FI80271C - FOERFARANDE FOER FRAMSTAELLNING AV NYA, THERAPEUTIC ANVAENDBARA 1-SULFO-2-AZETIDINONIDERIVAT, OCH NYA SOM MELLANPRODUKT ANVAENDBARA 2-AZETIDINONDERIVAT. - Google Patents

Description

1 80271

The invention relates to a process for the preparation of novel therapeutically useful 1-sulfo-2-azetidinone derivatives of the general formula I 'and to the preparation of new therapeutically useful 1-sulfo-2-azetidinone derivatives. -.— C-CO-NH - --R, _ji iF * 3 h *

HoN— ^ c / N 2 1 I *

1 b j o -N-SO -sH

Wherein R 9 and R 6 are the same or different and represent hydrogen, C 1-6 alkyl or C 1-6 cycloalkyl or one of R 9 and R 6 is hydrogen and the other is alkynyl-1-yl and R 6 is hydrogen, alkyl, cycloalkyl, alkylaminocarbonyl or alkyl substituted with one or. . with several halogens, cyano, nitro, amino, n-mercapto, alkylthio, carboxyl or its: sodium or potassium salt or its C 1-1 alkyl or benzyl ester, amido, alkoxycarbonyl, phenylmethoxycarbonyl, diphenylmethoxycarbonyl, diphenylmethoxycarbonyl, dihydroxyalkoxyphosphinyl, hydroxy (phenylmethoxy) phosphinyl or dialoxyphosphinyl, and pharmaceutically acceptable salts thereof.

In addition, the invention relates to new, e.g.

·; ·. 2-Azetidinone derivatives of the general formula • useful in the preparation of compounds of the formula I *, which are of the general formula \\ III '·' * · H R, • 35 H N --- 1_ v 2 III '

oA - ^ - sOjH

2 80271 wherein R 9 and R 2 are as defined in formula I ', and salts thereof.

The compounds of formula I1 form a group of novel β-lactam antibiotics and can be used as antibacterial agents. It has been found that β-lactam nuclei can be biologically activated by a sulfonic acid salt substituent attached to said. to the nitrogen atom of the nucleus.

The novel lactams of formula I *, in which 10a has a sulfonic acid salt (including "inner salt") substituent at the 1-position and an acylamino substituent at the 3-position, have activity against gram-negative and gram-positive bacteria.

Many compounds with a 2-azetide-15 non-backbone are known (see e.g. patent publications SE 76 11 103-8, CH 618 161, FI 75 1949, DE 28 30 077, DE 27 51 041 and DE 25 29 941). The most significant of these compounds are the nocardicins known from the Journal of Antiobiotics 29 (1976) 890-900, which have antibacterial activity. Nocardicins differ from the novel compounds of formula 1 'in that they have a phenylacetic acid group in the 1-position instead of a sulfonic acid group. However, according to Kirk-Othmer: Encyclopedia of Chemical Technology, Supplement, 3rd Edition, 1984, John Wiley £ 25 Sons, Inc., pages 131-144, especially page 133, some known nocardines have a very limited antibacterial spectrum.

The terms "alkyl" and "alkoxy" refer to both straight and branched chain groups. Groups of 1 to 10 carbon atoms are preferred.

The term "halogen" refers to fluorine, chlorine, bromine and iodine.

: The term "protected carboxyl" refers to a carboxyl group esterified with a conventional ester protecting group. These groups are known in the art; see, e.g., U.S. Patent 4,144,333. Preferred protected

II

3,80271 carboxyl groups include benzyl, benzhydryl and t-butyl esters.

Preferred are compounds of formula 1 'wherein R 2 is methyl, ethyl, carboxymethyl or 2-carboxyisopropyl.

Of the sulfonic acid salts of the novel compounds of formulas 1 'and III', pharmaceutically acceptable salts are preferred, although other salts are useful in the purification of the products of the invention or as intermediates in the preparation of pharmaceutically acceptable salts. The cationic moiety in the sulfonic acid salts of the invention can be obtained from either organic or inorganic bases. Cationic moieties include, without limitation, the following ions: ammonium, substituted ammonium such as alkylammonium (e.g., tetra-n-butylammonium, hereinafter referred to as tetrabutylammonium); alkali metal such as lithium, sodium and potassium; alkaline earth metals such as calcium and magnesium; pyridinium; dicyclohexylammonium; hydrabaminium; bentsatinium; 20 N-methyl-D-glucaminium.

Some of the compounds of formulas I * and III * may be crystallized or recrystallized from aqueous solvents. Hydration water may then form.

i i The compounds of the formulas I1 and III1 may be both: *: 25 as stoichiometric hydrates and compounds having: V: varying amounts of water which may be formed e.g.

during lyophilization.

The compounds of formulas I * and III 'have at least one chiral center - a carbon atom (in the 3-position of the β-lactam-30 core) to which an amino or acylamino substituent is present. support has joined. In the compounds of formulas 1 'and III', the stereochemistry of the β-lactam nuclei at the 3-position chiral center is the same as the configuration of naturally occurring penicillins (e.g. penicillin G) at the 6-position carbon atom and the configuration of naturally occurring penicillins. cefamycins (e.g. cefamycin C) * 80271 at the 7-position carbon atom.

Preferred γ-lactams of formulas 1 'and III * have the following stereochemistry at the 3-position chiral center: 5' * 3 -NH-C to C-R.

| 3 4! 4

JZ --N-SO-H

10 ^ 3 Xu 0

According to the nomenclature, such compounds of formulas 1 'and III' have the S-configuration.

The invention also includes racemic mixtures containing the β-lactams described above.

The β-lactams of the formula having the sulfonic acid salt substituent at the 1-position of the β-lactam core and the 3-position of the acylamino substituent β-lactam core have gram-negative and gram-positive activity. against organisms. The sulfonic acid salt substituent is essential for the activity of the compounds of formula 1 '. For compounds wherein and / or R 1 are; hydrogen or alkyl, especially methyl, is a particularly useful activity.

The compounds of formula I * may be used to treat bacterial infections (including urinary tract infections and respiratory infections) in mammals such as domestic animals (e.g. dogs, cats, cattle, horses and the like) and in humans.

The process according to the invention for the preparation of new 1-sulfo-2-azetidinone derivatives of the formula 1 'is characterized in that the corresponding 2-azetidinone derivative having a hydrogen substituent in the 1-position is sulfonated.

II

5 80271 H R3 N -.- C-CO-NH --- Ru 5 V-XT n J_I IV 'C ~ R13 <f or acylation of the corresponding 2-azetidinone derivative having an amino substituent in the 3-position 10 H R3 Η? Ν g ζ 1 ύ III '2 1

li-- N-SO, H

0 15 -1

The β-lactams of formulas 1 'and III' are generally prepared by attaching a sulfonic acid substituent (sulfo group -SO 2 -) to the nitrogen atom of the lactam nucleus at the 1-position. This sulfonation reaction is readily accomplished by treating the lactam with a sulfur trioxide complex or an equivalent sulfonating reagent such as chlorosulfonate.

The most commonly used sulfur trioxide complexes are pyridine sulfur trioxide; lutidine-sulfur trioxide; Dimethylformamide sulfur trioxide; and picoline sulfur trioxide. Instead of a preformed complex, the complex can be formed in situ, e.g., using chlorosulfonyl-trimethylsilyl ester and pyridine as reagents. Alternatively, sulfonation can be accomplished by an intermediate compound, such as, for example, first silylating the nitrogen atom of the β-lactam core and then subjecting the silylated compound to a silyl exchange reaction with trimethylsilyl chlorosulfonate or the like. Exemplary silylating agents include monosilyl 35 litrifluoroacetamide, trimethylsilyl chloride / triethylamine, and bis-trimethylsilyl trifluoroacetamide.

6 80271

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 polar solvents such as dimethylformamide and a halogenated hydrocarbon such as dichloromethane.

The product formed at the beginning of the suonization reaction is the salt of the sulfonated β-lactam.

When the sulfonating complex is pyridine-sulfur trioxide, the product initially formed is the β-lactam-sulfonated-pyridine salt of the sulfonated, β-lactam 10, where M + is the pyridium ion in the following formula ί— --N-SO "M + 15 0 These complexes can be converted to other sulfonic acid salts using conventional methods (e.g., ion exchange resins, crystallization, or ion pair extraction) These extraction methods are also useful in the purification of products. • with tetrabutylammonium hydrogen sulfate, or as an amphoteric ion (M + = hydrogen) using formic acid, are particularly useful.

It should be emphasized that a sulfonation reaction in which a sulfone group is attached to the nitrogen atom of a β-lactam nucleus can be performed at various stages of the synthesis, including coupling prior to β-lactam nucleus formation, such a method being performed as described below: in the presence and usually at room temperature. When an amino group is present, it is preferred that it be protected.

• - 35 When, for example, benzyloxycarbonyl-

II

7 80271 protecting groups The sulfonation reaction can be represented as follows:

i Ru R

i4 H R1 + 5 C6H5CH2OCO-NH --- c R3 H N_i_ - *. ά 1) sulfonate .0- NH 2) deprotection - ~ --N-SO "M + ^ ^ 3

II 0 III

10

Other protecting groups may be used to protect the amine group, for example, a t-butyloxycarbonyl group, a simple acyl group such as acetyl or benzoyl or phenylacetyl, a triphenylmethyl group, or the amino group is in the form of an azide group. The desired acyl group (R 1) can be obtained by a conventional acylation reaction.

In the process according to the invention, a 2-azetidinone of the formula IV having an acyl group (R1) 3-ac-mass is sulfonated to give a compound of the formula 1 '. "as follows: H h, H Rn 25 RrNH - t" R3 _, VNH - 1 - R3___H sulfonation --N-S0 "M + o"

Examples of acylation processes by which compounds of formula III 'can be converted to products of formula 1' are reactions with a carboxylic acid (R 1 -OH) or the corresponding carboxylic acid halide or carboxylic anhydride. The reaction with the carboxylic acid is best accomplished in the presence of a carbodiimide such as dicyclo-35-hexylcarbodiimide and an active ester-forming agent such as N-hydroxybenzotriazole. When the acyl group (R 1) contains a reactive group (such as an amino or carboxyl group), it may be necessary to first protect the functional groups, then perform the acylation reaction, and finally deprotect the product.

The β-lactam ring of the starting materials can be formed by a cyclization reaction, and any sulfonation reaction that can be performed before or after the cyclization.

V _> R

cyclization of acyl-NH ------ ^ acyl-NH --------- r_ ^ 15 J. NH-Sortk X - N-SO 3 - «+ cT °"

VII

The acyl group in this reaction may also be an easily removable group which is a protecting group and which, when removed, gives the -NH 2 - product.

Azetidinone starting materials in which at least one of the groups and R 1 is hydrogen can also be prepared from amino acids of formula - OH

25 r NH2 \ ^ ^ 4 'PH-C \

\ r3 XII

C- OH

0 '' 30 (at least one of R 1 and R 2 is hydrogen). The amino group is first protected with a classical protecting group, e.g., t-butoxycarbonyl (hereinafter referred to as "Boc"). The carboxyl group of the protected amino acid is then reacted with an amine salt of formula

II

9 80271

Y-O-NH * Cl "XIII

with formula wherein Y is alkyl or benzyl in the presence of carb-5-bodiimide to give a compound of formula 0H

Boc-NH-CH --- Q

10 I ^ R3 Xlv

C-NH-O-Y

(at least one of the groups and is hydrogen). The hydroxyl group of formula XIV is converted to a leaving group 15 (V) with a classical reagent, e.g. methanesulfonyl chloride (methanesulfonyl is hereinafter referred to as "Ms"). Other leaving groups (V) that can be used are benzenesulfonyl, toluenesulfonyl, chlorine, bromine and iodine.

A fully protected compound of the formula

V

I ^ / Ri +:; Boc-NH-CH-n ö 1

2 5 -NH-O-Y

O '' (at least one of R 1 and R 2 is hydrogen) is cyclized by treatment with a base, e.g. potassium carbonate. The reaction is preferably carried out in an organic solvent such as acetone under reflux conditions to give a compound of formula s

Boc-NH - j --— R3

35 I XVI

£ _N-O-Y

<Y '10 80271 (at least one of the groups and R 1 is hydrogen).

Alternatively, cyclization of a compound of formula XIV may be performed without first converting the hydroxyl group to a leaving group. Treatment of a compound of formula XIV 5 with triphenylphosphine and diethyl azodicarboxylate gives a compound of formula XIV wherein at least one of the groups is hydrogen.

Removal of the azetidinone protecting group 10 from the 1-position of formula XVI can be accomplished by reduction of sodium to give Y as alkyl to give an intermediate of formula 15 Boc-NH --- R3

XVII

-NH

(At least one of R 3 and R 2 is hydrogen).

If Y is benzyl, catalytic (e.g. palladium on carbon) hydrogenation initially gives the corresponding N-hydroxy compound which, after treatment with titanium chloride, gives an intermediate of formula XVII in which at least one of the groups and R 1 is hydrogen.

The synthesis comprising the ring closure described above gives an inversion of the stereochemical configuration of the R 1 and R 2 substituents.

As previously mentioned, the above azetidinone 30 can be sulfonated to a compound of the following formula - b

Boc-NH ----- R

35. XVIII

J-N-SO “M

0?

II

il 80271 (at least one of R 3 and R 2 is hydrogen).

In an alternative process for the preparation of a compound of formula 1 ', wherein at least one of R 3 and. is hydrogen, the starting material is an amino acid amide of formula

OH

I / "· nh2-ch-

^^ r3 XIX

10 0 ^ C-N »! (at least one of R 1 and R 2 is hydrogen).

Protection of the amino group with a classical protecting group, e.g. benzyloxycarbonyl (hereinafter referred to as Z) or Boc, and conversion of the hydroxyl group to a leaving group (V) such as Ms gives the compound OMs of formula XX; 20 ANH-CH-4

I XX

-NH 2 O 3 (at least one of R 1 and R 2 is hydrogen), wherein A is a protecting group.

Sulfonation of a compound of formula XX gives a compound of formula OMs 30 ANH-CH-

I R3 XXI

-NHSO "M

35 (at least one of R 3 and R 2 is hydrogen).

12 80271

The cyclization of a compound of formula XXI is carried out with a base, e.g. potassium carbonate. The reaction is preferably carried out in a mixture of water and an organic solvent (e.g. a halogenated hydrocarbon such as 1,2-dichloroethane) under reflux conditions to give a compound of formula XXII §4 ANH ---- R3

10 XXII

J-N-SO "M

O '3 (at least one of R 1 and R 2 is hydrogen).

Deprotection of a sulfonated azetidinone of formula XXII wherein A is a protecting group by catalytic hydrogenation gives a compound of formula H R 4 2 0 h 2n - i-r 3

-N-SO "M + XXIII

; Wherein at least one of R 1 and R 2 is hydrogen and which can be converted to the corresponding amphoteric ion of formula d r 4 nh 3 H-f- * 3

30 I XXIV

J-N-SO

O 3 (at least one of R 1 and R 2 is hydrogen) by treatment with an acid such as formic acid.

Il 13 80271

Deprotection of a sulfonated azetidinone of formula XXII wherein A is a Boc protecting group using acidic conditions (e.g. using formic acid) affords the corresponding amphoteric ion of formula XIV.

An excellent source for β-lactam starting materials are 6-amino-penicillanic acids and 7-amino-cephalosporanic acids, which may have a possible 6-alkoxy or 7-alkoxy substituent, respectively. These compounds have the formulas / CH3 vnh_-L- | ^ Nj ^ CH3 15 J-N-C-COOH XXVa

CT

and R3 ri_NH - S S v 2 0 _ J --- CHa XXVb / KY 3

o I

COOH

- - wherein R 2 is hydrogen and is hydrogen or acyl. Applying the methods described in the literature, 3-amino-2-azetidinones can be prepared, see e.g. Chem. Soc. Special Publication No. 28, pp. 288 (1977); The Chemistry of Penicillins, Princeton Univ. Press., P. 257, and Synthesis, 494 · (1977).

6-Amino-penicillanic acid or 7-amino-cephalosporanic acid is first desulfonated by reduction using

Raney-nickel. The reaction can be carried out in water under reflux conditions; the compound obtained has the formula R2 R1-NH--

--- N-CH-CH (CH „) 0 XXVI

0 ^. 32

^ COOH

35 14 80271

Substitution of the carboxyl group of the compound of formula XXVI with an acetate group followed by hydrolysis affords a 3-amino-2-azetidinone of formula 1 'having hydrogen or acyl and R 9 and R 6 are hydrogen.

Treatment of a compound of formula XXVI with copper acetate and lead tetraacetate in an organic solvent (e.g. acetonitrile) replaces the carboxyl group with an acetate group. The hydrolysis of the obtained compound can be carried out using potassium carbonate in the presence of sodium borohydride.

Coupling of the sulfo group to the 1-position of the 3-amino-2-azetidinone can be accomplished by reacting the intermediates with a complex of dimethylformamide and sulfur trioxide.

The starting material 3-azido-2-azetidinone can be prepared by first administering an olefin of formula

K

I XXVII

CH 2: --- C-R 3 20 to react with a halosulfonyl isocyanate (preferably chlorosulfonyl isocyanate) of formula

0 —C = N— SO2-halogen XXVIII

25 to give azetidinone of formula

S

- R3

30 XXIX

A - N-S02-halogen

QT

Reductive hydrolysis of an azetidinone of formula XXIX affords an N-unsubstituted lactam of formula 35.

-rR3

XXX

s ^ -NH

For a more detailed description of the reaction sequence described above, reference is made to the literature; see e.g. Chem. Soc. Rev., 5, 181 (1976) and J. Org. Chem., 35, 102043 (1970).

The azido group can be attached to the 3-position of an azetidinone of formula XXX (or a corresponding sulfonated compound) by reacting the compound with an arylsulfonyl azide (such as toluenesulfonyl azide) to give the starting azetidinone of formula N3 -hR3

on XXXI

2 0 Λ-tiH

CT

to obtain.

The reaction is best performed by first protecting the azetidinone nitrogen with a silyl residue (e.g., t-butyldimethylsilyl or t-butyldiphenylsilyl), then forming the anion at the 3-position of the core with a strong organic base (e.g., lithium diisopropylamine) at low temperature and treatment at low temperature. anion with toluenesulfonyl azide. The resulting intermediate is quenched with trimethylsilyl chloride and then acid hydrolysis or fluoride solvolysis of the N-protecting group affords the compound of formula XXXI.

Alternatively, a compound of formula XXXI can be obtained by first reacting a primary amine of formula 80271 H2N-CH2-O-O-alkyl or H2N- (O-alkyl) O-alkyl with an aldehyde of formula R1-CH2O to give the corresponding Schiff base. A / 2 + 2 / with the activated form of cycloaddithio-azido-acetic acid gives 3-azido-2-10 azetidinone of formula

RU

N3 - | -R3

15 J-N-Q

wherein Q is 20 H -O 2 CH 2 O -alkyl or O-alkyl O-alkyl Oxidative removal of the Q Q substituent gives a compound of formula XXXI.

3-Acylamino-2-azetidinones can be obtained by first reducing the 3-azido-2-azetidinone of formula XXXI to give the corresponding 3-amino-2-azetidinone and then acylating the 3-amino-2-azetidinone.

The compounds of the invention 1 'can be used to treat bacterial infections (including urinary tract infections and respiratory infections) in mammals, such as domestic animals (e.g., dogs, cats, cattle, horses, and the like) and humans.

