BE887428A - BETA-LACTAM CORE ANTIBIOTICS, AND PROCESS FOR PREPARING THEM - Google Patents

Description

       

  CORE & -LACTAM ANTIBIOTICS,

AND PROCESS FOR PREPARING THEM

  
The present invention relates to a new family

  
  <EMI ID = 1.1>

  
of these compounds as antibacterial agents. He was

  
discovered that the $ -lactam nucleus could be biologically

  
activated by a substituent formed by a sulfonic acid salt, attached to the nitrogen atom of the nucleus.

  
The 6 * lactams comprising a substituent formed by a

  
sulfonic acid salt (including an "internal salt") posi-

  
  <EMI ID = 2.1>

  
show active against a range of Gram negative bacteria

  
and Gram positive.

  
The preferred members of this new family of S-lactam antibiotics according to the invention are

  
those defined by the formula:

  

  <EMI ID = 3.1>


  
In addition to the 3-lactams described above comprising a salt sulfonic acid substituent in position 1 and a

  
substituting acylamine in position 3, the invention encompasses

  
also 6-lactams having an acid salt substituent

  
sulfonic (including an "internal salt") in position 1, and

  
an amine substituent in position 3. The preferred compounds of

  
  <EMI ID = 4.1>

  

  <EMI ID = 5.1>


  
  <EMI ID = 6.1>

  
  <EMI ID = 7.1>

  
  <EMI ID = 8.1>

  
and throughout the specification, the symbols have the definitions below:

  
  <EMI ID = 9.1>

  
  <EMI ID = 10.1>

  
1 to 4 carbon atoms;

  
R3 and R4 are the same or different and are each a hydrogen atom or an alkyl or cycloalkyl radical,

  
  <EMI ID = 11.1>

  
one hydrogen atom and the other is an alkoxy-

  
  <EMI ID = 12.1>

  
  <EMI ID = 13.1>

  
The terms "alkyl" and "alkoxy" refer to straight chain or reunified chain radicals. Radicals having from 1 to 10 carbon atoms are preferred.

  
The terms "cycloalkyl" and "cycloalkenyl" mean

  
  <EMI ID = 14.1>

  
carbon atoms.

  
The term ".alkenyl" denotes 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 dull "substituted phenyl" denotes a substituted phenyl radical

  
  <EMI ID = 15.1>

  
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 N [deg.] 4,144,333. The preferred protected carboxyl radicals are the benzyl, benzhydrylic and 1-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 $ -lactam nucleus antibiotics, including <EMI ID = 16.1>

  
Cephalosporins and Penicillins, edited by Flynnt Académie Press (1972), the German patent application published under the

  
  <EMI ID = 17.1>

  
  <EMI ID = 18.1>

  
N [deg.] 4,172,199, and the British patent N [deg.] 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:

  

  <EMI ID = 19.1>


  
  <EMI ID = 20.1>

  
alkoxy; alkenyl; cycloalkenyl; cyclohexadienyl; or alkyl or alkenyl respectively substituted by one or

  
  <EMI ID = 21.1>

  
cyano or nitro, or by one or more amino, mercapto, alkylthio or cyanomethylthio radicals.

  
(b) carbocyclic aromatic radicals having the formula:

  

  <EMI ID = 22.1>
 

  

  <EMI ID = 23.1>


  
  <EMI ID = 24.1>

  
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 radical; 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, phenylhydrazine radical , or [(alkylthio) thioxomethyl] thio.

  
Preferred carbocyclic aromatic acyl radicals include those having the formula:

  

  <EMI ID = 25.1>
 

  

  <EMI ID = 26.1>


  
  <EMI ID = 27.1>

  
pic) or

  

  <EMI ID = 28.1>


  
  <EMI ID = 29.1>

  
pic).

  
(c) Heteroaromatic radicals having the formula:

  

  <EMI ID = 30.1>


  
  <EMI ID = 31.1>

  
  <EMI ID = 32.1>

  
hexagonal or heptagonal, substituted or unsubstituted, containing 1, 2, 3 or 4 (preferably 1 or 2) nitrogen, oxygen or sulfur atoms. Exemplary heterocyclic rings are thienyl, furyl, pyrrolyl, pyridinyl,

  
  <EMI ID = 33.1>

  
tetrazolyl. Exemplary substituents are halogen atoms, hydroxyl and nitro groups, amine, cyano, trifluoromethyl, alkyl of 1 to 4 carbon atoms, or alkoxy of 1 to 4 carbon atoms.

  
Preferred heteroaromatic acyl radicals include radicals having the above formulas in

  
  <EMI ID = 34.1> <EMI ID = 35.1>
  <EMI ID = 36.1>
  <EMI ID = 37.1>

  
carbocyclic aromatic radicals such as those which

  
  <EMI ID = 38.1>

  

  <EMI ID = 39.1>


  
as well as heteroaromatic radicals as they are

  
  <EMI ID = 40.1>

  
alkyl, substituted alkyl (where the alkyl radical is substituted by one or more halogen atoms, or by one or more

  
  <EMI ID = 41.1>

  
amine (ie: -N = CH-R .., where R ,, has the same definition

  
0

  
) t as above), arylcarbonylamine (i.e.: -NH-C-R.

  
  <EMI ID = 42.1>

  
carbonylamine.

  
  <EMI ID = 43.1>

  
preferred carbonyl] amino] arylacetyl include those in which R12 is an ethyl, phenylmethyleneamine or 2-furylmethyleneamine radical.

  
Ce} Arylacetyl (substituted oxyimino) radicals having the formula:

  

  <EMI ID = 44.1>


  
  <EMI ID = 45.1>

  
is a hydrogen atom, an alkyl, cycloalkyl, alkylaminocarbonyl, arylaminocarbonyl radical (i.e. 0

  
he <EMI ID = 46.1> substituted alkyl radical (in which the alkyl radical is substituted by one or more halogen atoms or cyano, nitro, amine, mercapto, alkylthio, or aromati-

  
  <EMI ID = 47.1>

  
salts), amide, alkoxycarbonyl, phenylmethoxycarbonyl, diphenylmethoxycarbonyl, hydroxyalkoxyphosphinyl,

  
  <EMI ID = 48.1>

  
  <EMI ID = 49.1>

  
thiazolyl. We also prefer the radicals in which

  
  <EMI ID = 50.1>

  
carboxyisopropyl.

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

  

  <EMI ID = 51.1>


  
  <EMI ID = 52.1>

  
is a radical

  

  <EMI ID = 53.1>


  
  <EMI ID = 54.1>

  
amide, alkylamide, (cyanoalkyl) amide,
  <EMI ID = 55.1>
 
  <EMI ID = 56.1>
 The (acylamino) arylacetyl radicals preferred according to the

  
  <EMI ID = 57.1>

  
is an amine radical, or amide. Preference is also given to radicals in which R is a phenyl or 2-thienyl radical.

  
  <EMI ID = 58.1>

  
dinyl] carbonyl] amino] arylacetyl having the formula:

  

  <EMI ID = 59.1>


  
  <EMI ID = 60.1>

  
is a hydrogen atom or an alkylsulfonyl radical,

  
  <EMI ID = 61.1>

  
0

  
he

  
  <EMI ID = 62.1>

  
hydrogen or an alkyl or alkyl radical substituted by a halogen atom), an aromatic radical (according to the definition

  
  <EMI ID = 63.1>

  
which the alkyl radical is substituted by one or more halogen atoms or cyano, nitro, amine or mercapto groups).

  
The preferred [[[3-substituted-2-oxo-1-imidazolidinyl] carbonyl] amino] arylacetyl radicals according to the formula below.

  
  <EMI ID = 64.1>

  
phenyl or 2-thienyl. We also prefer radicals

  
  <EMI ID = 65.1>

  
  <EMI ID = 66.1>

  
amine.

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

  
  <EMI ID = 67.1>

  
lactams according to the invention encompasses all the sulfonic acid salts. It is preferred, of course, the pharmaceutically acceptable salts, although other salts are also usable for purifying the products of the invention or else as intermediates for the preparation of the 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 ions

  
  <EMI ID = 68.1>

  
after tetrabutylarnmonium); alkali metals, such as lithium, sodium and potassium; alkaline earth metals, such as calcium and magnesium; pyridinium; dicyclohexylammonium; hydrabaminium; benzathinium; N-methyl-Dglucaminium.

  
  <EMI ID = 69.1>

  
tions according to formula I, M can be a hydrogen ion.

  
As will be described below, the 3-lactams according to the invention can be prepared by synthesis. We can pre-

  
  <EMI ID = 70.1>

  
4 and not alkoxylated, that is to say the compounds of formula I

  
  <EMI ID = 71.1>

  
using 6-aminopenicillanic acid or 6-acylaminopenicillanic acid as starting material. The 0-lactams of formula I in which R2 is an alkoxy radical can be prepared from the corresponding non-alkoxylated 6-lactam. It is possible to crystallize or recrystallize certain of the compounds according to the invention in solvents containing water. In these cases, hydration water may form. The present invention does not exclude stoichiometric hydrates as well as compounds containing variable proportions of water which can be obtained by methods such as lyophilization.

  
  <EMI ID = 72.1>

  
been prepared by biological means. The culture of a strain of the microorganism Chromobacterium violaceum SC 11378

  
  <EMI ID = 73.1>

  
sulfonic in position 1 and an amine or acylamine substituent in position 3 contain at least one chiral center <EMI ID = 74.1> ration on the carbon atom occupying position 6 of the penicillins of natural origin (penicillin G for example), and that the configuration on the carbon atom occupying position 7 of the cephamycins of origin natural
(cephamycin C for example).

  
  <EMI ID = 75.1>

  
I and the, the formulas developed have been drawn so as to reveal the stereochemistry on the chiral center occupying position 3. For reasons of convention of

  
  <EMI ID = 76.1>

  
which R2 is a hydrogen atom have the S configuration, and the compounds of formulas 1 and la in which R2 is an alkoxy radical have the R configuration.

  
Also included in the scope of the invention are the racemic mixtures which contain the $ -lactams described above.

  
$ -Lactams having an acid salt substituent

  
  <EMI ID = 77.1>

  
amine or acylamine at position 3 of the B-lactam nucleus are active against a range of Gram-negative and Gram-positive organisms. The sulfonic acid salt substituent is essential for the activity of the compounds according to the invention. The

  
  <EMI ID = 78.1>

  
/ 1. or an alkyl radical, in particular the methyl radical, demonstrate 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 (for example 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 a rate about
14 mg / kg / day to approximately 100 mg / kg / day. All the modes of administration which have been used in the past to reach the site of infection with penicillins and cephalosporins are also envisaged with the new family of 0-lactams according to the invention. These modes of administration include oral, intravenous, intramuscular, and suppository forms.

  
We generally prepare 0-lactams according to the invention.

  
  <EMI ID = 79.1>

  
sulfonation reaction by treating 6-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, dimethylformamidetrioxide, 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 using an intermediate compound, such as by silylating first

  
  <EMI ID = 80.1>

  
to the silylated compound an exchange reaction of the silyl radical with trimethylsilylchlorosulfonate or a similar reagent. Exemplary silylating agents are monosilyl-trifluoroacetamide, the combination of trimethyl chloride

  
  <EMI ID = 81.1>

  
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 than dichloromethane.

  
The product initially formed in the sulfonation reaction is a sulfonated $ -lactam salt. When the sulfonation complex is the pyridine-sulfur trioxide complex, the product initially formed is the pyridinium salt of the sulfonated [cent] -lactam, M + being the pyridinium ion in the formula below:

  

  <EMI ID = 82.1>


  
These complexes can be transformed into other sulfonic acid salts using conventional 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 transformation of the pyridine salt into potassium salt using phosphate

  
  <EMI ID = 83.1>

  
tion of tetrabutylammonium salt using tetrabutyl ammonium hydrogen sulfate; or the transformation into a zwitterion (M = hydrogen) using formic acid;

  
It must be realized that the sulfonation reaction which introduces the sulfonic group on the nitrogen atom of the 6-lactam nucleus can take place at various stages of the synthesis, including introduction before the formation of a $ -lactam nucleus , 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.

  
1 When, for example, a protective group is used

  
  <EMI ID = 84.1>

  
represented as follows:,

  

  <EMI ID = 85.1>


  
Other protective groups can be used to protect the amine function, for example a t-batyloxycarbonyl group, a simple acyl radical such as the acetyl or benzoyl or phenylacetyl radical, a triphenylmethyl radical, or else put the amine function in the form an azide function. We can then fix the desired acyl radical

  
  <EMI ID = 86.1>

  
Exemplary acylation techniques allowing

  
  <EMI ID = 87.1>

  
  <EMI ID = 88.1>

  
or with a carboxylic acid halide or an anhydride

  
  <EMI ID = 89.1>

  
alkoxy radical, acylation is best done 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 that case,

  
  <EMI ID = 90.1>

  
tives (for example amine or carboxyl radicals), it may be necessary to protect these functional radicals first

  
  <EMI ID = 91.1>

  
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:

  

  <EMI ID = 92.1>


  
In the case where R2 is a lower alkoxy radical, there may be an additional variation in that one can introduce the lower R2-alkoxy radical after

  
sulfonation as well as before sulfonation using the 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:

  

  <EMI ID = 93.1>


  
In the above reaction, the acyl radicals also include easily removable radicals which judge the role of protective groups and which can be removed after the reaction in order to obtain the “deacylated” product (-NH2).

  
Furthermore, the O-lactam ring 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.

  

  <EMI ID = 94.1>


  
In this reaction, the acyl radical can also be an easy-to-remove group which acts as a protective group and which, after removal, makes it possible to obtain the product with substituent -NH2.

  
It is also possible to prepare the starting azetidinones in which R2 is a hydrogen atom and at least one

  
  <EMI ID = 95.1>

  
amino acids having the formula:

  

  <EMI ID = 96.1>


  
(at least one of R3 and R4 is a hydrogen atom). The amine radical is first of all protected using a conventional protective group, for example the t-butoxycarbonyl 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:

  

  <EMI ID = 97.1>


  
in which Y is an alkyl or benzyl radical, in the presence of a carbodiimide, to obtain a compound having the formula:

  

  <EMI ID = 98.1>


  
  <EMI ID = 99.1>

  
transforms the hydroxyl group of a compound of formula XIV 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) that can be used are the benzenesulfonyl and toluenesulfonyl radicals and the chlorine, bromine and iodine atoms.

  
The fully protected compound is cyclized, having the formula:

  

  <EMI ID = 100.1>


  
(at least one of R3 and R4 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:

  

  <EMI ID = 101.1>


  
  <EMI ID = 102.1>

  
Alternatively, it is possible to cyclize a compound of formula XIV without first transforming the hydroxyl group into a labile group. The treatment of a

  
  <EMI ID = 103.1>

  
diethylazodicarboxylate, gives a compound of formula XVI

  
  <EMI ID = 104.1>

  
hydrogen.

  
It is possible to carry out the elimination of the protecting group from position 1 of an azetidinone of formula XVI by using a reduction with sodium when Y is an alkyl radical, and this gives an intermediate of formula:

  

  <EMI ID = 105.1>


  
(at least one of R3 and R4 is a hydrogen atom). If Y is the benzyl radical, catalytic hydrogenation (using palladium on charcoal for example) will initially give the corresponding N-hydroxylated compound which, by treatment using titanium trichloride, will give an intermediate of formula XVII wherein at least one of R3 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:

  

  <EMI ID = 106.1>


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

  
Another operating mode for preparing a

  
  <EMI ID = 107.1>

  
  <EMI ID = 108.1>

  
use as ur starting material. amino acid amide having. for formula:

  

  <EMI ID = 109.1>


  
  <EMI ID = 110.1>

  
Protection of the amine group by a conventional protective group, the benzyloxycarbonyl radical for example
(hereafter 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:

  

  <EMI ID = 111.1>


  
  <EMI ID = 112.1>

  
a protective group.

  
The sulfonation of a compound of formula XX gives a

AT

  
composed of formula:

  

  <EMI ID = 113.1>


  
  <EMI ID = 114.1>

  
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

  
  <EMI ID = 115.1>

  
and we obtain a compound of formula:

  

  <EMI ID = 116.1>


  
  <EMI ID = 117.1>

  
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 R2-alkoxy radical, by catalytic hydrogenation, gives a compound of formula:

  

  <EMI ID = 118.1>


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

  
  <EMI ID = 119.1>

  
alkoxy, and this compound can be transformed into the corresponding zwitterion, having the formula:

  

  <EMI ID = 120.1>
 

  
  <EMI ID = 121.1>

  
treatment with an acid such as formic acid.

  
Removal of the protecting group from a sulfonated azetidinone of formula XXII in which A is a Boc protecting group using acidic conditions
(using for example formic acid} gives the corresponding zwitterion of formula Xlv.

  
An excellent source of starting p-lactams are

  
  <EMI ID = 122.1>

  
cephalosporanics which may carry an optional 6-alkoxy or 7-alkoxy substituent, respectively. These compounds have respectively the formulas:

  

  <EMI ID = 123.1>


  
  <EMI ID = 124.1>

  
is a hydrogen atom or an acyl radical. By adapting the procedures described in the literature, we can

  
  <EMI ID = 125.1>

  
page 257, and Synthesis, 494 (1977).

  
We start by desulfurizing the 6-amino acid

  
  <EMI ID = 126.1>

  
reduction using Raney nickel. The reaction can be carried out in water under reflux conditions; the resulting compound has the following structural formula:

  

  <EMI ID = 127.1>


  
The replacement of the carboxyl group of the compound of formula XXVI by an acetate group, followed by hydrolysis, gives a 3-amino-3-alkoxy-2-azetidinone of formula l in which R. is a hydrogen atone or a radical

  
  <EMI ID = 128.1>

  
XXVI with cupric acetate and lead tetraacetate in an 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.

  
We can realize the introduction of a sul-

  
  <EMI ID = 129.1>

  
by making the intermediary react with. a complex of dimethylformamide and sulfur trioxide.

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

  

  <EMI ID = 130.1>


  
with a halosulfonylisocyanate (preferably chlorosulfonylisocyanate) having the formula:

  

  <EMI ID = 131.1>


  
to obtain an azetidinone of formula:

  

  <EMI ID = 132.1>
 

  
The reductive hydrolysis of an azetidinone of formula XXIX gives an unsubstituted 3-lactam on nitrogen, having the formula:

  

  <EMI ID = 133.1>


  
For a more detailed description of the reaction sequence described above, one can refer to the literature; see, for example, Chem. Soc. Rev., 5, 181

  
  <EMI ID = 134.1>

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

  

  <EMI ID = 135.1>


  
The reaction is best done if the nitrogen of the azetidinone is first protected by a silyl residue (for example tbutyldimethylsilyl, 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) # low temperature, then treating this anion with toluenesulfonylazide. The resulting intermediate is stabilized using trimethylsilyl chloride, and then acid hydrolysis or solvolysis with a fluoride of the nitrogen protecting group gives the compound of formula XXXI.

  
Or, we can get the compound of formula XXXI

  
by first reacting a primary amine of formula:

  

  <EMI ID = 136.1>
 

  
with an aldehyde of formula R3CH = O, to obtain the corresponding Schiff base. A cycloaddition [2 + 2] with a

  
  <EMI ID = 137.1>

  
2-azetidinone having the formula:

  

  <EMI ID = 138.1>


  
in which Q is a radical of formula:

  

  <EMI ID = 139.1>


  
Removal 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 to obtain the corresponding 3-amino-2-azetidinone, then

  
  <EMI ID = 140.1>

  
  <EMI ID = 141.1>

  
lower alkoxy, the product can be prepared from the equivalent product in which R2 is a hydrogen atom. The chlorination of the nitrogen amide of a non-alkoxylated compound gives an intermediate of formula:

  

  <EMI ID = 142.1>


  
Reagents and procedures for chlorinating amides on the nitrogen atom are known in the art. Exemplary reagents are tert.-butyl hypochlorite, sodium hypochlorite, and chlorine. We

  
  <EMI ID = 143.1> in a two-phase mixed solvent (for example a mixture of water and methylene chloride) in the presence of 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 R2 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 to prepare the

  
  <EMI ID = 144.1>

  
consists in first alkoxylating an intermediate of formula VI

  
  <EMI ID = 145.1>

  
  <EMI ID = 146.1>

  
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:

  

  <EMI ID = 147.1>


  
Using the alkoxylation procedure described above (making it possible to transform a compound of formula XXXII into a product of formula I), and then adding a reducing agent such as trimethylphosphite, the compound of formula XXXIII can be transformed into an intermediate of formula:

  

  <EMI ID = 148.1>
 

  
in combination with its enantiomer,

  
  <EMI ID = 149.1>

  
formula I in which R2 is an alkoxy radical, under the
-orm of a racemic mixture. If necessary, the enantiomer having the R configuration can be isolated from this racemic mixture, using conventional techniques such as fractional crystallization of an appropriate salt with an optically active organic admir.e, or alternatively by d pair chromatography. 'ions, using an optically active cation.

  
Biological production of the antibiotic EM5117

  
The salts of acid (R) -3- can also be prepared

  
  <EMI ID = 150.1>

  
(formula I, R1 is an acetyl radical and R2 is a methoxy radical; designated below by the code EM5117) by culture of a strain of the microorganism Chromobacterium violaceum

  
SC 11.378 which was deposited in the American Type Culture Collection (A.T.C.C.) under N [deg.] 31.532.

  
The microorganism

  
The microorganism used for the production of EM5117 is a strain of Chromobacterium violaceum SC 11.378. We can get a subculture of this microorganism from

  
the A.T.C.C.'s permanent collection in Rockville,

  
  <EMI ID = 151.1>

  
collection is A.T.C.C. N [deg.] 31.532. In addition to the specific microorganism described and characterized here, it should be understood that mutant species of the microorganism (for example the mutant species produced using X-rays, ultraviolet radiation or "nitrogen mustard" ") can also be grown to produce EM5117.

  
We can isolate Chromobacterium violaceum SC 11.378, A.T.C.C. N [deg.] 31.532, of a wet soil sample containing the micro-organism, first applying the soil sample to a medium containing:

  

  <EMI ID = 152.1>
 

  
This medium is adjusted to pH 6.0 and sterilized in a

  
  <EMI ID = 153.1>

  
incubation at 25 [deg.] C, isolated from the soil applied to

  
  <EMI ID = 154.1>

  
We then grow these. colonies isolated on a medium containing:

  

  <EMI ID = 155.1>


  
Distilled water to make 1 liter.

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

  
Chromobacterium violaceum SC 11.378 is a stick

  
  <EMI ID = 156.1>

  
bipolar staining and lipid inclusions. It can move with a single polar flagellum, sometimes with side flagella that are smaller.

  
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 but no confluent film. The purple 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 [deg.] C no growth occurs at 4 [deg.] C or above 37 [deg.] 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
(Hugh & Leifson method, 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.

  
The fundamental characteristics described above form the basis for the identification of the microorganism as Chromobacterium violaceum and not Chromobacterium lividum, the only other species of the genus Chromobacterium which has been identified in the 8th edition of the "Manual of Determinative Bacteriology" from Bergey.

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

  
The antibiotic

  
The antibiotic EM5117 is obtained by growing Chromobacterium violaceum SC 11.378, A.T.C.C. N [deg.] 31.532, at

  
  <EMI ID = 157.1>

  
immersed in an aqueous nutrient medium containing a source of assimilable carbon and nitrogen. The fermentation is carried out until a substantial antibiotic activity has been imparted to the medium, that is to say usually for approximately 18 to 24 hours, preferably for approximately 20 hours.

  
Using the following procedure, EM5117 can be separated from the fermentation medium and purified. Once fermentation is complete, the

  
  <EMI ID = 158.1>

  
use of a filtratioh to separate the mycelium from the broth. After separating the mycelium from the broth ,. EM5117 can be extracted from the broth. We extract preferably

  
  <EMI ID = 159.1>

  
of cetyldimethylbenzylammonium chloride in methylene chloride, and it is reextracted in aqueous sodium iodide (adjusted to pH 5 with 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.

