceptable carrier for β-lactam. / λ (217 57. Process for the preparation of a β-lactam carrying a sulfonic acid salt substituent -SO ^ M, in which M is a hydrogen atom or a cation, in position 1, and a substituent amine -NE ^ or a protected form thereof, in position 3, characterized in that a corresponding β-lactam bearing a hydrogen substituent in position 1 and a protected amine substituent in position 3 is sulfonated, and in that 'said protective group of the amine function is eliminated. 58- A method according to claim 57, characterized in that the amine substituent is acylated in position 3, and in that said acylation is carried out either before or after the sulfonation reaction. 59. Method according to claim 57 or 58, characterized in that the product contains an alkyl substituent in position 4. 60. Method according to one of claims 57 to 59, characterized in that the product obtained has the formula: R2 | 4 20 Ξ = R. -NH-CC — R, 1 - 3 t J> 2 1 - +. CN — SO, M 25. in which R ^ is a hydrogen atom or an acyl group or else the group R ^ -NH- is a protected amine group; R2 is a hydrogen atom or a C ^ -C ^ alkoxy radical; R3 and R4 are the same or different and are each a hydrogen atom or an alkyl, cyclo-alkyl, phenyl or substituted phenyl radical, or one of R ^ and R ^ is a hydrogen atom and the another is an alkoxycarbonyl, alken-1-yl, alkyn-1-yl, 2-phenylethenyl or 2-phenylethynyl radical; and M + is a hydrogen atom or a cation, and in that said sulfonation is carried out on a compound of formula: A / 218 3a R4 'Ri-nb-c; -; c-r l. .1 -NH 0 or on a precursor compound of formula: V | / R4 VNH-CH —-- I r3 —-NH Ο 'λ which is cyclized, said sulfonation reaction being carried out either before or after said cyclization, the R 1 -NH group of the reactants being a protected amine group or else being an acyl radical; V being a labile group such as the methanesulfonayl, benzenesulfonyl or toluenesulfonyl radical, or a chlorine, bromine or iodine atom; and R2, R3 and R4 having the same definitions as above. · A method according to claim 60, characterized in that R4 is a hydrogen atom or else the group R ^ -NH- is a protected amine group. 62 · Method according to claim 60, characterized in that R ^ is an acyl radical. 63. Process according to claim 62, characterized in that the acyl radical is chosen from the group comprising: (a) an aliphatic group having the formula: O II r5-c- in which R, - is an alkyl radical; cycloalkyl; Alkoxy; alkenyl; cycloalkenyl; or cyclohexadienyl; or an alkyl or alkenyl radical substituted by one or more halogen atoms or cyano, nitro, 219 amine, mercapto, alkylthio or cyanomethylthio groups; (b) a carbocyclic aromatic group having the formula: R9 i / <P —CH2-0-C-:, 7 r ^^ i0.CB21 r ^ s.CH21, or R ^ cVSI in which n is equal to 0, 1, 2 or 3; Rg, R ^ and Rg are each, independently, a hydrogen or halogen atom, a hydroxyl, nitro, amine or cyano group, or a trifluoromethyl, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms, or aminomethyl; and Rg is an amine or hydroxyl group, a carboxylic salt, a protected carboxyl radical, formyloxy, / 220 μη sulfonic salt, a sulfoamine salt, an azide group, a halogen atom, or a hydrazine, alkylhydrazine, phenylhydrazine radical or [(alkylthio) thioxo-methyl] thio; 5 (c) a heteroaromatic group of formula: O rio-'ch2) „J- <0 Rio- ™ -c-. R9 î R10 '° -CH2-C-' R10-S-CU2-l-, or 0 O H10-c-L in which n is 0, 1, 2 or 3; Rg is an