I! I? 80271

For the treatment of bacterial infections in mammals, a compound of formula I * may be administered to a mammal at about 1.4 mg / kg / day to 50 mg / kg / day, preferably from about 14 mg / kg / day to 100 mg / kg / day. All of the forms of antam-5 used heretofore to deliver penicillins and cephalosporins to the site of infection have also been found to be suitable for use in the β-lactams of formula 1 '. Such forms of administration include oral, intravenous, intramuscular and suppository.

10 Biological activity

The following method is used to determine the minimum inhibitory concentration (hereinafter MIC) for the β-lactams of the invention. The test organisms are cultured in about 15-20 ml of antibiotic test broth (Difco) by inoculating (in tubes) the broth with a looped organism from a BHI (Difco) agar slant culture. The inoculated tubes are cultured at 37 ° C for 9 to 18 hours. These cultures are assumed to contain 10 column forming units (hereinafter CFU) per milliliter. Cultures are diluted 1:10 to 14.

20 bulging inocula to obtain 10 CFU; dilutions are made with K-10 broth *.

The compounds are dissolved in a suitable diluent at a concentration of 1000 ug per milliliter. Double dilutions are made in K-10 broth to give a range of 25 to 1000 pg / ml to 0.5 ug / ml. Place 1.5 ml of each dilution in separate square petri dishes to which 13.5 ml of K-10 agar ** is added. The final drug concentration in agar is 100 to 0.5 μg / ml. Growth control plates containing only agar and inoculated before and after the test plates are also prepared. The organisms are applied to the agar surface of each plate with a Denley Multipoint Inculator (which dispenses approximately 0.001 ml of each organism) to give a final inoculum level of 10 CFU on the agar surface.

The plates are cultured at 37 ° C for 18 hours and 80271 MIC are determined. The MIC is the lowest concentration of a compound to inhibit the growth of an organism.

* K-10 broth is a yeast broth containing: Meat extract 1.5 g 5 Yeast extract 3.0 g

Peptonia 6.0 g

Dextrose 1.0 g

Distilled water up to 1 liter ** K-10 agar contains 10 Meat extracts 1.5 g

Yeast extract 3.0 g

Peptonia 6.0 g

Dextrose 1.0 g

Agar 15.0 g 15 Distilled water up to 1 liter

The following tables table the results obtained when testing the β-lactams according to the invention against various organisms. The number indicated after each organism means the number of the organism in the collection: 20 mass E.R. Squibb S Sons, Inc., Princeton, New Jersey.

The dash (-) in the tables indicates that the test compound had no activity against a particular organism at 100 ug / ml. "N.T." means not studied.

Il 19 80271 M.I.C. (pg / ml) _Product by example_

Organismi_2_4_6_7_8_

Staphylococcus aureus 1276 6.3 12.5 12.5 25 12.5

Staphylococcus aureus 2399 6.3 6.3 6.3 25 6.3

Staphylococcus aureus 2400 3.1 12.5 6.3 12.5 6.3

Staphylococcus aureus 10165 3.1 25 -

Streptococcus faecalis 9011 - - 100 - 100

Streptococcus agalactiae 9287 25 6.3 1.6 12.5 1.6

Micrococcus luteus 2495 50 50 1.6 25 0.8

Escherichia coli 8294 - - 0.4 25 0.4

Escherichia coli 10857 - - 0.8 50 0.8

Escherichia coli 10896 - 100 1.6 50 3.1

Escherichia coli 10909 - 50 0.4 25 0.2

Klebsiella aerogines 10440 - - 0.4 25 0.4

Klebsiella pneumoniae 9527 - - 0.1 12.5 0.2

Proteus mirabilis 3855 - 100 0.2 12.5 0.1

Proteus rettgeri 8479 100 12.5 <0.05 1.6 <0.05; Proteus vulgaris 9416 - - 0.4 25 0.1 • .-. Salmonella typhosa 1195 - 100 0.1 12.5 <0.05 - · Shigella somnei 8449 - - 0.4 25 0.8

Enterobacter cloacae 8236 - - 0.4 - 0.4

Enterobacter aerogenes 10078 - - 0.8 100 0.8

Citrobacter freundii 9518 - - 3.1 - 3.1

Serratia marcescens 9783 - 50 12.5 - 3.1

Pseudomonas aeruginosa 9545 - - 0.8 50 0.4

Pseudomonas aeruginosa 8329 - - 12.5 12.5

Acinatobacter calcoaceticus 8333 - 100 25 - 100 20 80271 M. I. C. (pg / ml) _Product by example_

Organisms_9 10 11 12_14

Staphylococcus aureus 1276 12.5 12.5 - 12.5 12.5

Staphylococcus aureus 2399 12.5 25 - 12.5 6.3

Staphylococcus aureus 2400 12.5 25 - 25 6.3

Staphylococcus aureus 10165 100 100 - 100 25

Streptococcus faecalis 9011 100 25--

Streptococcus agalactiae 9287 1.6 0.4 12.5 0.4 1.6

Micrococcus luteus 2495 6.3 3.1 25 12.5 3.1

Escherichia coli 8294 0.8 12.5 0.4 12.5 1.6

Escherichia coli 10857 0.8 1.6 0.2 6.3 1.6

Escherichia coli 10896 1.6 3.1 0.8 12.5 6.3

Escherichia coli 10909 0.8 1.6 0.1 3.1 0.8

Klebsiella aerogines 10440 0.8 25 0.4 12.5 3.1

Klebsiella pneumoniae 9527 0.8 1.6 <0.05 6.3 0.8

Proteus mirabilis 3855 0.8 1.6 <0.05 6.3 1.6

Proteus rettgeri 8479 <0.05 0.2 <0.05 0.8 0.1

Proteus vulgaris 9416 1.6 1.6 0.1 12.5 3.1

Salmonella typhosa 1195 0.8 0.8 <0.05 3.1 0.8

Shigella somnei 8449 0.8 3.1 0.2 6.3 1.6 - - Enterobacter cloacae 8236 1.6 6.3 0, 8 6.3 3, 1

Enterobacter aerogenes 10078 0.8 50 0.8 12.5 3.1

Citrobacter freundii 9518 6.3 25 1.6 - 25

Serratia marcescens 9783 50 12.5 0.8 50 25

Pseudomonas aeruginosa 9545 3.1 3.1 0.8 12.5 6.3

Pseudomonas aeruginosa 8329 25 50 3.1 50

Acinatobacter calcoaceticus 8333 25 - 100 - 100

II

21 80271 M. I. C. (pg / ml) _Product by example_

Organismi_15_16 17_18_19

Staphylococcus aureus 1276 25 25 50 3.1

Staphylococcus aureus 2399 25 12.5 50 3.1 100

Staphylococcus aureus 2400 25 12.5 50 3.1 100

Staphylococcus aureus 10165 - 50

Streptococcus faecalis 9011 - 25 - 25 -

Streptococcus agalactiae 9287 3.1 1.6 12.5 0.1 25

Micrococcus luteus 2495 0.8 0.4 25 0.1 100

Escherichia coli 8294 12.5 12.5 1.6 12.5 1.6

Escherichia coli 10857 1.6 1.6 1.6 0.8 1.6

Escherichia coli 10896 12.5 6.3 6.3 12.5 3.1

Escherichia coli 10909 3.1 1.6 0.8 1.6 0.4

Klebsiella aerogines 10440 12.5 6.3 1.6 25 1.6

Klebsiella pneumoniae 9527 3.1 1.6 0.4 6.3 0.8

Proteus mirabilis 3855 0.8 0.4 0.4 1.6 1.6

Proteus rettgeri 8479 0.2 0.2 <0.05 0.2 <0.05

Proteus vulgaris 9416 0.1 0.2 0.8 0.4 0.8 - - Salmonella typhosa 1195 3.1 0.2 0.2 3.1 0.4

Shigella somnei 8449 12.5 6.3 1.6 6.3 0.8; Enterobacter cloacae 8236 12.5 6.3 0.8 12.5 0.8

Enterobacter aerogenes 10078 25 12.5 1.6 25 1.6 - Citrobacter freundii 9518 12.5 6.3 25 12.5 1.6

Serratia marcescens 9783 12.5 6.3 50 25 6.3

Pseudomonas aeruginosa 9545 25 6.3 3.1 1.6 1.6

Pseudomonas aeruginosa 8329 - 100 50 100 12.5

Acinatobacter calcoaceticus 8333 - - 22 80271 M. I. C. (pg / ml) _Product by example_

Organisms_2 0 21_23_2 <4 2 5

Staphylococcus aureus 1276 3.1 6.3 25 100 100

Staphylococcus aureus 2399 3.1 6.3 25 100 50

Staphylococcus aureus 2400 3.1 12.5 12.5 100 50

Staphylococcus aureus 10165 50 12.5 - - 100

Streptococcus faecalis 9011 50 - 100

Streptococcus agalactiae 9287 1.6 3.1 3.1 3.1 100

Micrococcus luteus 2495 3.1 6.3 1.6 3.1

Escherichia coli 8294 25 25 1.6 100

Escherichia coli 10857 3.1 25 0.4 25

Escherichia coli 10896 25 12.5 1.6 25

Escherichia coli 10909 12.5 12.5 0.4 25

Klebsiella aerogines 10440 50 25 3.1 100

Klebsiella pneumoniae 9527 12.5 25 0.8 100

Proteus mirabilis 3855 12.5 12.5 0.2 25

Proteus rettgeri 8479 1.6 1.6 <0.05 3.1

Proteus vulgaris 9416 3.1 25 0.2 50

Salmonella typhosa 1195 12.5 25 0.2 50

Shigella somnei 8449 25 12.5 1.6 50

Enterobacter cloacae 8236 50 25 1.6.! Enterobacter aerogenes 10078 100 25 6, 3

Citrobacter freundii 9518 50 25 3.1

Serratia marcescens 9783 100 25 3.1 100

Pseudomonas aeruginosa 9545 12.5 25 3.1

Pseudomonas aeruginosa 8329 - 100 25 -

Acinatobacter calcoaceticus 8333 - 50 - -

II

23 80271 M.I.C. (pg / ml) _Product by example_

Organisms_2 6 2 7 2 8_29_

Staphylococcus aureus 1276 100 100

Staphylococcus aureus 2399 100 100

Staphylococcus aureus 2400 100 50 -

Staphylococcus aureus 10165 ---

Streptococcus faecalis 9011 ---

Streptococcus agalactiae 9287 100 3.1 50 50

Micrococcus luteus 2495 100 3.1 12.5 100

Escherichia coli 8294 - 0.1 0.4 3.1

Escherichia coli 10857 - <0.05 <0.05 0.8

Escherichia coli 10896 - 0.1 0.2 3.1

Escherichia coli 10909 - <0.05 <0.05 0.8

Klebsiella aerogines 10440 - 0.2 0.4 3.1

Klebsiella pneumoniae 9527 - 0.1 <0.05 0.8

Proteus mirabilis 3855 - 0.1 <0.05 0.4

Proteus rettgeri 8479 - <0.05 <0.05 <0.05

Proteus vulgaris 9416 - <0.05 <0.05 0.4: · .: Salmonella typhosa 1195 - <0.05 <0.05 0.4

Shigella somnei 8449 - 0.1 0.2 1.6

Enterobacter cloacae 8236 - 0.2 0.4 0.8

Enterobacter aerogenes 10078 - 0.4 0.4 3.1

Citrobacter freundii 9518 - 0.1 0.4 3.1. , Serratia marcescens 9783 - 0,8 0,2 6,3 '' Pseudomonas aeruginosa 9545 - 0,8 0,8 6,3

Pseudomonas aeruginosa 8329 - 100 3.1 50

Acinatobacter calcoaceticus 8333 - 12.5 100 2.1 80271 M.I.C. (pg / ml) _Product by example_

Organismi_30_32_34_35_3 8

Staphylococcus aureus 1276 100 100 6.3 - 6.3

Staphylococcus aureus 2399 100 50 6.3 - 6.3

Staphylococcus aureus 2400 50 50 12.5 - 25

Staphylococcus aureus 10165 --50--

Streptococcus faecalis 9011 -----

Streptococcus agalactiae 9287 3.1 1.6 0.8 50 3.1

Micrococcus luteus 2495 6.3 6.3 1.6 25 3.1

Escherichia coli 8294 0.2 0.1 50 0.4 3.1

Escherichia coli 10857 <0.05 <0.05 0.4 0.2 <0.05

Escherichia coli 10896 0.2 0.1 6.3 0.2 0.8

Escherichia coli 10909 0.1 <0.05 6.3 <0.05 0.4

Klebsiella aerogines 10440 0.4 0.2 - 0.8 12.5

Klebsiella pneumoniae 9527 0.1 <0.05 12.5 0.1 0.4

Proteus mirabilis 3855 <0.05 <0.05 6.3 <0.05 1.6

Proteus rettgeri 8479 <0.05 <0.05 1.6 <0.05 1.6

Proteus vulgaris 9416 <0.05 <0.05 1.6 <0.05 <0.05

Salmonella typhosa 1195 <0.05 <0.05 6.3 <0.05 1.6

Shigella somnei 8449 0.2 0.1 25 0.4 1.6

Enterobacter cloacae 8236 0.2 0.1 25 0.8 1.6

Enterobacter aerogenes 10078 0.4 0.2 100 0.8 6.3

Citrobacter freundii 9518 0.2 0.1 25 0.8 3.1

Serratia marcescens 9783 0.4 0.2 50 0.4 3.1

Pseudomonas aeruginosa 9545 0.8 0.4 12.5 0.8 3.1

Pseudomonas aeruginosa 8329 - 100 - 3.1 100

Acinatobacter calcoaceticus 8333 50 25 - 50 50

II

25 80271 M. I. C. (Pg / ml)

Product by example_

Organisms_39 UI_42 44_4 5

Staphylococcus aureus 1276 - - 25

Staphylococcus aureus 2399 - - - 12.5

Staphylococcus aureus 2400 - - - 25

Staphylococcus aureus 10165 -

Streptococcus faecalis 9011 -

Streptococcus agalactiae 9287 12.5 - 12.5 6.3 50

Micrococcus luteus 2495 12.5 - 100 6.3 25

Escherichia coli 8294 0.2 50 6.3 3.1 0.4

Escherichia coli 10857 0.1 6.3 3.1 0.4 0.1

Escherichia coli 10896 0.2 25 3.1 12.5 0.8

Escherichia coli 10909 N.T. 25 1.6 0.8 <0.05

Klebsiella aerogines 10440 0.4 100 6.3 1.6 0.2

Klebsiella pneumoniae 9527 0.05 6.3 6.3 0.2 <0.05

Proteus mirabilis 3855 0.05 12.5 3.1 0.4 <0.05

Proteus rettgeri 8479 0.05 0, 8 0.4 0.4 <0.05

Proteus vulgaris 9416 0.05 12.5 6.3 0.2 <0.05

Salmonella typhosa 1195 0.05 12.5 3, 1 0.2 <0.05 • Shigella somnei 8449 0.1 25 3, 1 6.3 0.1

Enterobacter cloacae 8236 0.1 100 3.1 6.3 0.1 ---. Enterobacter aerogenes 10078 0.8 100 6.3 6.3 0.8

Citrobacter freundii 9518 0.4 25 3.1 3.1 0.4

Serratia marcescens 9783 0.2 100 3.1 3.1 0.8

Pseudomonas aeruginosa 9545 0.4 - 50 6.3 0.4

Pseudomonas aeruginosa 8329 1.6 - - 50 1.6

Acinatobacter calcoaceticus 8333 25 - 50 100 26 80271

Example 1

^ \ V

(S) -2-Oxo-3 - <{E (phenylmethoxy) carbonyl / amino] -1-azetidinesulfonic acid, pyridine (1: 1) salt Method I: 5 A) 1 - / (1R) -carboxy- 2-methyl (propyl) -7-2-oxo- (3S) -N- (phenylmethoxy) carbonyl-amine-4-azetidine

A slurry of 6-aminopenicillanic acid (12.98 g, 0.06 mol) in 140 ml of water containing 5.18 g of sodium bicarbonate (stirred for about 10 minutes without the reconstituted solution) is added in one portion to a well-stirred ( mechanically stirred) Raney nickel suspension (washed with water to pH 8.0; 260 ml slurry = 130 g) at 70 ° C in an oil bath. After 15 ml to 15 minutes, the slurry is cooled, filtered and the filtrate is treated with 5.18 g of sodium bicarbonate and a solution of 11.94 g (0.07 mol) 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, evaporated and triturated to dryness with ether-hexane to give a total of 6.83 g of the title compound.

B) 1- (C (acetyloxy) -2-methyl (propyl} -2-oxo-25 (3S) - (N- (phenylmethoxy) carbonyl) amino) 2) 'azetidine

A solution of 6.83 g (0.0213 moles) of the above acid in 213 ml of acetonitrile is treated with 1.95 g (0.0107 moles) of copper acetate mono-: hydrate and 9.5 g (0 , 0213 moles) of lead tetraacetate. The slurry is immersed in a 65 ° C oil bath and

II

27 80271 is mixed with a stream of nitrogen bubbling through the slurry until the starting material is consumed. The slurry is filtered and the solid is washed with ethyl acetate. The combined filtrate and washings are evaporated in vacuo and the residue is taken up in 100 ml of each of ethyl acetate and water and adjusted to pH 7. The ethyl acetate layer is separated, dried and evaporated to give 6.235 g of the title compound.

C) (S) - (2-Oxo-3-azedidinyl) carbamic acid, phenylmethyl ester

A solution of 3.12 g (0.0093 mol) of the above acetate in 70 ml of methanol and 7 ml of water is cooled to -15 ° C and 1.33 g of potassium carbonate and 349 mg of sodium borohydride are added.

The reaction mixture is stirred at -15 ° C to 0 ° C.

When the reaction is complete (about 2 hours), the mixture is neutralized to pH 7 with 2N HCl 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 obtained in a similar experiment and mixed with ether to give 3.30 g of the title compound.

D) (S) -2-Oxo-3-£ (phenylethoxy) carbonyl / amino-25-1-azedidine acid, pyridine salt

Method I:

A solution of 440 g (0.002 moles) of the above azetidinone in 2 ml of both dry methylene chloride and dry dimethylformamide is stirred for 2 hours under nitrogen with 350 g (0.0022 moles) of: pyridine-sulfur trioxide complex . Most of the solvent is removed in vacuo and the residue is mixed with ethyl acetate to give 758 mg of a solid which is (mainly) the title compound.

28 80271 NMR (D 2 q-CD 3 OD) 3.63 (1H, d, J = 6.4), 3.90 (1H, t, j = 6), 4.85 (1H, dd, J = 6.4) , 5.10 (2H, S) 7.27 (5H, S) 8-0-9. ppm (M's 5H).

Method II: Chlorosulfonyltrimethylsilyl ester (18.87 g) is added dropwise at -20 ° C to 7.9 g of anhydrous pyridine with stirring under a nitrogen atmosphere.

After the addition is complete, stirring is continued for 30 minutes at room temperature and the trimethylchlorosilane is then removed in vacuo. A solution of 20 g of the above azetidinone (Method I, Part C) in 120 ml of methylene chloride is added and stirring is continued at ambient temperature for 3.5 hours. The solvent is distilled off in vacuo and the oily residue is crystallized by adding ethyl acetate to give 31 g of the title compound. NMR values are identical to the product of Method I.