  
We can carry out the purification of the antibiotic

  
by dissolving the methanol concentrate in a 1: 1 mixture

  
methanol and water and passing it through a chromatography column, for example in a column comprising "Sephadex G-10" in the same mixture of solvents. After elution with this same mixture, the fractions are combined:
active 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 a 0.0 M sodium phosphate buffer of pH 5 and a 0.1 M sodium phosphate buffer of pH 5. The active fractions are combined, they are concentrated and the substances insoluble in methanol are separated and discarded.

  
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

  
  <EMI ID = 160.1>

  
the active fractions are concentrated. The concentrate is dissolved in water and passed through a column of "Dowex 50W-X2", in the potassium form, washing with water. The effluent can be concentrated to obtain a crystalline substance which is a relatively pure potassium salt 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 EM5210

  
  <EMI ID = 161.1>

  
  <EMI ID = 162.1>

  
  <EMI ID = 163.1>

  
glutamyl-D-alanyl and R2 is the methoxy radical; hereinafter designated by the code "EM5210") by culture of various bacteria of acetic acid.

  
/ 1 <EMI ID = 164.1>

  
The microorganism Gluconobacter species SC 11.435 can be used for the production of EM5210. 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 collection is A.T.C.C. N [deg.] 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, ultraviolet radiation or "nitrogen mustard") to produce EM5210.

  
Gluconobacter species SC 11.435, A.T.C.C. N [deg.] 31.581, of crushed foam containing the microorganism, by first incubating the crushed foam in a 10% aqueous pectin solution (pH 2.5) for 7 days at 25 [deg.] C. We can then isolate the microorganism

  
by applying it to a medium containing:

  

  <EMI ID = 165.1>


  
* We prepare the herb extract Spartina patens 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 [deg.] C for 20 minutes. After approximately

  
  <EMI ID = 166.1>

  
enrichment of pectin on a slide from colonies of Gluconobacter species SC 11.435. These isolated colonies are then grown on a medium containing:

  

  <EMI ID = 167.1>
 

  
The medium is adjusted to pH 7.3 and treated in an autoclave

  
  <EMI ID = 168.1>

  
, Gluconobacter species SC 11.435 is a pleomorphic Gram-negative rod which can move by means of 1 to 3 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 without pigment. 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 crowded muddy mud colonies. A similar growth is obtained on a malt agar - yeast extract

  
(1% each) adjusted to pH 4.5.

  
  <EMI ID = 169.1>

  
of glucose, 3% calcium carbonate and 2.5% agar, sufficient is produced from glucose to form a zone of disappearance of calcium carbonate around the growing 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 calcium yeast 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

  
1 to 1, middle 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.

  
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 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

  
A.T.C.C. 11894

  
The antibiotic

  
To obtain the antibiotic EM5210, one can grow Gluconobacter, species SC 11.435, A.T.C.C. N [deg.] 31581, or one of the micro-organisms listed above, at a temperature equal to or close to 25 [deg.] C, under immersed aerobic conditions, on an aqueous nutrient medium containing an assimilable source carbon and nitrogen. The fermentation is carried out until a substantial antibiotic activity has been imparted to the medium, that is to say usually for approximately 16 to 24 hours, preferably for approximately 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 eliminate 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 at about pH 4, and the eluate is concentrated, then passed through a charcoal 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 "Dowex 1-X2", and elution is carried out with a gradient of sodium chloride buffered to pH 4. The active fractions and they are desalted on a column of macroreticular styren-divinylbenzene copolymer resin ("Diaion HP20AG").

  
The active fractions eluted are concentrated with water and

  
  <EMI ID = 170.1>

  
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.

Example 1

  
  <EMI ID = 171.1>

  
phenylacetamide

  
Method I: <EMI ID = 172.1>

  
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.

  
  <EMI ID = 173.1>

  
500 ml of water, 54 g (90 ml) of Raney nickel are added. The balloon, equipped with a reflux condenser, is immersed in a

  
  <EMI ID = 174.1>

  
for 15 minutes. The flask is immediately cooled in an ice-water bath, then the Raney 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 it scrapes the walls of the container. After washing with water and drying under vacuum for <EMI ID = 175.1>

  
Nitrogen is bubbled through for 15 minutes in a stirred suspension of 608 mg (2 mmol) of the above compound in 20 ml of dry acetonitrile. A water bath at 40 - 45 [deg.] C is used for several minutes to dissolve all of the acid. The water bath is removed, and powdered cupric acetate monohydrate (182 mg, Immol) 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 is dried over sodium sulfate and concentrated to a residue (515 mg) which is used without further purification in the following reaction.

  
  <EMI ID = 176.1>

  
To a solution of 911 mg (2.86 mmol) of the above compound in 21 ml of methanol, 3.5 ml of water is added, then
383 mg (2.86 mmol) of potassium carbonate. The mixture is stirred under nitrogen for one minute, then 160 mg is added
(4.30 mmol) sodium borohydride. 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 is washed at pH 7.0 with aqueous sodium bicarbonate and then with a small volume of water, and finally, it is 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 amounts of product can be obtained by crystallization or chromatography.

  
/ 1 <EMI ID = 177.1>

  
1.35 mmol) to a stirred solution of 251 mg (1.23′-immole) of the above product in 2 ml of dry dimethylformamide and 2 ml of dry methylene chloride, 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. We adjust the pH

  
  <EMI ID = 178.1>

  
  <EMI ID = 179.1>

  
concentrates into 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 [deg.] C, decomposition.

  
  <EMI ID = 180.1>

  

  <EMI ID = 181.1>


  
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 acid salt is stirred in a hydrogen atmosphere for 2 hours.

  
  <EMI ID = 182.1>

  
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 the acetone and applied to a 200 ml column of "HP-20". Elution with water and then with a 9: 1 mixture 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

  
  <EMI ID = 183.1> mixture of methanol and ether gives 101 mg of the title compound, melting point 210 [deg.] C, decomposition.

  
  <EMI ID = 184.1>

  

  <EMI ID = 185.1>


  
Method III:

  
To a solution of tetrabutylammonium acid salt
(S) -3-amino-2-oxo-1-azetidinesulfonique (121 mg; see

  
  <EMI ID = 186.1>

  
add 40 mg of phenylacetic acid and 61 mg of dicyclohexylcarbodiimide. The mixture is 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 and, after drying, 48 mg of product are obtained, the spectral characteristics of which are in agreement with those of

  
  <EMI ID = 187.1>

  
Method IV:

  
A solution is stirred in a hydrogen atmosphere

  
  <EMI ID = 188.1>

  
methoxy) carbonyl] amino] -1-azetidinesulfonic (see Example 2) in 36 ml of water, with 707.5 mg of palladium at
10% on carbon (175 ml of hydrogen are absorbed). After 2 hours, the suspension is filtered and the filtrate is cooled to 0 [deg.] C and then diluted with 46 ml of acetone (initial pH = 4.25 adjusted to pH 6.7 with a cold solution of 10% potassium bicarbonate). 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

  
  <EMI ID = 189.1>

  
10% potassium bicarbonate. After 45 minutes, the suspension is diluted with 93 ml of potassium phosphate

  
  <EMI ID = 190.1>

  
filter the suspension and wash it with water. The filtrate and the washing waters 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

  
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

  
  <EMI ID = 191.1>

  
A suspension of 6-aminopenicillanic acid (12.98 g; 0.06 mole) in 140 ml of water containing 5.18 g of sodium bicarbonate (stirred for approximately 10 minutes without dissolution) is added all at once complete), to a well stirred suspension (mechanical stirrer) of Raney nickel
(washed with water at pH 8.0, 260 ml of suspension = 130 g)

  
in an oil bath at 70 [deg.] 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 a total of 6.83 g of the title compound.

  
  <EMI ID = 192.1>

  
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 per 9.5 g
(0.0213 mole) lead tetraacetate. The suspension is immersed in an oil bath at 65 [deg.] 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 acetate

  
n of ethyl. 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, dry and evaporate to obtain 6.235 g of the title compound.

  
C) Phenylmethyl ester of acid (S) - (2-oxo-3- <EMI ID = 193.1>

  
Cooled to -15 [deg.] C a solution of 3.12 g (0.0093 mole) of the above acetate in 70 ml of methanol and 7 ml of water, and 1.33 g of potassium carbonate and

  
349 mg sodium borohydride. The reaction mixture is stirred

  
  <EMI ID = 194.1>

  
complete reaction (approximately 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 is combined with the substance from a similar experiment and triturated with ether to obtain 3.30 g of the title compound.

  
  <EMI ID = 195.1>

  
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, with 350 mg (0.0022 mole) of a pyridine and sulfur trioxide complex. 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.

  
  <EMI ID = 196.1>

  
4.85 (1H, d of d,) = 6.4), 5.10 (2H, S), 7.27 (5H, S) 8.0-9.0ppm
(m's 5H).

  
Method II:

  
18.87 g of ester are added dropwise at -20 [deg.] C

  
  <EMI ID = 197.1>

  
stirring under a nitrogen atmosphere. Once the addition is complete, stirring is continued for 30 minutes at room temperature, then the trimethylchlorosilane is removed in vacuo. A solution of 20 g of the above azetidinone (Method I, part C) in 120 ml is added

  
  <EMI ID = 198.1>

  
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 to it. both ethyl acetate, which gives 31 g of the title compound. The NMR data are identical to that of the product obtained by Method I.

Example 3

  
  <EMI ID = 199.1>

  
  <EMI ID = 200.1>

  
Pyridine salt of (S) -2-oxo-3- acid is dissolved

  
  <EMI ID = 201.1>

  
(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 column of
25 ml 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.

  
  <EMI ID = 202.1>

  
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

  
  <EMI ID = 203.1>

  
(600 mg; see J. 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. It forms a solid, and we cool

  
  <EMI ID = 204.1>

  
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 "HP-20". This column is eluted with 200 ml of water and then with a 9: 1 mixture of water and acetone. The fractions are collected (50 ml); the evaporation of fraction 7 gives a solid. The crushing with

  
  <EMI ID = 205.1>

  

  <EMI ID = 206.1>


  
Method III:

  
  <EMI ID = 207.1>

  
(S) - (2-oxo-3-azetidinyl) carbamic acid phenylmethyl ester (20.0 g, see Example 2C), 21.6 ml are added

  
  <EMI ID = 208.1>

  
heats the mixture to 50 [deg.] C while stirring for one hour.

  
After cooling in an ice bath at 0 [deg.] 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 ethylhexanoate are added to the solution

  
of potassium in 100 ml of butanol and the stirring is continued for another hour. We pour the suspension in

  
  <EMI ID = 209.1>

  
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 proportions of potassium chloride 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- acid

  
  <EMI ID = 210.1>

  
Method I:

  
The pyridine salt (1: 1) is dissolved in 800 ml of water

  
  <EMI ID = 211.1>

  
azetidinesulfonic (34.3 g; see Example 2). The solution is made clear with charcoal, 30.7 g of tetrabutylammonium hydrogen sulfate in 80 ml of water are added,

  
  <EMI ID = 212.1>

  
solvent in vacuo until the volume is only 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 [deg.] C.

  
Method II:

  
Dissolve the potassium salt (1: 1) of the acid (S) -

  
  <EMI ID = 213.1>

  
that (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 under vacuum to approximately 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 From.

Example 5

  
Potassium salt (1: 1) of the phenylmethyl ester of

  
  <EMI ID = 214.1>

  
The phenylmethyl ester of the acid is hydrogenated
(S) - (2-oxo-3-azetidinyl) carbamic acid (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 separated

  
  <EMI ID = 215.1>

  
standing, 1.1 g of the title compound crystallizes.

  
  <EMI ID = 216.1>

  
The above azetidinone (3.0 g) is dissolved in 100 ml

  
  <EMI ID = 217.1>

  
add 4.5 g of N-methylmorpholine, then (drop by drop)

  
  <EMI ID = 218.1>

AT

  
  <EMI ID = 219.1>

  
of water to the residue. The aqueous suspension is extracted twice

  
  <EMI ID = 220.1>

  
the organic layers are combined, washed with sodium bicarbonate, 2N phosphoric acid and water, dried over sodium sulfate, filtered and evaporated to dryness. The residue is crystallized from ethyl acetate and petroleum ether to give 8.7 g of the title compound, melting point 164-166 [deg.] C.

  
  <EMI ID = 221.1>

  
6.9 g of the above compound are suspended in 150 ml of acetonitrile. 5.7 g of monotrimethyl are added.

  
  <EMI ID = 222.1>

  
minutes at 50 [deg.] C with stirring. The solution is cooled to 0 ° C. and 3.9 g of trimethylsilyl chlorosulfonate are added dropwise. Once the addition is complete, the mixture is heated to 50 [deg.] C for 5 hours. After cooling to
20 [deg.] C, 7.6 g of potassium ethylhexanoate are added to

  
10 ml of butanol and the 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 is separated by filtration, which gives 4.5 g of the

  
  <EMI ID = 223.1>

  
purification of the crude product by chromatography on "HP-20", followed by cryodessication, gives a pure substance of which

  
  <EMI ID = 224.1>

Example 6

  
  <EMI ID = 225.1> Example 4), and it is hydrogenated for approximately 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.

  
  <EMI ID = 226.1>

  
2.5 cps).

  
  <EMI ID = 227.1> acetyl] arnino] -2-oxo-l-azetidinesulforique

  
Stirred at 0 [deg.] 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 dicyclohexylcarbodiimide is added dropwise in 10 ml of dimethylformamide. Once the addition is complete, the stirring is continued for 12 hours at 20 [deg.] 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 filtered off, dried and purified on a chromatography column containing 300 ml of "HP-20", using water as the eluting. The yield is 850 mg of the compound

  
  <EMI ID = 228.1>

  
see Example 6A) in 100 ml of dimethylformamide and 2 ml of propylene oxide. A solution of O-formylmandelic acid chloride in

  
10 ml of acetonitrile. The temperature is maintained for

  
1 hour 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, and is filtered, giving 1.5 g of product. We

  
/ 1 purifies the product by chromatography on. "HP-20", m.p.

  
  <EMI ID = 229.1>

  
Following the procedure of Example 7, but replacing the O-formylmandelic acid chloride by the D-O-formylmandelic acid chloride, the compound is obtained

  
  <EMI ID = 230.1>

  
Following the procedure of Example 7 but replacing the 0-formylmandelic acid chloride by LO-formylmandelic acid chloride, the title compound is obtained, containing 1 mole of water, melting point 203- 205 [deg.] C. After careful drying, the product melts at 228-230 [deg.] C.

Example 10

  
  <EMI ID = 231.1>

  
see Example 6A) in 100 ml of dimethylformamide and 2 ml of prepylene 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

  
  <EMI ID = 232.1>

  
potassium 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 resin "HP-20" using as eluent a 9: 1 mixture of water and acetone, which gives 0.9 g of the compound

  
  <EMI ID = 233.1>

  
Stir for 24 hours at room temperature

  
he <EMI ID = 234.1>

  
  <EMI ID = 235.1>

  
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, giving 500 mg of product, melting point 210-215 [deg.] C, with decomposition.

Example 12

  
  <EMI ID = 236.1>

  
The tetrabutylammonium salt of (S) -3-amino-2-oxo1-azetidinesulfonic acid (1.5 g; see Example 6A) is stirred at room temperature for 24 hours.

  
100 ml of dimethylformamide, 0.6 g of hydroxybenzotriazole,

  
  <EMI ID = 237.1>

  
solvent by distillation and the residue is dissolved in 30 ml of acetone. The urea is separated by filtration 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 filtered off and dried. The purification is carried out by column chromatography on "HP-20" using water as eluent, which gives 1.1 g of the title compound, melting point 180-185 [deg.] C, with decomposition.

Example 13

  
  <EMI ID = 238.1>

  
fuck

  
Following the procedure of Example 12, but

  
  <EMI ID = 239.1>

  
thiazoleacetic, we get the title compound, point of

  
17 <EMI ID = 240.1>

  
sulfonic

  
Following the procedure of Example 12, but replacing (Z) -2-amino-a- (ethoxyimino) -4thiazoleacetic acid with (Z) -2-amino-a - [(2, 2,2-trifluor-

  
  <EMI ID = 241.1>

  
title, melting point 160-170 [deg.] C after cryodessication.

Example 15

  
  <EMI ID = 242.1>

  
1-azetidinesulfonic

  
  <EMI ID = 243.1>

  
  <EMI ID = 244.1>

  
  <EMI ID = 245.1>

  
(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 and is filtered, which gives 0.8 g of product, melting point.

  
  <EMI ID = 246.1>

  
Method II:

  
The tetrabutylammonium salt of the acid is hydrogenated

  
  <EMI ID = 247.1>

  
sulfonic (4 g; see Example 4) in 100 ml of diglyme with 1 g of palladium on carbon as catalyst. The hydrogenation is complete after 2.5 hours. We separate

  
  <EMI ID = 248.1>

  
propylene. After cooling to 0 [deg.] C, 0.5 g of propionic acid chloride is added with stirring in 10 ml of dry diglyme. 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 title compound crystallizes, it is filtered and it is recrystallized from a mixture of water and acetone, which gives 0.9 g of product,

  
  <EMI ID = 249.1>

  
About 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 is filtered off and the mother liquor is treated with an equivalent quantity 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 recrystallization from water, the product has a melting point of

  
  <EMI ID = 250.1>

  
example 17

  
Potassium salt of acid (S) -3 - [[[(cyanomethyl) thio] -

  
  <EMI ID = 251.1>

  
The tetrabutylammonium salt of the acid is dissolved in 70 ml of acetonitrile <S) -3-amino-2-oxo-1-azetidinesulfonic (1.5 g; see Example 6A) and 0.72 g of acid

  
  <EMI ID = 252.1>

  
a solution of 1.04 g of dicyclohexylcarbodiimide in acetonitrile. The mixture is stirred for about 16 hours at 0 [deg.] C and the precipitated dicyclohexylurea is filtered off, the filtrate is evaporated and the oily residue is dissolved in acetone. By adding a solu-

  
  <EMI ID = 253.1>

Example 18

  
(S) -2-oxo-3 acid potassium salt - [(1H-tetrazol-1-yl-

  
  <EMI ID = 254.1> (see Example 6A) in 70 ml of dimethylformamide, we

  
  <EMI ID = 255.1>

  
  <EMI ID = 256.1>

  
about 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 mol of a solution of potassium perfluorobutane sulfonate in acetone, which

  
  <EMI ID = 257.1>

  
with decomposition.

Example 19

  
(S) -2-oxo-3 acid potassium salt - [(2H-tetrazol-2-yl-

  
  <EMI ID = 258.1>

  
Following the procedure of Example 18, but

  
  <EMI ID = 259.1>

  
melting point 175 - 177 [deg.] C with decomposition.

Example 20

  
  <EMI ID = 260.1>

  
The tetrabutylammonium salt of (S) -3-amino-2-oxo-1-azetidinesulfonic acid (cooled as described in Example 6A) is cooled to 0 [deg.] C from 7.9 g of salt (S) -2-oxo-3 acid tetrabutylammonium - [[(phenylmethoxy) -

  
  <EMI ID = 261.1>

  
(Z) -2-amino-a- (methoxyimino) -4-thiazoleacetic acid, then a solution of 3.27 g of dicyclohexylcarbodiimide in 10 ml of dimethylformamide. The mixture is stirred for

  
  <EMI ID = 262.1>

  
empty. The residue is dissolved in acetone and filtered. By adding 60 ml of a 10% solution of potassium perfluorobutane sulfonate in acetone,

  
1) crystallize 4.7 g of crude product. This crude product is purified by chromatography on "HP-20", of particle size

  
  <EMI ID = 263.1>

  
of fusion 235 [deg.] C.

Example 22

  
  <EMI ID = 264.1>

  
Dissolve in 20 ml of anhydrous methanol the potassium salt of the phenylmethyl ester of the acid (3S) -a-

  
  <EMI ID = 265.1>

  
(100 mg; see Example 5). Palladium on carbon is added (10 mg, 10%), and the mixture is treated with hydrogen. 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

  
  <EMI ID = 266.1>

  
gross product. 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 [deg.] C.

Example 23

  
  <EMI ID = 267.1>

  
azetidinesulfonic

  
  <EMI ID = 268.1> A) Potassium salt of (S) -3-amino-2-oxo-1azetidinesulfonic acid

  
Dissolve the potassium salt of acid (S) -3-

  
  <EMI ID = 269.1>

  
see Example 3) in 4.0 ml of water, and hydrogenated on
37 mg of 10% palladium on charcoal for 1 hour 40 minutes. The catalyst is filtered off and washed with 1 ml of 50% aqueous acetone.

  
  <EMI ID = 270.1> azetidinesulfonic

  
The above solution of the free amine is diluted with 3.5 ml of acetone and stirred in an ice bath. We

  
  <EMI ID = 271.1>

  
in small fractions, alternating the addition of solid potassium bicarbonate to maintain the pH between <EMI ID = 272.1>

  
thin layer on silica gel in a mixture (19: 1). of acetone and acetic acid and the Rydon test indicate that the reaction is almost complete. 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

  
  <EMI ID = 273.1>

  
This solid is digested with methanol to obtain

  
282 mg of an extractable substance which contains, still a little salt. The product is subjected to a further purification by passage through an "IRC-50" column, then by acidification to pH 3.8 with subsequent concentration to dryness under vacuum. Trituration with acetone gives 64 mg of the desired product containing about 0.5 equivalent of mineral potassium salts. The final passage in a column of
200 ml 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 which is dried under vacuum for 2 hours at <EMI ID = 274.1>
100 [deg.] C.

  

  <EMI ID = 275.1>


  
Method II:

  
A solution of 2.0 g of pyridine salt is hydrogenated

  
  <EMI ID = 276.1>

  
  <EMI ID = 277.1>

  
on 500 mg of 10% palladium on carbon. At the end of

  
2 hours, the solution is filtered, cooled to 0 [deg.] 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 bicarbonate of

  
  <EMI ID = 278.1>

  
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 fractions of

  
not <EMI ID = 279.1>

  
13 and 14, contaminated with a little potassium acetate. A new chromatography on "HP-20 AG" gives an analytical sample, melting point 205 - 210 [deg.] C.

  

  <EMI ID = 280.1>

Example 24

  
  <EMI ID = 281.1>

  
  <EMI ID = 282.1>

  
(S) -3-amino-2-oxo-1-azetidinesulfonic acid (1.5 g;

  
  <EMI ID = 283.1>

  
  <EMI ID = 284.1>

  
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 separated by filtration and dried. After recrystallization from boiling water, the title compound has a melting point of 176-180 [deg.] C.

Example 25

  
  <EMI ID = 285.1>

  
sulfonic (3 g; see Example 4) in 100 ml of dimethylformamide with 1.5 g of palladium on carbon as catalyst. After half an hour, the catalyst is filtered off and 1.8 g of dicyclohexylcarbodiimide are added,

  
  <EMI ID = 286.1>

  
hydroxybenzotriale. After 3 hours, the reaction mixture is concentrated to dryness, dissolved in 50 ml of dry acetone and the precipitated urea is filtered off. An equivalent amount of potassium perfluorobutane sulfonate in 20 ml of acetone is added, which precipitates the title compound. 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 [deg.] C, with decomposition.

Example 26

  
  <EMI ID = 287.1>

  
sulfonic (3 g; see Example 4) in 100 ml of dry dimethylformamide with 1.5 g of palladium on carbon as catalyst; the catalyst is separated by filtration after 30 minutes. 1.8 g of (R) -a - [[(2-oxo-1imidazolidinyl) carbonyl] amino] benzeneacetic 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 separated by filtration after adding 200 ml of ether, which gives 1.8 g of product, melting point 210 -215 [deg.] C after recrystallization from a mixture of water and acetone.