amine, hydroxyl group, a carboxylic salt, a protected carboxyl or formyloxy radical, a sulfonic salt, a salt of. sulfoamine, an azide radical, a halogen atom or a hydrazine, alkylhydrazine, phenylhydrazine or [(alkylthio) thioxomethyl] thio radical; and R1Q is a substituted or unsubstituted pentagonal, hexagonal or heptagonal heterocyclic aromatic ring containing 1, 2, 3 or 4 nitrogen, oxygen or sulfur atoms; (D) a group of formula: î S r ~ \ -C-CH-NH-CN NR ,, I \ _ / 12 Rii in which is (1) a group of formula: 22 1 in which Rg, and Rg are each, independently, a hydrogen or halogen atom, a hydroxy, nitro, amine or cyano group, a trifluoromethyl radical, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms, or aminomethyl or alternatively (2) a substituted or unsubstituted pentagonal, hexagonal or heptagonal heterocyclic ring containing 1, 2, 3 or 4 nitrogen, oxygen or sulfur atoms; and R ^ 2 is a group of 0 II 10 formula -N = CH-R ^ 1 or-NH-CR ^, or an alkylcarbonyl-amine, alkyl, or alkyl radical substituted by one or more halogen atoms or cyano groups , nitro, amine or mercapto; (e) a group of formula: O II -CC = N-0-R ,, I 13 Rn 15 in which R ^ is (1) a group of formula: R7 R6YtVR8 in which Rg, R7 and Rg are each, independently , a hydrogen or halogen atom, a hydroxyl, nitro, amine, or cyano group, or a trifluoromethyl, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms, or aminomethyl, or well (2) a substituted or unsubstituted pentagonal, hexagonal or heptagonal heterocyclic ring containing 1, 2, 3 or 4 nitrogen, oxygen or sulfur atoms; and R ^ g is a hydrogen atom, or an alkyl, cycloalkyl, alkylaminocarbonyl radical, 25 O II -C-NH-Rou "alkyl substituted by one or more halogen atoms or cyano, nitro, amine, mercapto, alkylthio, R ^, carboxyl, carboxylic salt, amide, alkoxycarbonyl, phenylmethoxycarbonyl, diphenylmethoxy-222 car .bonyl, hydroxyalkoxyphosphinyl, dihydroxyphosphinyl, hydroxy (phenylmethoxy) phosphinyl or dialkoxyphosphinyl; (f) a group of formula: »f -c-CH-NH-CR,. I 14 R11 in which R. ^ is (1) a group of formula: * 7 w + y 5 in which Rg, R ^ and Rg are each, independently, a hydrogen or halogen atom, a hydroxyl, nitro, amine or cyano group, or a trifluoromethyl or alkyl radical from 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms, or aminomethyl, or (2) a heterocyclic ring 5 pentagonal, hexagonal or heptagonal, substituted or unsubstituted, containing 1, 2, 3 or 4 nitrogen atoms, d 'oxygen or sulfur; and R ^ is a group of formula: (CH2) n- ° - in which n is equal to 0, 1, 2 or 3, o u well R ^ is an amine, alkylamine, (cyanoalkyl) amine, amide, 15 alkylamide, (cyanoalkyl) amide radical, or a group of formula: NH NH9 O il / 7 “Λ I 2 II -CHpNH-CN, -CH -CH2-C-NH-CH3, HO * O_S02'N (CH2-CH2-0H> 2 --¾-¾ OH t 223 0H OH ÎÔ. „TCLoi. (g) a group of formula 0 II 0 0 II II \ -C-CH-NH-C-N N-R._ 'I 15 R11 CH2-CH2 in which R ^ is (1) one. group of formula: in which Rg, R ^ and Rg are each, independently, a hydrogen or halogen atom, a hydroxyl, nitro, amine or cyano group, or a trifluoromethyl or alkyl radical of 1 to 4 atoms carbon, alkoxy of 1 to 4 carbon atoms, or aminomethyl, or alternatively (2) a heterocyclic pentagonal, hexagonal or heptagonal ring, substituted or unsubstituted, containing 1, 2, 3 or 4 atoms of nitrogen, 10 of oxygen or sulfur? and R15 is a hydrogen atom, O 11 or an alkylsulfonyl radical, -N = CH-R · ^, -C-R ^ g, in which is a hydrogen atom, or an alkyl radical. or alkyl substituted by a halogen atom, R-q 'alkyl, or alkyl substituted by one or more halogen atoms or cyano, nitro, amine or mercapto groups. 