Example 2 (S) -2-Oxo-3- [N- (phenylmethoxy) carbonyl] amino] 7-20 azetidinesulfonic acid, potassium salt

Method I: •: (S) -2-Oxo-3 - [(1-phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid, pyridine salt (135 mg; see Example 1) is dissolved in 2 ml of 0.5 M mono- to 25-basic potassium phosphate ( adjusted to pH 5.5 with 2N potassium hydroxide) and applied to a 25 ml HP-20AG column. The column is eluted using 100 ml of buffer solution, 200 ml of water and 100 ml of 1: 1 acetone-water. Fractions (25 mL) 14-15 are very Rydon-posi-30 tight. Evaporation gives 80 mg of the substance, which is primarily the title compound (spectral values are identical to the product prepared below).

Il 29 80271

Method II: (S) -2-Oxo-3- (phenylmethoxy) carbonylamino] -1-azetidinesulfonic acid, pyridine salt (600 mg; see Example 1) is dissolved in 2 ml of water and stirred in 15 ml of water. (pH 5.5) with monobasic potassium phosphate buffer solution. The solids and slurry are cooled to 0 ° C, filtered, washed with cold buffer, cold 50% ethanol, ethanol and ether to give 370 mg of the title compound (containing excess potassium ion by analysis). A solution of 280 mg of salt in 10 ml of water is applied to a 100 ml HP-20 column. The column is eluted with 200 ml of water and then with aqueous acetone (9: 1). Fractions (50 ml) are collected; evaporation of fraction 7 gives a solid. Mixing with acetone, filtering and drying in vacuo gives 164 mg of the title compound, mp 193-196 ° C.

Anal. Calculated for OgSK * 1 / 2H2O: C, 38.02; 20 H, 3.48; N, 8.06; S, 9.23; K, 11.25 // Found: C, 38.19; H, 3.24; N, 8.15; S, 9.12; ;; γ K, 11.53.

NMR (D 2 O) 3.69 (1H, d, d, J = 6.4), 3.91 (1H, 5, J = 6), 4.76 (1H, m), 5.16 (2H, S), 7.43 ppm (5H, S).

:; Method 5: (S) - (2-Oxo-3-azetidinyl) -carbamic acid phenylmethyl ester (20.0 g, see Example 1C) is suspended in 200 ml of acetonitrile, 21.6 ml of monotrimethylsilyltrifluoroacetamide (25.3 g) are added. ) and the mixture is heated to 50 ° C with stirring for 1 hour. After cooling in an ice bath to 0 ° C, 17.2 g of trimethylsilyl chlorosulfonate are added dropwise and the solution is stirred at ambient temperature for 6 hours. To the solution is added 24.2 g of potassium methyl 35 hexanoate in 100 ml of butanol and stirring is continued for 1 hour. The slurry is poured into 1 liter of dry diethyl ether and the precipitate is filtered off and dried in vacuo. The compound is dissolved in 500 ml of water, the pH is adjusted to 5, with potassium carbonate, the insoluble matter is filtered off and the mother liquor is cooled to dryness. The yield of the crude product is 19.4 g. The compound contains a small amount of potassium chloride which is removed by K-chromatography, the spectrum being identical to that of Method II.

Example 3 · 10 (S) -2-Oxo-3 - [[4- (phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid, tetrabutylammonium salt (1: 1)

Method I: (S) -2-Oxo-3 - [(phenylmethoxy) carbonyl) amino] -15-1-azetidinesulfonic acid, pyridine salt (1: 1) (34.3 g; see Example 1) is dissolved in 800 ml of water. The solution is clarified with charcoal, 30.7 g of tetrabutylammonium hydrogen sulphate in 80 ml of water are added and the pH is adjusted to 5.5 with 1N potassium hydroxide. The solvent 20 is removed in vacuo until a volume of about 200 ml is reached. The precipitated tetrabutylammonium salt is directly dated and dried in vacuo. The compound can be recrystallized from water or dissolved in methylene chloride, filtered and precipitated by the addition of ether. Yield 34.3 g, melting point 108-110 ° C.

Method II: (S) -2-Oxo-3- [1- (phenylmethoxy) carbonyl] -7-amino] -1-azetidinesulfonic acid, potassium salt (1: 1) (20.2 g; see Example 2) is dissolved in 500 ml of hydrogen. Filter and add 20.3 g of tetrabutylammonium hydrogen sulphate in 100 ml of water. The pH is adjusted to 5.5 with 1N potassium hydroxide. The volume is reduced in vacuo to 100 ml and the precipitated tetrabodyylammonium salt is filtered off. The compound is dissolved in 30 ml of methylene chloride, filtered and; precipitating by the addition of ether to give 21 g of the title compound; melting point 109-111 ° C.

Il 31 80271

Example H

(S) -3- (N- (2-amino-4-thiazolyl) acetylamino] -2-oxo-1-azetidinesulfonic acid, potassium salt A) (S) -3-Amino-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt (S) -2-oxo-3fenyylimetoksi) carbonyl / amino? ~-1-Azetidinesulfonic acid, tetrabutylammonium salt (2 g; see Example 3) is dissolved in 100 ml of dimethylformamide and hydrogenated for 30 minutes using 1 g of palladium on carbon as catalyst (10%). The catalyst is filtered off and the dimethylformamide is removed, leaving the title compound as an oil.

NMR (CDCl 3) 3.82 (1H, t, j = 5.5), 4.05 (d, 1H, d d, j = 5.5, 2.5 cps).

B) (S) -3 - [((2-amino-1 + -thiazolyl) acetyl) 'amino)' - 2-oxo-1-azetidinesulfonic acid, potassium salt

The above compound (2 g), 0.5 g of aminothiazole acetic acid and 0.4 g of hydroxybenzotriazole o are stirred at 0 ° C in 100 ml of dry dimethylform-20 amide while a solution of 0.7 g of dicyclohexylcarbodi -imide in 10 ml of dimethylformamide is added dropwise. After the addition is complete, stirring is continued for 12 hours at 20 ° C. Insoluble t * '. the urea is filtered off and the solvent is evaporated off under vacuum, the oily residue is treated with a solution of potassium perfluorobutanesulphonate in 20 ml of acetone at room temperature for 15 minutes. After addition of 200 ml of dimethyl ether, the title compound precipitates and is filtered, dried and purified by 300 ml of 30 HP-20 column chromatography using water as eluent. The yield is 850 mg of the title compound, melting point> 300 ° C.

32 80271

Example 5 (3S (Z1) -3- (E2-Amino-4-thiazolyl) -N-hydroxy (phenylmethoxy) phosphinyldx) (methoxy)) imin (4) -acetyl) -amino--2-oxo-1- azetidinesulfonic acid, potassium salt 5 (S) -3-Amino-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt (0.8 g; see Example 4A) in 30 ml of dimethylformamide, 0.9 g of (Z) -2-amino- <5 (N, N-hydroxy (phenylmethoxy) phosphinyl) -methoxy-imine?-7-4-thiazoleacetic acid, 0.3 g of hydroxybenzo-10-triazole and 0.7 g of dicyclohexylcarbodiimide are stirred for 24 hours at room temperature. The mixed urea is filtered off and the solvent is removed The residual oil is treated with an equivalent amount of potassium perfluorobutanesulfonate in 10 ml of acetone.

The title compound is filtered and purified using HP-20 resin and water as eluent to give 500 mg, mp 210-215 ° C, dec.

Example 6 E 3S (Zi) -3 - (N- (2-Amino-4-thiazolyl) (ethoxyimino) -20-acetylamino] -2-oxo-1-azetidinesulfonic acid, potassium salt (S) -3-amino 2-Oxo-1-azetidinesulfonic acid tetrabutylammonium salt (1.5 g; see Example · * 4A) in 100 ml of dimethylformamide, 0.6 g of hydroxybenzotriazole, 1 g of dicyclohexylcarbodiimide and 0.8 g of (Z) 2-Amino-N- (ethoxyimino) -4-thiazoleacetic acid is stirred at room temperature for 24 hours, the solvent is distilled off and the residue is dissolved in 30 ml of acetone, the urea is filtered off and the mother liquor is treated with a solution of 2 g of potassium perfluorobutanesulfonate in 20 ml. After addition of 200 ml of ether, the title compound precipitates, is filtered off and dried, and purified by chromatography on an HP-20 column and water as eluent to give 1.1 g of the title compound, m.p. 180-185 °. C, haj.

li 33 80271

Example 7 E 3 S (E) -3- [N- (2-amino-4-thiazolyl) (ethoxyimino) acetyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt Following the procedure used in Example 6 , but substituting (Z) -2-amino- <* - (ethoxyimino) -4-thiazoleacetic acid with (E) -2-amino-O- (ethoxyimino) -4-thiazoleacetic acid gives the title compound, m.p. 160-170 ° C after freeze-drying.

10 Example 8

ClSiZ) J-3- (C <· 2-amino-4-thiazolyl) / (2,2,2-trifluoroethoxy) imino? acetyl? amino β-oxo-1-azetidine sulfonic acid, potassium salt

Following the procedure used in Example 6, but substituting (Z) -2-amino-4- (2,2,2-trifluoroethoxy) imino-4-thiazoleacetic acid for (Z) -2-amino- ( ethoxyimino) -4-thiazoleacetic acid, the title compound is obtained, m.p. 169-170 ° C after freeze-drying.

Example 9 ^ * 3S (Z, R-3- (2-amino-4-thiazolyl) (methoxyimino) acetylamino] -2-oxo-1-azetidinesulfonic acid, potassium salt -L (S) -3 -amino-2-oxo-1-acetidinosulfonic acid, tetrabutylammonium salt (prepared as described in Example 4A from 7.9 g of (S) -2-oxo-3 - [(phenylmethoxy) carbonyl] amino) 1 -1-acetidinosulfonic acid tetrabutylammonium salt) is cooled to 0 ° C and 3.53 g of (Z) -2-amino-# - (methoxyimino) -4-thiazo-30-acetic acid are added and then a solution is added, wherein: is 3.27 g of dicyclohexylcarbodiimide in 10 ml of di-N: methylformamide. The mixture is stirred for 16 hours at 5 ° C, filtered and the solvent removed in vacuo. The residue is dissolved in acetone and filtered. Addition of 60 ml of a 10% solution of potassium perfluorobutanesulfonate in acetone crystallizes 4.7 g of crude product. The crude product is purified by chromatography on HP-20, 100-200 mesh to give 3.0 g of the title compound, mp 235 ° C.

3 "80271

Example m ^ 3S (Z ^ -3 - ^ (2-amino-4-thiazolyl) - ^ 2- (diphenylmethoxy) -1,1-dimethyl-2-oxoethoxyimination acetylamino-2H-oxo -1-azetidinesulfonic acid, potassium salt 5 (1: 1)

A solution of 0.005 moles of the tetrabutylammonium salt of S-3-amino-2-oxo-1-azetidinesulfonic acid (see Example 4A) and 0.006 moles of (Z) -2-amino-C (-? 2 - (diphenylmethoxy) -1 1,1-Dimethyl-2-oxoethoxy-10-imin-4-thiazoleacetic acid in 60 ml of dimethylformamide are treated 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 solution of 10% potassium perfluorobutanesulfonate in acetone. After addition of petroleum ether, the title compound precipitates and is treated with ether and filtered to give 3.3 g of product, m.p. 190 ° C, dec.

Example 11 E 3S (Z 2 -amino-4-thiazolyl) -1-carboxy-1-; methylethoxy) imino (acetyl) amino] -2-oxo-1-azetidinesulfonic acid, dipotassium salt 25 (3S) -3- [[(2-amino-4-thiazolyl)] - 2- (diphenylmethoxy) - The potassium salt of 1,1-dimethyl-2-oxoethoxy (imino) acetyl / amino] -2-oxo-1-azetidinesulfonic acid (2 g; see Example 10) is suspended in 5 ml of anisole and 25 ml of trifluoroacetic acid are added. . 30 acid acids at -10 ° C. The mixture is stirred for 10 minutes at -10 ° C. Ether (100 ml) is added slowly at -10 ° C and then 50 ml of petroleum ether are added. The precipitate is filtered to give 1.6 g of the trifluoroacetic acid salt. This is suspended in 20 ml of 3 water at 0 ° C, adjusted to pH 5.5 with dilution

II

35 80271 with potassium hydroxide and purified using an HP-20 column. The title compound is eluted with water and has a melting point of 225 ° C, dec.

Example 12 5 (3S (Z) 4) -3- (2-Amino-2-thiazolyl) - (2- (diphenylmethoxy) -2-oxoethoxy (imino) acetyl) amino) -2-oxo-1- azetidinesulfonic acid, potassium salt

Following the procedure described in Example 10, but substituting (Z) -2-amino-Λ -) - (2- (diphenylmethoxy) -10 2-oxoethoxy) imin) -1 + -thiazoleacetic acid with (Z) -2-amino-ηο- * - (2- (diphenylmethoxy) -1,1-dimethyl-2-oxoethoxy) imino) -4-thiazoleacetic acid gives the title compound, m.p. 180 ° C, dec.

Example 13 3-Methoxy-2-oxo-3- (phenylmethoxy) carbonyl) amine) -1-azetidinesulfonic acid, tetrabutylammonium salt Method I: A) 2-Oxo-3- (N-chloro-N - (phenylmethoxy) carbonyl) - (1) amino) -1-azetidinesulfonic acid, tetra-20-butylammonium salt - (S) -2-oxo-3- (phenylmethoxy) carbonyl) -; amino) -1-azetidinesulfonic acid, tetrabutylammonium salt (0.9 g; see Example 3) dissolved in 80 ml of methylene chloride, is added to a mixture (cooled to 0-5 ° C) of 3.17 g of sodium borate decahydrate. and 11.8 g of a 5.25% sodium hypochlorite solution in 70 ml of water. The reaction mixture is stirred vigorously for 55 minutes and cooled in an ice bath. After diluting with 0.5 M monobasic potassium phosphate solution, the product is extracted with methylene chloride 36 80271 (three 15 ml portions). Combining the extracts, drying (sodium sulfate) and removing the solvent in vacuo gave the title compound as an oil (0.94 g).

B) 3-Methoxy-2-oxo-3 - [(phenylmethoxy) carbonyl-5-amine) -1-azetidinesulfonic acid, tetrabutylammonium salt

To a stirred solution of lithium methoxide (667 mg) in anhydrous methanol (10 mL) at -78 ° C is added a solution of 2-oxo-3 -, β-chloro-N - β-phenylmethoxy ) carbonyl-4-amino-1-azetidinesulfonic acid tetrabutylammonium salt (0.94 g) in dry dimethylformamide (10 ml). After stirring for 1 hour at -78 ° C, the mixture is poured into 0.5 M monobasic potassium phosphate solution and extracted with methylene chloride (three 150 ml portions). The combined extracts are dried (sodium sulfate) and the solvent removed in vacuo to give an oil (0.83 g). The desired product is obtained by chromatography of the oil on silica gel (100 g) and eluting with 4-5% methanol in methylene chloride to give an oil (513 mg): max (pure) 1767> 1720 cm-1; NMR (CDCl 3) δ 3.40 (S, CH 3), 3.03 (ABq, J = 6.5 Hz, H 2), 5.08 (S, CH 2), 6.00 (S, NH), 7.27 (S, aromatic).

Method II: To a solution of the potassium salt of 3-benzyloxycarbonyl-: amino-3-methoxy-2-oxo-1-azetidinesulfonic acid (400 mg) in water is added a 0.1 M solution of tetrabutylammonium bisulfate (10.9 ml, pH adjusted). to 4.3 with potassium hydroxide). Relationship to new: ·. 30 times with methylene chloride, the extracts are combined, dried (Ν3230 ^) and the solvent is removed in vacuo. . to give a foam (625 mg) having similar spectral properties compared to the product of Method I.

Il 37 80271

Example 1β / 3S (Z)) - 3- (2-Amino-4-thiazolyl) (X 2-methoxy-2-oxoethoxy) iminoacetylamino) -2-oxo-1-azetidinesulfonic acid, potassium salt 5 (Z) - 2-Amino-.beta .- (2-methoxy-2-oxoethoxy) imino-4-thiazoleacetic acid (1.3 g) and (S) -3-amino-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt (2.03 g, see Example MA) is dissolved in 50 ml of acetonitrile and 1.03 g of dicyclohexylcarbodiimide dissolved in 5 ml of acetonitrile are added dropwise at 0 ° C. After stirring for 15 hours and filtering the dicyclohexylurea, the solvent is distilled off. The remaining oily residue is dissolved in acetone and treated with an equivalent amount of potassium perfluorobutanesulfonate. The title compound is isolated and purified by column chromatography using HP-20 and water as eluent to give the title compound m.p. 195-198 ° C.

Example 15 20 years *. 32-Amino-4-thiazolyl) - ((ethoxyphosphinyl) methoxy (imino) acetyl) amino) -2-oxo-1'-azetidinesulfonic acid, potassium salt: (S) -3-amino-2- oxo-1-azetidinesulfonic acid ***: tetrabutylammonium salt (2.25 g; see Example 4A) in 100 ml of dry dimethylformamide is treated with 1.87 g of (Z) -2-amino- <X-) ) "(diethoxyphosphinyl) methoxy) iminq) -M-thiazoleacetic acid, 0.7 g of hydroxybenzotriazole and 2.29 g of dicyclohexylcarbodiimide with stirring for 12 hours. The precipitated urea is filtered off and the solvent is removed in vacuo. .

The residual oil is treated with an equivalent amount of 1-1 potassium perfluorobutanesulfonate in 20 ml of acetone. After addition of ether, the title compound; precipitates and is filtered to give 2.7 to 7 g of crude product. Purification of the crude product by column chromatography using HP-20 and water / acetone (9: 1) as eluent gives the title compound, m.p. 155-160 ° C ...: dec. * 38 8 0 2 71

Example 16 (3S (Z 1 - -3 - [(2-amino-4-thiazolyl) (N- (1,1-dimethylethoxy) -2-oxo-1-phenylethoxy) imino) acetyl) amino 2-Oxo-1-azetidinesulfonic acid, potassium salt 5 (S) -3-Amino-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt (2.25 g; see Example 4A) in 60 ml of dimethylformamide is stirred at room temperature for 12 hours 2, With 4 g of (Z) -2-amino-3 - [(2- (1,1-dimethylethoxy) -2-oxo-1-phenylethoxy) -imino] -3-thiazoleacetic acid, with 1 g of hydroxybenzotriazole and 1.5 g of dicyclohexylcarbodiimide. The solvent is removed in vacuo and the residue is dissolved in 50 ml of acetone. The precipitated urea is filtered off and the mother liquor is treated with an equivalent amount of potassium perilobutanesulfonate. After addition of ether, the title compound crystallizes and is filtered. Purification of the compound is accomplished by HP-20 column chromatography using water / acetone (7: 3) as eluent to give 1 g of product, m.p. > 250 ° C dec.

20 Example 17 (3S (Z 2-amino-4-thiazolyl) - (1H-tetrazolyl: 5-ylmethoxy) imino) acetyl) amino) -2-oxo-1-acetylsulfonic acid, potassium salt ': (S) -3-Amino-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt (1.9 g; see Example 4A) in 60 ml of dimethylformamide is treated with 1.4 g of. 11a (Z) -2-Amino-tert - [(1H-tetrazol-6-ylmethoxy) -imino) -4-thiazoleacetic acid, 0.7 g of hydroxybenzotriazole and 1.4 g of dicyclohexylcarbo-30-di- imide with stirring for 24 hours. After removal of 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 amount of potassium perfluorobutanesulfone. ‘Sodium in 10 ml of acetone. The title compound is precipitated by the addition of 200 ml of ether. Purification • * is accomplished by HP column chromatography using HP-20 resin and water as eluent to give 1.05 g of product, m.p. 250 ° C, if j.