Example 27

  
  <EMI ID = 288.1> .azetidinone

  
3.58 g of (Z) -a- (methoxyimino) acid are dissolved -

  
  <EMI ID = 289.1>

  
at 5 [deg.] C and treated with a solution of 4.53 g of dicylohexylcarbodiimide in 50 ml of methylene chloride. We

  
  <EMI ID = 290.1>

  
solution of 1.72 g of 3-amino-2-azetidinone (see Example 5A) in 100 ml of methylene chloride. 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,

  
  <EMI ID = 291.1>

  
substance by chromatography on 750 g of silica gel, using a mixture (7: 3) of methylene chloride and ethyl acetate as eluent, which gives 2.9 of product.

  
  <EMI ID = 292.1>

  
0.5 ml of pyridine is dissolved in 5 ml of absolute methylene chloride, cooled to -30 [deg.] 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 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 salt.

  
  <EMI ID = 293.1>

  
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.

  
Treatment of tetrabutylammonium salt with an equimolar amount of potassium perfluorobutane sulfonate in acetone, evaporation, and treatment

  
  <EMI ID = 294.1>

  
title, which is purified by chromatography on "HP-20". The elution is carried out with a 90:10 mixture of water and acetone, which gives a product whose melting point is 220 [deg.] C, with decomposition.

Example 28

  
  <EMI ID = 295.1>

  
with 0.7 g of hydroxybenzotriazole and 1.13 g of dicyclohexylcarbodiimide. 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 10% solution of potassium perfluorobutane sulfonate in acetone. After addition of petroleum ether, the title compound precipitates and it is treated with ether and then filtered to obtain 3.8 g of product, melting point 190 [deg.] C, with decomposition.

Example 29

  
  <EMI ID = 296.1>

  
The potassium salt of [3S (Z)] - 3 - [- [[(2-amino-4-thiazolyl)] [[2-

  
  <EMI ID = 297.1>

  
and 25 ml of trifluoroacetic acid are added at -10 [deg.] C. The mixture is stirred for 10 minutes at -10 [deg.] C. 100 ml of ether is added slowly at -10 [deg.] C, then 50 ml of petroleum ether. The precipitate is filtered to obtain 1.6 g of the trifluoroacetic acid salt. We put this salt in suspension in

  
  <EMI ID = 298.1>

  
diluted and purified on a column of "HP-20". The title compound is eluted with water, and it has a melting point of 225 [deg.] C, with decomposition.

Example 30

  
  <EMI ID = 299.1>

  
ethoxy] imino] -4-thiazoleacetic, the title compound is obtained, melting point 180 [deg.] C, with decomposition.

Example 31

  
  <EMI ID = 300.1>

  
(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. We add

  
  <EMI ID = 301.1>

  
acetyl dissolved in 10 ml of acetone, while maintaining the temperature between 0 and 5 [deg.] C and the pH at 6.8 with sodium bicarbonate. 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 fractions 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,

  
  <EMI ID = 302.1>

  
trifluoracetamide in 20 ml of acetonitrile. 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-ethylhexanoate in n-butanol is added and stirring is continued for 45 minutes. We pour the solution into
300 ml of ether and the precipitate is separated by filtraLion. A filtered solution of 1.2 g of precipitate is subjected to

  
in a phosphate buffer (pH 5.5) using chromatography

  
on 100 ml of "HP-20". The elution is carried out with: 1) 20 ml 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 test on silica). 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 [deg.] C, with decomposition.

  
Method II:

  
The tetrabutylammonium salt of (S) -3-amino-2-oxo-1-azetidine-sulfonic acid (2.03 g; see Example 6A) and 0.9 g is dissolved in 30 ml of acetonitrile acid

  
  <EMI ID = 303.1>

  
1.03 g of dicyclohexylcarbodiimide in 10 ml of acetonitrile. The temperature is maintained at 0 [deg.] 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:

  
  <EMI ID = 304.1>

  
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 oily residue is treated with an equivalent amount of potassium perfluorobutane sulfonate in acetone. After addition of ether, the title compound crystallizes and is filtered, melting point 148 - 149 [deg.] C, with decomposition.

Example 32

  
  <EMI ID = 305.1>

  
azetidinesulfonic

  
To a solution of 2.03 g of tetrabutyl salt

  
  <EMI ID = 306.1>

  
of acetonitrile, a solution of 1.03 g of dicyclohexylcarbodiimide in 10 ml of acetonitrile is added dropwise. The temperature is maintained at 5 [deg.] C for one hour and at room temperature for 6 hours. The

  
  <EMI ID = 307.1>

  
solvent allow 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, point of

  
  <EMI ID = 308.1>

  
Examples 33 to 37

  
Following the procedure of Example 32, but

  
  <EMI ID = 309.1>

  
carbonyl] amino] benzeneacetic by the compound indicated in column I below, the compound indicated in column II below is obtained.

  

  <EMI ID = 310.1>
 

Example 38

  
  <EMI ID = 311.1>

  
of tetrabutylammonium salt of 3-amino-2-oxo-lazetidinesulfonic acid (see Example 6A) in 30 ml of acetonitrile, per 0.8 g of dicylcohexylcarbodiimide dissolved in
10 ml of acetonitrile at -5 [deg.] C. After shaking for

  
About 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.

  
  <EMI ID = 312.1>

  
  <EMI ID = 313.1>

  
see Example 38), and treat the solution with an equivalent amount of potassium perfluorobutane sulfonate. The addition of ether crystallizes the title compound, which is separated by filtration, which gives 0.18 g of product, melting point 157 [deg.] C, with decomposition.

Example 40

  
  <EMI ID = 314.1>

  
2-bxo-1-azetidinesulfonic

  
3.25 g of acid are dissolved in 25 ml of acetonitrile <EMI ID = 315.1> 2-thiopheneacetic, 4.20 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 [deg.] C takes 10 minutes and the stirring is continued for approximately 16 hours. The formed waste is filtered off, 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 crystallizes, melting point 185-187 [deg.] C, with decomposition.

  
  <EMI ID = 316.1>

  
  <EMI ID = 317.1>

  
Add 3 drops 1.4 g of abromophenylacetyl chloride in 10 ml of acetonitrile to a solution

  
  <EMI ID = 318.1>

  
oxo-1-azetidinesulfonique, and 3 g of propylene oxide

  
  <EMI ID = 319.1>

  
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 [deg.] C, with decomposition.

Example 42

  
  <EMI ID = 320.1>

  
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, the following is added

  
  <EMI ID = 321.1>

  
oxazolone. After stirring for three hours at 0 [deg.] 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 on "HP-20", using water as the eluent, gives the title compound, melting point 218-222 [deg.] C, with decomposition.

  
Method II:

  
To a suspension of 2 g of acid (&#65533;) - a - [(amino-

  
  <EMI ID = 322.1>

  
1 <EMI ID = 323.1>

  
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. The addition of the 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, melting point
220-223 [deg.] C.

  
Example 43

  
  <EMI ID = 324.1>

  
sulfonic

  
0.54 g of acid is dissolved in 20 ml of acetonitrile

  
  <EMI ID = 325.1>

  
add 0.5 g of dicyclohexylcarbodiimide at a temperature

  
  <EMI ID = 326.1>

  
filtration the dicyclohexylurea and, after having removed the solvent by distillation, the oily residue is dissolved in
50 ml of acetone and the equivalent amount of potassium perfluorobutane sulfonate is added. The crystalline product is separated by filtration and a purification is carried out on "HP-20" using water as the eluent, period

  
  <EMI ID = 327.1>

  
Following the procedure of Example 42, method II, but replacing the acid (&#65533;) - a - [(amino-

  
  <EMI ID = 328.1>

  
composed of the title, melting point 218-222 [deg.] C.

Example 45

  
  <EMI ID = 329.1>

  
oxo-1-azetidinesulfonic

  
Following the procedure of Example 40, but

  
  <EMI ID = 330.1>

  
benzeneacetic, the title compound is obtained, melting point 155-157 [deg.] C, with decomposition.

Example 46

  
  <EMI ID = 331.1>

  
oxo-1-azetidinesulfonic

  
  <EMI ID = 332.1> A) Mixed sodium and potassium salt of acid 3 - [(N- <EMI ID = 333.1>

  
  <EMI ID = 334.1>

  
sodium borate decahydrate cooled between -15 [deg.] and -10 [deg.] c,

  
  <EMI ID = 335.1>

  
the mixture cold for one hour and 45 minutes, it is poured into 50 ml of a solution of potassium phosphate monobasic 0.5 M, and the pH is lowered to 5.5. The solvent is removed in vacuo and the residue is dissolved in a minimum amount of water, then subjected to chromatography.

  
  <EMI ID = 336.1>

  
Elution with water gives an oil (63 mg) which crystallizes on standing. Trituration with a mixture of methanol and ether; then with ether alone gives a

  
  <EMI ID = 337.1> 2-oxo-1-azetidinesulfonic

  
It is dissolved in 1.5 ml of dry dimethylformamide

  
  <EMI ID = 338.1>

  
solution of 50 mg of lithium methylate in 1 ml of methanol at -78 [deg.] C. After stirring for 15 minutes at
-78 [deg.] C, 10 ml of a 0.5 M solution of monobasic potassium phosphate is added, then the solution is acidified to pH 4 with IN hydrochloric acid. To this solution, 70 mg of tetrabutylammonium hydrogen sulfate are added 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 a

  
  <EMI ID = 339.1>

  
  <EMI ID = 340.1>

  
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 of a colorless powder (11 mg); melting point 182-183 [deg.] C, decomposition.

  
Method II:
A) Tetrabutylammonium salt of 3-amino-3-methoxy- acid <EMI ID = 341.1>

  
4% sodium borate solution is added

  
  <EMI ID = 342.1>

  
30 mg of 10% palladium on carbon in 2 ml of methanol, and the mixture is stirred under an atmosphere of hydrogen for 15 minutes. Tetrabutylammonium salt is added

  
  <EMI ID = 343.1>

  
1-azetidinesulfonic (60 mg; see Example 49) in 2 ml of methanol, and the mixture is stirred vigorously for
15 minutes under a hydrogen atmosphere. The catalyst is separated by filtration on "Celite" on a filter

  
  <EMI ID = 344.1>

  
under vacuum, then the residue is extracted with methylene chloride. Removal of the solvent under reduced pressure gives 35.mg of the title compound, in the form of an oil.

  
  <EMI ID = 345.1> <EMI ID = 346.1>

  
of propylene oxide and 74 μl of acetyl chloride. After 2 hours, the solvent is removed under reduced pressure and the residual oil is subjected to chromatography on 4 g of silica gel. Elution with 6 to 8% of

  
  <EMI ID = 347.1>

  
oil which is dissolved in water and passed through an ion exchange resin (3 ml of "AG 50W-X2", form K &#65533;, 0.6 meq / ml ). Removal of water from the eluate in vacuo gives the desired product (10 mg).

Example 47

  
N- (3-methoxy-2-oxo-1-sulfo-3-) tetrabutylaminonium salt

  
  <EMI ID = 348.1> azetidinyl) -2-phenylacetamide

  
To a solution of potassium salt of (S) -N- (2-oxo1-sulfo-3-azetidinyl) -2-phenylacetamide (50 mg; see Example 1) in methanol containing 4% sodium borate decahydrate (5 ml) cooled in a bath at -5 [deg.] C, we

  
  <EMI ID = 349.1>

  
stirred for 32 minutes, pour the mixture into a

  
  <EMI ID = 350.1>

  
The pH of the resulting solution (5.9) is adjusted to 4.5, concentrated in vacuo to remove the methanol, and it is

  
  <EMI ID = 351.1>

  
(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. Concentration in vacuo gives 50 mg of the title compound.

  
B) Tetrabutylammonium salt of N- (3-methoxy-2-oxo-1- <EMI ID = 352.1>

  
  <EMI ID = 353.1>

  
lithium in 5 ml of methanol cooled to -78 [deg.] C, a solution of 149 mg of potassium salt of N-chloro-N- is added

  
  <EMI ID = 354.1>

  
dry dimethylformamide. When the addition is complete, the mixture is stirred for 15 minutes at -78 ° C., poured into 100 ml of a 0.5M monobasic potassium phosphate solution, and washed three times with methylene chloride. 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 under

  
  <EMI ID = 355.1>

  
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.

Example 48

  
  <EMI ID = 356.1>

  
2-phenylacetamide

  
The tetrabutylammonium salt is dissolved in water

  
  <EMI ID = 357.1>

  
acetamide (91 mg; see Example 47), and the solution is passed through an ion exchange column (10 ml of

  
"AG 50W-X2", form K. The eluate is concentrated under vacuum and, by scraping the walls of the container, the residual oil is solidified in a mixture of methanol, acetone and ether. After having triturated twice with ether, the product is obtained in the form of a solid (53 mg):

  
  <EMI ID = 358.1>

  
3.59 (S, 2H, CH2), 3.84 (ABq, J = 6.3 Hz, 2H, H4), 7.30 (m, 5H, aromatic).

  

  <EMI ID = 359.1>

Example 49

  
  <EMI ID = 360.1>

  
Method I:
A) Tetrabutylammonium salt of 2-oxo-3- [N-chloro- acid

  
  <EMI ID = 361.1>

  
sulfonic (0.9 g; see Example 4) dissolved in 80 ml of methylene chloride, to a mixture (cooled to 0 - 5 [deg.] C) of 3.17 g of sodium borate decahydrate and 11 , 8 ml of a 5.25% sodium hypochlorite solution in 70 ml of water. The reaction mixture is stirred vigorously for 55 minutes while cooling in an ice bath. After diluting the mixture with a 0.5 M monobasic potassium phosphate solution, the product is extracted with three 150 ml fractions of methylene chloride. Grouping the extracts, drying over sodium sulfate and removing the solvent in vacuo gives the title compound as an oil

  
(0.94 g).

  
B) Tetrabutylammonium salt of 3-methoxy-2-oxo-3- acid <EMI ID = 362.1>

  
To a stirred solution of 667 mg of methylate

  
lithium in 10 ml of anhydrous methanol at -78 [deg.] C, a solution of 0.94 g of tetrabutylammonium acid salt is added

  
  <EMI ID = 363.1>

  
After stirring the mixture at -78 ° C. for one hour, it is poured into a solution of 0.5 M monobasic potassium phosphate and extracted with three fractions of 150 ml 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. We get the desired product

  
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:

  

  <EMI ID = 364.1>


  
Method II:

  
To a solution of 400 mg of potassium acid salt

  
  <EMI ID = 365.1>

  
that in water, a 0.1 M tetrabutylammonium bisulfate solution (10.9 ml, adjusted to pH 4.3 with potassium hydroxide) is added. The mixture is extracted three times with methylene chloride, the extracts are combined, dried over sodium sulphate and the solvent is removed in vacuo to obtain 625 mg of a foam whose spectral characteristics are roughly those of the product obtained by method I.

Example 50

  
  <EMI ID = 366.1>

  
Method I:
A) Potassium salt of 2-oxo-3- [N-chloro-N- <EMI ID = 367.1>

  
(1.00 g; see Example 3) in 90 ml of methanol contained

  
  <EMI ID = 368.1>

  
  <EMI ID = 369.1>

  
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 having concentrated the solution under vacuum at 30 ml, the

  
  <EMI ID = 370.1>

  
149 p (200 ml). After passing a solution of 50 g of monobasic potassium phosphate in 1000 ml of water, then

  
  <EMI ID = 371.1>

  
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 [deg.] C.

  
B) Potassium salt of 3-methoxy-2-oxo-3 - [[(phenyl- <EMI ID = 372.1>

  
To a solution of 874 mg of lithium methylate in
10 ml of dry methanol at -78 [deg.] C, 857 mg of potassium salt of 2-oxo-3- [N-chloro-N - [(phenylmethoxy) - are added

  
  <EMI ID = 373.1>

  
the mixture in 200 ml of a 0.5 M monobasic potassium phosphate solution and the pH is adjusted to 5.5 with IN hydrochloric acid. The aqueous mixture is washed with three 100 ml fractions of methylene chloride, and 1.169 g of tetrabutylammonium bisulfate is added. The product is extracted with three 200 ml fractions of methylene chloride, dried over sodium sulfate, filtered and concentrated in vacuo. The residual oil is subjected to chromatography on 150 g of silica gel and the product is eluted with 2 to 4% methanol in methylene chloride, which gives the tetrabutylammonium salt of the product in the form of a oil (701 mg).

  
Part of this oil (51 mg) is dissolved in water and this solution is passed through an exchange column.

  
  <EMI ID = 374.1>

  
ml). The concentration of the eluate under vacuum gives 30 mg of an oil which is made to crystallize by scraping the areas of the container with acetone.

  
  <EMI ID = 375.1>

  
than ; melting point 196-198 [deg.] C.

  
Method II: <EMI ID = 376.1> 2-azetidinone

  
  <EMI ID = 377.1>

  
  <EMI ID = 378.1>

  
Example 2C) in 40 ml of 4% methanolic borax, and 0.5 ml of t-butyl hypochlorite is added. At the end of

  
  <EMI ID = 379.1>

  
cold and extracted with two 100 ml portions 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.

  
  <EMI ID = 380.1> azetidinone

  
Cooled to -78 [deg.] C a solution of 730 mg (0.0025

  
  <EMI ID = 381.1>

  
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 [deg.] C, allowed to warm to room temperature and stirred for 30 minutes. The resulting solution is diluted with ethyl acetate, washed with bicarbonate of

  
  <EMI ID = 382.1>

  
water, saturated saline, and dry it.

  
  <EMI ID = 383.1>

  
on four 20 x 20 cm x 1 mm silica gel plates.

  
Development with a (1: 1) mixture of benzene and ethyl acetate and isolation of the main active band in the ultraviolet, of Rf = 0.25, gives 91 mg of an oil as it is crystallized from ether to obtain a solid. Recrystallization from ether gives the title compound, melting point 112-114 [deg.] C.

  
  <EMI ID = 384.1>

  
in 0.175 ml of dichloromethane and 0.175 ml of dimethylformamide, with 55.4 mg of a complex of pyridine and sulfur trioxide. The resulting suspension is diluted with 5 ml of cold 0.5 M potassium phosphate mnobasic
(adjusted to pH 4.5) and it is 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

  
  <EMI ID = 385.1>

  
difier slowly. Crystallization from acetone gives the title compound, melting point 196 -198 [deg.] C, with decomposition.

Example 51

  
  <EMI ID = 386.1>

  
crude sulfonic azetidine (431 mg, see Example 74),

  
  <EMI ID = 387.1>

  
the solution is carefully stirred at -10 [deg.] C under dry nitrogen. 568 mg of a- (benzyloxycarbonyl) phenylacetyl chloride in 3 ml of dry acetonitrile are added dropwise. Thin layer chromatography indicates that the reaction is complete 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 with equal volumes of methylene chloride. The extract is dried over anhydrous sodium sulfate and concentrated in vacuo to give 1.032 g of crude product, which is dissolved in 3 ml of methylene chloride and which is subjected to chromatography on a silica column.

  
using as eluant a mixture of methylene chloride and methanol, to obtain 470 mg of the title compound.

  
  <EMI ID = 388.1> sulfonic

  
Dissolve in 15 ml of a mixture of 30% acetone and water 470 mg of tetrabutylammonium salt of acid 3 - [[1,3dioxo-2-phenyl-3- (phenylmethoxy) propyl] amino] - 3-methoxy-2oxo-1-azetidinesulfonic, and the solution is passed through a column of "Dowex SOWX2" (form K +), using

  
  <EMI ID = 389.1>

  
eluate in vacuo to obtain 345 mg of an amorphous solid which is lyophilized into an amorphous powder, melting point 100 - 120 [deg.] C.

  

  <EMI ID = 390.1>

Example 52

  
  <EMI ID = 391.1>

  
Example 74) in 50 ml of dry acetonitrile at -20 [deg.] C, we

  
  <EMI ID = 392.1>

  
cyanomethylthioacetyl. After 10 minutes, the solvent is removed under reduced pressure and a 0.1 M solution ( <2> <2>, 8 ml) of tetrabutylammonium sulfate adjusted to pH 4.3 with potassium hydroxide, then the product is extracted

  
  <EMI ID = 393.1>

  
&#65533;

  
dry, filter and concentrate 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 is passed through an ion exchange resin (16 ml of

  
"AG 50W-X2" (form K +), 0.6 mequiv. / Ml, 37 - 74 p. Elimination of water gives 164 mg of a product partially

  
  <EMI ID = 394.1>

  
tion on 100 ml of "Diaion AG HP 20" resin 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,

  
  <EMI ID = 395.1>

  

  <EMI ID = 396.1>


  
Examples 53 to 56

  
Following the procedure of Example 42, Method II, but replacing the acid (&#65533;) -a- [(amino-

  
  <EMI ID = 397.1>

  
  <EMI ID = 398.1>

  
in Column II below.

  

  <EMI ID = 399.1>
 

Example 57

  
  <EMI ID = 400.1>

  
obtains the title compound, melting point 173 [deg.]. C, with decomposition.

Example 58

  
  <EMI ID = 401.1>

  
(0.8 g; see Example 6A), While stirring, 0.4 g of dicylohexylcarbodiimide is added dropwise at 0 [deg.] 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 treated with potassium perfluorobutane sulfonate to precipitate the title compound. Purification by column chromatography of "HP-20" using

  
  <EMI ID = 402.1>

  
fusion 193-194 [deg.] C.

Example 59

  
  <EMI ID = 403.1>

  
azetidinesulfonic

  
1.3 g of acid are dissolved in 50 ml of acetonitrile

  
  <EMI ID = 404.1>

  
  <EMI ID = 405.1>

  
hexylcarbodiimide dissolved in 5 ml of acetonitrile. After having stirred for 15 hours and separating the dicyclohexylurea by filtration, the product is removed. solvent by distillation. The remaining oily residue is dissolved in acetone and

  
1 treats it with the equivalent amount of potassium perfluorobutane sulfonate. We isolate the title compound and

  
it is purified by chromatography on a column of "HP-20"

  
using water as the eluent, which gives the compound of

  
  <EMI ID = 406.1>

  
  <EMI ID = 407.1>

  
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, which gives 1.4 g of product, melting point 217 - 222 [deg.] C.

Example 61

  
  <EMI ID = 408.1>

  
Stir together at room temperature for
12 hours the tetrabutylammonium salt of (S) -3amino-2-oxo-1-azetidinesulfonic acid (2.25 g; see Example 6A) in 100 ml of dry dimethylformamide (DMF), an equivalent amount of dicyclohexylcarbodiimide , 2.5 g of acid (R) -

  
  <EMI ID = 409.1>

  
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 perfluorobutane sulfonate in 20 ml of acetone. After adding 100 ml of ether, the title compound precipitates and is separated by filtration. The purification is carried out by dissolving the compound in a mixture of DMF and acetone and by causing it to precipitate with water, which gives 1.5 g of product, melting point 224-226 [deg.] C, with decomposition.

  
  <EMI ID = 410.1>

  
  <EMI ID = 411.1>

  
The tetrabutylammonium salt of (S) -3-amino-2-oxo-1-azetidinesulfonic acid (2.25 g;

  
see Example 6A) in 60 ml of dimethylformamide, with

  
  <EMI ID = 412.1>

  
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 [deg.] C , with decomposition.

Example 63

  
  <EMI ID = 413.1>

  
6A) in 100 ml of dry dimethylformamide, for about 16 hours with 1.5 g of dicyclohexylcarbodiimide, 0.5 g

  
  <EMI ID = 414.1> acetic. 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

  
  <EMI ID = 415.1>

  
tion.

Example 64

  
  <EMI ID = 416.1>

  
Following the procedure of Example 63, but replacing the (R) -a-hydroxybenzene acetic acid with the acid
(S) -a-hydroxybenzeneacetic, the title compound is obtained, melting point 195-197 [deg.] C.

Example 65

  
  <EMI ID = 417.1>

  
The tetrabutylammonium salt of (S) -3-amino-2-oxo-1-azetidinesulfonic acid (2.25 g) is cooled to 0 ° C.
see, Example 6A) in 100 ml of dry diglyme, 2.4 g of triethylamine and 0.3 g of dimethylaminopyridine. We add

  
  <EMI ID = 418.1>

  
methanesulfonic acid in 20 ml of diglyme. The temperature is maintained for 2 hours, the solvent is distilled off under vacuum 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; melting point 240-245 [deg.] C with decomposition, after recrystallization from a mixture of water and acetone.