64. The method of claim 63, wherein R11 is a 2-amino-4-thiazolyl radial. 65. Process for the preparation of an acid salt (R) -3-20 (acetylamino) -3-methoxy-2-oxo-1-azetidinesulfonic, characterized in that a strain of the microorganism Chromobacterium violaceum, A.T.C.C. 31532, in an aqueous medium comprising yeast extract, 224 beef extract, NZ amine A, glucose and agar. 66. Process for the preparation of a salt of (R) -3- [[N- (D - & - glutamyl) -D-alanyl] amino] -3-methoxy-2-oxo ~ 5 azetidinesulfonic acid, characterized in that we cultivate a strain of the microorganism Gluconobacter, ATTC 31581 in an aqueous medium comprising yeast extract, beef extract, NZ amine A, glucose and agar. 67. Compound of formula: V U®4 VN “"! - <R3: -NH-SO3V 15 0 in which RI is a hydrogen atom or an acyl group or else the group R1-NH- is a protected amine group; R3 and R4 are the same or different and are each a hydrogen atom or an alkyl, cycloalkyl, phenyl or substituted phenyl radical, or one of R3 and R4 is a hydrogen atom and the other is an alkoxycarbonyl, alken-1-yl, alkyn-1-yl, 2-phenylethylenyl or 2-phenylethynyl radical; M + is a hydrogen atom or a cation; and V is a labile group such as the methanesulfonyl, benzenesulfonyl or toluenesulfonyl radical, or a chlorine, bromine or iodine atom. 68. Compound of formula: R2 R2 R4; ‘4—-7-k3 QJ-N-SO3“ in which R2 is a hydrogen atom or a C -C4 alkoxy radical; R3 and R4 are the same or different and are each a hydrogen atom or an alkyl, cycloalkyl, phenyl or substituted phenyl radical, or one of R3 and R4 is a hydrogen atom and the other is a alkoxycarbonyl, alken-1-yl, alkyn-1-yl, 2-phenylethenyl or 2-225 radical 69. Compound of formula: Ci H R4 RrN - f R3 J-N-S03'M + in which is a hydrogen atom or an acyl group; R ^ and R ^ are the same or different and are each a hydrogen atom or an alkyl, cyclo-alkyl, phenyl or substituted phenyl radical, or either of and is a hydrogen atom and the other is an alkoxycarbonyl, alken-1-yl, alkyn-1-yl, 2-phenylethenyl or 2-phenylethynyl radical; and M is a hydrogen atom or a cation. 70. Process according to claim 57, characterized in that it consists in sulfonating a compound of formula: VIR 4 ANH - CH --— IR 20 --NH2 3 in which at least one of R ^ and R ^ is a hydrogen atom and the other, optionally, is an alkoxycarbonyl, alken-1-yl, alkyn-1-ylë, 2-phenylethenyl or 2-phenylethynyl radical; A is a protective group for the amine function; and V is a labile group such as the methanesulfonyl, benzenesulfonyl or toluenesulfonyl radical, or a chlorine, bromine or iodine atom, to obtain a compound of formula: V I / r4; 30 ANH-f— ^ R3 çjJc-NHS03 “M +. in which M + is a hydrogen atom or a cation. h 226 71. Compound of formula: V I ^ R4 ANH-CH-C | R3 + 5, çjc- · NHS03-M + wherein R and R ^, A and V have the same definitions as in claim 70. 72. Compound of formula: 10 §2 §4 N c-C-R3 '· -Lso®M® “Λ. Wherein R 2 is a hydrogen atom or a C 1 -C 4 alkoxy radical; R3 and R ^ are the same or different and are each a hydrogen atom or an alkyl, cycloalkyl, phenyl or substituted phenyl radical, or one of R3 and R ^ is a hydrogen atom and the other is an alkoxycarbonyl, alken-1-yl, alkyn-1-yl, 2-phenylethenyl or 2- + phenylethynyl radical; and M is a hydrogen atortia or a cation.