Il 39 80271

Example 18 (3S) - (2S) -3- ([(2-Amino-4-thiazolyl) enylmethoxy) -imin (2) acetyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt 5 (S) - 3-amino-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt (1.5 g; see Example 4A), 1.23 g of (Z) -2-amino-β- (phenylmethoxy) imino} -4-thiazoleacetic acid, 0 57 g of hydroxybenzotriazole and 1.14 g of dicyclohexylcarbodiimide are stirred in 60 ml of dimethylformamide for 24 hours at room temperature. The precipitated urea is filtered off, the solvent is removed and the residue is treated with an equivalent amount of potassium perfluorobutanesulfonate in 10 ml of acetone. When 200 ml of ether are added, the title compound 15 precipitates, is filtered and purified by HP-20 column chromatography using water / acetone (9: 1) as eluent to give 1 g of material, m.p. 200 ° C dec.

Example 19 (3S (Z)) 2-Amino-4-thiazolyl) (carboxymethoxy) -210 imino (acetyl) amino) -2-oxo-1-azetidinesulfonic acid, potassium salt (1: 1) 3S (Z) -4- [3- (2-Amino-2-thiazolyl) -N- (2- (diphenylmethoxy) -2-oxoethoxyimino)? amino--2-oxo-1-azetidinesulfonic acid, potassium salt (1: 1) 25 (1.3 g; see Example 12) is mixed with 5 ml of anisole. At -15 ° C, 25 ml of trifluoroacetic acid are added and the mixture is stirred for 10 minutes. Ether (100 ml) is added slowly at -10 ° C and then 50 ml of petroleum ether are added. The precipitate is suspended by cooling in 20 ml of water and adjusted to pH 5.0 with dilute potassium hydroxide. The product is purified by chromatography on an HP-20 column to give 3.0 g of the title compound, m.p. 230-235 ° C, dec.

80271

Example 20 (3S (Z) Y-2-Amino-4-thiazolyl) -N, 2-oxo-2- (phenylmethoxy) ethoxyiminoacetyl) amino] -2-oxo-1-azetidinesulfonic acid, potassium salt Following the procedure used in Example 10, but substituting (Z) -2-amino-<* - [1,1'-oxo-2- (phenylmethoxy) ethoxy] imino] -4-thiazoleacetic acid (Z) -2-amino - # 2- [2- (Diphenylmethoxy) -1,1-dimethyl-2-oxoethoxy] -4-thiazoleacetic acid to give the title compound, m.p. about 170 ° C, dec.

Example 21 E 3S (Z -3- (N - (, 2-amino-4-thiazolyl) (hydroxyimino) acetyl-amino] -2-oxo-1-azetidinesulfonic acid, potassium salt 15 Solution of 0.6 g 90% hydroxybenzotriazole in 100 ml of dimethylformamide is stirred for 1 hour with 10 g of 4A molecular sieves, filtered and the filtrate is added to a solution of 0.004 mol of (S) -3-amino-2-oxo-1-azetidinesulfonic acid tet-20 rabutylammonium salt (see Example 4A) in dimethylformamide (Z) -2-amino-O - (- (hydroxyimino) -4-thiazoleacetic acid (0.89 g) is added followed by 0.91 g of dicyclohexylcarbodiimide. after stirring for 16 hours, evaporating in vacuo and dissolving the residue in 20 ml of acetone and filtering, adding a solution of potassium perfluorobutanesulphonate to give the title compound as a precipitate Chromatography using HP-20 resin to give 0.44 g of product, mp> 240 ° C.

ti 41 ¢) 0071 V * w «- / i

Example 22 (3S (Z) 7-3-, 7/7 (2-Amino-4-thiazolyl) ((carboxyimidoxy) imino) acetyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt 5 £ 3S (Z) 7 (2-Amino-H-thiazolyl) -γ- (2-oxo-2- (phenylmethoxy) ethoxyimino] acetylamine -2-oxo-1-azetidinesulfonic acid potassium salt (0.1 g; see Example 20) is dissolved in a mixture of 5 ml of ethanol and 5 ml of water and hydrogenated at 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 freeze-dried. to give the title compound, mp 235 ° C (dec.)

Example 23 15S (Z) 7-3- [72-Amino-4-thiazolyl] -7- (1,1-dimethylethoxy) -1- (methylthio) -2-oxoethoxyimination / acetyl? -amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

Following the procedure used in Example 21, but substituting (Z) -2-amino - # - /? /? - (1,1-dimethyl-20-ethoxy) -1- (methylthio) -2-oxoethoxy. With 4-thiazole-acetic acid (Z) -2-amino-β- (hydroxy-quinino) -4-thiazo-acetic acid, the title compound is obtained, m.p. 130 ° C

- hair.

Example 24 25β- (2R, 3 - [(2-Amino-4-thiazolyl) methoxyimino) acetyl learning] -3-methoxy-2-oxo-1-azetidinesulfonic acid. acid, potassium salt AJ 3-Amino-3-methoxy-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt 30 4% Sodium borate decahydrate in methanol is added to a suspension of 10% palladium on carbon (30 mg) in methanol (2 ml) and a mixture of stirred under a hydrogen atmosphere for 15 minutes. 3-Methoxy-2-oxo-3 - [(phenylmethoxy) carbonyl] amino] -4-35 1-azetidinesulfonic acid, tetrabutylammonium salt (60 mg; see Example 13) in methanol (2 ml) is added and the mixture is stirred vigorously for 15 minutes under an atmosphere of hydrogen. The catalyst is removed by filtration * + 2 80271

Celite through a millic pore filter (0.5 sec) and the solvent is removed from the filtrate in vacuo and the residue is extracted with methylene chloride. Removal of the solvent under reduced pressure gives 35 mg of the title compound as an oil.

5 ([(2) (2-Amino-4-thiazolyl) methoxyimino) acetyl] amino) -3-methoxy-2-oxo-1-azetidinesulfonic acid, potassium salt 3-amino-3-methoxy-2- The tetrabutylammonium salt of oxo-1-azetidinesulfonic acid (see Part A) is dissolved in aceto-10-nitrile (20 ml) and pyridine (1 ml) and added with vigorous stirring to a suspension of (Z) -dk, -methoxyimino) -2- amino-4-thiazole acetyl chloride in acetonitrile (20 mL) cooled to 0-5 ° C. After stirring in the cold for 1 hour, the mixture is diluted with 0.5 M one-15 basic potassium phosphate solution (100 ml) (pH 4.8) and the solvent is removed in vacuo. The residue is taken up in a small amount of water containing little acetone. Chromatography using ion exchange resin (AG 50W-X2, 100-200 mesh, K + form 200 ml) gives the crude product 20 as the potassium salt eluting with water. Further purification with HP-20 resin (200 ml) using water as eluent gives 59 mg of product as a powder after trituration with acetonitrile ether. The product is an amorphous powder that melts slowly and decomposes above 150 ° C.

Analysis calculated for O 2 SK: C, 28.7 7; H, 2.90: N, 16.78; S, 15.36; K, 9.37 Found: C, 27.77; H, 2.82; N, 15.87; S, 13.63; K, 10.11.

Example 25 3 (O-cLs) -4-Methyl-2-oxo-3 - [(phenylmethoxy) carbonyl] -amino} -1-azetidinesulfonic acid, potassium salt A) N-benzyloxy- boc-allothreonine amide ("boc" means butoxycarbonyl)

A solution of 6.9 g of d, 1-t-boc-allothreonine 35 and the free amine obtained from 5.3 g of o-benzylhydroxylamine. HCl (ru0.033 moles, ethyl acetate-sodium bicarbonate is released) in 80 ml of tetrahydrofuran, treated with 4.82 g of N-hydroxybenzotriazole and 6.5 g of dicyclohexylcarbodiimide in 20 ml of tetra-

II

43 802 71 Hydrofuran. After stirring for 16 hours at room temperature, the slurry is filtered, concentrated in vacuo and chromatographed on a * 400 ml silica gel column. Elution with 5-10% ethyl acetate in chloroform 5 gives 6.8 g of the title compound in fractions 7-22 (200 ml each).

B) (+ -cis) -N-Benzyloxy-3-t-butoxycarbonylamino-4-methylazetidinone

A solution of 6.8 g of N-benzyloxy-t-boc-10 allothreoninamide in 200 ml of tetrahydrofuran is stirred for 16 hours with 5.2 * + g of triphenylphosphine and with 3.2 ml of diethyl azodicarboxylate. thia. The solvents are evaporated off under vacuum and the residue is chromatographed on a 500 ml silica gel column.

Elution with methylene chloride followed by crystallization from ether gives 2.65 g of azetidinone. Rechromatography of the mother liquors and mixed fractions gives an additional 0.6 g. Crystallization of the portion twice from ether (-20 ° C) gives 20 analytical samples of the title compound, m.p. l * l + 0 + + 2 ° C.

C) (+ -cis) -3-t-butoxycarbonylamino-1-hydroxy-; 4-methylazetidinone •: A solution of 3.2 g of cis-N-benzyloxy-3-t-butoxycarbonylamino-U-methylazetidinone in 200 ml of 95% ethanol is stirred under a hydrogen atmosphere with 07 g of 10% palladium on carbon. After 40 minutes, the slurry is filtered (249 ml recovery) and the filtrate is evaporated and triturated with ether to give 2.05 g of solid in two yields, 30 m.p. 13 4-13 6 ° C.

D) (+ -cis-3-t-butoxycarbonylamino-4-methylazetidinone

A solution of 2.05 g of cis-3-t-butyloxycarbonylamino-1-hydroxy-4-methylazetidinone in 60 ml of 35 methanol is treated with a total of 95 ml of 4.5 M ammonium acetate (40.20 and 30 ml doses) and 45 ml ... · 1.5 M titanium trichloride (20.10 and 15 ml doses); the second and third additions are 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. The combined organic layer is washed with a mixture of 100 ml of 5% sodium bicarbonate and 100 ml of saturated salt, dried and evaporated. Trituration with ether gives 1.65 g of solid in two portions. A portion of the first crop is crystallized from ether to give an ana-10 lytic sample, m.p. 176-178 ° C.

E) (+ -cis) -3-Benzyloxycarbonylamino-4-methylazetidinone

A solution of 1.55 g of cis-3-t-butoxycarbonylamino-4-methylazetidinone in 4 ml of methylene chloride and 4 ml of anisole is cooled to 0 ° C and 50 ml of cold trifluoroacetic acid are added. After 90 minutes, the solvents are evaporated in vacuo (benzene is added and evaporated 3 times).

The residue is dissolved in 25 ml of acetone, starting pH (2.5); 20 is raised to 7 with 5% sodium bicarbonate and 2 ml of benzyl chloroformate are added. The solution is maintained at 0 ° C and pH 7 for four hours and the acetone is removed in vacuo to give a slurry which is filtered. The filtrate is saturated with salt and extracted with methylene chloride. The solid is dissolved in methylene chloride and dried. The organic layers are combined, concentrated and the residue is chromatographed on a 200 ml silica gel column. Elution with 3: 1 chloroform and ethyl acetate gives 850 mg of the title compound in fractions (100 ml each) 4-11.

Crystallization of a small sample from ether gives an analytical sample, m.p. 165-166 ° C.

(80-Methyl-2-oxo-3-phenylmethoxy) -carbonylamino? -1-azetidinesulfonic acid, potassium salt

To a suspension of cis-3-benzyloxycarbo-5-ylamino-4-methylazetidinone (0.75 g) in 7 ml of dimethylformamide (dried on 4A screens activated at 320 ° C for 15 hours under a stream of argon) and 7 ml of: in methylene chloride (dried over basic Al 2 O 4), 1.66 g of pyridine-sulfur trioxide complex are added.

After stirring for 3 hours at room temperature under nitrogen, more pyridine-sulfur trioxide complex (1.66 g) is added. The reaction mixture is stirred at room temperature under nitrogen for 16 hours. The dimethylformamide is removed in vacuo to give 4.6 g of residue-15, which is dissolved in 300 ml of 0.5 M monobasic potassium phosphate solution (40 ° C, 10-15 min). Cool the solution, apply the HP-20 resin (3 cm x 60 cm) to the column with 400 ml of 0.5 M monobasic potassium phosphate, 1 liter of distilled water and (14: 1) 20 water: acetone to give 280 mg of analytical sample, m.p. 214-215 ° C, dec.

Analysis calculated for C 'H, 3.7 2; N, 7.95; S, 9.10; K, 11.10 Found: C, 40.43; H, 3.60; N, 7.89; S, 8.69; 25 K, 10.82.

Example 26 (3S-trans) -4-Methyl-2-oxo-3 - [(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid, potassium salt Following the procedure used in Example 25, but substituting d, 1-t -boc-allothreonine 1-t-boc-threonine gives the title compound, m.p. 133-135 ° C. Analysis calculated for 2 ° 6SK: C, 40.90: H, 3.72; N, 7.95; S, 9.10; K, 11.10 Found: C, 40.72; H, 3.60; N, 7.99; S, 8.80; 35 K, 10.82 46 80271

Example 27 (3S- [3a (Z), 4- [7-3 - [(2-amino-4-thiazolyl)] - (methoxyimino) acetylamino} -1'-methyl-2-oxo-1 -azetidine sulfonic acid, potassium salt 5 A) (3S-trans) -4-methyl-2-oxo-3- (T} phenylmethoxy) carbonyl} amino? -1-azetidinesulfonic acid, tetrabutylammonium salt (3S-trans) The salt (352.4 mg; see Example 26) of the 4-methyl-2-oxo-3- [N- (phenylmethoxy) - (N-carbonyl) amino] -1-azetidinesulfonic acid, potassium) is dissolved in 20 ml: To this is added water and tetrabutylammonium hydrogen sulfate (373.5 mg). The aqueous solution is extracted 3 times with methylene chloride and the combined extracts are dried over sodium sulfate. After removal of the solvent, 534.6 mg of the title compound are obtained.

B) (3 '- (3) (4)} - 3 - [({(2-amino-4-thiazolyl) methoxyimino) acetyl); Amino-4-methyl-2-oxo-1-azetidine sulfonic acid, potassium salt Solution of 534.6 mg of (3S-trans) -4-methyl-20-oxo-3-//} (phenylmethoxy) ) carbon li li? amino} -1-azetidinesulfonic acid, tetrabutylammonium salt in 20 ml of dimethylformamide, is hydrogenated with 20 mg of 10% palladium on carbon at atmospheric pressure for 2.75 hours; hydrogen uptake is 26.3 ml. The mixture is filtered and washed twice with 2.5 ml of dimethylformamide. The filtrate and washings (total about 25 ml) are mixed with 161 mg of (Z) -Ot- (methoxyimino) -2-amino-4-thiazoleacetic acid, 136 mg of N-hydroxybenzotriazole and 164 mg of nitrogen. , With 8 mg of dicyclohexylcarbo-30 diimide. The mixture is stirred under nitrogen for about 16 hours.

• Dimethylformamide is removed in vacuo and the gummy residue is dissolved in acetone and filtered to remove urea. A solution of 272 mg (0.8 mmol) of potassium salt of perfluorobutanesulfonic acid in 0.8 ml of acetone is added to the filtrate. The slurry is diluted with an equal volume of ether and filtered to give 325.5 mg of crude product, which is purified by chromatography using 75 ml of HP-20AG. Elution

II

uv 80271 400 ml of water and 400 ml (9: 1) of a water: acetone mixture (50 ml fractions) give 335 mg in fractions 3-10. After trituration with acetone-hexane, 97.3 mg of an analytical sample are obtained from fractions 5 to 3-5. Similar trituration of fractions 6-10 gives an additional 90.4 mg of product as a solid.

Analysis calculated for C 10 H 12 N 5 O 6 S 2 K: C, 29.92; H, 3.01; N, 17.45; S, 15.97; K, 9.74 Found: C, 30.32; H, 3.49; N, 15.8 ?; S, 13.95; 10 K, 10.45.

NMR (D 2 O) 1.57 (3H, d, J = 7), 3.97 (3H, S), 4.30 (1H, dg,> = 7.3), 4.7 δ (1H, d. J = 7), 6.95 ppm (1H, S).

Example 28 {3S- [3c] (Z), 4 -3- [4- (2-amino-4-thiolyl) - [(1-carboxy-1-methylethoxy) amino] acetyl} amino - methyl-2-oxo-1-azetidinesulfonic acid, dipotassium salt A) N-Benzyloxy-t-boc-threoninamide

A solution of 7.76 g of t-boc-threonine and the free amine obtained from 6.4 g of O-benzyl-· ·· 'hydroxylamine. HCl (ethyl acetate-sodium bicarbonate is released) in 100 ml of tetrahydrofuran, treated 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 chromatographed on a 300 g silica gel column (eluting with chloroform and chloroform-ethyl acetate (3: 1)) to give 7.2 g of the title compound. Crystallization from ether-hexane gives 4.18 g of the title compound.

:: B) (3S-trans) -N-Benzyloxy-3-t-butoxycarbonyl-2-aminoamino-4-methylazetidinone A solution of 12.67 g of N-benzyloxy-5-boc-. . 35 threoninamide, 11.5 g of triphenylphosphine and 6.23 ml of diethyl azodicarboxylate in 380 ml of tetrahydrofuran are stirred under nitrogen for about 16 hours. Solution

U ft O r \ r> r-, A

O U ^ / I

evaporated and chromatographed on a 900 g s1-gel gel column. Elution with chloroform-ethyl acetate (3: 1) gives 13.69 g of compound which crystallizes from ether-hexane to give 9.18 g of the title compound.

B) (3S-trans) -3-t-butoxycarbonylamino-1-hydroxy-4-methylazetidinone

A solution of 9.18 g of (3S-trans) -N-benzyloxy-3-t-butoxycarbonylamino-4-methylazetidinone in 300 ml of 95% ethanol is stirred under a hydrogen atmosphere with 1.85 g of 10% palladium on carbon. After 14 ml / min, the slurry is filtered and evaporated in vacuo. The residue is crystallized from ether-hexane to give 5.12 g of the title compound.

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

A solution of 4.98 (3S-trans) -3-t-butoxycarbonylamino-1-hydroxy-4-methylazetidinone in 200 ml of methanol is treated with 132 ml of 4.5 M ammonium acetate and then 66 ml of 1.5 M 'titanium trichloride and stirred for 4.5 hours.

:: The aqueous solution is diluted with an equal volume of 8% sodium chloride and extracted with ethyl acetate to give 3.48 g of crude product. Crystallization of the extract from ether-hexane gives 3.3 g of the title compound.

E) (3S-trans) -3-Benzyloxycarbonylamino-4-methylazetidinone

A solution of 3.3 g of (3S-trans) -3-t-butoxy-30-carbonylamino-4-methylazetidinone in 10 ml of dichloromethane and 10 ml of anisole is cooled to 0 ° C and 112 ml are added. trifluoroacetic acid. The solution is stirred for 90 minutes and evaporated in vacuo (benzene is added and evaporated 3 times).