Example 66

  
  <EMI ID = 419.1>

  
thiazoleacetic, 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 separated by filtration, which gives 2.77 g of crude product. Purification of this crude product by chromatography on a column of "HP-20" using as eluent a mixture (9: 1) of water and acetone, gives the title compound, melting point
155 - 160 [deg.] C, with decomposition.

Example 67

  
  <EMI ID = 420.1>

  
sulfonic (2.25 g; see Example 6A) in 60 ml of dimethylformamide, with 2.4 g of (Z) -2-amino-a - [[2-

  
  <EMI ID = 421.1>

  
solvent 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. 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 "HP-20" column using as eluent a mixture (7: 3) of water and acetone, <EMI ID = 422.1>

  
decomposition.

  
  <EMI ID = 423.1>

  
  <EMI ID = 424.1>

  
oxo-1-azetidinesulfonic

  
The tetrabutylammonium salt of the acid is treated
(S) -3-amino-2-oxo-1-azetidinesulfonique (1.9 g; see

  
  <EMI ID = 425.1>

  
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 as eluent, and 1.05 g of product is obtained, melting point 250 [deg.] C, with decomposition.

Example 69

  
  <EMI ID = 426.1>

  
azetidinesulfonic

  
The tetrabutylammonium salt of the acid is stirred

  
  <EMI ID = 427.1>

  
methoxy) imino] -4-thiazoleacétique, 0.57 g of hydroxybenzotriaole 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.

  
"HP 20" using as eluent a mixture (9: 1) of water and acetone, which gives 1 g of substance, melting point <EMI ID = 428.1>

  
azetidinesulfonic

  
The potassium salt is mixed with 5 ml of anisole

  
  <EMI ID = 429.1>

  
sulfonic (1: 1) (1.3 g; see Example 30). At -15 ° C., 25 ml of trifluoroacetic acid are added and the mixture is stirred for 10 minutes. 100 ml of ether are added slowly at -10 [deg.] 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, which gives 3.0 g of the title compound, melting point
230-235 [deg.] C, with decomposition.

Example 71

  
Potassium salt of [3S (Z)] - 3 - [[(2-amino-4-

  
  <EMI ID = 430.1>

  
Following the procedure of Example 28, but replacing the acid (Z) -2-amino-a - [[2- (diphenylmethoxy) -

  
  <EMI ID = 431.1>

  
azetidinesulfonic

  
  <EMI ID = 432.1>

  
  <EMI ID = 433.1>

  
thiazoleacetic, the title compound is obtained, melting point 2 05-210 [deg.] C, with decomposition.

Example 73

  
  <EMI ID = 434.1>

  
sulfonic

  
A solution of 0.6 g of hydroxybenzotriazole is stirred for one hour with 10 g of a 4A molecular sieve. 90% in 100 ml of dimethylformamide, filtered, and the filtrate is added to a solution of 0.004 mole of tetrabutylammonium salt of (S) -3-amino-2-oxo-1azetidinesulfonic acid (see Example 6A) in dimethylformamide. 0.89 g of (Z) -2-amino-a- acid is added

  
  <EMI ID = 435.1>

  
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 [deg.] C.

Example 74

  
  <EMI ID = 436.1> A) Tetrabutylammonium salt of 3-amino-3-methoxy2-oxo-1-azetidinesul [pound] onic acid

  
Dissolve in 15 ml of dry methanol the salt of

  
  <EMI ID = 437.1>

  
This mixture is hydrogenated under a pressure of one atmosphere for 15 minutes. The catalyst is separated by filtration

  
  <EMI ID = 438.1>

  
114 mg of the title compound.

  
B) Tetrabutylammonium salt of 3-methoxy-2-oxo-3- acid <EMI ID = 439.1>

  
  <EMI ID = 440.1>

  
dry and stir the solution carefully at -10 [deg.] C under <EMI ID = 441.1>

  
  <EMI ID = 442.1>

  
is completed after 15 minutes, as shown by thin layer chromatography. Add a potassium phosphate buffer (0.5 M; pH 5.5; 4.2 ml) and 8.5 mg of tetrabutylammonium sulphate (0.1 equiv.), Then remove most of the vacuum. 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 obtain 107 mg of a gum. The crude product is purified by chromatography on silica gel using as eluent a mixture of methylene chloride and methanol, which gives 66 mg of the title compound.

  
  <EMI ID = 443.1>

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

  

  <EMI ID = 444.1>

Example 75

  
  <EMI ID = 445.1>

  
azetidinesulfonic

  
Dissolve the potassium salt of the acid [3S (Z)] - 3-

  
  <EMI ID = 446.1>

  
imino] acetyl] amino] -2-oxo-1-azetidinesulfonic (0.1 g; see Example 71) in a mixture of 5 ml of ethanol and 5 ml of water, and the solution is hydrogenated with room temperature in the presence of 0.2 g of 10% palladium on carbon. After 2 hours, the catalyst is filtered off and the remaining solution is subjected to freeze drying, which gives the title compound, m.p.
235 [deg.] C with decomposition.

Example 76

  
  <EMI ID = 447.1>

  
that, isomer A

  
To a solution of tetrabutylammonium acid salt

  
  <EMI ID = 448.1>

  
thienyl) -5- (4H) -oxazoline. This mixture is stirred for

  
  <EMI ID = 449.1>

  
pyridine and 166 mg of oxazoline. 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 +, 37 - 74 u). Elimination of water from fractions 2 and 3 gives a mixture of diastereoisomers (248 mg).

  
This product is purified and the diastereoisomers are separated on "Diaion AG HP 20" (130 ml), eluting with water. Isomer A is eluted in fractions 23 to 28

  
(30 mg) and the isomer B in fractions 35 to 45 (30 mg)
(8 ml fractions). We join the intermediate fractions
(10 mg) to fractions from other passages and a total of 35 mg of isomer A and 50 mg of isomer B are isolated.

  
Analysis for isomer A, m.p. 158-165 [deg.] C

  

  <EMI ID = 450.1>


  
Analysis for isomer B, m.p. 160-170 [deg.] C (decomposition)

  
  <EMI ID = 451.1>

  

  <EMI ID = 452.1>
 

Example 77

  
  <EMI ID = 453.1>

  
crude (366 mg see Example 74A) and 38 mg of borax in 35 ml of dry acetonitrile at -10 [deg.] C, under nitrogen, 0.53 ml of dry pyridine is added, then, in two minutes , a solution of ao-sulfophenylacetyl chloride monoetherate (348 mg) in 8 ml of acetonitrile. After 20 minutes, the solvent is removed in vacuo and the residue is treated with 35 ml of a 0.5 M potassium phosphate buffer at pH 5.5. 383 mg of tetrabutylammonium 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.

  
This crude product is combined 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 as the eluent, then 2.4 methanol. , 6, 8 and 10% in methylene chloride. The product, formed from a mixture (approximately 6: 1) of racemic diastereoisomers in the form of the tetrabutylammonium salt, is transformed into the potassium salt by passing it through

  
  <EMI ID = 454.1>

  
20% acetone solvent in water. The product is lyophilized, giving 171 mg of the title compound, melting point 205-210 [deg.] C, with decomposition.

  

  <EMI ID = 455.1>

Example 78

  
  <EMI ID = 456.1>

  
Example 51). 3.9 mg of anhydrous potassium carbonate 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

  
  <EMI ID = 457.1>

  
vitreous (34 mg) which is lyophilized into an amorphous powder,

  
  <EMI ID = 458.1>

  

  <EMI ID = 459.1>

Example 79

  
  <EMI ID = 460.1>

  
thiazoleacetic, the title compound is obtained, melting point 130 [deg.] C, with decomposition.

Example 80

  
  <EMI ID = 461.1>

  
Cooled to -78 ° C. a solution of 185 mg of tetrabutylammonium salt of 2-oxo-3- [N-chloro-N - [(phenyl-

  
  <EMI ID = 462.1>

  
3.8 ml of 0.73 M lithium n-butoxide in n-butanol. 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

  
  <EMI ID = 463.1>

  
of the title compound.

  
To a solution of 109 mg of this tetrabutylammonium salt in acetone is added 60 mg of potassium salt of perfluorobutylsulfonic acid in acetone. The solvent is removed in vacuo and ethyl acetate is added. the product crystallizes, and is collected and dried to obtain 66 mg of the title compound, period <EMI ID = 464.1>

  
(Example 46, Method II, Part A) (0.175 mmol) and 0.0175 mmol of sodium borate in. 2 ml of dichloro-

  
  <EMI ID = 465.1>

  
  <EMI ID = 466.1>

  
carbonyl] amino] -4-thiazolylacetyl. After one hour, the mixture is diluted with dichloromethane and stabilized.

  
  <EMI ID = 467.1>

  
with saturated saline solution, dry, and evaporate in vacuo. The residue is purified on silica gel.

  
Mallinckrodt "SilicAR CC-4" (20 ci) yard get 43 ma from

  
Corresponding tetrabutylammonium salt of the title compound.

  
This tetrabutylammonium salt (43 mg) is dissolved in

  
  <EMI ID = 468.1>

  
potassium sulfonate 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 [deg.] C, with decomposition.

Example 82

  
  <EMI ID = 469.1>

  
acetyl] amino] -2-oxo-1-azetidinesulfonic

  
Following the procedure of Example 81, but

  
  <EMI ID = 470.1>

  
(Z) -a- (methoxyimino) -2 - [[(phenylmethoxy) carbonyl] amino] -4-thiazolylacetyl chloride, the title compound is obtained, melting point 168 - 172 [deg.] C, with decomposition.

Example 83

  
  <EMI ID = 471.1>

  
2-methoxy-oxo-1-azetidinesulfonic A stirred solution of 0.69 mmol of tetrabutylammonium salt of 3-amino-3-methoxy-2-oxo-1-azetidinesulfonic acid (Example 46, Method II, Part A) in 30 ml

  
  <EMI ID = 472.1>

  
dry pyridine, then a solution of 352 mg of chloride

  
  <EMI ID = 473.1>

  
phenylacetyl in 4 ml. of acetonitrile. After a

  
  <EMI ID = 474.1>

  
acid chloride 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 in vacuo to remove acetonitrile. The aqueous residue is extracted three times with methylene chloride and the combined extracts are dried over sodium sulfate and then they are evaporated, which leaves 546 mg of residue. Passing this residue through a column of silica gel using as

  
  <EMI ID = 475.1>

  
4% and 6% in methylene chloride, gives two fractions (285 mg and 173 mg) of the tetrabutylammonium salt

  
  <EMI ID = 476.1>

  
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. Successive elution with water, acetone to

  
5% in water and 10% acetone in water gives 60 mg

  
  <EMI ID = 477.1>

  
decomposition.

Example 84

  
N- (3-butoxy-2-oxo-1-sulfo-3- tetrabutylammonium salt

  
  <EMI ID = 478.1>

  
Following the procedure of Example 80, but replacing the tetrabutylammonium salt of 2oxo-3- [N-chloro-N - [(phenylmethoxy) carbonyl] amino] -1azetidinesulfonic acid with the tetrabutylammonium salt of

  
1 <EMI ID = 479.1>

  
(see Example 47A for the preparation of the corresponding potassium salt), the title compound is obtained in the form of an oil:

  
  <EMI ID = 480.1>

  
C-4 CH2), 6.98 (s, 1H, NH) and 7.30 ppm (S, 5H, C6H5).

Example 85

  
  <EMI ID = 481.1>

  
A 1M solution of dimethylformamide-sulfur trioxide complex is prepared by slowly adding trimethylsilyl chlorosulfonate to dimethylforinainide

  
  <EMI ID = 482.1>

  
0.2 ml of anhydrous dimethylformamide and cooled to 0 [deg.] C.

  
A cold solution of dimethyl complex is added.

  
1M sulfur formamide trioxide (0.428 ml), the mixture is stirred for 2 hours and poured into 15 ml of 0.5M monobasic potassium phosphate. The solution is extracted twice with dichloromethane (discarded) and added 73 mg tetrabutylammonium bisulfate. The extrac-. tion with dichloromethane (three 10 ml fractions) gives a viscous oil, after drying and evaporation under vacuum. Mallinckrodt "CC-4" silica gel chromatography
(50: 1) using 2% methanol in dichloromethane as eluent, gives 34 mg of tetrabutylammonium salt

  
  <EMI ID = 483.1>

  
azetidinesulfonic. An ion exchange on. "Dowex 50W-X2"

  
  <EMI ID = 484.1>

  
In <EMI ID = 485.1>

  
fusion 233 - 236 [deg.] C, with decomposition.

Example 87

  
  <EMI ID = 486.1> 2-methoxy-oxoazetidine

  
To a 650 mg solution of (6R-cis) -7-acetyl- acid

  
  <EMI ID = 487.1>

  
oct-2-ene-2-carboxylic acid and 191 mg of sodium bicarbonate in water, 11 ml of a commercially available Raney nickel suspension (0.6 g / ml) are added, neutralized by washing with water. The mixture is lowered into an oil bath preheated to 170 [deg.] C and refluxed in 2 to 3 minutes, while maintaining the bath temperature at 150-170 [deg.] C. After 15 minutes of reflux, we

  
stabilizes the reaction by cooling in an ice bath. The catalyst is separated by filtration on "Celite" and the pH of the filtrate (11) is adjusted to pH 2 with 1N hydrochloric acid. After extracting the aqueous solution

  
  <EMI ID = 488.1>

  
of sodium the combined extracts and the solvent is removed in vacuo to obtain 487 mg of an oil. Chromatography on silica gel gives the product (eluted in chloroform) in the form of an oil (381 mg).

  
  <EMI ID = 489.1> 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. 359 mg of copper acetate are added, the mixture is stirred for one minute to dissolve the salt, and 797 mg of lead tetraacetate are added. While the argon continues to dabble in the mixture, the reaction temperature is raised by lowering the flask into an oil bath, preheated and maintained at 55 - 65 [deg.] C for

  
15 minutes. The mixture is allowed to cool to room temperature, filtered through "Celite" and washed

  
  <EMI ID = 490.1>

  
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 obtain the desired product in the form of an oil (382 mg).

  
  <EMI ID = 491.1>

  
The preceding oil is dissolved in a mixture of

  
10 ml of methanol and 1 ml of water, the mixture is cooled in an ice-methanol bath at -10 to -15 * 0, and added
194 mg of potassium carbonate, then 53 mg of sodium borohydride. After stirring at a temperature of -15 [deg.] To
- 8 [deg.] C for 110 minutes, the solvent is removed under vacuum, the residue is taken up in water and the pH of the solution is adjusted to 6 with 1N hydrochloric acid. An exhaustive extraction at 1 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 [deg.] C (sintering

  
  <EMI ID = 492.1> 2-oxo-1-azetidinesulfonic

  
Under an argon atmosphere, we place in a balloon and

  
  <EMI ID = 493.1>

  
oxo-1-azetidine. Then add a 1 M solution of

  
  <EMI ID = 494.1>

  
of dimethylformamide, and the solution is stirred for 15 minutes. Then pour the contents of the flask into 40 ml

  
  <EMI ID = 495.1>

  
with 10 ml of methylene chloride. 1.2 equivalents of tetrabutylammonium sulphate are 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. We transform the

  
111 <EMI ID = 496.1>

  
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

  
  <EMI ID = 497.1>

  
The tetrabutylammonium salt of 3-amino-3-methoxy-2-oxo-lazetidinesulfonic acid (202 mg; 202 mg; see Example 74A) is dissolved in 20 ml of dry acetonitrile. To the well-stirred solution, at -20 [deg.] C, under dry nitrogen, the following is added

  
  <EMI ID = 498.1>

  
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% of methanol, which gives 231 mg of the composed of the title.

  
B) Potassium salt of acid (&#65533;) - 3 - [(azidophênylacétyl) - <EMI ID = 499.1>

  
azetidinesulfonique in 15 ml of 30% aqueous acetone, in a column of "Dowex 50W-X2" (form K; 3 ml), and elution is carried out with water. The total eluate is evaporated in vacuo to obtain a colorless glass (168 mg) in the form of a

  
  <EMI ID = 500.1>

  
  <EMI ID = 501.1> and 10% aqueous acetone as eluent gives 71 mg

  
  <EMI ID = 502.1>

  
a 1: 3 mixture of racemic diastereoisomers. The mixture is lyophilized 1: 1 and dried under vacuum at 40 [deg.] C to obtain the dry product in the form of a hemihydrate,

  
  <EMI ID = 503.1>

  

  <EMI ID = 504.1>


  
  <EMI ID = 505.1>

  
  <EMI ID = 506.1>

  
mics obtained in Example 88B, and the A isomer crystallizes. After refrigeration, the mother liquor is separated and the crystals are dried (28 mg) under vacuum at 40 [deg.] C, melting point 130 [deg.] C, with decomposition, in the form of a monodeuterate.

Example 90

  
  <EMI ID = 507.1>

  
2-methoxy-oxo-1-azetidinesulfonic

  
To a stirred solution of tetrabutylammonium salt

  
  <EMI ID = 508.1>

  
4 ml of acetonitrile. After 1 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 0.5 M monobasic potassium phosphate buffer at pH 5.5, and concentrated in vacuo to remove 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 obtain

  
546 mg of a residue. The passage of this substance in a 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, '1 under the

  
  <EMI ID = 509.1>

  
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: 1 approximately) of diastereoisomers and 21 mg of product as

  
  <EMI ID = 510.1>

  
sation of the 60 mg fraction gives the title compound, melting point 171-172 [deg.] C with decomposition.

  

  <EMI ID = 511.1>


  
Lyophilization of the 21 mg fraction gives the title compound, melting point 171-172 [deg.] C, with decomposition.

  

  <EMI ID = 512.1>


  
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 of 119 mg, of

  
potassium obtained by treatment with "Dowex" resin. The

  
passage of this substance through a resin "HP 20-AG", as already described, provides an additional 31 mg of product in the form of a mixture (approximately 1: 1) of diastereoisomers and an additional 42 mg of product under the form

  
a mixture (about 9: 1) of diastereoisomers. The total quantity of the diastereoisomeric mixture (1: 1) is i

  
  <EMI ID = 513.1>

  
isomers is 63 mg.

Example 91

  
  <EMI ID = 514.1>

  
2 - [[(phenylmethoxy) carbonyl] amino] -4-thiazoleacétyle. 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.

  
  <EMI ID = 515.1>

  
dissolution in a mixture of acetone and water, in an ion exchange resin (8 ml, "AG 50W-X2", form K +, 74 to

  
  <EMI ID = 516.1>

  
fractions 1 and 2 give 40 mg of the title compound, melting point 172 -174 [deg.] C, with decomposition.

  

  <EMI ID = 517.1>

Example 92

  
  <EMI ID = 518.1>

  
azetidinesulfonic (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 of sodium hypochlorite and 20 ml of water, at 0 [deg.] 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. 4. The organic extracts are dried over sodium sulfate, filtered and concentrated in vacuo to obtain 344 mg of the title compound.

  
B) Tetrabutylammonium acid salt (&) - 3-butoxy-2-

  
  <EMI ID = 519.1>

  
A solution of 344 mg of tetrabutylammonium salt of 3 - [[chloro (phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid in 5 ml of dimethylformamide is added to 0.73 M lithium n-butoxide in 6 ml of nbutanol, in 1 ml of dimethylformamide at -78 [deg.] C under an inert atmosphere. 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 "

  
  <EMI ID = 520.1>

  
methanol in methylene chloride.

  
C) Potassium salt of the acid (#) - 3-butoxy-2-oxo-3- <EMI ID = 521.1>

  
azetidinesulfonic in a mixture (9: 1) of water and acetone and the solution is placed in a column of exchange of

  
  <EMI ID = 522.1>

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

  

  <EMI ID = 523.1>

Example 93

  
  <EMI ID = 524.1>

  
The tetrabutylammonium salt of the acid is added <EMI ID = 525.1>

  
of ethanol, at -78 [deg.] C under an inert atmosphere. After 10 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 salt is eluted

  
  <EMI ID = 526.1>

  
2% in methylene chloride.

  
The tetrabutylammonium salt is dissolved in a mixture (9: 1) of water and acetone and it is passed through a cation exchange column ("Dowex AGMP 50W-X2", 74

  
  <EMI ID = 527.1>

  

  <EMI ID = 528.1>

Example 94

  
  <EMI ID = 529.1>

  
azetidinesulfonic

  
The tetrabutylammonium salt of 3-amino-3-methoxy-2-oxo-1-azetidinesulfonic acid (see Example 46, Method II, Part A) is dissolved in 20 ml of acetonitrile and 1 ml of pyridine in suspension

  
  <EMI ID = 530.1>

  
amino-4-thiazoleacétyle in 20 ml of acetonitrile cooled to 0 - 5 [deg.] C. After having been stirred cold for one hour, the mixture is diluted with 100 ml of a 0.5 M monobasic potassium phosphace 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. Resin chromatography

  
  <EMI ID = 531.1>

  
gives the crude product in the form of the potassium salt, after elution with water. A new purification on

  
  <EMI ID = 532.1>

  
gives 59 mg of the product in the form of a powder after trituration with a mixture of acetoninitrile and ether then twice with ether. The product is an amorphous powder which slowly melts and decomposes above

  
  <EMI ID = 533.1>

  

  <EMI ID = 534.1>

Example 95

  
  <EMI ID = 535.1>

  
ple 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 acid f3R (R *) and 3S (S *)] - 3- <EMI ID = 536.1>

  
oxo-1-azetidinesulfonic

  
271 mg of the salt are dissolved in 6.5 ml of water.

  
  <EMI ID = 537.1>

  
under vacuum to about 2 ml and subjected to chromatography in a 100 ml column of "HP 20-AG" using water as eluent. Isomer A is isolated (29 mg) after lyophilization, melting point 160 [deg.] C, with decomposition. Analysis for C13H15N407SK monohydrate:

  

  <EMI ID = 538.1>
 

Example 96

  
  <EMI ID = 539.1>

  
azetidinesulfonic

  
With the potassium salt of the acid [3R (R *) and

  
  <EMI ID = 540.1>

  
in the form of a lyophilisate, melting point 160 [deg.] C, with decomposition.

  

  <EMI ID = 541.1>

Example 97

  
  <EMI ID = 542.1>

  
(306 mg of crude substance supposed to contain 274 mg of organic substance, prepared as described in the Example

  
74) in 20 ml of dry acetonitrile at -20 [deg.] C, under nitrogen, 0.30 ml of dry pyridine (3.72 mmol) is added, 484 mg

  
  <EMI ID = 543.1>

  
ment dissolved and suspended in 10 ml of dry acetonitrile. The reaction mixture is stirred and allowed to settle.

  
  <EMI ID = 544.1>

  
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 methylene chloride then 2% and 4% methanol as eluent.

  
n in methylene chloride, to obtain 251 mg of the tetrabutylammonium salt of the title compound.

  
  <EMI ID = 545.1>

  
solution of 107 mg of potassium salt of perfluorobutanesulfonic acid in several milliliters of acetone. ethyl acetate is added and the precipitate is washed three times with ethyl acetate by centrifugation, then dried under vacuum at 40 [deg.] C under 1 mm for 2 hours to obtain 95 mg of the desired potassium salt in the form of a mixture (about 1: 2) of diastereoisomers having a melting point of 200 [deg.] C, with decomposition.

  

  <EMI ID = 546.1>

Example 98

  
Acid potassium salt (& -cis) -4-methyl--2-oxo-3-

  
  <EMI ID = 547.1> A) N-Benzyloxy-t-boc * -allothreonine amide

  
A solution of 6.9 g of d, lt-bocallothreonine and free amine of 5.3 g of o-benzylhydroxylamine hydrochloride (approximately 0.033 mol, release of ethyl acetate and sodium bicarbonate) are treated ) in 80 ml of tetrahydrofuran with 4.82 g of N-hydroxybenzotriazole and 6.5 g of dicyclohexylcarbodiimide in 20 ml of tetrahydrofuran. After shaking for about 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 titled compound in fractions 7 to 22

  
(200 ml each).