The residue is dissolved in 70 ml of acetone and the solution is adjusted to pH 7 with 5% sodium bicarbonate solution. A total of 5.33 g of benzyl chloroformate is added over 1 hour at pH 6.5-7.5. The mixture is stirred

II

roOTi 49 υ υ c. / »30 minutes at pH 7, dilute with 100 ml of saturated salt and extract with ethyl acetate (three 400 ml portions). The residue obtained by evaporation is chromatographed on a 1 liter silica gel-5 column. Elution with chloroform-ethyl acetate (4: 1) gives 2.19. Crystallization from ether-hexane gives 1.125 g of the title compound.

F) (3S-trans) -4-Methyl-2-oxo-3 - [(phenylmethoxy) carbonyl) amine) -1-azetidinesulfonic acid, tetrabutylammonium salt

A solution of 600 mg of (3S-trans) -3-benzyloxycarbonylamino-4-methylazetidinone in 2 ml of dimethylformamide is cooled to 0 [deg.] C. and 4 ml of 0.8 M sulfur trioxide in dimethylformamide are added.

The solution is stirred at room temperature under nitrogen for 1 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 portions of methylene chloride (discarded) and 868 mg of tetrabutylammonium-20 bisulfate are added. The remaining solution is extracted with four 75 ml portions of methylene chloride. The combined organic layer is washed with 8% aqueous sodium chloride, dried and evaporated in vacuo to give 1.54 g of the title compound.

25 G) [(3S- (trans) (2) amino-4-thiazolyl '- [(1-diphenylmethoxycarbonyl-1-methylethoxy) imino) ethyl) amino) 4-Methyl-2-oxo-1-azetidinesulfonic acid, potassium salt A solution of 1.54 g of (3S-trans) -4-methyl-2-oxo-3 - ((5S-phenylmethoxy) carbonyl) amino] -1- tetrabutylammonium salt of cetidine sulfonic acid in 45 ml - dimethylformamide is stirred under an atmosphere of hydrogen with 800 g of 10% palladium on carbon for 2 hours. The catalyst is filtered off and the filtrate is stirred for 16 hours-35 hours with 1.24 g of (Z) -2-amino - (-, E - (1-diphenylmethoxycarbonyl-1-methylethoxy) imination) -4-t ages s was acetic acid, 0.4 g: r. with N-hydroxybenzotriazole and with 580 mg of dicyclohexylcarbodiimide. The broth οϋ 2 / ί te is evaporated in vacuo and the residue is triturated with 20 ml of acetone and filtered. The filtrate (and 2 ml of washings) is treated with 868 mg of potassium perfluorobutanesulfonate in 3 ml of acetone. Dilution with 7 mL of 5 ethers gives a solid which is isolated by decantation of the mother liquor, trituration with ether and filtration to give 0.91 g of the title compound. The mother liquor is further diluted with 100 ml of ether to give a second crop of the title compound 0.45 g.

10 H) ([3S- (f3a) (Z) 2-Amino-4-thiazolyl) - E '- (1-carboxy-1-methylethoxy) imino) -acetyl? -Amino--4-methyl-2-oxo-1-azetidinesulfonic acid, dipotassium salt

Slurry with 140 mg of (3S -? - 3a (Z), "3 - 15 (? 2-amino-4-thiazolyl)?" (1-diphenylmethoxycarbonyl-1-methylethoxy) imino) acetylamino) -'4-methyl-2-oxo-1-azetidinesulfonic acid, potassium salt (first crop) in 0.5 ml of anisole, stirred at -12 ° C under nitrogen and 2.5 ml of cold (-10 ° C) trifluoroacetic acid are added . After 10 minutes, 10 ml of ether and 5 ml of hexane are added and the resulting slurry is stirred for 5 minutes at -12 ° C and allowed to warm to room temperature. The solid is isolated by centrifugation and washed twice with ether. A solution of this solid in 5 ml of cold water is immediately adjusted to pH 5.5 with 0.4 N potassium hydroxide and then applied to an 80 ml HP-20AG column. Elution with water gives 72 mg of the title compound in fractions (10 ml) after 7-11 evaporations (acetones-30 grams are added and evaporated 3 times) and trituration with ether, m.p. / ^ 250¾ (dec.).

Analysis calculated for O 5 S 2 K 2: C, 30.51; H, 2.95; N, 13.69; S, 12.53; K, 15.28 Found: C, 29.63; H, 3.20; N, 12.96; S, 11.94; 35 K, 12.78 NMR (D 2 O) 1.46 (S, 6H), 1.5 δ (1H, d, J = 7) 4.28 (1H, dg, J = 7, 2.5), δ , 67 (1H, d, j-2), 6.95 ppm (b, 1H9.

Il si 80271 The residual 1.22 g of (3S- (3a) (N, N) -3-CC2-amino-4-thiazolyl-1-diphenylmethoxycarbonyl-1-methyl (oxy) imino) acetyl ) amino) -4-methyl-2-oxo-1-azetidinesulfonic acid, potassium salt 5 (yields 1 and 2) are treated as above (M, 2 ml of anisole, 16 ml of trifluoroacetic acid, 13 minutes at -15 ° C). you). Chromatography on a 300 ml HP-20AG column gives 649 mg of the title compound in fractions 6-9 (60 ml) after the above treatment.

10 Example 29

Following the procedure used in Example 19, but substituting A3S (Z) J-3-7) (2-amino-4-thiazolyl) -N - ((2- (diphenylmethoxy) -2-oxoethoxy) imino) acetylamino] -2- Potassium salt of oxo-1-azetidinesulfonic acid β 3 S (S) 7-3 - ((T (2-amino-4-thiazolyl)) - 2- (1,1-dimethylethoxy) -1- (methylthio) -2-oxoethoxy The potassium salt of 4-amino-2-oxo-1-azetidinesulfonic acid (see Example 23) gives ε-3S (Z, β-β, β-β (2-amino-4-thiazolyl) {T-carboxy). (methyl) methoxy) amino) acetylamino) -2-oxo-1-20 azetidinesulfonic acid, potassium salt (1: 2), mp 165 ° C, dec.

• - Example 30 {3oc (Z), 4λ) -3 - [((2-amino-4-thiazolyl) (methoxyimino) - (acetyl) amino) -4-methyl-2-oxo-1-azetidinesulfone - 25 acids, potassium salt •: A solution of 51.8 mg of (cis) -4-methyl-2-oxo-3 - [(phenylmethoxy) carbonyl) 'amino-1-azetidinesulfonic acid potassium salt and 51 mg of tetra -n-butyl ammonium bisulfate in 5 ml of water is extracted with methylene chloride (four 10 ml portions) to give 81 mg of an oil. It is stirred under a hydrogen atmosphere for 2 hours with 40 mg of 10% palladium on carbon in 4 ml of dimethylformamide. The catalyst is filtered off and washed with 1 ml of dimethylformamide. The filtrate and washings are combined and stirred for 16 hours with 31 mg of (Z) -2-amino- (methoxyimino) -4-thiazoleacetic acid, 27 mg of N-hydroxybenzotriazole 52 80271 and 31.5 mg with dicyclohexylcarbodiimide. The solution is evaporated in vacuo and the residue is triturated with 3 ml of acetone. The resulting slurry is centrifuged and the liquid is treated with 51 mg of potassium perfluorobutane-5 sulfonate. Dilution with 5 ml of ether and filtration gives a solid. Chromatography using HP-20 AG (40 mL) gives Rydon-positive material in fractions (20 mL) of 3-5 (eluted with water). Evaporation and trituration with ether gives 23 mg of product 10 as a hygroscopic solid.

Analysis calculated for qH12N2O8Sk2: C, 29.91; H, 3.01; N, 17.44 Found: C, 29.30; H, 3.31; H, 16.66.

NMR (D 2 O) 1 0 0 (3H, d, J -7), 3.97 (3H, S), 4.46 (1H, 15 Apparent pentet, J-7), 5.37 (1H, d, J = 7), 6.97 ppm (1H, S).

Example 31 (3S-cis) -3-Amino-4-methyl-2-oxo-1-azetidinesulfonic acid 20 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 and diluted with 70 ml of water and 140 ml of ethyl acetate. Stir thoroughly and separate the layers and wash the organic layer with 30 mL (2: 1) of aqueous brine. The combined aqueous layers are then back-extracted with 70 ml of ethyl acetate. The aqueous layer is cooled in an ice bath and 10% potassium bisulfite solution is added to pH 2.3. The acidified solution is extracted with ethyl acetate (four 150 ml portions). The combined organic layers are dried over anhydrous sodium sulfate and the solvent is evaporated to give 9.13 g of the title compound.

Il 53 802 71 B) N-methoxy-t-boc-1-allothreonine amide t-boc-1-allothreonine (9.13 g) is dissolved in 85 ml of water and 41 ml of 1N potassium hydroxide solution.

Methoxyamine hydrochloride (5.22 g) and 8.67 g of 5'-ethyl-3,3- (dimethylaminopropyl) carbodiimide are added. HCl.

The mixture is stirred at room temperature for 4 hours and then saturated with sodium potassium tartrate. The resulting mixture is extracted with ethyl acetate (four 150 ml portions) and the organic layer is dried over anhydrous sodium sulfate and the solvent is removed to give 7.38 g of the title compound as a solid.

C) C-Methanesulfonyl-N-methoxy-t-boc-1-allothreoninamide N-methoxy-t-boc-1-allothreoninamide (7.32 g) is dissolved in 40 ml of pyridine and cooled - To 20 ° C under nitrogen. Add methanesulfonyl chloride (3 mL) dropwise with a hand syringe over 5 minutes. The resulting mixture is slowly warmed to 0 ° C and stirred at said temperature for 3 hours. Ethyl acetate-20 (500 mL) is added and the solution is washed with 250 mL of ice-cold 3N HCl solution, then with 100 mL of 5% NaHCO 3 solution. The ethyl acetate layer is dried over anhydrous sodium sulfate and the solvent is removed to give 8.64 g of the title compound as a white solid.

D) (3S-cis) -3-t-butoxycarbonylamino-1-methoxy-4-methylazetidinone O-methanesulfonyl-N-methoxy-t-boc-1-allo-threoninamide (8.64 g) is dissolved in 530 ml acetone and 11 g of solid potassium carbonate are added. The mixture 30 is slowly heated to 65 ° C under nitrogen and stirred at this temperature for 1 hour. The reaction mixture is then filtered through Celite and the filter cake is washed with ethyl acetate. The filtrate is concentrated and the residue is taken up in 250 ml of ethyl acetate. The ethyl acetate solution 35 is washed with 100 ml of 5% sodium bicarbonate solution.

The ethyl acetate layer is dried over anhydrous sodium sulfate and the solvent is removed to give 6.63 g of crude product.

54 ϋ ϋ 2 7 i E) (3S-cis) -3-t-butoxycarbonylamino-4-methylazetidinone

Sodium (1.35 g) is dissolved in about 300 ml of liquid ammonia at -50 ° C and 5.87 g of (3S-cis) -3-t-butoxycarbonylamino-1-methoxy- 4-methylazetidinone in 35 ml of tetrahydrofuran. An additional 10 ml of tetrahydrofuran is used for rinsing. At the end of the addition, an additional 100 mg of sodium is added. The mixture is stirred for a further 5 minutes 10 and then cooled by adding 3.35 g of solid ammonium chloride in one portion. The ammonia is removed under a stream of nitrogen and 250 ml of ethyl acetate are added to the residue. After filtration and washing of the solid, the combined filtrate was removed with ethyl acetate to give 4.82 g of the title compound.

F) (3S-cis) -3-t-butoxycarbonylamino-4-methyl-2-oxo-1-azetidinesulfonic acid, tetrabutyl ammonium salt 20 (3S, 4R) '- 3-t-butoxycarbonylamino-4-methylazetidinone (4, 98 g) is dissolved in 30 ml of dimethylformamide. 11.9 g of pyridine-sulfur trioxide complex are added and the mixture is stirred at room temperature under types. After stirring for 14 hours, 1.8 g of li-25 weather pyridine-sulfur trioxide complex are added and stirring is continued for 80 hours. The reaction mixture is poured into 700 ml of 0.5 M monobasic potassium phosphate solution and washed with methylene chloride (three 300 ml portions). Tetra-n-butylammonium bisulfate (8.45 g) is added to the aqueous solution and the mixture is extracted with methylene chloride (four 300 ml portions). The combined methylene chloride layers are dried over anhydrous sodium sulfate and the solvent is removed to give 10.76 g of the title compound as a gum.

Il 55 80271 G) (3S-cis) -3-Amino-4-methyl-2-oxo-1-azetidine sulfonic acid (3S-cis) -3-t-butoxycarbonylamino-4-methyl-2-oxo-1-azetidinesulfonic acid the tetrabutylammonium-5 salt (10.76 g) is dissolved in 50 ml of 95-97% formic acid and stirred for 4 hours under nitrogen. A small amount of the product of the previous reaction is added and the mixture is stirred for another 1 hour. The mixture is stored frozen for 16 hours and the frozen mixture is warmed to room temperature for 10 hours and stirred for an additional 1 hour. The solid formed is filtered and washed with methylene chloride to give 982 mg of the title compound. The filtrate is diluted with 1 liter of methylene chloride and kept at -20 ° C for 4 hours. The precipitate formed is recrystallized from water-methanol-acetone to give an additional 167 mg of the title compound.

Example 32 (3S- (3 (X (Z), 4 (E (-3- (5-amino-4-thiazolyl) methoxyimino) acetyl) amino] -4-methyl-2-oxo-1- at s et idine-20 sulfonic acid, potassium salt

A solution of 201 mg of (Z) -2-amino-α- (methoxy-: imino) -4-thiazoleacetic acid and 153 mg of N-hydroxybenzotriazole monohydrate in 3 ml of dimethylformamide is treated with 206 mg of dicyclohexylcarbode -25 imides. 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 of triethylamine in 2 ml of dimethylformamide (1 ml more dimethylformamide is used for 30 rinses) * and the mixture is stirred for 16 hours. The slurry is evaporated in vacuo, triturated with 12 ml of acetone, centrifuged and the liquid treated with 338 mg of potassium perfluorobutanesulfonate. Dilution with 10 mL of ether and filtration gives a solid product which is chromatographed using 200 mL of HP-20 resin and eluting with water. Fractions (20 ml each) 18-30 are combined and lyophilized to give 274 mg of the title compound as a hygroscopic solid.

V Π 'V7 ·)

5 6 u \ j c. / I

Anal Calcd for O 9 S 2 K: C, 29.91; H, 3.01; N, 17.44 Found: C, 30.03; H, 3.21; N, 17.06 NMR (D 2 O) 1.40 (3H, d, J = 6.5), 3.98 (3H, S), 4.48 1H, t 5 d, J = 6.4, 5.5), 5.36 (1H, d, J-5.5) 6.97 (1H, S).

Example 33 (33-Trans) -3-Amino-4-methyl-2-oxo-1-azetidine-sulfonic acid A) Threonine, methyl ester hydrochloride 10 A flask containing 500 ml of methanol is cooled to -5 ° C under a nitrogen atmosphere. (ice / brine and add 130 mL (greater than necessary) of thionyl chloride at such a rate that the reaction temperature remains between 0 ° C and 10 ° C. After recooling to 59 ° C, 59.5 g of 1 threonine and the mixture is allowed to reach room temperature and stirred for 16 hours, concentrated and evacuated at 10 ° C for 2 hours to give a viscous oil, which is used in the next 20 steps.

B) Threoninamide

The crude product from Part A is dissolved in 2.5 liters of methanol and cooled to -5 ° C (ice / brine).

The solution is saturated with ammonia gas, the cooling bath is removed and the sealed flask is allowed to stand for 3 days. After most of the unreacted ammonia has been removed using a vacuum cleaner, 100 g of sodium bicarbonate and 50 ml of water are added and the mixture is evaporated to a viscous oil.

30 c) Benzyloxycarbonylthreoninamide

The crude product from Part B (already containing the required amount of sodium bicarbonate) is diluted to a volume of 1 liter with water. To this rapidly stirred solution is added 94 g (88 ml of 90% pure 35) benzyloxycarbonyl chloride as a solution in 80 ml of tetrahydrofuran over 1 hour. The reaction mixture was stirred; then for a further 16 hours and extracted with ethyl acetate (one portion of 500 ml, two portions of 250 ml). The combined extracts are dried over magnesium sulfate

II

57 80271 and concentrated. The crystalline residue is then dissolved in 250 ml of hot ethyl acetate and 300 ml of hexane are added and then boiled until a clear solution is obtained. Cooling, filtration and drying of the crystalline mass gives 104 g of the title compound.

D) Benzyloxycarbonylthreoninamide, O-mesylate 100 g of benzyloxycarbonylthreoninamide 10 are dissolved in 400 ml of anhydrous pyridine under argon and cooled in an ice / salt bath. To this stirred solution is added methanesulfonyl chloride (36.8 mL, 54.5 g) over 15 minutes. After stirring for 2 hours, 0.3 equivalent of additional methanesulfonyl chloride is added. The reaction is stirred for 1 hour and poured into a mixture of 1.5 L of ice and 2 L of water. The resulting slurry is stirred for 30 minutes and filtered. Drying of the crude product at 60 ° C in a vacuum oven for about 16 hours gives 109 g of the title compound.

E) N-Sulphonyl-benzyloxycarbonyl-threonine amide, O-mesylate, tetrabutylammonium salt A solution of 2-picoline (17.8 ml) in 90 ml of ... · methylene chloride is cooled to -5 ° C (ice-salt). Solution) and 5.97 ml of chlorosulfonic acid are added at such a rate that the internal temperature of the reaction remains below 5 ° C. The resulting solution is added via tube to a suspension of 7.56 g of benzyloxycarbonyl-threoninamide O-mesylate in 120 ml of methylene chloride. The resulting heterogeneous mixture is refluxed for 16 hours to give a clear solution. The solution is poured into 500 ml of phosphate buffer solution (5.0 M, pH 4.5) and further diluted with 120 ml of methylene chloride. The separated organic layer is washed once with 100 ml of 35 buffer solutions and the combined aqueous phases are treated with 10.2 g of tetra-n-butylammonium hydrogen sulphate and extracted with methylene chloride (one 300 ml portion and two 150 ml portions). n doses). After the combined organic extracts have been dried over sodium sulfate, the solution is concentrated to give 12.7 g of a foam.

Γ) (3S-trans) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid

A mixture of 5.52 g of potassium carbonate in 20 ml of water and 160 ml of 1,2-dichloroethane is refluxed and 15.5 mmol of the tetrabutylammonium salt of N-sulfonyl-benzyloxycarbonylthreonamide O-mesylate in 20 ml of 1.2 -dichloroethane (20 ml is used for rinsing). After cooling to reflux for 30 minutes, the mixture is poured into a separatory funnel, diluted with 50 ml of water and 100 ml of methylene chloride and the phases are separated. The resulting organic phase is dried over sodium sulfate and concentrated to give crude (3S-trans) -3-benzyloxycarbonyl-20-amino-4-methyl-2-oxo-1-azetidinesulfonic acid tetra-butylammonium salt. The crude azetidinone in 250 ml of ethanol is treated with 0.8 g of 5% palladium on carbon as a catalyst and hydrogen is bubbled through the solution. After 90 minutes, the mixture is filtered through Celite; 50 ml of ethanol are used for rinsing. Upon addition of 1.2 ml of formic acid to this solution, the title amphoteric ion precipitates immediately and is filtered after stirring for 1 hour to give, after drying at 10 torr for 1 hour, 1.1 g of product.

A second crop of product is obtained by concentrating the filtrate and adding formic acid to give 1.3 g of the title amphoteric ion.

Sp. > 218 ^ (dec.); {<* J ^ = -41.1 (C = 1, H 2 O).

NMR (D 2 O) 1.58 (3H, d = 7), 4.80 (2H, M).

fl 59 8027 j

Example 3H.

£ 3 S3Λ (ζ), 4- [2,3-3- [7-2-amino-ethyl] dimethyl-2- (1H-nitrophenyl) n-ethoxy) -2-oxoethoxy) imine} Acetyl} amine} -4-methyl-2-oxo-1-azetidine-5-sulfonic acid, potassium salt

To a slurry of (3S-trans) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid (0.36 g; see Example 33) in dry dimethylformamide (30 mL) under nitrogen at 26 ° C is added triethylamine (309 L).

After about 5 minutes a clear solution is obtained and (Z) -2-amino-Gt-1-dimethyl-2 - ((4-nitrotenyl) -methoxy] -2-oxoethoxy-4-iming} -4-thiazoleacetic acid ( 0.816 g) and then N-hydroxybenzotriazole (0.334 g) and dicyclohexylcarbodiimide (0.453 g) are added, the mixture is stirred for 20 hours at 26 ° C, the solvent is removed in vacuo and the residue is triturated with acetone. After stirring for 5 minutes, the solids are removed and the filtrate is treated with potassium perfluorobutanesulfonate (3.680 g) in acetone (5 ml). Addition of ether (about 40 ml) gives a precipitate which is collected and dried in vacuo (1.073 g; second yield 0.066 g, total 1.14 g) Anal, calculated for E 20 ^ 21 ^ 6 ^ 10 ^ 2 ^ · 1 H 2 O C, 38.33; H, 3.70; N, 13.41; Found: C, 38.30; H, 3.63; N, 13.41; S, 9.88; K, 5.98.

Example 35 / 3a (Z), 4 '- (3 - [[(2-amino-4-thiazolyl)] - (1-carboxy-1-methylethoxy) imino} acetyl learning} -4-methyl-2-oxo -1-azetidinesulfonic acid, potassium salt (1: 2) 30 (3) (3 ((Z), 4 ') - 3- (2-amino-4-thiazolyl) / (1-diphenylmethoxycarbonyl-1-methylethoxy) -imino) acetyl } amine-4-methyl-2-oxo-1-azetidinesulfonic acid, potassium salt Solution of (cis) -4-methyl-2-oxo-3 - [(3-phenylmethoxy) carbonyl} amine] -1-azetidinesulfonic acid - the acid, tetrabutylammonium salt (201 mg; prepared from 60 80271 of the corresponding potassium salt of Example 25 (as described in Example 30) in 5 ml of dimethylformamide, is mixed with 90 mg of 10% palladium on calcium carbonate under an atmosphere of hydrogen for 2 hours. the filtrate is stirred for about 16 hours with 146 mg of (Z) -2-amino-O-trans-diphenylmethoxycarbonyl-1-methylethoxy) imin-4-thiazoleacetic acid, 73 mg of dicyclohexylcarbodiimide and 51 mg with N-hydroxybenzotriazole under nitrogen. The slurry is evaporated in vacuo and triturated with 4 ml of acetone. The slurry is filtered and the solid is washed twice with 2 ml portions of acetone. The filtrate and washings are combined and treated with 113 mg of potassium perfluorobutanesulfonate. Dilution 2 with 4 ml of ether gives a solid which is isolated by centrifugation and washed 3 times with ether to give 186 mg of the title compound.

B) (3 (* (Z), 4a) -3 - ((2-amino-4-thiazolyl) (1-carboxy-1-methylethoxy) (imino) acetyl) amino) -2-methyl-20 4-Oxo-1-azetidinesulfonic acid, potassium salt (1: 2)

Slurry of 186 mg of (3dU Z), 4 (R) -3 - [(2-amino-4-; thiazolyl) (t1-diphenylmethoxycarbonyl-1-methylethoxy) imino) acetylamino] -4-methyl- The potassium salt of 2-oxo-1-25 azetidinesulfonic acid in 0.6 ml of distilled anisole is cooled to -12 ° C and 3.0 ml of distilled trifluoroacetic acid (at -10 ° C) are added. The solution is stirred for 10 minutes and 12 ml of ether are added, followed by 6 ml of hexane. After 5 minutes at -10 ° C and 15 minutes after stirring at ambient temperature, the solid is isolated by centrifugation and washed 4 times with ether to give 141 mg of material. This is dried in vacuo, ground, dissolved in 5 ml of cold water and immediately adjusted to pH 5.6 with 0.4 N potassium hydroxide. Solution

II

61 80271 is applied to a 100 ml HP-20AG column and eluted with water. Fractions (10 ml) 8-12 are combined and evaporated in vacuo (acetonitrile is added 3 times and evaporated. The residue is triturated with ether to give 101.7 mg of product as a hygroscopic solid.

Anal. Calculated for '^ 0.51; H, 2.95; N, 13.69; S, 12.53; K, 15.28 Found: C, 30.11; H, 3.26; N, 13.35; S, 12.12; 10 K, 15.02.

Example 36 (3S - [3 ((Z), - 3 - [(2-amino-4-thiazolyl) -] - (1-N, carboxy-1-methylethoxy) imino) acetyl) amino] -4-methyl-2-oxo- 1-Azetidinesulfonic acid 15 / 3S- (Z), 4H-N- (3- (2-amino-4-thiazolyl) - (1-carboxy-1-methylethoxy) imino) acetyl} amino) -4-methyl The dipotassium salt of 2-oxo-1-azetidinesulfonic acid (87.3 mg; see Example 28) is dissolved in 1.38 ml of water, cooled to 0 ° C, treated with 0.34 ml of 20 N hydrochloric acid and the crystals obtained separated by centrifugation. The wet solid is dissolved in methanol, filtered, concentrated to about 0.5 mL and mixed with 1 mL of water to give 55.9 mg of the title compound.

Example 37 / 3S- [1a (Z), H / &) - 3 - (({7-amino-4-thiazolyl) - (1-carboxy-1-methylethoxy) imino] acetylamino] -4-methyl-2- oxo-1-azetidinesulfonic acid, sodium salt 99.7 mg sample (3S- (Sct (Z), 4'-y-3 2-amino-4-thiazolyl) -1H-carboxy-1-methylethoxy) imino ) - Acetyl-amino} -4-methyl-2-oxo-1-azetidinesulfonic acid is mixed with 0.207 ml of 1N sodium hydroxide and the resulting mixture is gradually heated to dissolve the remaining solid. The water is azeotroped with acetonitrile and the residue is crystallized from a mixture of 0.5 ml of methanol to dissolve the residue) and 1 ml of acetonitrile to give 81.8 mg of a solid. A second recrystallization from 0.8 ml of methanol gives 47.9 mg, a third from 0.24 ml of methanol and 0.24 ml of absolute ethanol gives 44.8 mg and a fourth from 0.225 ml of methanol and 0.225 ml of per ml of absolute ethanol gives 38.8 mg. The solid is dried at 20 ° C and 0.01 mm Hg for 18 hours and then equilibrated at atmospheric humidity for 24 hours to give 40.9 mg of the title compound.

Example 38

Following the procedure used in Example 32, but substituting (R) - (3-) - ({3- [3-dioxo-4- (phenylmethoxy) carbonyl) amino) -1-piperazinyl / carbonyl] amino] benzoic acid (Z) - 2-amino-α- (methoxyimino) -4-thiazoacetic acid, gives S 2 -N-Ct (R *), 4 <x7> 7 - ^ - ^ y (2- / C (4-methyl- (2-oxo-1-sulfo-3-azetidinyl) amino] -2-oxo-1-phenylethylamino] carbonyl] -2,3-dioxo-1-piperazinyl-urea, phenylmethyl ester, potassium salt; 191 ° C, dec.

Example 39 E (3S- [3 ((Z), 4- [3- (3- (2-amino-4-thiazolyl) (carboxy-1-methylethoxy) imino] acetyl) amino) -4-methyl-2 -oxo-1-azetidinesulfonic acid, potassium salt (1: 2) A) / (3S- (3) (Z), 4 - ((3)) - 3 - ((2-amino-4-thiazolyl)) (1,1-diphenylmethoxycarbonyl-1-methylethoxy) -3-imino) acetyl) amine) -4-methyl-2-oxo-1-azetidinesulfonic acid, potassium salt Solution with 440 mg (Z) 2-amino-N- [1- (1-carboxylmethyloxy) imino} -4-thiazoleacetic acid and 153 mg of N-hydroxybenzotriazole monohydrate; 35 in 3 ml of dimethylformamide, treated with 206 mg of dicyclohexylcarbodiimide. The mixture is stirred at room temperature for 30 minutes under nitrogen and a solution of 180 mg of (3S-cis) -3-amino-4-methyl-

II

ro - f * 6 3 0 0 4. [i 2-oxo-1-azetidinesulfonic acid (see Example 31) and 0.14 ml of triethylamine in 2 ml of dimethylformamide (1 ml of dimethylformamide is used for rinsing) and the mixture is stirred for 16 hours .

5 The slurry is evaporated in vacuo and triturated with 12 ml of acetone. The slurry is filtered and the solid is washed with acetone (two 3 mL portions). The combined filtrate and washings are treated with 338 mg of potassium perfluorobutanesulfonate. Dilution with 30 ml of eet-10 blades gives a gummy solid which solidifies slowly. The solid is filtered and washed with ether to give 656 mg of the title compound.

B) (3S-A3a (Z) 2-Amino-4-thiazolyl) - (1,1-carboxy-1-methylethoxy) imino} acetyl) -15-amino--4-methyl-2-oxo-1-azetidinesulfonyl acid, potassium salt (1: 2)

Slurry with 656 mg of (3S- [3®) (Z), 40β-3 - ((X2-amino-4-thiazolyl)) - [(1-diphenylmethoxycarbonyl-1H-1-methylethoxy) imino] -1-acetyl} amino--4-methyl-2-20-oxo-1-azetidinesulfonic acid, potassium salt in 2.3 ml of distilled anisole, cooled to -12 ° C and 11.5 ml of trifluoroacetic acid (precooled to -10 ° C) are added ). The solution is stirred for 15 minutes and; 46 ml of ether are added followed by 23 ml of hexane. After 5 minutes at -10 ° C and stirring for 15 minutes at room temperature, the solid is filtered and washed with ether to give 457 mg of a highly hygroscopic gum. This is dissolved in 6 ml of cold water and immediately adjusted to pH 30 with 5.6 M potassium hydroxide solution. The solution is taken up in 200 ml of HP-20 resin and eluted with water. Fractions (50 ml each) 7-11 are combined and lyophilized to give 239 mg of the title compound as a solid.

r n * - * 64 w u l. / j

Anal. Calculated for C 13 H 15 N 8 N 5 S 2 K 2 • 1/2 H 2 O: C, 29.99; H, 3.10; N, 13.45; S, 12.32 Found: C, 29.94; H, 3.30; N, 13.30; S, 11.93. NMR (D 2 O) 1.44 (3H d, J = 7), 1.46 (6H, S), 4.48 (1H, dt δ J-75, 5.5), 5.34 (1H, d, J = 5.5) 6.96 ppm (1H, S).

Example 40 (+) - 3-Amino-4,4-dimethyl-2-oxo-1-azetidinesulfonic acid A) (+) - 4,4-dimethyl-2-oxo-1-azetidine-tert-butyl-10-diphenylsilane

A solution of 40.5 ml of t-butylchlorodiphenylsilane in 112 ml of dimethylformamide is cooled to 0 ° C. To this is added 22 ml of triethylamine. A solution of 12.87 g of 4,4-dimethyl-2-acetyl-15-dinone in 25 ml of dimethylformamide is added dropwise over 10 minutes to a cooled solution of triethylamine. The resulting cloudy solution is stirred for 18 hours at 5 ° C under argon. This mixture is poured into 400 ml of ice water and extracted with three 15 ml portions (2: 1) of 20 ether / ethyl acetate. The combined extracts are washed with four 100 ml portions of 0.5M monobasic potassium phosphate buffer solution, one 150 ml portion of sodium bicarbonate solution, two 150 ml portions of water and one 150 ml portion of saturated solution. sodium chloride solution. The solution is dried over sodium sulfate and concentrated in vacuo to give 33.03 g of the title compound as a solid.

B) (+) - 3-Azido-4,4-dimethyl-2-oxo-1-azetidine-30-tert-butyldiphenylsilane

A solution of 4.25 ml of 1.6M (in hexane) n-butyllithium and 11 ml of dry tetrahydrofuran is prepared at -50 ° C under argon in a 100 ml three-necked flask. A solution of 0.083 g of triphenylmethane in 1 ml of tetrahydrofuran is added. The resulting solution is cooled to -60 ° C and 1.0 ml of diisopropylamine is added dropwise using a hand syringe. This is stirred for 15 minutes and then cooled

II

cc r r * r r ~> * OU ^ / | to -78 ° C. A solution of 2.3 g of (+) - 4,4-dimethyl-2-oxo-1-azetidine-tert-butyldiphenylsilane in 8 ml of tetrahydrofuran is slowly added using a hand syringe. The resulting solution is stirred for 20 ml to 5 minutes at -78 ° C and at the same time heavy precipitation occurs and uniform mixing becomes difficult. A solution of 1.33 g of p-toluenesulfonyl azide in 5 ml of tetrahydrofuran is added dropwise. The resulting mixture is allowed to stir at -78 ° C for 20 minutes and 2 ml of trimethylsilyl chloride are added dropwise. The reaction mixture is warmed to ambient temperature and stirred for 1 hour. The mixture is then cooled to 0 ° C and poured into 150 ml of 0 ° C ethyl acetate. Add sufficient 0,5 M monobasic potassium phosphate buffer solution to make the organic and aqueous layers clear. The layers are separated and the organic layer is washed with three 150 ml portions of 0.5 M monobasic potassium phosphate solution, one 150 ml portion of sodium chloride solution, one 150 ml portion of saturated sodium chloride solution and dried over sodium sulfate. The solution is concentrated in vacuo to 2.83 g of an oil which, on trituration with hexane, gives 1.67 g of the title compound as a solid.

.. · 25 C) (+) - 3-azido-4,4-dimethyl-2-oxo-1-azetidine In a 50 ml three-necked flask dissolve 1.52 g of: (+) - 3-azido-4 , 4-dimethyl-2-oxo-1-azetidine tert-butyldiphenylsilane 25 any acetonitrile.

To this stirred solution is added 0.25 ml of 48% hydrofluoric acid. Stir at ambient temperature:. and a 48 ml portion of 48% hydrofluoric acid is added every 60 minutes until, after 6.5 hours, a total of 3.25 ml of 48% hydrofluoric acid has been added.

The reaction mixture is then cooled to 0 [deg.] C., neutralized with saturated sodium bicarbonate and extracted with 120 ml of ethyl acetate. The organic layer is washed with 100 ml of water, 100 ml of saturated sodium chloride solution and dried over sodium chloride. The dry solution is concentrated in vacuo to give 1.34 g of an oil, the crude oil is chromatographed on 27 g of silica gel with hexane, followed by 33% ethyl acetate in hexane to give 0.358 g of the title compound as a solid.

E) (+) - 3-azido-4,4-dimethyl-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt to 0.100 g of (+) - 3-azido-4,4-dimethyl-2-oxo-1 -acetidine at 0 ° C under argon is added 2.8 ml of 0.5M dimethylformamide-sulfur trioxide complex. The mixture is allowed to warm to ambient temperature and stirred for 45 minutes. The solution is poured into 20 ml of 0.5 M pH 5.5 monobasic potassium phosphate buffer solution. This is washed with three 20 ml portions of methylene chloride (discarded) and 0.237 g of tetrabutylammonium hydrogen sulphate is added to the aqueous solution.

It is extracted with four 20 ml portions of methylene chloride and the combined organic extracts are washed with 20 ml of 8% sodium chloride solution. The methylene chloride solution is dried (sodium sulfate) and concentrated in vacuo to give 0.31 g of an oil which, according to nmr, is 50% dimethylformamide and 50% of the title compound.

E) (+) - 3-Amino-4,4-dimethyl-2-oxo-1-azetidinesulfonic acid 25 Solution of 0.155 g of (+) - 3-azido-4,4-dimethyl-2-oxo 1-Azetidinesulfonic acid tetrabutylammonium in 0.6 ml of methanol is hydrogenated using 10% palladium on carbon for 20 minutes under 1 atmosphere. The catalyst is filtered off and rinsed with methylene chloride, which is combined with the methanol solution.

This clear solution is treated with 0.123 ml of 97% formic acid. Upon addition of acid, the solution immediately becomes cloudy. After standing for 1 hour at 5 ° C, the solution is filtered to give 0.0664 g of the title compound, m.p. 200-202 ° C (dec.).

NMR (D 2 q) 1.6 δ (3H, S), 1.68 (3H, S), 4.42 (1H, S), IR (KBr) 1765 cm -1.

Il g 7 o π n n λ

Ό 'υ υ £ / I

Example 41 (3- (Z) - (32-Amino-4-thiazolyl) (methoxyimino) -acetyl-amino--4,4-dimethyl-2-oxo-1-azetidinesulfonic acid, potassium salt 5 is N-hydroxybenzotriazole hydrate (50 mg) and (Z) -2-amino-N- (methoxyimino) -4-thiazoleacetic acid in 0.5 ml of dimethylformamide, treated with 67 mg of dicyclohexylcarbodiimide under argon at ambient temperature. The resulting mixture was stirred for 1 hour while (+) - 3-amino-4,4-dimethyl-2-oxo-1-azetidinesulfonic acid (57 mg; see Example 40) was added as a solid, followed by dropwise addition of triethylamine. (0.05 mL). The reaction is stirred at ambient temperature for 16 hours to 15 hours. The dimethylformamide is removed under high vacuum at 30 ° C and the residue is slurried in 4 ml of acetone and filtered. The filter cake is washed with 4 ml of more acetone and potassium perfluorobutane sulfonate (85 mg) is added to the filtrate, followed by ether.

Trituration of the gummy substance with ether gives 40 mg of a dark solid which is chromatographed on a 70 ml HP-20AG column. Elution with water gives 20 mg of the title compound in fractions (5 ml) 16-40 after evaporation, trituration with 25 (1: ¾ acetone-hexane) and drying.

Sp. 22 5 ° C (dec.).

Anal. Calculated for C 11 H 11 N 2 O 5 So.K: C, 31.80;

11 14 o b Z

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

Example 42; (3S-trans) -3- (2-Amino-4-thiazolyl) oxoacetyl-4-amino--4-methyl-2-oxo-1-azetidinesulfonic acid, potassium salt

To a solution of diphenylphosphinyl-chloride (1.85 g) in dry dimethylformamide (15 ml) cooled in an ice-methanol bath (-15 ° C to -20 ° C) is added (2-amino-4- thiazolyl) glyoxalic acid triethylamine salt (2.14 g). After stirring for 0.5 h, a solution of (3S-trans) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid (3S-trans) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid is added to the cold stirred anhydrous solution. 1.08 g, see Example 33) and triethylamine (1.92 ml) in dry dimethylformamide (5 ml) and the reaction mixture is stirred at 5 ° C for 24 hours. The solvent is removed in vacuo, the remaining dark oil is dissolved in water and chromatographed using Dowex 50 X 2-400 resin (K + -me-th, 200 ml). Eluting with water (15 ml fractions), 10 crude product is collected in fractions 13-27 (3.37 g).