  
  <EMI ID = 548.1> azetidinone

  
A solution of 6.8 g of the amide of Nbenzyloxy-t-boc-allothreonine in 200 ml of tetrahydrofuran is stirred for approximately 16 hours with 5.24 g of triphenylphosphine and 3.2 ml of diethyl azodicarboxylate. The solvents are evaporated in vacuo and subjected to

  
* "boc" is an abbreviation for "butoxycarbonyle".

  
there was 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 azetidfnone. A new chromatography of the mother liquors and mixed fractions gives an additional 0.6 g. The crystallization of a fraction twice in ether (-20 [deg.] C) gives the analytical sample of the title compound, melting point 140 - 142 [deg.] C.

  
  <EMI ID = 549.1> azetidinone

  
A solution of 3.2 g of cis-N-benzyloxy- is stirred

  
  <EMI ID = 550.1>

  
95% ethanol, under a hydrogen atmosphere, with 0.7 g of 10% palladium on charcoal. 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 [deg.] C.

  
  <EMI ID = 551.1>

  
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 ammonium acetate

  
4.5 M (fractions of 40, 20 and 30 ml respectively) and by
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 300 ml portions of ethyl acetate. We wash

  
  <EMI ID = 552.1>

  
5% sodium bicarbonate and 100 ml of saturated salt solution, dry and evaporate. Trituration with ether gives, in two harvests, 1.65 g of solid. Part of the first harvest is recrystallized from ether to obtain the analytical sample, melting point 176-178.5 [deg.] C.

  
  <EMI ID = 553.1>

  
/ 7 methylene chloride and 4 ml of anisole, 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 chloro-

  
  <EMI ID = 554.1>

  
pH 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 [deg.] C.

  
  <EMI ID = 555.1>

  
To a suspension of cis-3-benzyloxycarbonylamino-4methylazetidinone (0.75 g) in 7 ml of dimethylformamide
(dried with a 4A molecular sieve activated at 320 [deg.] C for
15 hours under an argon stream) and 7. ml of methylene chloride (dried over basic alumina), 1.66 g of a pyridine and sulfur trioxide complex are added. 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 obtain 4.6 g of a residue which is dissolved in 300 ml of a phosphate solution

  
  <EMI ID = 556.1>

  
The solution is cooled, passed through a column of "HP 20" resin (3 cm x 60 cm) with 400 ml of 0.5 M monobasic potassium phosphate, 1 liter of distilled water and a mixture (14: 1) water and acetone, to 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 [deg.] C with decomposition.

  

  <EMI ID = 557.1>

Example 99

  
  <EMI ID = 558.1>

  
Following the procedure of Example 98 but replacing the d, l-t-boc-allothreonine by 1-t-bocthreonine, the title compound is obtained, melting point
133 - 135 [deg.] C.

  

  <EMI ID = 559.1>

Example 100

  
Potassium salt of (3S-trans) -4-methyl-2-oxo-3- acid,

  
  <EMI ID = 560.1>

  
azetidinesulfonic (352.4 mg; see Example 99) in

  
20 ml of distilled water, and the solution is treated with 373.5 mg

  
  <EMI ID = 561.1>

  
having stirred for 10 minutes at room temperature, the solution is extracted three times with fractions of

  
10 ml 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 fractions of

  
2.5 ml of dimethylformamide to obtain the title compound in solution.

  
B) Potassium salt, (3S-trans) -4-methyl-2-oxo- acid <EMI ID = 562.1>

  
The crude tetrabutylammonium salt of (3S) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid obtained in part A (the combined filtrate and washing water) is treated at 0 [deg .] C, per 206 mg of dicyclohexylcarbodiimide,

  
  <EMI ID = 563.1>

  
  <EMI ID = 564.1>

  
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

  
  <EMI ID = 565.1>

  
aqueous solution of this substance 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, melting point 211-13? Deg.

  

  <EMI ID = 566.1>

Example 101

  
Potassium salt of (cis) -4-methyl-2-oxo-3- acid

  
  <EMI ID = 567.1>

  
ammonium, and the solution is adjusted to pH 5.5. Extraction with six 25 ml fractions of methylene chloride gives
517.3 mg of an oil. We stir a solution of this <EMI ID = 568.1>

  
10% palladium on charcoal in a hydrogen atmosphere for 90 minutes. The catalyst is filtered and the filtrate is stirred with 150 mg of phenylacetic acid,
169 mg of N-hydroxybenzotriazole and 247 mg of dicyclohexylcarbodiimide, for 7.5 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. 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. with 200 ml of water, then with a 1: 9 mixture of acetone and water, gives a substance positive for the Rydon test in fractions 6 to 10 (50 ml). The evaporator

  
  <EMI ID = 569.1>

  
recrystallization from a mixture of acetonitrile and water gives 46 mg of the title compound, which decomposes at more than 205 [deg.] C. A second harvest (6 mg) is obtained from the filtrate. An additional 5 mg is obtained from fractions 6 and 10, by evaporation and recrystallization.

  

  <EMI ID = 570.1>

Example 102

  
  <EMI ID = 571.1>

  
azetidinesulfonic A) Tetrabutylammonium salt of (3S-trans) -4- acid <EMI ID = 572.1>

  
tetrabutylammonium. 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.

  
  <EMI ID = 573.1> oxo-1-azetidinesulfonic

  
A solution of 534.6 mg of tetrabutylammonium salt of (3S-trans) -4-methyl-2-oxo-3- acid is hydrogenated.

  
  <EMI ID = 574.1>

  
20 ml of dimethylformamide, with 220 mg of 10% palladium on carbon, under atmospheric pressure, for 2 hours 45 minutes; the hydrogen absorption is 26.3 ml.

  
The mixture is filtered and washed twice with 2.5 ml

  
dimethylformamide. The filtrate and the washing water (about 25 ml total) are stirred under nitrogen with 161 mg of acid

  
  <EMI ID = 575.1>

  
N-hydroxybenzotriazole and 164.8 mg of dicyclohexylcarbodiimide. The mixture is stirred under nitrogen for approximately 16 hours. Dimethylformamide is removed in vacuo and the rubbery residue is dissolved in acetone, then filtered to remove the urea. To the filtrate, a solution containing 272 mg (0.8 mmol) of potassium salt of perfluorobutanesulfonic acid in 0.8 ml of acetone is added. The suspension is diluted with an equal volume of ether and

  
  <EMI ID = 576.1>

  
purifies 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 (50 ml fractions) 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. Likewise, the trituration of fractions 6 to 10 gives an additional 90.4 mg of product in the form of a solid.

  
  <EMI ID = 577.1>

  

  <EMI ID = 578.1>

Example 103

  
  <EMI ID = 579.1> 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-benzy] hydroxylamine hydrochloride (release of ethyl acetate and sodium bicarbonate) are treated, in 100 ml of tetrahydrofuran, with 6.12 g of N-hydroxybenzotriazole and 8.24 g of dicyclohexylcarbodiimide 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 compound. crystallization from a mixture of ether and hexane gives 4.18 g of the title compound.

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

  
A solution of 12.67 g of N-benzyloxy-t-boc-threonine amide is stirred under nitrogen for approximately 16 hours.
11.5 g of triphenylphosphine and 6.23 ml of diethyl azodicarboxylate in 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 composed of the title.

  
  <EMI ID = 580.1> azetidinone

  
A solution of 9.18 g of (3S-trans) -Nbenzyloxy-3-t-butoxycarbonylamino-4-methylazetidinone is stirred in
300 ml of 95% ethanol, in a hydrogen atmosphere,

  
  <EMI ID = 581.1>

  
141 minutes, the suspension is filtered and evaporated in vacuo. The residue is recrystallized from a mixture of ether and hexane to obtain 5.12 g of the title compound.

  
  <EMI ID = 582.1>

  
200 ml of methanol with 132 ml of 4.5 M ammonium acetate, then with 66 ml of 1.5 M titanium trichloride, and the mixture is stirred

  
  <EMI ID = 583.1>

  
with an equal volume of 8% sodium chloride and extracted with ethyl acetate to obtain 3.48 g of product

  
  <EMI ID = 584.1>

  
  <EMI ID = 585.1>

  
trans) -3-t-butoxycarbonylamino-4-methylazetidinone in
10 ml of dichloromethane and 10 ml of anisole, and added
112 ml of trifluoroacetic acid. 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 is stirred for 30 minutes at pH 7, diluted with 100 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) Tetrabutylammoniurn acid salt (3S-trans) -4- <EMI ID = 586.1>

  
azetidinesulfonic

  
Cooled to 0 [deg.] C a solution of 600 mg of (3S-

  
  <EMI ID = 587.1>

  
2 ml of dimethylformamide, 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 monobasic potassium phosphate (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.

  
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.

  
  <EMI ID = 588.1>

  
dimethylformamide, in a hydrogen atmosphere, with
800 mg of 10% palladium on charcoal, 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-

  
  <EMI ID = 589.1>

  
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 washing 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.

  
  <EMI ID = 590.1>

  
10 minutes, 10 ml of ether and 5 ml of hexane are added and the resulting suspension is stirred for 5 minutes at -12 [deg.] C, then it is allowed to warm to room temperature. The solid is isolated by centrifugation and washed twice with ether. 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 a column of
80 ml of "HP-20 AG". Elution with water gives 72 mg

  
  <EMI ID = 591.1>

  
rer three times) and trituration with ether, m.p. 250 [deg.] C approximately with decomposition.

  

  <EMI ID = 592.1>


  
The remaining 1.22 g of salt is treated as above

  
  <EMI ID = 593.1>

  
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 the acid (Z) -2-amino-a - [[[hydroxy (phenyl-

  
  <EMI ID = 594.1>

  
  <EMI ID = 595.1>

  
compound indicated in column II below.

  

  <EMI ID = 596.1>
 

  

  <EMI ID = 597.1>
 

  

  <EMI ID = 598.1>
 

  

  <EMI ID = 599.1>
 

  

  <EMI ID = 600.1>
 

  
Examples 134 and 135

  
  <EMI ID = 601.1>

  
compound indicated in column II below.

  

  <EMI ID = 602.1>
 

Example 136

  
  <EMI ID = 603.1>

  
azetidinesulfonic

  
Extracted with methylene chloride (four

  
  <EMI ID = 604.1>

  
potassium (cis) -4-methyl-2-oxo-3 - [[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic potassium and 51 mg of tetra-n-butylammonium bisulfate in 5 ml of water for

  
  <EMI ID = 605.1>

  
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 acid.

  
  <EMI ID = 606.1>

  
N-hydroxybenzotriazole and 31.5 mg of dicyclohexylcarbodiimide. 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. The

  
  <EMI ID = 607.1>

  
positive for the Rydon test in fractions 3 to 5 (20 ml) (elution with water). Evaporation and crushing with. ether give 23 mg of product in the form of a hygroscopic solid.

  

  <EMI ID = 608.1>


  
  <EMI ID = 609.1> A) t-boc-1-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 hours, then diluted with 70 ml of water and 140 ml of ethyl acetate. After having stirred thoroughly, the layers are separated and the organic layer is washed with

  
30 ml of a 2: 1 mixture of water and brine. The combined aqueüses layers are then re-extracted with 70 ml of ethyl acetate. The aqueous layer is cooled in an ice bath and a potassium bisulfite solution is added.

  
at 10% up 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.

  
  <EMI ID = 610.1>

  
diimide. The mixture is stirred at room temperature for 4 hours and then saturated with sodium and potassium tartrate. The resulting mixture is extracted at

  
  <EMI ID = 611.1>

  
the organic layer over anhydrous sodium sulfate and freed from the solvent to obtain 7.38 g of the title compound, in the form of a solid.

  
  <EMI ID = 612.1> allothreonine

  
7.32 g of N-methoxy-t-boc-1allothreonine amide are dissolved in 40 ml of pyridine and the solution is cooled to -20 [deg.] C under nitrogen. 3 ml of methanesulfonyl chloride are added dropwise using a syringe over 5 minutes. slowly warms up the resulting mixture

  
  <EMI ID = 613.1>

  
500 ml of ethyl acetate are added and the solution is washed with 250 ml of an ice-cold 3N hydrochloric acid solution, 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.

  
  <EMI ID = 614.1> azetidinone

  
8.64 g of 0-methanesulfonyl-N- amide is dissolved

  
  <EMI ID = 615.1>

  
add 11 g of solid potassium carbonate. We heat

  
  <EMI ID = 616.1>

  
this temperature. The reaction mixture is then filtered:
on "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 solution

  
  <EMI ID = 617.1>

  
5% sodium bicarbonate. The ethyl acetate layer is dried over anhydrous sodium sulfate and the solvent is freed to obtain 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

  
  <EMI ID = 618.1>

  
using a syringe, 5.87 g of (3S-cis) -3-t-butoxycarbonylamino-1-methoxy-4-methylazetidinone in 35 ml of

  
  <EMI ID = 619.1>

  
tetrahydrofuran. 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 to it at once. 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.

  
  <EMI ID = 620.1>

  
4.98 g of [3S, 4R] -3-t-butoxycarbonylamino-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 700 ml of a 0.5 M 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 fractions of
300 ml 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.

  
  <EMI ID = 621.1>

  
sulfonic

  
10.76 g of tetrabutylammonium salt of (3S-cis) -3-t-butoxycarbonylamino-4-methyl-2-oxo-1azetidinesulfonic acid are dissolved in 50 ml of 95 -97% formic acid, 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 an additional hour. The mixture is stored in a freezer for
About 16 hours 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 [deg.] 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.

  
  <EMI ID = 622.1>

  
a- (methoxyimino) -4-thiazoleacétique and 153 mg of Nhydroxy-benzotriazole monohydrate in 3 ml of dimethylformamide, 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 ml is added.

  
/ 1 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

  
  <EMI ID = 623.1>

  
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. Fractions 18 to 30 are combined (20 ml each) and lyophilized to obtain 274 mg of the title compound, in the form of a hygroscopic solid.

  

  <EMI ID = 624.1>


  
NMR (D20) 1.40 (3H, d, j = 6.5), 3.98 (3H, S), 4.48 (1H, d of t,

  
  <EMI ID = 625.1>

  
sulfonic A) Threonine methyl ester hydrochloride

  
  <EMI ID = 626.1>

  
(ice / brine) a flask containing 500 ml of methanol, and 130 ml (excess) of thionyl chloride are added, at a rate suitable for maintaining the reaction temperature between 0 [deg.] and 10 [deg.] C. After having cooled again to -5 [deg.] 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. Concentrate the mixture and put it under vacuum
10 1 torr for 2 hours to obtain a viscous oil. This substance is used directly in the next step.

  
B) Threonine amide

  
The crude product of part A is dissolved in 2.5 l of methanol and the mixture is cooled to -5 [deg.] 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 most of the unreacted ammonia by suction, add 100 g of sodium bicarbonate and

  
50 ml of water and the mixture is reduced to a viscous oil.

  
C) Benzyloxycarbonyithreonine amide

  
The crude product from part B (already containing the required quantity of sodium bicarbonate) is diluted to a volume of 1 liter with water. To this rapidly stirred solution, 94 g (88 ml of pure substance to
90%) of benzyloxycarbonyl chloride, in the form of a solution in 80 ml of tetrahydrofuran, in 1 hour. The reaction mixture is then stirred for an additional 16 hours and extracted with ethyl acetate (one fraction of 500 ml and two fractions of 250 ml). 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, over 15 minutes,
36.8 ml (54.5 g) of methanesulfonyl chloride. After

  
2 hours of stirring, an additional 0.3 equivalent of methanesulfonyl chloride is added. Then shake

  
the reaction mixture for 1 hour and it is 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 [deg.] C for about 16 hours in a vacuum oven gives 109 g of the title compound.

  
E) N- amide O-mesylate tetrabutylammonium salt <EMI ID = 627.1>

  
Cool to -5 [deg.] C (ice-brine) a solution of
17.8 ml of 2-picoline in 90 ml of methylene chloride, and 5.97 ml of chlorosulfonic acid are added at a rate suitable for maintaining the internal reaction temperature <EMI ID = 628.1>

  
using a cannula, to a suspension of 7.56 g of benzyloxycarbonylthreonine amide meside in 120 ml of methylene chloride. The resulting heterogeneous mixture is refluxed for approximately 16 hours, which gives a clear solution. We pour the solution into

  
  <EMI ID = 629.1>

  
further dilute with 120 ml of methylene chloride. The separated organic layer is then washed, once with 100 ml of a buffer solution, and the aqueous pnases are treated.

  
  <EMI ID = 630.1>

  
ammonium, 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.

  
  <EMI ID = 631.1> sulfonic

  
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 are added. N-sulfonyl benzyloxycarbonylthreonine amide in 20 ml of 1,2-dichloroethane
(20 ml of the latter are used for rinsing). After having maintained at reflux for 30 minutes, pour the mixture into a separating funnel, dilute with 50 ml of water and 100 ml of methylene chloride and separates the phases. The resulting organic phase is dried over sodium sulfate and concentrated to obtain the crude tetrabutylammonium salt of (3S-trans) -3-benzyloxy- acid.

  
  <EMI ID = 632.1>

  
treats crude azetidinone, in 250 ml of ethanol, with

  
0.8 g of 5% palladium on carbon serving 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 immediate precipitation of the title zwitterion, which is filtered after stirring for one hour to obtain, after <EMI ID = 633.1>

  
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.

  
  <EMI ID = 634.1>

  
Examples 140 to 143

  
  <EMI ID = 635.1>

  
(methoxyimino) -4-thiazoleacetic acid indicated in

  
  <EMI ID = 636.1>

  
column II below.

  

  <EMI ID = 637.1>
 

Example 144

  
  <EMI ID = 638.1>

  
sulfonic

  
To a suspension of (3S-trans) -3-amino-4- acid

  
  <EMI ID = 639.1>

  
139) in 30 ml of dry dimethylformamide, under nitrogen at 26 [deg.] C

  
  <EMI ID = 640.1>

  
approximately, we obtain a clear solution and add

  
  <EMI ID = 641.1>

  
then 0.334 g of N-hydroxybenzotriazole and 0.453 g of dicyclohexylcarbodiimide. The mixture is stirred for twelve hours at 26 [deg.] 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 approximately 40 ml of ether gives a precipitate which is collected and

  
  <EMI ID = 642.1>

  
  <EMI ID = 643.1>

  

  <EMI ID = 644.1>

Example 145

  
Potassium salt (1: 2) of acid [3a (Z), 4a] -3 - [[(2-amino-

  
  <EMI ID = 645.1>

  
aminol-1-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- 'acid.

  
  <EMI ID = 646.1>

  
dicyclohexylcarbodiimide and 51 mg of N-hydroxybenzotriazole, 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 washing waters are combined and treated with 113 mg of potassium perfluorobutanesulfonate. 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.

  
  <EMI ID = 647.1>

  
Cooled to -12 [deg.] C a suspension of 186 mg of potassium salt of acid [3a (Z), 4a] -3 - [[2-amino-4-

  
  <EMI ID = 648.1>

  
solution 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 under vacuum, it is pulverized, it is dissolved in 5 ml of cold water and it is immediately adjusted to pH 5.6 with 0.4 N potassium hydroxide. The solution is passed through

  
  <EMI ID = 649.1>

  
and we evaporate). The residue is triturated with ether to obtain 101.7 mg of product in the form of a hygroscopic solid.

  

  <EMI ID = 650.1>
 

Example 146

  
  <EMI ID = 651.1>

  
1-azetidinesulfonic

  
The dipotassium salt is dissolved in 1.38 ml of water

  
  <EMI ID = 652.1>

  
1-Azetidinesulfonic (87.3 mg; see Example 103). we

  
  <EMI ID = 653.1>

  
  <EMI ID = 654.1>

  
gation. 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

  
  <EMI ID = 655.1>

  
ment the resulting mixture 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, which gives 81 , 8 mg of solid. A second recrystallization in 0.8 ml of methanol gives 47.9 mg, a third in 0.24 ml of methanol and 0.24 ml of absolute ethanol gives 44.8 mg, and a fourth in 0.225 ml of methanol and 0.225 ml of absolute ethanol gives 38.8 mg. The solid is dried at 20 [deg.] C and under), 01 mm Hg, for 18 hours, then equilibrated with atmospheric humidity for 24 hours, which gives
10.9 mg of the title compound.

  
example 148

  
  <EMI ID = 656.1>

  
A sample is suspended in 30 ml of water <EMI ID = 657.1>

  
oxo-1-azetidinesulfonique (see Example 146), and the suspension is treated with IN sodium hydroxide, and used

  
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, giving 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-a- (methoxyimino) -4thiazoleacetic acid with the compound indicated in column I below, the compound indicated in column II below.

  

  <EMI ID = 658.1>
 

Example 152

  
  <EMI ID = 659.1> sulfonic

  
A solution of 440 mg of acid (Z) -2- is treated

  
  <EMI ID = 660.1>

  
dicyclohexylcarbodiimide. The mixture is stirred at room temperature for 30 minutes, under nitrogen, and

  
  <EMI ID = 661.1>

  
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 water 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.

  
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.

  
  <EMI ID = 662.1>

  
The solution is stirred for 15 minutes and 45 ml of ether are added, then 23 ml of hexane. After 5 minutes at -10 [deg.] C

  
and 15 minutes of stirring at room temperature, the solid is filtered and 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.

  

  <EMI ID = 663.1>

Example 153

  
  <EMI ID = 664.1> diphenylsilane

  
  <EMI ID = 665.1>

  
butylchlorodiphenylsilane in 112 ml of dimethylformamide. 22 ml of triethylamine are added to this solution. 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 [deg.] 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 fraction of
150 ml of sodium bicarbonate solution, two fractions of 150 ml of water and a fraction of 150 ml of a saturated solution of sodium chloride.

   The solution is dried over sodium sulfate and concentrated in vacuo to give 33.03 g of the title compound as a solid.

  
  <EMI ID = 666.1> diphenylsilane

  
A solution of 4.25 ml of 1.6M n-butyl lithium (in hexane) and 11 ml of is prepared at -50 [deg.] C under argon, in a 100 ml flask with three necks. dry tetrahydrofuran. A solution of 0.083 g of triphenylmethane in 1 ml of tetrahydrofuran is added. We cool

  
  <EMI ID = 667.1>

  
drop, using a syringe, 1.0 ml of diisopropylamine.

  
The whole is stirred for 15 minutes, then cooled to
-78 [deg.] C. One slowly adds, using a syringe, a <EMI ID = 668.1>

  
tert-butyldiphenylsilane in 8 ml of tetrahydrofuran.

  
The resulting solution was stirred for 20 minutes at -78 [deg.] C, during which time heavy precipitation occurs and uniform stirring becomes difficult. A solution of 1.33 g of p-toluene sulfonyl 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 chloride are added dropwise.

  
  <EMI ID = 669.1>

  
room temperature and stirred for 1 hour. Then

  
  <EMI ID = 670.1>

  
ethyl acetate at 0 [deg.] 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 0.5 M monobasic potassium phosphate solution, a fraction of
150 ml of sodium chloride solution, a fraction of
150 ml of a saturated solution of sodium chloride, 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 in the form of a solid.

  
  <EMI ID = 671.1>

  
stirred solution, 0.25 ml of hydrofluoric acid is added to
48%. The whole is stirred at room temperature, and 0.5 ml fractions are added every 60 minutes

  
  <EMI ID = 672.1>

  
1) 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 [deg.] C, neutralized with bicarbonate of

  
  <EMI ID = 673.1>

  
ethyl. The organic layer is then washed with 100 ml of water, 100 ml of a saturated solution of sodium chloride, 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 d 'a solid.