Chromatography on HP-20 resin (200 ml), eluting with water (15 ml fractions) gives the desired product in fractions 18-26. Removal of water in vacuo gives the title compound as an amorphous powder.

Anal, calculated for C 9 H 9 N 2 O 3 S 2 K 3 (372.42); C, 29.02; H, 2.44; N, 15.04; S, 17.22; K, 10.50 Found: C, 28.87; H, 2.62, N, 14.85; S, 15.09; K, 10.81.

Example 43 20 S - [(3) - (Z), 4H & (X) -3 - [(2-amino-4-thiazolyl) ({2- (diphenylmethoxy) -2-oxoethoxy] acetyl) amino N-2-methyl-4-oxo-1-azetidinesulfonic acid, potassium salt

Following the procedure used in Example 32, but substituting (Z) -2-amino-Q (- (N, 2- (diphenylmethoxy) -2-oxoethoxy) imino) -4-thiazoleacetic acid: (Z) -2 -amino - (- (methoxyimino) -4-thiazoleacetic acid and first treatment of (3S-cis) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid with triethylamine to give the title compound, m.p. 155-160 ° C, dec.

S

il 69 80271

Example UU

S (R *) (-3-CCO2 ** - 3 (1) - (2 (3-dioxo-1-piperazinyl) carbonyl) amino} phenylacetylamino} -4-methyl-2-oxo-1-azetidinesulfonic acid, potassium salt 4 -methyl-2-oxo-1-sulfo-3-azetidinyl) amino 2-oxo-1-phenylmethyl,) amino? The potassium salt of the phenylmethyl β-2,3-dioxo-1-piperazinyl-urea acid phenylmethyl ester (see Example 38) is hydrogenated using gaseous hydrogen and 10% palladium on carbon as a catalyst to give the title compound, m.p. 165 ° C> dec.

Example M-5- (3S (Z), (-3-Amino-4-thiazolyl) - (E (1-carboxy-1-methylethoxy) imyl) acetyl) amino) -2-oxo-1-acetyl-15-sulfonic acid , sodium salt (1: 2) ^ 3:; (7,) 4- [3- (2-amino-1-ylcolyl) -N- (2- (diphenylmethoxy) -1,1-dimethyl-2- The tetrabutylammonium salt of oxoethoxy-imino) -acetyl} -amethyl) -2-oxo-1-azetidinesulfonic acid (see Example 10) is deprotected using trifluoroacetic acid and anisole to give the title compound, m.p. 185 ° C; : decomposes with aqueous sodium hydroxide after conversion to the disodium salt and after purification using HP-20.

25 Example 4 6

Following the procedure used in Example 5, but substituting (Z) -2-amino - # -, ((hydroxy (phenylmethoxy) phosphinyl) -methoxy) imin) -4-thiazoleacetic acid (Z) -2-amino-α- - (Diphenylmethoxy-N-oxobutoxy-imin} -β-thiazoleacetic acid gives ^ 3S (Z - * - & 2-amino-H-thiol-solyl) -4- (diphenylmethoxy) -4- oxobutoxy *) imino-acetyl-amino--2-oxo-1-azetidinesulfonic acid, potassium salt. ‘La; mp. 125-130 ° C, dec.

70 pno ^ -i

ν / \ J L. I I

Example 47 (trans, Ζ) -3- (2-Amino-4-thiazolyl) - [κ (1-carboxy-1-methyl-ethoxy) -imino] -acetyl] -amino-4-ethyl-2-oxo-1 -azetidinesulfonic acid, dipotassium salt 5 A) (trans, Z) -3- [N- (2-amino-4-thiazolyl) -N- (1-diphenylmethoxycarbonyl-1-methylethoxy) -im (g) acetyl] -amino] -4-ethyl 2-Oxo-1-acetylsulfonic acid, dipotassium salt (trans) -3-amino-4-ethyl-2-oxo-1-azetidine-10-sulfonic acid (0.55 g) and 335 mg of triethylamine are dissolved in 50 ° C. to 1 ml of dry dimethylformamide. (Z) -2-Amino-N - [(1-diphenylimidecarbonyl-1-methylethoxy) -imino] -4-thiazoleacetic acid (1.14 g) is added with stirring at 0 ° C and then 450 mg of hydroxy-15 benzotriazole and then 0.69 g of dicyclohexylcarbodiimide. After stirring for 16 hours at 0 ° C, the flask is evacuated. Dry acetone (25 mL) is added to the solid with stirring. The mixture is filtered and 0.94 g of potassium perfluorobutanesulfonate is added to the filtrate and then 100 ml of ether are added. After standing at 0 ° C for 1 hour, the solid is filtered, washed with ether and dried in vacuo to give 1.58 g of the title compound.

B) (trans, Z) -3-Amino-4-thiazolyl) - (1-carboxy-1'-methylethoxy) iminylacetyl-amino-4-ethyl-2-oxo-1-azetidinesulfonic acid, dipotassium salt is 1.31 g of (trans, Z) -3- [C (2-amino-4-thiazolyl) / (1-diphenylmethoxycarbonyl-1-methylethoxy) iminoacetyl] amino] -4-ethyl-2-30 oxo-1 -azetidinesulfonic acid dipotassium salt in 10 ml of anisole, 5 ml of trifluoroacetic acid are added for 10 minutes at -15 ° C. After stirring for 2 hours at -10 ° C, a clear solution is obtained. At -30 ° C, 80 ml of dry ether are added and the precipitate obtained is filtered off and then treated with 5 ml of water. The pH is adjusted to 5.5 with potassium hydroxide at 0 ° C and the mixture is filtered to remove unchanged starting material. Swamp-

II

7i 80271 dos are chromatographed using HP-20 and water as eluent. Lyophilization gives 185 mg of the title compound, m.p. 160 ° C, dec.

Example H8 5 (3S (Z) - (3-O-S (2-amino-4-thiazolyl)) - ((4-hydroxy-4-oxobutoxy) imine) and -2-oxo-1-azetidine sulfonic acid , potassium salt (3S (Z)> - (3- (2-amino-4-thiazolyl)) - [4- (diphenylmethoxy) -H-oxobutoxy-imino] acetyl} amino] -10 2-oxo-1-azetidinesulfonic acid potassium salt (see Example 46) deprotection using trifluoroacetic acid and anisole gives the title compound m.p. > 20 ° C.

Example 49 15 S-trans-3 '- 3-amino-4-cyclohexyl-2-oxo-1-azetidinesulfonic acid A) t-butoxycarbonylamino) - (3-cyclohexyl) -N-hydroxy-threo-propionic acid Λ-cyclohexyl- Amino-hydroxy-threo-propionic acid (15 g) is suspended in 150 ml of acetonitrile and 70 ml of water. Triethylamine (17.8 g) is added and the mixture is heated to 60 ° C with stirring. At this temperature a clear solution is obtained and: 21.0 g of di-t-butyl pyrocarbonate are added and stirring at 60 [deg.] C. is continued for 1.5 hours. The solvent is removed in vacuo and 50 ml of water are added. The aqueous layer is extracted with ethyl acetate at pH 2, adjusted by the addition of 3N HCl. The organic layer is separated, dried over Na 2 O and evaporated to dryness.

The remaining crystalline material is filtered through petroleum ether to give 20.4 g of the title compound, m.p. 113-115 ° C.

·: B) N - (t-Butoxycarbonylamino) - [4-cyclohexyl] -N- (hydroxy-N-methoxy-threo-propionamide) 35- (t-Butoxycarbonylamino) - [? - cyclohexyl] “Hydroxy-threo-propionic acid (20.2 g) and 7.6 g of O-methylhydroxylamine hydrochloride are suspended in 350 ml of water and 175 ml of t-butanol. The pH of the mixture is adjusted to * 72 .mu.l with potassium carbonate. 4. Add 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (16.4 g) and maintain the pH at 4 with stirring for 1.5 hours, remove the t-butanol in vacuo and saturate the remaining aqueous solution with sodium chloride. and extracted twice with 100 ml portions of ethyl acetate The organic controls are combined, dried over £ 30 and evaporated to dryness, the remaining crystals are filtered off with petroleum ether to give 18.6 g of the title compound, mp 125-127 ° C.

C) Cf- (t-Butoxycarbonylamino) - β-cyclohexyl- [N- (methanesulfonyloxy) -N-methoxy-threo-propionamide t-butoxycarbonylamino) - β-cyclohexyl-15H-hydroxy-N-methoxy -treo-propionamide (18.3 g) is dissolved in 100 ml of dry pyridine with stirring. The solution is cooled to 0 ° C with stirring and 9.3 g of methanesulfonyl chloride are added dropwise.

After 1 hour at 0 [deg.] C., 3.3 g of methanesulphon-20-yl chloride are added and stirring is continued for a further 1 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 over Na 2 SO 4 and the solvent is removed in vacuo.

The residual solid is taken up in petroleum ether to give 19.0 g of the title compound, m.p. 150-15 2 ° C.

D) [3S-trans] -3- (t-butoxycarbonylamino) -4-cyclohexyl-1-methoxy-2-azetidinone Qi- (t-butoxycarbonylamino) - [β-cyclohexylmethanesulfonyloxy) -N-methoxy-threo- propionamide (18.7 g) is dissolved in 500 ml of dry acetone. Potassium carbonate (9.8 g) is added and the suspension is heated to reflux with stirring for 5 hours.

**: 3 5 The insoluble inorganic substance is filtered off and the solvent is removed in vacuo and the residual oil is dissolved in 30 ml of ethyl acetate. Upon addition of petroleum ether, the title compound precipitates and is filtered (12.9 g), m.p. 110-112 °. C.

I! 80271 73 E) -3S-trans-3- (t-butoxycarbonylamino) -4-cyclohexyl-2-azetidinone E3S-trans--3- (t-butoxycarbonylamino) -H-cyclohexyl-1-methoxy-2 -azetidinone (1 g) is added to 50 ml of liquid ammonia with stirring. Sodium (0.154 g) is added in 5-6 portions over 5 minutes. After this time, an additional 0.025 g of sodium is added and stirring is continued for 5 minutes. Ammonium chloride (0.89 g) is added and ammonia-10 ki is removed. The residue is extracted with warm ethyl acetate. The organic extract is evaporated to dryness and the remaining crystals of the title compound are filtered off with petroleum ether to give 0.5 g, m.p. 130-132 ° C.

F) [3S-trans] -3- (t-butoxycarbonylamino) -4-15 cyclohexyl-2-oxo-1-azetidinesulfonic acid, pyridine salt N, S-3S-trans-3- (t-butoxycarbonylamino) -4 -cyclohexyl-2-azetidinone (5.3) is dissolved in 20 ml of methylene chloride and 80 ml of dimethylformamide.

After the addition of 60 mmol of pyridine-sulfur trioxide complex, the solution is stirred for 6 hours at room temperature. Removal of the solvent in vacuo gives 11.3 g of the title compound as an oil.

! G.) / [3S-Tranyl-3- (t-butoxycarbonylamino) -4-25-cyclohexyl-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt - ε-3S-trans] -3- (t-butoxycarbonylamino) -4 The pyridine salt of cyclohexyl-2-oxo-1-azetidinesulfonic acid (11.3 g) is dissolved in 250 ml of water. Tetrabutylammonium hydrogen sulfate (9.0 g) is added with stirring and the pH is adjusted to 6.5 with potassium hydroxide. The aqueous solution is extracted twice with 200 ml portions of methylene chloride. The organics are dried over Na 2 SO 4, filtered and the solvent is distilled off to give 8 g of the title compound, m.p. 135-138 ° C.

pno7i 71 H) 43S-Trans-3-3-amino-4-cyclohexyl-2-oxo-1-azetidinesulfonic acid 43S-trans-3- (t-butoxycarbonylamino) -4-cyclohexyl-2-oxo-1-azetidinesulfonic acid tetra- The 5 butylammonium salt (3.8 g) is stirred in 20 ml of formic acid for 3 hours and then 20 ml of methylene chloride are added. The precipitated title compound is filtered off, m.p. 217-219 ° C.

Example 50 10 (3S- £ 30t (Z), 4 (R (-3- (R (2-Amino-4-thiazolyl)) - (methoxyimino) acetyl) -amino] -4- (4-cyclohexyl-2-oxo- 1-Azetidinesulfonic acid, potassium salt 43S-Trangy-3-amino-4-cyclohexyl-2-oxo-1-azetidinesulfonic acid (0.25 g) is dissolved in 30 ml of dry dimethylformamide and 0.12 g of triethylamine with stirring. . After a clear solution is obtained, (Z) -2-amino-α- (methoxyimino) -4-thiazoleacetic acid (0.2 g), 0.16 g of hydroxybenzotriazole and 0.42 g of dicyclohexylcarbodiimide are added. Stirring is continued for 48 hours at ambient 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 addition of 50 ml of ether *: the title compound precipitates and is filtered off. Column chromatography using HP-20 and water / acetone (9: 1) as eluent gives 0.36 g of product, m.p. 200-205 ° C (after lyophilization).

Example 51 (5S- [3 (* (Z), 4 (ft))) - 3- (p2-amino-4-thiazolyl) -4- (1-30 carboxylmethylethoxy) imino) acetyl} amino} -4- cyclohexyl-2-oxo-1-azetidinesulfonic acid, dipotassium salt A) E 3 S- (3 * (Z), 4H)) - 3- (E (2-amino-4-thiazolyl)) - N, N-diphenylmethoxycarbonyl-1- methylethoxy) -3 imino (acetyl) amino] -4-cyclohexyl-2-oxo-1-azetidinesulfonic acid, potassium salt: 1-3S-trans7-3-amino-4-cyclohexyl-2-oxo-1-azetidinesulfonic acid (0 , 2 g, see Example 49) is dissolved in 30 ml of dimethylformamide and 0.09 g

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75 80271 triethylamine with stirring. 0.30 g of (Z) -2-amino-O- [N- (1-diphenylmethoxycarbonyl-1-methylethoxy) imino] -1t-thiazoleacetic acid and 0.33 g of dicyclohexylcarbodiimide are added and the mixture is stirred at ambient temperature. continued for 12 hours. The precipitated urea is filtered off and the mother liquor is evaporated to dryness. The residual oil is dissolved in 5 ml of acetone, treated with 0.3 g of potassium perfluorobutanesulfonate and poured into 100 ml of ether with stirring.

10 (3S- (3) - (Z), 4 (R) - (3 -, (2-amino-1-thiazolyl)) - ((1-diphenylmethoxycarbonyl-1-methyloxy)) - imino / acetyl) amino--4-cyclohexyl-2-oxo-1-azetidinesulfonic acid potassium salt (0.61 g) precipitates and is filtered.

B) (3S- (3R) (Z), (32-Amino-4-thiazolyl) ((1-carboxy-1-methylethoxy) imino) acetyl] amino) -4-cyclohexyl-2-oxo-1- azetidine sulfonic acid, dipotassium salt (3S- (3) (4R, 3 - ((3- (2-amino-4-thiazolyl) -2-1-diphenylmethoxycarbonyl-1-methylethoxy) imino) -. ; the potassium salt of acetylamine} -4-cyclohexyl-2-oxo-1-azetidinesulfonic acid (0.61 g) is suspended in 6 ml of anisole, cooled to -15 ° C and added dropwise; 5 ml of trifluoroacetic acid with stirring.

The temperature is maintained for 1 hour and then lowered to -30 ° C. Add about 100 ml of dry ether at such a rate that the temperature does not exceed -30 ° C. The precipitated compound is filtered and chromatographed using HP-20 resin and water / acetone (9: 1) as eluent to give 0.3 g of the title compound, m.p. 5.200 ° C dec. (after lyophilization).

76 80271

Example 52 (trans, Z) -3 - [(2-Amino-4-thiazolyl) (methoxyimino) acetyl] amino] -4-ethyl-2-oxo-1-azetidinesulfonic acid, potassium salt Following the procedure used in Example 47, part Δ but substituting (Z) -2-amino-α- (methoxyimino) -4-thiazoleacetic acid with (Z) -2-amino-N - [(1-diphenylmethoxycarbonyl-1-methylethoxy) ) iminc-N-thiazoleacetic acid to give the title compound, 10 m.p. 190 ° C dec.

Example .5 3 (+) - (trans) -3-Amino-2-oxo-4-phenyl-1-azetidine sulfonic acid A) (+) - (trans) -2-oxo-4-phenyl-1-azetidine - 15 tert-butyldiphenylsilane

A solution of tert-butylchlorodiphenylsilane (20.56 g) in dimethylformamide (45 ml) is cooled to 0 ° C under argon and treated with triethylamine (10.4 ml) and then (+) - 2-oxo-4 -phenyl-1-20 with azetidine. After several hours at 0 ° C, the mixture is treated with additional triethylamine (1 ml) and tert-butylchlorodiphenylsilane (2.11 g) and allowed to stir for 65 hours at 5 ° C. The reaction mixture is poured into ice water (300 ml) and extracted (3: 1) with ether-ethyl acetate (three 125 ml portions). The organic extracts are washed with pH 4.5 phosphate buffer solution (three 50 ml portions), saturated with sodium bicarbonate solution (50 ml), water (two 50 ml portions), saturated sodium chloride solution and dried (Na 2 SO 4). Filtration and concentration in vacuo gives a solid which is washed with hexane to give, after drying (high vacuum), 15 g of the title compound as a solid.

Il 77 80271 B) (+) - (trans) -3-azido-2-oxo-4-phenyl-1-azetidine-tert-butyldiphenylsilane 50 ml flask equipped with a stir bar, a gas inlet and a partitioned, flame-dried under argon and n-butyllithium (0.65 ml of a 1.6 M solution in hexane) is added, which is cooled to -40 ° C and dissolved in tetrahydrofuran (2 ml). Diisopropylamine (0.16 ml) is added dropwise and the resulting mixture is stirred for 30 minutes and cooled to -78 ° C.

A solution of (+) - (trans) -2-oxo-4-phenyl-1-azetidine-tert-butyldiphenylsilane (400 mg) in tetrahydrofuran (1.5 ml) is added dropwise over about 5 minutes. After stirring for an additional 20 minutes, the solution is treated with p-toluenesulfonyl azide (204 mg) in tetrahydrofuran (0.5 mL). The resulting mixture was stirred for 10 minutes at -78 ° C and treated dropwise with chlorotrimethylsilane (0.4 mL). After stirring for 10 minutes, the cooling bath is removed and the reaction mixture is stirred at ambient temperature for 2.5 hours.

Ethyl is then added while cooling to 0 ° C. acetate (20 ml) followed by the addition of pH 4.5 phosphate buffer solution (8 ml). The organic layer is washed with buffer solution (two 8 ml portions), 5% sodium bicarbonate solution (three 10 ml portions), 50% sodium chloride solution (10 ml), saturated sodium chloride solution (10 ml) and dried (Na 2 SO 4). Filtration and concentration in vacuo gives 500 mg of an oil which is chromatographed on 5% ethyl acetate-hexane to give the title compound (253 mg).

30 C) (+) - (trans) -3-azido-2-oxo-4-phenyl-1-azetidine

A solution of 17 g of crude (+) - (trans) -3-azi-. do-2-oxo-4-phenyl-1-azetidine-tert-butyldiphenylsilane, dissolved in methanol (24 mL) and treated dropwise at 0 ° C with concentrated HCl (35 mL).