  
  <EMI ID = 674.1>

  
azetidine at 0 [deg.] C, 2.8 ml of a dimethylformamide-sulfur trioxide comolex 0.5 M are added under argon. This mixture is allowed to warm to ambient temperature and is stirred for 45 minutes. The solution is then poured into 20 ml of potassium phosphate buffer

  
  <EMI ID = 675.1>

  
20 ml fractions of methylene chloride (discarded) and 0.237 g of tetrabutylammonium hydrogen sulfate 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 a

  
  <EMI ID = 676.1>

  
in methylene chloride (on sodium sulfate) and concentrated in vacuo to obtain 0.31 g of an oil which, according to the NMR spectrum, consists of 50% of <EMI ID = 677.1> E) Acid (i) -3-amino-4,4-dimethyl-2-oxo-1-azetidinesulfonic

  
A solution of 0.155 g of tetrabutylammonium salt of acid (& - 3-azido-4,4-dimethyl-2-oxo-) is hydrogenated.

  
  <EMI ID = 678.1>

  
10% palladium on charcoal, for 20 minutes under an atmosphere. The catalyst is filtered off and rinsed with methylene chloride, and the rinses are combined with methanolic solution. This clear solution is treated with 0.123 ml of 97% fornic acid. When the acid is added, the solution immediately becomes cloudy.

  
After standing for 1 hour at 5 [deg.] C, the solid is separated by filtration to obtain 0.0664 g of the title compound,

  
m.p. 200-202 [deg.] C, with decomposition.

  
NMR (D20) 1.64 (3H, S), 1.68 (3H, S), 4.42 (1H, S)

  
  <EMI ID = 679.1>

Example 154

  
  <EMI ID = 680.1>

  
sulfonic

  
A solution of 50 mg of Nhydroxybenzotriazole hydrate and 0.323 mmol of (Z) -2-amino- acid is treated.

  
  <EMI ID = 681.1>

  
formamide, with 67 mg of dicyclohexylcarbodiimide, under argon, at room temperature. The resulting mixture is stirred for 1 hour, at the end of which is added

  
  <EMI ID = 682.1>

  
the reaction mixture at room temperature for
16 hours. Dimethylformamid e is removed under high vacuum at 30 [deg.] 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 to the filtrate is added 85 mg of potassium perfluorobutane sulfonate, then ether. Trituration of the resulting gum with ether gives
40 mg of a solid buff color which is subjected to chromatography on a column of 70 ml of "HP-20 AG".

  
  <EMI ID = 683.1>

  
  <EMI ID = 684.1>

  
ration with a 1: 1 mixture of acetone and hexane and drying. m.p. 225 [deg.] C, with decomposition.

  

  <EMI ID = 685.1>
 

Example 155

  
Acid potassium salt (&#65533;) - 4,4-dimethyl-2-oxo-3-

  
  <EMI ID = 686.1>

  
A 45 mg solution of N- hydrate is treated

  
  <EMI ID = 687.1>

  
in 0.5 ml of dimethylformamide, per 61 mg of dicyclohexylcarbodiimide under argon at room temperature.

  
The resulting mixture is stirred for 1 hour and added

  
  <EMI ID = 688.1>

  
sulfonic (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. Dimethylformamide is removed in a high vacuum at 30 [deg.] 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.

  

  <EMI ID = 689.1>

Example 156

  
  <EMI ID = 690.1>

  
sulfonia

  
2.14 g of salt are added to a solution of 1.85 g of diphenylphosphinyl chloride in 15 ml of dry dimethylformamide, cooled in an ice and methanol bath (-15 to -20 [deg.] C) (2-amino) -4thiazolyl) glyoxylic acid triethylamine. After stirring for half an hour, a solution of (3S-trans) -3-amino-4-methyl-2-oxo-1azetidinesulfonic acid (1.08 g; to the cold solution of mixed anhydride (1.08 g; see Example 139) and 1.9.2 ml of triethylamine in 5 ml of dry dimethylformamide, then the reaction mixture is stirred at 5 [deg.] C for 24 hours. The solvent is removed in vacuo, the residual dark oil is dissolved in water, and chromatography on resin

  
  <EMI ID = 691.1>

  
water (15 ml fractions), the crude product is collected in fractions 13 to 27 (3.37 g). Chromatography on 200 ml of "HP-20" resin, eluting with water
(15 ml fractions), gives the desired product in fractions 18 to 26. Removal of water in vacuo gives the title compound as an amorphous powder.

  

  <EMI ID = 692.1>

Example 157

  
Trifluoroacetate (1: 1) potassium salt of acid

  
  <EMI ID = 693.1>

  
oxo-1-azetidinesulfonic (see Example 127), using trifluoroacetic acid and anisole, gives the title compound, melting point 165 [deg.] C, with decomposition.

Example 158

  
  <EMI ID = 694.1> carbonyl] amino] azetidine

  
  <EMI ID = 695.1>

  
mole) of (3R - trans) -4-methyl-2-oxo-3 - [[(phenylmethoxy) -

  
  <EMI ID = 696.1>

  
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 3.5 ml of t hypochlorite are added -butyle. After 20 minutes, the solution is poured into 1 liter of cold water and extracted 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'-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. We add in
30 seconds, using a syringe, a solution of

  
  <EMI ID = 697.1>

  
(-78 [deg.] C). 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 fractions 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-methoxy-4- acid <EMI ID = 698.1>

  
sulfur formamide trioxide. After one hour at 0 [deg.] 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 it is extracted with two 50 ml fractions of methylene chloride (discard). The aqueous layer is treated with 1.04 g of tetrabutylammonium sulphate and extracted with dichloromethane to obtain 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 extensive 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).

   The crushing of
114 mg of this substance with ether gives 104 mg of an analytical sample.

  

  <EMI ID = 699.1>

Example 159

  
Potassium salt of (3S-trans) -3-methoxy-4-methyl- acid

  
  <EMI ID = 700.1>

  
  <EMI ID = 701.1>

  
ammonium. Following the procedure of Example 88,

  
  <EMI ID = 702.1>

  
composed of the title.

  

  <EMI ID = 703.1>

Example 160

  
  <EMI ID = 704.1>

  
amino] -2-methyl-4-oxo-1-azetidinesulfonic

  
Following the procedure of Example 138, but

  
  <EMI ID = 705.1>

  
thiazoleacetic with (Z) -2-amino-a - [[2- (diphenylmethoxy) -2-oxo-ethoxy] imino] -4-thiazoleacetic acid, and

  
  <EMI ID = 706.1>

  
tion.

Example 161

  
  <EMI ID = 707.1>

  
160), using trifluoroacetic acid and anisole, gives the title compound, which decomposes at> 250 [deg.] C.

Example 162

  
  <EMI ID = 708.1>

  
potassium perfluorobutanesulfonate, gives the title compound, melting point 212-214 [deg.] C.

Example 163

  
  <EMI ID = 709.1>

  
carbamic (see Example 149) using hydrogen gas and 10% palladium on carbon as catalyst, to give the title compound, melting point 165 [deg.] C, with decomposition.

Example 164

  
  <EMI ID = 710.1> <EMI ID = 711.1>

  
decomposition, after transformation into disodium salt using aqueous sodium hydroxide, and purification on "HP-20".

  
  <EMI ID = 712.1>

  
Following the procedure of Example II, but

  
  <EMI ID = 713.1>

  
  <EMI ID = 714.1>

  
compound shown in Column II below.

  

  <EMI ID = 715.1>
 

Example 169

  
  <EMI ID = 716.1>

  
  <EMI ID = 717.1>

  
t 2.46 g of di-t-butyl pyrocarbonate, the mixture is stirred in two phases for 4 hours at room temperature. 1.25 g of 0-methylhydroxylammonium are added and

  
  <EMI ID = 718.1>

  
  <EMI ID = 719.1>

  
propyl) carbodiimide and the pH is again adjusted to 4. After stirring for one hour, the reaction mixture is saturated with sodium chloride 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.

  
  <EMI ID = 720.1>

  
After stirring for 3 hours at room temperature, the reaction mixture is poured into 200 g of ice and

  
  <EMI ID = 721.1>

  
concentrated hydrochloric acid. After extracting with three 85 ml portions of ethyl acetate, the combined extracts are dried over magnesium sulfate and dried. 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.

  
  <EMI ID = 722.1> azetidinone

  
4.15 g of anhydrous potassium carbonate and 125 ml of dry acetone are brought to reflux and 3.4 g of t- are added.

  
  <EMI ID = 723.1>

  
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 to obtain 2.2 g of the title compound.

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

  
  <EMI ID = 724.1>

  
ethyl-1-methoxy-2-azetidinone to 170 ml of liquid ammonia, at -78 [deg.] C under nitrogen, and 1.68 g of sodium are added in five batches, with stirring, over 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- acid <EMI ID = 725.1>

  
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 are added. The addition is made at -30 [deg.] C, under nitrogen, in 10 minutes. After stirring at room temperature for 30 minutes, the flask is placed under vacuum to obtain a pyridine and sulfur trioxide complex. 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 [deg .]VS. At the end of
15 minutes, a clear solution is obtained which is poured into 200 ml of a 1M solution of dibasic potassium phosphate. After adding 27 g of phosphate

  
  <EMI ID = 726.1>

  
clear. This solution is extracted with two fractions of
60 ml of ethyl acetate. Add to the aqueous layer

  
  <EMI ID = 727.1>

  
the aqueous solution with three fractions of 100 ml of dichloromethane, then the combined organic layers are dried over magnesium sulfate. Concentration in vacuo gives 6.9 g of the title compound.

  
  <EMI ID = 728.1> sulfonic

  
Stir for 3 hours at room temperature <EMI ID = 729.1>

  
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 [deg.] C, with decomposition.

Example 170

  
  <EMI ID = 730.1>

  
oxo-1-azetidinesulfonique and 335 mg of triethylamine in
50 ml of dry dimethylformamide. 1.14 g of acid are added

  
  <EMI ID = 731.1>

  
imino] -4-t hiazoleacetic, stirring at 0 [deg.] C, then adding
450 mg of hydroxybenzotriazole and 0.69 g of dicyclohexylcarbodiimide. After having stirred for approximately 16 hours at 0 [deg.] C, the flask is placed under vacuum. We add to the solid

  
25 ml of dry acetone, stirring. The mixture is filtered and 0.94 g of potassium perfluorobutanesulfonate is added to the filtrate, followed by 100 ml of ether. After rest for 1

  
  <EMI ID = 732.1>

  
it is dried under vacuum, which gives 1.58 g of the title compound.

  
B) Dipotassium salt of (trans, Z) -3 - [[(2-amino-4- <EMI ID = 733.1>

  
ethyl-2-oxo-1-azetid inesu lfon ic

  
To a solution of 1.31 g of dipotassium acid salt

  
  <EMI ID = 734.1>

  
of trifluoroacetic acid, in 10 minutes, at -15 [deg.] C. After stirring for 2 hours at -10 [deg.] C, a clear solution is obtained. At -30 [deg.] C, 80 ml of dry ether are added and the resulting precipitate is filtered and then treated with 5 ml

  
1 <EMI ID = 735.1>

  
and filtering the mixture 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 [deg.] C, with decomposition.

Example 171

  
  <EMI ID = 736.1>

  
  <EMI ID = 737.1>

  
azetidinesulfonic (see Example 166), using trifluoroacetic acid and anisole, gives the title compound, melting point> 200 [deg.] C.

Example 172

  
Internal salt of acid (S) -3 - [[2- (aminomethyl) benzoyll-

  
  <EMI ID = 738.1>

  
using hydrogen gas, palladium on carbon and hydrochloric acid, gives the title compound, melting point 162-165 [deg.] C.

Example 173

  
Potassium salt of acid (S) -3 - [[[2- (4-formyl-l-

  
  <EMI ID = 739.1>

  
aminol-2-oxo-1-azetidinesulfonic

  
The tetrabutylammonium salt of the acid is condensed

  
  <EMI ID = 740.1>

  
compound resulting from potassium perfluorobutanesulfonate in acetone, to obtain the title compound, melting point 290 [deg.] C, with decomposition.

Example 174

  
  <EMI ID = 741.1>

  
Condensed (3S-trans) -3-amino-4-methyl-2oxo-1-azetidinesulfonic acid (see Example 139) with the

  
  <EMI ID = 742.1>

  
resulting compound with triethylamine and potassium perfluorobutane to obtain the title compound, period

  
  <EMI ID = 743.1>

  
nitrile and 70 ml of water. 17.8 g of triethylamine are added

  
  <EMI ID = 744.1>

  
temperature, we obtain a clear solution and add
21.0 g of di-t-butyl pyrocarbonate, and the 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 material is filtered with petroleum ether to give 20.4 g of the title compound, melting point
113-115 [deg.] C.

  
  <EMI ID = 745.1>

  
7.6 g of 0-methylhydroxylamine hydrochloride 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 maintained at 4, with stirring, for one and a half hours. The t-butanol is removed in vacuo and the solution is saturated <EMI ID = 746.1>

  
extract it twice with fractions of 100 ml 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, melting point 125-127 [deg.] C.

  
  <EMI ID = 747.1>

  
stirring, in 100 ml of dry pyridine. We cool the

  
  <EMI ID = 748.1>

  
9.3 g of methanesulfonyl chloride. After one hour at 0 ° C., an additional 3.3 g of methanesulfonyl chloride are 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 compound

  
  <EMI ID = 749.1>

  
500 ml of dry acetone. 9.8 g of potassium carbonate are added and the suspension is heated to reflux temperature, 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 separated by filtration (12.9 g), melting point 110-112 [deg.] C.

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

  
1 g of [3S-trans] -3- (t-butoxycarbonyl-

  
  <EMI ID = 750.1> liquid, with stirring. 0.154 g of sodium is added in 5 to

  
6 fractions, in 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 are 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.

  
  <EMI ID = 751.1>

  
methylene 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.

  
  <EMI ID = 752.1>

  
butoxycarbonylamino) -4-cyclohexyl-2-oxo-l-azetidinesulfonique

  
11.3 g of pyridine salt of [3S-trans] -3- (t-butoxycarbonylamino) -4-cyclohexyl-2-oxo-1-azetidinesulfonic acid are dissolved in 250 ml of water. 9.0 g of tetrabutylammonium hydrogen sulphate are added with stirring, and

  
  <EMI ID = 753.1>

  
aqueous solution twice with 200 ml fractions of methylene chloride. The organic fractions are dried with sodium sulfate, filtered and the solvent is distilled off, giving 8 g of the title compound, melting point 135-138 [deg.] C.

  
  <EMI ID = 754.1>

  
sulfonic

  
3.8 g of tetrabutylammonium acid salt are stirred

  
  <EMI ID = 755.1>

  
azetidinesulfonic 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, melting point 217-219 [deg.] C.

  
r \ <EMI ID = 756.1>

  
  <EMI ID = 757.1>

  
azetidinesulfonic

  
0.25 g of [3S-trans] -3-amino-4-cyclo- acid is dissolved

  
  <EMI ID = 758.1>

  
dry formamide and 0.12 g of triethylamine, with stirring. When a clear solution has been obtained, 0.2 g of acid is added

  
  <EMI ID = 759.1>

  
of hydroxybenzotriazole and 0.42 g of dicyclohexylcarbodiimide. 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 perfluorobutanesulfonate is added. After adding 50 ml of ether, the title compound precipitates, and is filtered. Column chromatography using the resin "HP-20" and a 9: 1 mixture of water and acetone as eluent, gives 0.36 g of product, melting point 200-205 [deg.] C (after lyophilization).

  
  <EMI ID = 760.1>

  
  <EMI ID = 761.1> sulfonic

  
[3S-trans] -3-amino-4-cyclohexyl2-oxo-1-azetidinesulfonic acid is dissolved (0.2 g; see Example 175)

  
  <EMI ID = 762.1>

  
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, the solution is treated with 0.3 g of perfluorobutanesulfonate

  
/ 1 potassium and poured into 100 ml of ether, with stirring. The

  
  <EMI ID = 763.1>

  
4-cyclohexyl-2-oxo-1-azetidinesulfonic acid in 6 ml of anisole, the mixture is cooled to -15 [deg.] C, and 5 ml of trifluoroacetic acid is added dropwise, with stirring. The temperature is maintained for one hour and then lowered to -30 [deg.] C.

  
About 100 ml of dry ether are added, at a rate such that the temperature does not exceed -10 [deg.] 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

  
  <EMI ID = 764.1>. sition (after lyophilization).

Example 178

  
  <EMI ID = 765.1>

  
sulfonic

  
[3S-trans] -3-amino-4-cyclohexyl2-oxo-1-azetidinesulfonic acid (0.1 g; see Example 175) is dissolved in a mixture of 30 ml of dry dimethylformamide and

  
  <EMI ID = 766.1>

  
benzotriazole and 0.17 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 separated by filtration. The mother liquor is stirred with 0.15 g of potassium perfluorobutanesulfonate and diluted with 50 ml of ether.

AT

  
The precipitate is filtered and subjected to chromatography with "HP-20" resin using a 9: 1 mixture of water and acetone as eluent, giving 0.14 g of

  
  <EMI ID = 767.1>

  
(after lyophilization).

Example 179

  
  <EMI ID = 768.1>

  
Dissolve [3S-trans] -3-amino-4-cyclohexyl2-oxo-1-azetidinesulfonic acid (0.1 g; see Example 175)

  
  <EMI ID = 769.1>

  
  <EMI ID = 770.1>

  
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. We separate the

  
  <EMI ID = 771.1>

  
written on resin "HP-20", using as eluent a 9: 1 mixture of water and acetone, which gives 0.15 g of the

  
  <EMI ID = 772.1>

  
station).

Example 180

  
Potassium salt of acid (trans, Z) -3 - [[(2-amino-4-

  
  <EMI ID = 773.1>

  
azetidinesulfonic

  
Following the procedure of Example 170,

  
  <EMI ID = 774.1>

  
acetic with (Z) -2-amino-a- (methoxyimino) -4thiazoleacetic acid, the title compound is obtained, melting point 190 [deg.] C, with decomposition.

Example 181

  
  <EMI ID = 775.1>

  
sulfonic <EMI ID = 776.1> diphenylsilane

  
  <EMI ID = 777.1>

  
of tert-hutylchlorodiphenylsilane in 45 ml of dimethyl-

  
  <EMI ID = 778.1>

  
hours at 0 [deg.] C, the resulting mixture is treated with an additional 1 ml of triethylamine and with 2.11 g of tert-butylchlorodiphenylsilane and allowed to stir for 65 hours at 5 [deg.] 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 fractions of

  
50 ml of water, with a saturated solution of 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 in the form of a solid.

  
  <EMI ID = 779.1> butyldiphenylsilane

  
A 50 ml flask equipped with a stirring bar, a gas inlet and a partition wall is dried with a torch under argon, and charged with n-butyl lithium (0.65 ml of a 1.6 M solution in hexane) which is cooled to -40 [deg.] 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 it is cooled to -78 [deg.] C. A solution of (&#65533;) - ((trans) - is added dropwise over about 5 minutes.

  
  <EMI ID = 780.1>

  
in 1.5 ml of tetrahydrofuran. 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 shaking for another

  
10 minutes, remove the cooling bath and stir

  
n the reaction mixture at room temperature for

  
  <EMI ID = 781.1>

  
add 20 ml of ethyl acetate, then 8 ml of a phosphate buffer at pH 4.5. Wash the organic layer with two

  
  <EMI ID = 782.1>

  
10 ml fractions of a 5% sodium bicarbonate solution, with 10 ml of a sodium chloride solution at

  
  <EMI ID = 783.1>

  
sodium, then dried over sodium sulfate. Filtration and concentration in vacuo gives 500 mg of oil,

  
which is subjected to flash chromatography with a mixture of 5% ethyl acetate and hexane, which gives
253 mg of the Compound of the Liter.

  
  <EMI ID = 784.1>

  
Dissolve in 240 ml of methanol a solution of
17 g of (&#65533;) - (trans) -3-azido-2-oxo-4-phenyl-1-azetidine-tert-butyldiphenylsilane, then the solution is treated drop by drop at 0 [deg.] C per 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 sodium chloride at 50%, then with a saturated solution of sodium chloride, and dried over sodium sulfate.

   Filtration and concentration under vacuum 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. <EMI ID = 785.1> phenyl-1-azetidinesulfonic

  
Cool to 0 [deg.] C under argon a solution of 300 mg of (&#65533;) - (trans) -3-azido-2-oxo-4-phenyl-1-azetidine in 3 ml

  
  <EMI ID = 786.1>

  
a complex of dimethylformamide and sulfur trioxide

  
1-n (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 monobasic potassium phosphate at pH 5.5. The solution is extracted with dichloromethane (disposable) and 541 mg of tetrabutylammonium bisulfate are added. The resulting mixture is extracted with dichloromethane and the organic extracts are washed with a 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 rest being formed by dimethylformamide. This mixture is used without purification in the next step.

  
  <EMI ID = 787.1> sulfonic

  
A solution of tetrabutyl salt is hydrogenated

  
  <EMI ID = 788.1>

  
sulfonic acid in 4 ml of methanol, on 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 under vacuum to 18 ml and 0.2 ml of acid is added

  
  <EMI ID = 789.1>

  
several hours, the resulting solid is filtered and washed with dichloromethane to obtain, after drying,
150 mg of the title compound as a solid.

  
  <EMI ID = 790.1>

  

  <EMI ID = 791.1>

Example 182

  
  <EMI ID = 792.1> A) Tetrabutylammonium salt of acid (&#65533;) - trans-2-oxo1-IN <EMI ID = 793.1>

  
A solution of 52 mg of N-hydroxybenzotriazole monohydrate and 46 mg of phenylacetic acid in 0.3 ml of dimethylformamide is treated with 70 mg of solid dicyclohexylcarbodiimide under argon at 0 [deg.] C. The cooling bath is removed and the mixture is stirred. Resulting at room temperature for one hour. After diluting with an additional 0.3 ml of dimethylformamide,

  
  <EMI ID = 794.1>

  
sulfonic in the form of a solid (75 mg, see Example
181), then 0.05 ml of triethyl- is added dropwise

  
  <EMI ID = 795.1>

  
ambient 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

  
  <EMI ID = 796.1>

  
ethyl acetate (discard), and 105 mg (0.31 mmol) of tetrabutylammonium bisulfate is added. We extract the mixture

  
  <EMI ID = 797.1>

  
The extracts are washed with two 15 ml fractions of 10% sodium chloride solution, then with 10 ml of a saturated sodium chloride solution, and dried over sodium sulfate. Filtration and concentration under vacuum give (after heating to 32 [deg.] C under high vacuum)

  
165 mg of an oil, comprising approximately 40% of the desired compound, and 60% of dimethylformamide.

  
  <EMI ID = 798.1>

  
azetidinesulfonic acid in 1.5 ml acetone, per 41 mg

  
(0.121 mmol) of potassium perfluorobutanesulfonate. A dilution with 12 ml of ether gives a glass which, after trituration with ether, provides 43 mg of a solid which contains approximately 20% of an impurity itself containing a tetrabutylammonium part. The solid is dissolved in 50% aqueous acetone and the solution is passed through

  
  <EMI ID = 799.1>

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

  

  <EMI ID = 800.1>

Example 183

  
  <EMI ID = 801.1>

  
(methoxyimino) -4-thiazoleacetic in 0.3 ml of dimethylformamide, with 70 mg of solid dicyclohexylcarbodiimide, under argon, at room temperature. We stir the mixture

  
  <EMI ID = 802.1>

  
  <EMI ID = 803.1>

  
(75 mg; see Example 181) as 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 in a high vacuum at 30 ° C., and the residue is triturated with 2 ml of acetone, then filtered. The filter cake is washed with two additional 3 ml fractions of acetone and 86 mg of potassium perfluorobutanesulfonate are 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.

  

  <EMI ID = 804.1>

Example 184

  
Potassium salt of (cis) -2-oxo-4-phenyl-3 - [(phenyl-

  
  <EMI ID = 805.1> A) N-benzylidene-2,4-dimethoxybenzylamine

  
12.0 g of 2,4-dimethoxy hydrochloride are added.

  
  <EMI ID = 806.1>

  
extract the mixture with 125 ml of ethyl acetate. The organic layer is dried over anhydrous sodium sulfate and freed from the solvent to obtain 10.2 g of 2,4dimethoxybenzylamine in the form of 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) (&#65533;) - (cis) -4-phenyl-1- (2,4-dimethoxybenzyl) -2-oxo-3azidoazetidine

  
1.62 g of a-azidoacetic acid are dissolved in 25 ml

  
  <EMI ID = 807.1>

  
solution in 10 ml of methylene chloride. The mixture is cooled in an ice bath and 3.36 g of trifluoroacetic anhydride are slowly added; 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

  
  <EMI ID = 808.1>

  
the organic layer is dried over anhydrous sodium sulfate and the solvent is freed to obtain 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. Fractions

  
It combined give crystals by rapid cooling in a bath of dry ice and acetone. Using these

  
  <EMI ID = 809.1>

  
in 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.

  
  <EMI ID = 810.1>

  
of acetonitrile and the mixture is heated to 80 - 83 [deg.] 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

  
  <EMI ID = 811.1>

  
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

  
  <EMI ID = 812.1>

  
removes the solvent 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 acid (&) - (cis) -4- <EMI ID = 813.1>

  
phenyl-2-oxo-3-azidoazetidine, and dropwise added,

  
  <EMI ID = 814.1>

  
a complex of dimethylformamide and sulfur trioxide.

  
  <EMI ID = 815.1>

  
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

  
n 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

  
  <EMI ID = 816.1>

  
nique 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";

  
certain particles of 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. 81 mg of N-hydroxybenzotriazole 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. We

  
  <EMI ID = 817.1>

  
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 for &#65533; obtain 26 mg of the title compound.

  

  <EMI ID = 818.1>

Example 185

  
Potassium salt of (cis, Z) acid -3 - [[(2-amino-4-

  
  <EMI ID = 819.1> azetidinesulfonic

  
Dissolve the potassium salt of the acid (cis) -2-

  
  <EMI ID = 820.1>

  
of platinum serving 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) acid are added -

  
  <EMI ID = 821.1>

  
the mixture is stirred for approximately 16 hours under 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.

  
When 15 ml of ether are added, an eraser separates. The liquid is eliminated and the gum is washed with ether.

  
  <EMI ID = 822.1>

  
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.

  

  <EMI ID = 823.1>

Example 186

  
  <EMI ID = 824.1>

  
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 a

  
  <EMI ID = 825.1>

  
stir the mixture under nitrogen for 2 hours then filter. The filtrate is evaporated to obtain a solid.

  
r'1 The crude product is recrystallized from a mixture of methylene chloride and petroleum ether to obtain
20.96 g of the title compound as a solid.

  
  <EMI ID = 826.1>

  
24.26 g of 2-azidoacetic acid are dissolved in
100 ml of methylene chloride and the solution is cooled in an ice bath. To the solution, we add
48.57 g of triethylamine and 14.24 g of N- (3-phenyl-2-

  
  <EMI ID = 827.1>

  
methylene chloride. In the resulting solution, we

  
  <EMI ID = 828.1>

  
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. -

  
  <EMI ID = 829.1>

  
anhydrous sodium and the solvent is freed 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.

  
  <EMI ID = 830.1>

  
azetidine dissolved in 65 ml of acetonitrile, in 15 minutes
(an additional 10 ml of acetonitrile is used for rinsing). The mixture is stirred for another 15 minutes

  
at 0 [deg.] C, it is diluted with 750 ml of ethyl acetate, it is

  
  <EMI ID = 831.1>

  
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 11 were combined to 16 (50 ml each) and evaporated to obtain 802 mg of the title compound as an oil.

  
  <EMI ID = 832.1>

  
868 mg of a pyridine and sulfur trioxide complex are added. The mixture is stirred at room temperature

  
  <EMI ID = 833.1>

  
of a 0.5 M monobasic potassium phosphate solution, then washing with 30 ml of methylene chloride. 530 mg of tetra-n-butylammonium bisulfate are added to the aqueous solution and the mixture is extracted with four fractions of

  
  <EMI ID = 834.1>

  
nics combined with two fractions of 100 ml 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.

  
  <EMI ID = 835.1> azetidinesulfonic

  
300 mg of the tetra-n-butylammonium salt are dissolved

  
  <EMI ID = 836.1>

  
shakes quickly. To this mixture, 600 mg of zinc powder is added, then 0.8 ml of a 1M 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 fractions 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 is

  
  <EMI ID = 837.1>

  
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.

  
  <EMI ID = 838.1>

  
/ 7 nitrile to dissolve the precipitate and pass the

  
  <EMI ID = 839.1>

  
eluting first with 150 ml of water, then with a mixture of
10% acetone and water. We join fractions 2 to 13
(15 ml each) and evaporated to obtain 101 mg of the title compound in the form of a foam.

Example 187

  
Acid potassium salt (&) - (cis, Z) -3 - [[(2-amino-4-

  
  <EMI ID = 840.1>

  
formamide, per 69 mg of dicyclohexylcarbodiimide. The mixture is stirred at room temperature for 30 minutes under nitrogen. (Cis) -3-amino-2oxo-4- (2-phenylethenyl) -1-azetidinesulfonic acid (90 mg;

  
see Example 186) and 34 mg of triethylamine and the mixture is stirred for 20 hours under nitrogen. The suspension is evaporated in vacuo and triturated with 10 ml

  
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 is passed over 34 ml of "HP-20" resin, first eluting with 150 ml of water, then with a mixture of 10% acetone and water. We join the fractions 16 to
19 (15 ml each) and freed from the solvent to obtain 110 mg of the title compound as a solid.

  

  <EMI ID = 841.1>

Example 188

  
  <EMI ID = 842.1>

  
sulfonic A) Methyl ester of [(4-m & thoxyphenyl) imino] acetic acid We load a 1 liter flask with three necks, dry, equipped with a nitrogen inlet and a bar stirring, with 56.88 g of magnesium sulfate, 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 hemiacetal is added over an hour and a half.

  
methyl glyoxylate in 50 ml of dichloromethane.

  
After having stirred for a further 20 minutes at 0 [deg.] 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 dissolves

  
  <EMI ID = 843.1>

  
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 methyl acid ester [(4-

  
  <EMI ID = 844.1>

  
and cooled to 0 [deg.] C. 19.2 ml are added dropwise
(0.14 mole) of triethylamine, then a solution of 28.4 g of (N-phthalimido) acetyl chloride in 150 ml of dichloromethane, in one hour. The resulting mixture is stirred

  
  <EMI ID = 845.1>

  
2.5 1 of dichloromethane. The organic solution is washed with two 500 ml fractions of monobasic potassium phosphate! pH 4.5, two 500 ml fractions of bicarbo-

  
  <EMI ID = 846.1>

  
sodium chloride, then dried 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.

  
  <EMI ID = 847.1>

  
We load a 500 ml dry flask equipped with a nitrogen inlet, a stirring bar and a partition

  
  <EMI ID = 848.1> <EMI ID = 849.1>

  
azetidine and 325 ml of dichloromethane. We cool to
-30 [deg.] C the resulting suspension, and 3.52 ml of methyl hydrazine are added dropwise. We heat the <EMI ID = 850.1>

  
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, it is redissolved in 225 ml of dichloromethane, and it is left to stand at room temperature for approximately 16 hours, during which time a considerable amount solid precipitates. The mixture is filtered under nitrogen, the filtrate is cooled to 0 [deg.] C (nitrogen atmosphere) and treated with
17 ml of diisopropylethylamine, then with 7 ml of benzyl chloroformate, drop by drop. We stir the mixture

  
  <EMI ID = 851.1>

  
room for 1 hour and a half. The mixture is washed with two 300 ml fractions of a monobasic potassium phosphate buffer at pH 4.5, with two fractions of

  
300 ml of 5% sodium bicarbonate with 300 ml of a saturated solution of sodium chloride, 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.

  
  <EMI ID = 852.1>

  
A solution of 8.59 g of ammonium nitrate is treated

  
  <EMI ID = 853.1>

  
of water with a suspension of 2 g of (cis) -4- (methoxy-

  
  <EMI ID = 854.1>

  
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

  
  <EMI ID = 855.1>. 9% <EMI ID = 856.1>

  
and the combined organic extracts are washed with three fractions

  
  <EMI ID = 857.1>

  
the basic washing waters 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 chloride solution sodium 5%, two 50 ml fractions of a saturated sodium chloride solution, then stirred on charcoal "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) (cis) -4- (methoxy-) tetrabutylammonium salt <EMI ID = 858.1>

  
1 ml of pyridine. The reaction mixture is poured into

  
  <EMI ID = 859.1>

  
and extracted with four 30 ml fractions of dichloromethane (discard). 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 a solution of

  
  <EMI ID = 860.1>

  
we filter. Concentration in vacuo gives 186 mg of the title compound as a viscous oil.

  
  <EMI ID = 861.1>

  
A solution of 186 mg of tetrabutylammonium salt of (cis) -4- (methoxycarbonyl) -2-oxo-3- acid is hydrogenated.

  
  <EMI ID = 862.1>

  
in 2 ml of methanol, on 95 mg of 10% palladium on carbon, for one and a half hours under 1 atmosphere. The catalyst is filtered off and rinsed with

  
\ dichloromethane, then the filtrate is treated with 97% formic acid and cooled to -50 [deg.] C (the presence of a primer crystal at this stage is necessary to induce crystallization). Once the crystallization has started, the mixture is left to stand for 16 hours

  
  <EMI ID = 863.1>

  
dichloromethane, hexane, and dried in vacuo to give 50 mg of the title compound.

Example 189

  
Potassium salt of (cis) acid -3 - [[2-amino-4-thiazolyl) -

  
  <EMI ID = 864.1>

  
acetic acid 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-

  
  <EMI ID = 865.1>

  
  <EMI ID = 866.1>

  
0.03 ml of triethylamine dropwise. The reaction mixture is stirred at room temperature for approximately 16 hours. Dimethylformamide is removed in a high vacuum at 30 [deg.] 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.

Example 190

  
  <EMI ID = 867.1>

  
A suspension of potassium salt of (cis) acid -3 - [[2-amino-4-thiazolyl) - [[1-] is stirred at -12 ° C. under argon.

  
  <EMI ID = 868.1>

  
4- (methoxycarbonyl) -2-oxo-1-azetidinesulfonic in 0.4 ml of anisole, and 0.9 ml of cold trifluoroacetic acid (-10 [deg.] C) is added. After an hour and a half, 4 ml are added

  
1 <EMI ID = 869.1>

  
ether and 2 ml hexane and the resulting suspension is stirred for. 15 minutes at -10 [deg.] C, then for 15 minutes

  
  <EMI ID = 870.1>

  
fugation and washed with ether. The pH of a suspension of this substance in 0.5 ml of cold water is adjusted to

  
  <EMI ID = 871.1>

  
30 ml column of "HP-20 AG". Elution with water gives
30 mg of the title compound after evaporation (added

  
  <EMI ID = 872.1>

  

  <EMI ID = 873.1>

Example 191

  
  <EMI ID = 874.1>

  
sulfonic A) 2- (trimethylsilyl) ethynylmagnesium bromide

  
20 ml are introduced into a 50 ml flask dried with a torch and maintained under a positive nitrogen pressure.

  
  <EMI ID = 875.1>

  
and 5.05 ml of a 3.06 M solution of methylmagnesium bromide in ether. The mixture is stirred for 140 minutes to give the title compound.

  
  <EMI ID = 876.1> [(triphenylmethyl) amino] azetidine

  
6.00 g of (S) - (cis) -4- (methylsulfonyl) -2- are added to a 250 ml three-necked flask

  
  <EMI ID = 877.1>

  
with nitrogen and then it is 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 1 ml is added dropwise using a syringe. ether, shaking quickly. Add the 2- bromide solution

  
  <EMI ID = 878.1>

  
using a "Teflon" tube, under positive nitrogen pressure
(the flask containing the reagent is rinsed with 7 ml of tetrahydrofuran). Once the addition is complete, the cooling bath is removed. After 45 minutes, a solution of 3.5 g of potassium bisulfate in 20 ml is added <EMI ID = 879.1>

  
furan 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. Elimination of

  
  <EMI ID = 880.1>

  
matography on a silica column. Elution with 2 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.

  
  <EMI ID = 881.1>

  
in 30 ml of dichloromethane and 330 mg of tetrabutylammonium fluoride (containing 20 to 25% water) are added. 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 a saturated aqueous solution of sodium chloride, 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).

  
  <EMI ID = 882.1> sulfonic

  
404 mg of (S) - are added to a 25 ml flask

  
  <EMI ID = 883.1>

  
and 560 mg of a pyridine and sulfur trioxide complex. After having purged the flask with nitrogen, 4.0 ml of

  
1 dry pyridine and the mixture is heated to 80-85 [deg.] C for

  
  <EMI ID = 884.1>

  
stirred 4.0 ml of concentrated hydrochloric acid, 50 ml of water and 50 ml of ethyl acetate. The pH is adjusted to 3.15 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 solvent under vacuum gives a foam which is taken up in 10 ml

  
  <EMI ID = 885.1>

  
8 ml), 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

  
  <EMI ID = 886.1>

  
azetidinesulfonic

  
Weigh in a 10 ml flask 100 mg of acid (Z) -

  
  <EMI ID = 887.1>

  
hydroxybenzotriazole monohydrate and 113 mg dicyclohexylcarbodiimide. The flask is purged with nitrogen and cooled 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- acid is added

  
  <EMI ID = 888.1>

  
191) in the form of a solid with 1.0 ml of dimethyl-

  
  <EMI ID = 889.1>

  
cooling and the mixture is stirred for 22 hours. 3 ml of acetone are added and the solids present are filtered off, then washed 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, melting point> 230 [deg.] C.

Example 193

  
Potassium salt of (S) -3 - [[[(2,5-dichlorophenyl) thio] acetyl] amino] -2-oxo-1-azetidinesulfonic acid

  
100 mg of 3-amino-2-oxo-lazeti inesulfonic acid is dissolved in 2 ml of dry dimethylformamide with 0.083 ml of triethylamine. 123 mg (0.602) are added

  
  <EMI ID = 890.1>

  
of N-hydroxybenzotriazole hydrate and 124 mg of dicyclohexylcarbodiimide, the permeable mixture is stirred for 2 hours at room temperature, then for 2 days at 5 [deg.] 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 an IN potassium solution, and after having extracted a total of three times with dichloromethane, it is dried over sulfate of sodium of the combined extracts and the solvent is removed in vacuo, which gives 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 is collected by filtration, washed several times with acetone and dried to obtain 206 mg of a powder.

  

  <EMI ID = 891.1>

Example 194

  
  <EMI ID = 892.1>

  
sulfonic

  
Dissolve (3S-trans) -3-amino-4-methyl-2-oxo1-azetidinesulfonic acid (250 mg; see Example 139) in

  
  <EMI ID = 893.1>

  
  <EMI ID = 894.1>

  
N-hydroxybenzotriazole hydrate and 287 mg of dicyclohexylcarbodiimide. After shaking for 16 hours

  
n at room temperature, the mixture is filtered and the solvent is removed in vacuo. The residue is taken up in the water and filtered. The filtrate is washed with ethyl acetate, it is decanted with dichloromethane and 4.2 mmol of tetrabutylammonium bisulfate are 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,

  
946 mg of potassium perfluorobutanesulfonate dissolved in acetone are added. 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 water and methanol. Trituration of the residue with acetone gives 233 mg of a powder, melting point 212-213 [deg.] (With decomposition).

  

  <EMI ID = 895.1>


  
Examples 195 and 196

  
Following the procedure of Example 138,: born by replacing the (3S-cis) -3-amino-4-methyl-2-oxo-1azetidinesulfonic acid with (3S-trans) -3-amino acid -4-methyl2-oxo-1-azetidinesulfonic and (Z) -2-amino-a- acid

  
  <EMI ID = 896.1>

  
column I below, the compound indicated in column II below is obtained.

  

  <EMI ID = 897.1>
 

Example 197

  
  <EMI ID = 898.1>

  
sulfonic

  
  <EMI ID = 899.1>

  
  <EMI ID = 900.1>

  
melting point 128 [deg.] C, with decomposition.

Example 198

  
Potassium salt of [3S (R *)] - 3 - [[(2-amino-4-

  
  <EMI ID = 901.1>

  
azetidinesulfonic

  
Following the procedure of Example 6, but replacing the aminothiazoleacetic acid with the acid

  
  <EMI ID = 902.1>

  
imidazolidinyl] carbonyl] amino] -4-thiazoleacétique, the title compound is obtained, melting point> 250 [deg.] C;

Example 195

  
Organic production of EM5117

  
9 liter fermentation

  
Chromobacterium violaceum SC 11.378, A.T.C.C. N [deg.] 31532 on the following sterilized agar medium

(AT) :

Grams

  

  <EMI ID = 903.1>


  
Distilled water to make 1 liter.

  
The pH is adjusted to 7.3 before sterilization at 121 [deg.] C for 30 minutes.

  
The contents of a microorganism superficial growth "magnifier" are used to inoculate each of three 500 ml Erlenmeyer flasks, each containing
100 ml of the following sterilized medium (B):

  

  <EMI ID = 904.1>


  
Tap water to make 1 liter.

  
The pH is adjusted to 7.0 before sterilizing at 121 [deg.] C for
15 minutes.

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

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

  

  <EMI ID = 905.1>

Tap water to make 1 liter.

  
The pH is adjusted to 7.0 before sterilizing at 121 [deg.] C for
15 minutes.

  
After inoculation, in fact incubate the flasks at 25 [deg.] C on a rotating shaking device (300 rpm, stroke of

  
5 cm) for approximately 18 to 24 hours. At the end of this time, the contents of the vials are collected and the broth is centrifuged, which gives approximately 9 liters of supernatant broth.

  
250 liter fermentation

  
Using a surface growth "magnifying glass" of an inclined agar culture (medium A) of Chromobacterium violaceum SC 11.378, A.T.C.C. N [deg.] 31532, to inoculate each of five 500 ml Erlenmeyer flasks each containing 100 ml of sterilized medium (B). These flasks are then incubated at 25 [deg.] C on a rotating shaker (300 rpm; 5 cm stroke) for approximately 24 hours. After the appropriate incubation as described above, transfers of 1% (volume / volume) are carried out from the culture flasks grown in five 4-liter Erlenmeyer flasks each containing 1.5 liter of sterilized medium (B). After

  
  <EMI ID = 906.1>

  
  <EMI ID = 907.1>

  
for about 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 continues

  
  <EMI ID = 908.1>

  
stirring 155 rpm. "Ucon" is added as necessary as an anti-foaming agent. At the end of approximately 18 to 24 hours, the fermentation is finished. The fermentation broth is then adjusted to pH 5.0 using hydrochloric acid and the contents of the broth in the tank are centrifuged, giving approximately 230 liters of supernatant broth.

  
Isolation and purification

  
The 250 ml fermentation supernatant broth is adjusted to pH 5 using sulfuric acid and filtered using 3 - 5% diatomaceous earth.

  
  <EMI ID = 909.1>

  
0.005 M ammonium in methylene chloride.

  
The combined lower phase is extracted with 6 liters of 0.05 M sodium iodide which has previously been adjusted to pH 5 with acetic acid. The lower phase is discarded and the upper phase is concentrated under vacuum at

  
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. We dissolve the residue
(if possible) in 150 ml of methanol. The insoluble substance is discarded and the methanolic solution is concentrated to dryness under vacuum, which gives 38.6 g of crude antibiotic.

  
The crude product is dissolved in 10 ml of a mixture
(1: 1) of methanol and water and it is subjected to chromatography on a column of 500 ml of crosslinked dextran gel ("Scphadex G-10") in the same mixture of solvents, eluting with 2 ml / minute and collecting fractions of
20 ml. 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 to

  
pH 5 and passed through a column of DEAEcellulose ("Whatman DE52" cellulose) packed and balanced in the same buffer. The column is eluted at 5 ml / minute with a linear gradient prepared from 4 liters

  
  <EMI ID = 910.1>

  
4 liters of 0.1 M sodium phosphate buffer at pH 5, 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 10 ml fractions. The active fractions are combined (38 to 44) and concentrated, which gives 459 mg of residue.

  
348 mg of this residue are dissolved in water and the solution is passed through a column of macroreticular styrene-divinylbenzene copolymer resin. ("Diaion HP 20AG"); 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 (36 to

  
43) and they are concentrated, which gives 186.4 mg of substance. A chromatography (similarly) of another 100 mg of
459 mg of residue from the previous operation gives

  
not <EMI ID = 911.1>

  
this and the 51.5 mg of substance consist of almost pure EM5117, as shown by thin layer chromatography and nuclear magnetic resonance spectra.

  
The 186.4 mg fraction of EM5117 from the previous operation is dissolved in water and passed through a column of ion exchange resin

  
  <EMI ID = 912.1>

  
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, which gives
145 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 [deg.] C with decomposition.

  

  <EMI ID = 913.1>


  
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

  

  <EMI ID = 914.1>


  
Distilled water to make one liter.

MIDDLE E

  

  <EMI ID = 915.1>


  
Distilled water to make 1 liter.

  
Adjust the pH to 7.3 before sterilization. MIDDLE F

  

  <EMI ID = 916.1>


  
Tap water to make 1 liter. MIDDLE G

  

  <EMI ID = 917.1>


  
Distilled water to make 1 liter.

  
Adjust the pH to 7.0. MIDDLE H

  

  <EMI ID = 918.1>


  
Distilled water to make 1 liter.

MIDDLE I

  

  <EMI ID = 919.1>


  
Distilled water to make 1 liter.

MIDDLE J

  
Grams

  

  <EMI ID = 920.1>


  
Grams

  

  <EMI ID = 921.1>


  
  <EMI ID = 922.1>

  
Glucose 34 MIDDLE M

  
Grams

  

  <EMI ID = 923.1>


  
Tap water to make 1 liter.

Example '196

Organic production of EM5210

  
Gluconobacter, - species SC 11.435, A.T.C.C. N [deg.] 31581 on the following sterilized agar medium (A):

  

  <EMI ID = 924.1>


  
Distilled water to make 1 liter.

  
The pH is adjusted to 7.3 before sterilization at 121 [deg.] C for
30 minutes.

  
A "growth loop" 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);

  

  <EMI ID = 925.1>


  
Distilled water to make 1 liter.

  
  <EMI ID = 926.1>

  
15 minutes.

  
After inoculation, the vials are incubated at
25 [deg.] C on a rotating stirring device (300 rpm; 5 cm stroke) for approximately 24 hours. After the appropriate incubation as described above,

  
  <EMI ID = 927.1>

  
culture flasks developed in one hundred 500 ml Erlenmeyer flasks each containing 100 ml of the following sterilized medium (C):

  

  <EMI ID = 928.1>

Distilled water to make 1 liter.

  
This medium is sterilized at 121 [deg.] C for 15 minutes.

  
After inoculation, the vials are incubated at
25 [deg.] C on a rotating stirring device (300 rpm;

  
  <EMI ID = 929.1>

  
ble the contents of the vials and the broth is centrifuged, which gives 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 quaternary ammonium groups attached to a network of styrene-divinylbenzene copolymer ["Dowex AG1- X2 "(CI-)],

  
  <EMI ID = 930.1>

  
5% in 0.01 M aqueous NaH2PO4 The eluate is concentrated in vacuo.

  
The residue is adsorbed with 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.

  
The crude EM5210 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 for 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 is absorbed from a fer-

  
  <EMI ID = 931.1>

  
SC 11.435, with 10.8 kg of a strongly basic anion exchange resin ["Dowex 1-X8" (Cl-)]. This resin is washed with water and eluted with 5% sodium chloride in 0.01 M sodium diacid phosphate. 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 carbon column

  
  <EMI ID = 932.1>

  
wash the column with water and elute the EM 5210 with a 1: 1 mixture of methanol and water. The active fractions are combined and concentrated. We dissolve the residue
(16 g) in water and it is subjected to chromatography on a column (600 ml) of a strongly basic ion exchange resin ["Bio. Rad AG 1-X2" (CI-), 37 -
74 p], 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 under vacuum to a small volume, and the precipitated salts are filtered off. We pass the filtrate through a

  
  <EMI ID = 933.1>

  
macroréticulalre. The column is eluted with water. The active fractions are combined, concentrated to a small volume and subjected to cryodessication, which gives 120 mg of the sodium salt of EM5210.

  
  <EMI ID = 934.1>

  
an ion exchange resin column is used

  
("Dowex 50 W-X2", lithium form). 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. For example, about 2 , 5 mg of the lithium salt in a column of "Bio.Rad Bio-Rex 70" (H)

  
and elute with water to obtain 1.45 mg of the free acid

  
(internal salt).

  
Chemical properties of EM5210

  
1) Positive "Ninhydrin" test (N inhydrin is the

  
amino acid analytical reagent).

  
  <EMI ID = 935.1>

  
for 16 hours) gives two major positive spots to Ninhydrin by chromatography on paper ("Whatman"

  
  <EMI ID = 936.1>

  
of water), and a weak spot positive for Ninhydrin which quenches the fluorescence excited by ultraviolet rays.

  
The two major positive spots for Ninhydrin are D-glutamic acid and D-alamine.

  
Physical characteristics of EM5210

  
1) UV spectrum of sodium salt in water: absorption

  
final.

  
2) IR - Major peaks of the lithium salt in bromide

  
potassium:

  
  <EMI ID = 937.1>

  
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 [deg.] C (C = 0.15%) (pH 2.7);

  

  <EMI ID = 938.1>


  
The following fermentation media have been shown to be effective in producing EM5210 and can be

  
  <EMI ID = 939.1>

MIDDLE D

  
  <EMI ID = 940.1>

  

  <EMI ID = 941.1>


  
Tap water to make 1 liter.

  
Adjust the pH to 7.0 before sterilization at 121 [deg.] C for 15 minutes.

MIDDLE E

  
Grams

  

  <EMI ID = 942.1>


  
Tap water to make 1 liter.

  
Sterilization at 121 [deg.] C for 15 minutes.

MIDDLE F

  

  <EMI ID = 943.1>


  

  <EMI ID = 944.1>


  

  <EMI ID = 945.1>


  
Distilled water to make 1 liter.

  
Sterilization at 121 [deg.] C for 30 minutes. MIDDLE H

  

  <EMI ID = 946.1>


  
Tap water to make 1 liter.

  
Sterilization at 121 [deg.] C for 15 minutes.

  

  <EMI ID = 947.1>

Distilled water to make 1 liter.

Sterilization at 12l [deg.] C for 30 minutes.

  
Biological activity

  
The following methodology is used to determine the minimum inhibitory concentration (hereinafter referred to as

  
  <EMI ID = 948.1>

  
The test microorganisms are grown in about 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 [deg.] 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 inoculum level of 10 4 CFU; dilutions are made with K-10 * broth.

  
The compounds are dissolved in the diluent suitable for

  
  <EMI ID = 949.1>

  
of each dilution 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

  
  <EMI ID = 950.1>

  
microorganism growth control plates containing only agar and inoculated before and after the test plates. We apply microorganisms

  
on the agar surface of each plate with the Denley Multipoint inoculator (which delivers approximately 0.001 ml of each microorganism), which allows a final inoculum level of 10 4 CFU to be obtained on the agar surface.

  
  <EMI ID = 951.1>

  
and the MICs are determined. The ICD is the minimum concentration of compound that inhibits the growth of the microorganism.

  
* K-10 broth is a beef and yeast broth containing:

  

  <EMI ID = 952.1>


  
Distilled water in q.s. to make 1 liter ** K-10 Agar

  

  <EMI ID = 953.1>


  
  <EMI ID = 954.1>

  
The tables which follow are the results obtained when the efficacy of the 3-lactams according to the invention is tested against various microorganisms. The number following each microorganism designates the number following each microorganism designates 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 indicates that the compound tested did not show any activity vis-à-vis the particular microorganism to

  
  <EMI ID = 955.1>

  
has not been tried.

  

  <EMI ID = 956.1>


  

  <EMI ID = 957.1>
 

  

  <EMI ID = 958.1>


  

  <EMI ID = 959.1>
 

  

  <EMI ID = 960.1>


  

  <EMI ID = 961.1>
 

  

  <EMI ID = 962.1>


  

  <EMI ID = 963.1>
 

  

  <EMI ID = 964.1>


  

  <EMI ID = 965.1>
 

  

  <EMI ID = 966.1>


  

  <EMI ID = 967.1>
 

  

  <EMI ID = 968.1>


  

  <EMI ID = 969.1>
 

  

  <EMI ID = 970.1>


  

  <EMI ID = 971.1>
 

  

  <EMI ID = 972.1>


  

  <EMI ID = 973.1>
 

  

  <EMI ID = 974.1>


  

  <EMI ID = 975.1>
 

  

  <EMI ID = 976.1>


  

  <EMI ID = 977.1>
 

  

  <EMI ID = 978.1>


  

  <EMI ID = 979.1>
 

  

  <EMI ID = 980.1>


  

  <EMI ID = 981.1>
 

  

  <EMI ID = 982.1>


  

  <EMI ID = 983.1>
 

  

  <EMI ID = 984.1>


  

  <EMI ID = 985.1>
 

  

  <EMI ID = 986.1>


  

  <EMI ID = 987.1>
 

  

  <EMI ID = 988.1>


  

  <EMI ID = 989.1>
 

  

  <EMI ID = 990.1>


  

  <EMI ID = 991.1>
 

  

  <EMI ID = 992.1>


  

  <EMI ID = 993.1>
 

CLAIMS

  
  <EMI ID = 994.1>

  
is a hydrogen atom or a cation, in position 1, and

  
an amine substituent -NH2 or a protected form thereof

  
in position 3.

  
2. &#65533; -lactam, characterized in that it carries a

  
  <EMI ID = 995.1>

  
is a hydrogen atom or a cation, in position l, and

  
an azide-N3 substituent in position 3.


    

Claims (1)

3. 8-lactam according to claim '1, characterized in that it carries a substituent sulfonic acid salt <EMI ID = 996.1> cation, in position 1, an amine substituent -NH2 in position 3, and an alkyl substituent in position 4. 4. (3-lactam according to claim 1, characterized in that it carries a sulfonic acid salt substituent <EMI ID = 997.1> cation, in position 1, an amine substituent -NH2 in position 3, and a cycloalkyl, phenyl or substituted phenyl substituent, in position 4. <EMI ID = 998.1>  <EMI ID = 999.1> 3, and an alkoxycarbonyl, alken-1-yl, alkyn-1- substituent  <EMI ID = 1000.1> 6. 0-lactam according to claim 1, characterized in that it carries a salt sulfonic acid substituent  <EMI ID = 1001.1>  <EMI ID = 1002.1> alkyl substituent in position 4.  <EMI ID = 1003.1> in that it carries a sulfonic acid salt substituent 6? <EMI ID = 1004.1> in position 1, an azide-N3 substituent in position 3, and a cycloalkyl, phenyl or substituted phenyl substituent, in position 4. <EMI ID = 1005.1>  <EMI ID = 1006.1> in that it carries a sulfonic acid salt substituent  <EMI ID = 1007.1> <EMI ID = 1008.1>  <EMI ID = 1009.1> 3, and a methyl substituent in position 4.  <EMI ID = 1010.1> formula :  <EMI ID = 1011.1> in which R2 is a hydrogen atom or a radical  <EMI ID = 1012.1> 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 radical -yle,  <EMI ID = 1013.1> is a hydrogen atom or a cation.  <EMI ID = 1014.1>  <EMI ID = 1015.1> in which R2 is a hydrogen atom or a radical  <EMI ID = 1016.1> 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 radical -yle,  <EMI ID = 1017.1> is a hydrogen atom or a cation. <EMI ID = 1018.1> R2 is a hydrogen atom. 13. 8-lactam according to claim 10 or II ,. in which]  <EMI ID = 1019.1> 14. 6-lactam according to claim 10 or 11, wherein  <EMI ID = 1020.1> 15. B-lactam according to claim 10. or 11, wherein  <EMI ID = 1021.1> 16. 6-lactam according to claim 10 or 11, wherein  <EMI ID = 1022.1> 17. B-lactam according to one of claims 10 to 16, in which the amino substituent (NH2-) in position 3 is protected.  <EMI ID = 1023.1> the amine substituent is in the form of an azide group.  <EMI ID = 1024.1> formula :  <EMI ID = 1025.1> wherein R3 and R4 are the same or different and are each a hydrogen atom or an alkyl radical, and M is a hydrogen atom or a cation. 20. S-lactam according to claim 19, wherein R3 and R4 are the same or different and are each a hydrogen atom or a methyl radical.  <EMI ID = 1026.1>  <EMI ID = 1027.1> is a hydrogen atom or a cation, in position 1, and an acylamine substituent in position 3.  <EMI ID = 1028.1> in that it carries a sulfonic acid salt substituent <EMI ID = 1029.1> cation, in position 1, an acylamine substituent in position 3, and an alkyl substituent in position 4. 23. e-lactam according to claim 21, characterized in that it carries a salt sulfonic acid substituent  <EMI ID = 1030.1> in position 1, an acylamine substituent in position 3, and a methyl substituent in position 4 <EMI ID = 1031.1> formula :  <EMI ID = 1032.1>  <EMI ID = 1033.1>  <EMI ID = 1034.1>  <EMI ID = 1035.1> hydrogen atom, the other may be an alkenyl, styryl, alkynyl, alkoxy, alkylthio, carboxyl radical or an alkyl ester thereof, hydroxymethyl, lower alkylsulfonylmethyl, phenylsulfonylmethyl, in which the phenyl radical may be substituted by a methyl radical or a halogen, halomethyl, mercaptomethyl atom or a benzyl or triphenylmethyl- derivative  <EMI ID = 1036.1> is a hydrogen atom or a cation. 25. The 8-lactam according to claim 24, wherein R2 is a hydrogen atom. 26. The 8-lactam according to claim 25, wherein R3 and R4 are methyl radicals.  <EMI ID = 1037.1> <EMI ID = 1038.1> 28. 6-lactam according to claim 25, in which R3 is a hydrogen atom and R4 is a methyl radical 29. 8-lactam according to claim 25, in which R3 is a methyl radical and R4 is a hydrogen atom. 30. 0-lactam according to one of claims 24 to 29, in which R1 is an aliphatic group having the formula:  <EMI ID = 1039.1>  <EMI ID = 1040.1> <EMI ID = 1041.1> alkyl or alkenyl substituted by one or more halogen atoms or cyano, nitro, amine, mercapto, groups. alkylthio or cyanomethylthio. 31. (3-lactam according to one of claims 24 to 29, in  <EMI ID = 1042.1> for formula:  <EMI ID = 1043.1>   <EMI ID = 1044.1> independently, a hydrogen or halogen atom or a hydroxyl, nitro, amine, cyano, trifluoro- group  <EMI ID = 1045.1> 4 carbon atoms, or aminomethyl; and Rg is an amine, hydroxyl group, a carboxylic salt, a protected carboxyl radical, formyloxy, a sulfonic salt, a salt sulfoamine, an azide group, a halogen atom, a hydrazine, alkylhydrazine, phenylhydrazine radical or  <EMI ID = 1046.1>  <EMI ID = 1047.1> formula :  <EMI ID = 1048.1> where n is 0, 1, 2 or 3; Rg is an amine, hydroxyl group, a carboxylic salt, a protected formyloxy carboxyl group, a sulfonic salt, a sulfoamine salt, an azide group, a halogen atom, or a hydrazine, alkylhydrazine, phenylhydrazine or [(alkylthio) radical -  <EMI ID = 1049.1> cyclic pentagonal, hexagonal or heptagonal, substituted or unsubstituted, containing 1, 2, 3 or 4 atoms of nitrogen, oxygen or sulfur. 33. P-lactam according to one of claims 24 to 29, in which R. is a group of formula:  <EMI ID = 1050.1>  <EMI ID = 1051.1>  <EMI ID = 1052.1>  <EMI ID = 1053.1> 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 0 <EMI ID = 1054.1> alkyl or alkyl substituted by one or more halogen atoms or cyano, nitro, amine or mercapto groups. 34. $ -lactam according to one of claims 24 to 29, in  <EMI ID = 1055.1>  <EMI ID = 1056.1>  <EMI ID = 1057.1>  <EMI ID = 1058.1>  <EMI ID = 1059.1> hydrogen or halogen atom, a hydroxyl, nitro, amine or cyano group, or a trifluoromethyl radical, alkyl of 1 to 4 carbon atoms, aicoxy 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 atoms  <EMI ID = 1060.1> 0  <EMI ID = 1061.1> alkyl substituted by one or more halogen atoms or  <EMI ID = 1062.1> carboxyl, carboxylic salt, amide, alkoxycarbonyl, phenylmethoxycarbonyl, diphenylmethoxycarbonyl, hydroxyalkoxyphosphinyl, dihydroxyphosphinyl, hydroxy (phenylmethoxy) phosphinyl or dialcoxyphosphinyl. 35. p-lactam according to one of claims 24 to 29, in  <EMI ID = 1063.1>  <EMI ID = 1064.1>  <EMI ID = 1065.1>  <EMI ID = 1066.1>  <EMI ID = 1067.1> hydrogen or halogen atom, a hydroxy, nitro, amine or cyano group, 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 nitrogen atoms,  <EMI ID = 1068.1> formula :  <EMI ID = 1069.1>  <EMI ID = 1070.1> amine, alkylamine, (cyanoalyl) amine, amide, alkylamide, (cyanoalkyl) amide, or a group of formula:  <EMI ID = 1071.1>  <EMI ID = 1072.1> which R is a group of formula:  <EMI ID = 1073.1>  <EMI ID = 1074.1>  <EMI ID = 1075.1>  <EMI ID = 1076.1> 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 (2 ) a substituted or unsubstituted pentagonal, hexagonal or heptagonal heterocyclic ring, - containing 1, 2, 3 or 4 nitrogen atoms,  <EMI ID = 1077.1> 0  <EMI ID = 1078.1> R16 is a hydrogen atom, or an alkyl radical, or <EMI ID = 1079.1> alkyl substituted by one or more halogen atoms or cyano, nitro, amine or mercapto groups. 37. A 3-lactam according to claim 34, wherein  <EMI ID = 1080.1> 38. $ -lactam according to claim 37, which is a  <EMI ID = 1081.1> azetidinesulfonic. 41. $ -lactam according to claim 37, which- is a  <EMI ID = 1082.1> methoxyimino) acetyl] amino] -4-methyl-2-oxo-1-azetidinesulfonic. 42. 5-lactam according to claim 37, which is a  <EMI ID = 1083.1> azetinesulfonic. 43-. 3-lactam according to claim 37 which is a  <EMI ID = 1084.1>  <EMI ID = 1085.1> 45 - B-lactam according to claim 37, which is a salt of the acid [3S- [3a (Z), 4a]] - 3 - [[2-amino-4-thiazolyl) -  <EMI ID = 1086.1> sulfonic. 46 - $ -lactam according to claim 37 which is a  <EMI ID = 1087.1> oxo-1-azetidinesulfonic. <EMI ID = 1088.1> R. is a group of formula:  <EMI ID = 1089.1> 48. A 6-lactam according to claim 47, wherein  <EMI ID = 1090.1>  <EMI ID = 1091.1> 49. $ -lactam according to claim 47, which is a  <EMI ID = 1092.1> 52. B-lactam according to claim 33, in which R. is a group of formula:  <EMI ID = 1093.1>   <EMI ID = 1094.1> azetidinesulfonic. 54. S-lactam according to claim 52, which is a  <EMI ID = 1095.1> 2-oxo-1-azetidinesulfonic.  <EMI ID = 1096.1> 2-oxo-1-azetidinesulfonic. 56. Composition comprising an effective amount  <EMI ID = 1097.1>  <EMI ID = 1098.1> in which R is an acyl radical; R2 is an atom  <EMI ID = 1099.1> R3 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 R3 or R4 is a hydrogen atom, the other can be an alkenyl, styryl, alkynyl, alkoxy, alkoxy, alkylthio, carboxyl radical or an alkyl ester of the latter, hydroxymethyl , alkylsulfonyl-  <EMI ID = 1100.1> phenyl radical may be substituted by a methyl radical or a halogen, halomethyl, mercaptomethyl atom or a benzyl or triphenylmethylthio derivative thereof,  <EMI ID = 1101.1> gene or cation, as well as a pharmaceutically acceptable carrier for 8-lactam. <EMI ID = 1102.1>  <EMI ID = 1103.1> is a hydrogen atom or a cation, in position 1, and a  <EMI ID = 1104.1> in position 3, characterized in that a corresponding 0-lactam carrying a hydrogen substituent in position 1 and a protected amine substituent in position 3 is sulfonated, and in that said protective group is removed from the amine function. 58. Process 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:  <EMI ID = 1105.1>  <EMI ID = 1106.1> acyl or else the group RI-NH- is a protected amine group; R2 is a hydrogen atom or an alkoxy radical  <EMI ID = 1107.1> 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 & l-yl, 2-phenylethenyl radical  <EMI ID = 1108.1> cation, and in that said sulfonation is carried out on a compound of formula:  <EMI ID = 1109.1> or on a precursor compound of formula:  <EMI ID = 1110.1> which is cyclized, said sulfonation reaction being carried out either before or after said cyclization, the R.-NH- group of the reactants being an amine group  <EMI ID = 1111.1> 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. 61. Method according to claim 60, characterized  <EMI ID = 1112.1>  <EMI ID = 1113.1> 62. Method according to claim 60, characterized in that R1 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:  <EMI ID = 1114.1>  <EMI ID = 1115.1> alkoxy; alkenyl; cycloalkenyl; or cyclohexadienyl; or an alkyl or alkenyl radical substituted by one or more halogen atoms or cyano, nitro, amine, mercapto, alkylthio or cyanomethylthio groups; (b) a carbocyclic aromatic group having the formula:  <EMI ID = 1116.1>  <EMI ID = 1117.1> are each, independently, a hydrogen or halogen atom, a hydroxyl, nitro, amine or cyano group, or a trifluoromethyl radical, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms, or aminomethyl ;  <EMI ID = 1118.1> carboxylic, a protected carboxyl radical, formyloxy, a sulfonic salt, a sulfoamine salt, an azide group, a halogen atom, or a hydrazine, alkylhydrazine, phenylhydrazine or [(alkylthio) thioxomethyl] thio radical; <EMI ID = 1119.1>  <EMI ID = 1120.1> where n is 0, 1, 2 or 3; Rg is an amine, hydroxyl group, a carboxylic salt, a protected or formyloxy carboxyl radical, a sulfonic salt, a sulfoamine salt, an azide radical, a halogen atom or a hydrazine, alkylhydrazine, phenylhydrazine or  <EMI ID = 1121.1> substituted or unsubstituted pentagonal, hexagonal or heptagonal heterocyclic tick containing 1, 2, 3 or 4 nitrogen, oxygen or sulfur atoms; (d) a group of formula: '  <EMI ID = 1122.1>  <EMI ID = 1123.1>  <EMI ID = 1124.1>   <EMI ID = 1125.1> hydrogen or halogen atom, a hydroxy, nitro, amine or cyano group, a trifluoromethyl, alkyl radical  <EMI ID = 1126.1> carbon, or aminomethyl, or alternatively (2) a heterocyclic pentagonal, hexagonal or heptagonal ring, substituted or unsubstituted, containing 1, 2, 3 or 4 atoms of nitrogen, oxygen or sulfur; and R12 is a group of O he  <EMI ID = 1127.1> amine, alkyl, or alkyl substituted by one or more halogen atoms or cyano, nitro, amine or mercapto groups; (e) a group of formula:  <EMI ID = 1128.1>  <EMI ID = 1129.1>  <EMI ID = 1130.1>  <EMI ID = 1131.1> hydrogen or halogen atom, a hydroxyl, nitro, amine, or cyano group, or a trifluoromethyl radical, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms, or aminomethyl, or else (2 ) a pentaqonal, hexagonal or heptagonal heterocyclic ring, substituted or unsubstituted, containing 1, 2, 3 or 4 nitrogen atoms,  <EMI ID = 1132.1> or an alkyl, cycloalkyl, alkylaminocarbonyl radical, 0 &#65533; I  <EMI ID = 1133.1> halogen or cyano, nitro, amine, mercapto groups,  <EMI ID = 1134.1> alkoxycarbonyl, phenylmethoxycarbonyl, diphenylmethoxycarbonyl, hydroxyalkoxyphosphinyl, dihydroxyphosphinyl, hydroxy (phenylmethoxy) phosphinyl or dialkoxyphosphinyl; (f) a group of formula:  <EMI ID = 1135.1>  <EMI ID = 1136.1>  <EMI ID = 1137.1>  <EMI ID = 1138.1> 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 (2) a substituted or unsubstituted pentagonal, hexagonal or heptagonal heterocyclic ring containing 1, 2, 3 or 4 nitrogen, oxygen atoms  <EMI ID = 1139.1>  <EMI ID = 1140.1>  <EMI ID = 1141.1> amine radical, alkylamine, (cyanoalkyl) amine, amide, alkylamide, (cyanoalkyl) amide, or a group of formula:  <EMI ID = 1142.1>    <EMI ID = 1143.1> (g) a group of formula:  <EMI ID = 1144.1>  <EMI ID = 1145.1>  <EMI ID = 1146.1>  <EMI ID = 1147.1> 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 else (2) a substituted or unsubstituted pentagonal, hexagonal or heptagonal heterocyclic ring containing 1, 2, 3 or 4 nitrogen atoms,  <EMI ID = 1148.1> 0 ii  <EMI ID = 1149.1> which R16 is a hydrogen atom, or an alkyl radical.  <EMI ID = 1150.1> or alkyl substituted by one or more halogen atoms or cyano, nitro, amine or mercapto groups. 64. The method of claim 63, wherein  <EMI ID = 1151.1> 65. Process for the preparation of an acid salt (R) -3-  <EMI ID = 1152.1> characterized in that a strain of the microorganism Chromobacterium violaceum, A.T.C.C. 31532, in an aqueous medium comprising yeast extract, beef extract, amino NZ A, glucose and agar. 66. Process for the preparation of an acid salt  <EMI ID = 1153.1> strain of the microorganism Gluconobacter, A.T.T.C. 31581 in an aqueous medium comprising yeast extract, beef extract, amino NZ A, glucose and agar. 67. Compound of formula:  <EMI ID = 1154.1>  <EMI ID = 1155.1>  <EMI ID = 1156.1> protected; 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 radical  <EMI ID = 1157.1> or 2-phenylethynyl; 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:  <EMI ID = 1158.1> in which R2 is a hydrogen atom or a radical  <EMI ID = 1159.1> 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 radical -1-yl, 2-phenylethenyl or 2-  <EMI ID = 1160.1> 69. Composed of formula:  <EMI ID = 1161.1> in which R is a hydrogen atom or an acyl group;  <EMI ID = 1162.1> each a hydrogen atom or an alkyl, cycloalkyl, phenyl or substituted phenyl radical, or one of R3 and R &#65533; is a hydrogen atom and the other is an alkoxycarbonyl, alken-1-yl, alkyn-1-yl, 2-phenylethenyl radical  <EMI ID = 1163.1> cation. 70. Process according to claim 57, characterized in that it consists in sulfonating a compound of formula:  <EMI ID = 1164.1>  <EMI ID = 1165.1> gene and the other, optionally, is an alkoxycarbonyl, alken-1-yl, alkyn-1-yl, 2-phenylethenyl or 2phenylethynyl radical; A is a protective group for the amine function; and V is a labile group such as the radical  <EMI ID = 1166.1> a chlorine, bromine or iodine atom, to obtain a compound of formula:  <EMI ID = 1167.1>  <EMI ID = 1168.1> 71. Composed of formula:  <EMI ID = 1169.1> wherein R3 and R4, A and V have the same definitions as in claim 70. 72. Compound of formula:  <EMI ID = 1170.1> in which R2 is a hydrogen atom or an alkoxy radical  <EMI ID = 1171.1> 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- L-yl, 2-phenylethenyl or 2-  <EMI ID = 1172.1>