78 80271 The cooling bath is removed, the reaction is stirred at ambient temperature for 1 hour and re-cooled to 0 ° C while saturated sodium bicarbonate solution is added until neutral. The resulting mixture 5 is extracted with ethyl acetate (one portion of 300 ml and four portions of 100 ml) and the organic extracts are washed with 1: 1 5% sodium bicarbonate solution and 50% sodium chloride solution and saturated sodium chloride solution. and dried (Na 2 SO 4). Filtration and concentration in vacuo gave 15 g of a heavy oil which was chromatographed on 100 g of silica gel with 20% ethyl acetate-hexane to give 460 mg of the title compound.

D) (+) - (trans) -3-Azido-4-phenyl-1-azetidine-sulfonic acid, tetrabutylammonium salt

A solution of (+) - (trans) -3-azido-2-oxo-4-phenyl-1-azetidine (300 mg) in dimethylformamide (3 ml) is cooled to 0 ° C under argon and treated dropwise with dimethylformamide. and sulfur trioxy-20 din complex (4.78 ml of a 0.5 M solution in dimethylformamide). The cooling bath is removed, the reaction mixture is stirred at ambient temperature for 2 hours and poured into 80 ml of 0.5 M monobasic potassium phosphate (pH 5.5). The solution is extracted with dichloromethane (discarded) and 541 mg of tetrabutylammonium bisulfate are added. The resulting mixture is extracted with dichloromethane and the organic extracts are washed with 10% sodium chloride solution and dried (£30%). Filtration and concentration in vacuo gives 800 mg of an oil; about 40% of the desired product, residual dimethylformamide. This mixture is used crude in the following method.

E) (+) - (trans) -3-Amino-4-phenyl-1-azetidinesulfonic acid

Solution of the tetrabutylammonium salt of (+) - (trans) -3-amino-4-phenyl-35 1-azetidinesulfonic acid

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79 80271 in 4 ml of methanol, hydrogenated using 30 mg of platinum oxide in one atmosphere and at room temperature. After 15 minutes, the system is degassed and fresh hydrogen is added. After 45 minutes, reaction 5 is complete and the system is purged with nitrogen. After several days at room temperature in dichloromethane-methanol (4: 1, 200 ml) the catalyst aggregate is complete and filtered. The filtrate is concentrated in vacuo to 18 ml and 0.2 ml of 97% formic acid is added.

After cooling for several days at 5 ° C, the solid obtained is filtered and washed with dichloromethane to give, after drying, 150 mg of the title compound as a solid.

Analysis calculated for C 9 H 9 N 2 O 2 S: C, 44.62; 15 H, 4.17; N, 11.57; S, 13.23 Found: C, 43.36; H, 4.31; N, 11.09; S, 13.02. Example 54 (+) - (Trans, Z) -3-Amino-4-thiazolyl) (methoxyimino) -acetylamino} -2-oxo-4-phenyl-1-azetidine sulfonyl 20 acid, potassium salt

A solution of N-hydroxybenzotriazole hydrate (52 mg) and (Z) -2-amino-N- (methoxyimino) -4-thiazoacetic acid (69 mg) in dimethylformamide (0.3 ml) is treated with solid. with dicyclohexylcarbodiimide 25 (70 mg) under argon at ambient temperature. The resulting mixture was stirred for 1 hour, V (+) - (trans) -3-amino-2-oxo-4-phenyl-1-azetidinesulfonic acid (75 mg; see Example 53) was added as a solid, followed by dropwise addition of triethylamine ( 0.05 ml). The reaction is stirred at ambient temperature for 23 hours. The dimethylformamide is removed under high vacuum at 30 ° C and the residue is triturated with 2 ml of acetone and filtered. The filter cake is washed with additional acetone (two 3 ml portions) and potassium perfluorobutane-35-sulfonate (86 mg) is added to the filtrate. Dilution with 10 mL of ether gives a gummy solid which is triturated, washed with acetone and hexane to give, after drying, 82 mg of the title compound as a solid.

802/1 80 '1

Analysis for C 1-4 H 2 N 2. OgS ^. K

Calculated: C, 40.25; H, 3.16; N, 15.65; S, 14.33; K, 8.74 Found: C, 38.60; H, 3.19; N, 15.07; S, 13.87; 5 K, 7.5.

Example 55 (cis) -2-Oxo-4-phenyl-3- (phenylacetyl) amino] -1-acetidinesulfonic acid, potassium salt A) N-Benzylidene-2,4-dimethoxybenzylamine 10 12 0.0 g of 2,4-dimethoxybenzylamine hydrochloride is added to 100 ml of 1N sodium hydroxide solution and the mixture is extracted with 125 ml of ethyl acetate. The organic layer is dried over anhydrous sodium sulfate and the solvent is removed to give 10.2 g of 2,4-dimethoxybenzylamine as an oil. This amine is dissolved in 150 ml of benzene; 6.47 g of benzaldehyde and 0.6 g of p-toluenesulfonic acid monohydrate are added. The mixture is heated to reflux, which removes water using a Dean-Stark separator, and the calculated amount of water (1.1 ml) separates in two hours. The mixture is cooled to room temperature. Upon further cooling in the benzene solution, a small precipitate forms.

1: The benzene is removed under reduced pressure and 60 ml of petroleum ether are added to the residue, an oily layer separates together with an additional precipitate. Benzene (10 mL) is added to make the layers homogeneous and the remaining precipitate is filtered. The filtrate is removed from the solvent to give 14.2 g of the title compound as an oil.

B) (+) - (cis) -4-Phenyl-1- (2,4-dimethoxybenzyl) -2-oxo-3-azidoazetidine N-azidoacetic acid (1.62 g) is dissolved in 25 ml of methylene chloride under nitrogen. To this solution are added 3.24 g of triethylamine and 1.02 g (4.0 mmol) of imine-35 N-benzylidene-2,4-dimethoxybenzylamine dissolved in 10 ml of methylene chloride. The mixture is cooled in an ice bath and 3.36 g of trifluoro-

II

n n o n λ

81 ou ^ f I

acetic anhydride; the solution turns dark. After stirring for 1 hour in an ice bath, the mixture is warmed to room temperature and stirred for an additional 15 minutes. The solution is then washed with water (60 mL), 5% NaHCO 3 solution (two 50 mL portions), and 1N HCl solution (60 mL). The organic layer is dried over anhydrous sodium sulfate and the solvent is removed to give 1.72 g of crude product as a dark gum. The gum is treated with charcoal several times and the resulting brown mixture is chromatographed on 40 g of silica gel and eluted with (1: 1) petroleum ether: ethyl acetate. The combined fractions give a crystal rapidly on freezing in a dry ice-acetone bath. Using this as the initial crystal, the product is recrystallized from petroleum ether-ethyl acetate to give the title compound as needles which melt when warmed to room temperature.

C) (+) - (cis) -4-phenyl-2-oxo-3-azedoazetidine (+) - (cis) -4-phenyl-1- (2,4-dimethoxybenzyl) -20 2-oxo-3- azedoazetidine (737 mg) is dissolved in 25 ml of acetonitrile and heated to 80-83 ° C under nitrogen. To the resulting solution are added 943 mg of potassium persulfate and 57 mg of potassium monohydrogen phosphate, each dissolved in 25 ml of water, over 1 hour. After the addition, the se-J 25 is further heated at 80 ° -83 ° C for 7 hours. Seos ··; is cooled and the pH is adjusted to 6-7 by adding solid potassium monohydrogen phosphate. Most of the acetonitrile is removed under reduced pressure and the resulting mixture is extracted with 60 ml of chloroform. The chloroform layer 30 was washed with water (60 ml), dried over anhydrous sodium sulfate and the solvent was removed to give the crude product as an oil. The crude product is chromatographed on 40 g of silica gel and eluted with (1: 1) petroleum ether-ethyl acetate. The combined fractions give 35 crystals and the product is recrystallized from petroleum ether-ethyl acetate to give 267 mg of the title compound.

82 80271 D) (+) - (cis) -4-Phenyl-2-oxo-3-azedo-1-azetidinesulfonic acid, tetrabutylammonium salt (+) - (cis) -4-phenyl-2-oxo-3-azidoazetidine (162 mg) is cooled to 0 ° C under nitrogen and 3.5 ml of a solution of about 0.5M dimethyl-formamide-sulfur trioxide complex in dimethylformamide is added dropwise via a hand syringe. The resulting clear solution is stirred at 0 ° C for 15 minutes. The mixture is then poured into 50 ml of 0.5M monobasic potassium phosphate solution and washed with methylene chloride (three 50 ml portions). To the aqueous solution is added tetra-n-butylammonium bisulfate (292 mg) and the mixture is extracted with methylene chloride (six 50 ml portions). The combined methylene chloride layers are dried over anhydrous sodium sulfate and the solvent is removed to give 272 mg of the title compound as a gum.

E) (+) - (cis) -2-oxo-4-phenyl-3-phenylacetyl) -N ....

Amino--1-azetidinesulfonic acid, potassium salt (+) - (cis) -4-phenyl-2-oxo-3-azido-1-azidinesulfonic acid tetrabutylammonium salt (293 mg) is dissolved in 4 ml of ethanol and hydrogenated to 80 mg: 11a platinum oxide as a catalyst in one atmosphere.

:: after stirring for 1 hour, the catalyst is filtered through a millic pore filter using Celite; 2 5 some catalyst particles pass through the filter to give a black filtrate. The ethanol is removed under reduced pressure and the residue is dissolved in 4 ml of dimethylformamide. N-hydroxybenzotriazole monohydrate (81 mg), 78 mg of phenylacetic acid 30 and 117 mg of dicyclohexylcarbodiimide are added and the mixture is stirred for about 16 hours under nitrogen. The slurry is evaporated in vacuo and triturated with 10 ml of acetone. The resulting slurry is filtered through a millic pore filter with a Celite supernatant and the brown filtrate is treated with 193 mg *: 35 potassium perfluorobutanesulfonate. Upon addition of 20 ml of ether, a gummy substance separates. The liquid is removed and the gummy substance is washed with ether. The gummy substance is dissolved in 10 ml of methanol. When adding ether

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83 80271 a small amount of precipitate is formed. The mixture is filtered and the colored filtrate is treated with additional ether. The precipitate formed is filtered off and recrystallized twice from ether-methanol to give 26 mg of the title compound.

Analysis calculated for C 1-4 H 2 O 2 ^ SK. 2H 2 O: C, 46.99; H, 4.41; N, 6.45 Found: C, 47.24; H, 4.19; N, 6.34.

Example 56 10 (cis, Z) -3- [4- (2-Amino-4-thiazolyl) (methoxyimino) acetyl] amino] -2-oxo-4-phenyl-1-azetidinesulfonic acid, potassium salt (cis ) The potassium salt of -2-oxo-4-phenyl-3 - [(phenylacetyl) -amino] -1-azetidinesulfonic acid (560 mg; see Example 55, Part E) is dissolved in 5 ml of ethanol and hydrogenated with 110 mg of platinum oxide as a catalyst. in one atmosphere. After stirring for 1 hour, the catalyst is filtered through a millic pore filter using Celite; the catalyst particles pass through the filter to give a black filtrate. Ethanol. . is removed under reduced pressure and the residue is dissolved in 4 ml of dimethylformamide. N-Hydroxy-γ-benzotriazole monohydrate (168 mg), 221 mg of (Z) -2-amino- (- (methoxyimino) -4-thiazoleacetic acid and 227 mg of 2-dicyclohexylcarbodiimide are added and the mixture is stirred for 16 hours under nitrogen. The slurry is evaporated in vacuo and triturated with 15 ml of acetone. The gum is dissolved in 5 ml of water and taken up in 150 ml of HP-20 resin and eluted with water, fractions (30 ml each) of compound 16-34 are combined and lyophilized to give 201 mg of the title compound. compound as a solid.

84 80271

Analysis calculated for 1 1/2 H 2 O: C, 36.73; H, 3.49; N, 14.28; S, 13.07; K, 7.79 Found: C, 36.65; H 3.00; N 13.99; S, 13.48; K, 8.30 Example 57 (eis) -3-Amino-4- (methoxycarbonyl) -2-oxo-1-azetidinesulfonic acid - (S) - (4-methoxyphenyl) imino / acetic acid, methyl ester 10 Dry 1 A 1 liter three-necked flask equipped with a nitrogen inlet and a stir bar is filled with 56.88 g of MgSO 4 and then a solution of recrystallized anisidine (19.43 g) in dichloromethane (250 ml) is added. After stirring for an additional 20 minutes at 0 ° C, the reaction mixture is filtered off with suction, dried over sodium sulfate, filtered and concentrated to an oil which solidifies in half under high vacuum at 5 ° C.

B) (cis) -3- (1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl) -4-methoxycarbonyl-2-oxo-1- (4-methoxyphenyl) azetidine. A 1,500 ml three-necked flask equipped with a stir bar, addition funnel, septum and nitrogen inlet is filled with a solution of IJ. 4-Methoxyphenyl) iminate / acetic acid methyl ester (21.09 g) in dichloromethane (150 ml) and cooled to 0 ° C. Triethylamine (19.2 mL, 0.14 mol) is added dropwise, followed by a solution of (N-phthalimido) acetylic acid chloride (28.4 g) in dichloromethane (150 mL) over 1 hour. The resulting mixture is stirred for 1.5 hours at 0 ° C and diluted with 2.5 liters of dichloromethane. The organic solution is washed with pH 4.5 monobasic potassium phosphate (two 500 ml portions), 5% sodium bicarbonate (two 500 ml portions), saturated sodium chloride solution (500 ml) and dried over sodium sulfate.

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es 80271 la. Filtration and concentration in vacuo gives a solid which is washed with ethyl acetate, cold acetone and hexane to give 18.65 g of product.

5 C) (cis) -4- (Methoxycarbonyl) -1- (4-methoxyphenyl) -2-oxo-3- [trans-phenylmethoxy] carbonyl] amino] -7-azetidine

A dry 500 ml flask equipped with a nitrogen inlet, a stir bar and a septum is filled with 18.65 g of (cis) -3- (1,3-dioxo-2H-isoindol-2-yl) -4-methoxycarbonyl 2-oxo-1- (4-methoxyphenyl) azetidine and 325 ml of dichloromethane. The resulting suspension is cooled to -30 ° C and methylhydrazine (3.52 ml) is added dropwise. The reaction is warmed to 0 ° C and stirred for 1 hour. An additional 0.4 mL of methylhydrazine is added and the mixture is stirred for 10 minutes. This treatment is repeated after a total of 2.9 equivalents of methylhydrazine (7.7 ml) have been added. The solvent is removed in vacuo; 200 ml of fresh 20 dichloromethane are added and the mixture is reconcentrated. This treatment is repeated twice more, the resulting foam is dried in a large volume for 20 minutes, redissolved in 225 ml of dichloromethane and allowed to stand at ambient temperature for 16 hours, during which time: a considerable amount of solid precipitates. The mixture is filtered under nitrogen, the filtrate is cooled to 0 ° C (nitrogen atmosphere) and treated with diisopropylethylamine (17 ml) followed by dropwise addition of benzyl chloroformate (7 ml). The reaction is stirred at 0 ° C: The mixture is washed with two 300 ml portions of pH 4.5 monobasic potassium phosphate buffer solution, 5% sodium bicarbonate (two 300 ml portions), saturated sodium chloride solution. (300 ml), dried (sodium sulphate) and filtered, and concentrated in vacuo to give a foam which, on trituration with ether, gives 9.9 g of the title compound as a solid.

86 80271 D) (cis) -4- (Methoxycarbonyl) -2-oxo-3 - [(phenylmethoxy) carbonyl] amine] -1-acetidine Solution of cerium ammonium nitrate (8.59 g) in 60 ml: in 1: 1 acetonitrile-water, treated for 10 minutes with a slurry of 2 g of (cis) -4- (methoxycarbonyl) -1- (4-methoxyphenyl) -2-oxo-3 - [( phenylmethoxy) carbonyl-amine-4-azetidine in 50 ml of acetonitrile.

The reaction mixture is stirred for an additional 10 minutes at ambient temperature and diluted with ethyl acetate (100 mL). The separated aqueous layer is washed with ethyl acetate (three 40 mL portions) and the combined organic extracts are washed with 50% sodium bicarbonate (three 70 mL portions). The basic washings were washed back with ethyl acetate (50 mL) and the combined organic extracts were washed with aqueous sodium sulfite, 5% aqueous sodium carbonate (100 mL), 5% sodium chloride solution (two 100 mL portions), saturated sodium chloride (two 50 mL). n doses) and. : 20 Stir using Darco G-60 charcoal for 30 minutes.

Sodium sulfate is added and the mixture is filtered and concentrated in vacuo to give an oil which is triturated. trituration with ether gives 685 mg of the title compound as a solid.

E) (cis) -4- (Methoxycarbonyl) -2-oxo-3- (phenylmethoxy) carbonyl-amino-1-azetidinesulfonic acid, tetrabutylammonium salt

A mixture of (cis) -4- (methoxycarbonyl) -2-oxo-3- (phenylmethoxy) carbonylamino] -1-acetidine 30 (100 mg) and 172 mg of pyridine-sulfur trioxide complex 1 in 1 ml of pyridine, stirred under argon for 3 hours at 80 ° C. ; The reaction mixture is poured into 70 ml of 0.5 H monobasic; potassium phosphate (pH 5.5) and extracted with four 30 ml portions of dichloromethane (discard). Add tetra-3 butylammonium hydrogen sulfate (122 mg) to the aqueous solution

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87 80271 to a layer which is then extracted with dichloromethane (four 30 ml portions). The organic extracts are washed with 8% sodium chloride solution, dried over sodium sulfate and filtered. Concentration in vacuo gives 186 mg of the title compound as a viscous oil.

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

A solution of 186 mg of (cis) -4- (methoxycarbonyl) -2-oxo-3 - [(phenylmethoxy) carbonyl] amine-Ι-et azetidinesulfonic acid tetrabutylammonium salt in 2 ml of methanol is hydrogenated using 10% palladium on carbon (95 mg) for 1.5 hours in one atmosphere. The catalyst is filtered and rinsed with dichloromethane and the filtrate is treated with 97% formic acid and cooled to -50 ° C (the presence of an initial crystal at this stage is appropriate to effect crystallization). After crystallization begins, the mixture is allowed to stand at 10 ° C for 16 hours. The resulting solid is washed with dichloromethane, hexane and dryness; The mixture is concentrated in vacuo to give 50 mg of the title compound.

Example 58; (cis) -3- (2-amino-4-thiazolyl) - (1- (diphenylmethoxycarbonyl) -1-methylethoxy) iminoacetyl] -25-amino-4- (methoxycarbonyl) -2-oxo- 1-azetidinesulfonic acid, potassium salt

A solution of N-hydroxybenzotriazole hydrate (34 mg) and 101 mg of 2-amino-N- (1- [1- (diphenylmethoxycarbonyl) -1-methyl ethoxyimino] -4-thiazole-30 acetic acid 0.5 in 1 ml of dimethylformamide, treated with solid dicyclohexylcarbodiimide (45 mg) V. and the mixture is stirred under argon for 45 minutes (ambient temperature), then solid (cis) -3-amino-4- (methoxycarbonyl) -2-oxo-1 -azetidinesulfonic acid-35 poa (45 mg; see Example 57) followed by dropwise addition of triethylamine (0.03 ml) Reaction 88 80271 is stirred at ambient temperature for 16 hours The dimethylformamide is removed under high vacuum at 30 ° C and the residue is triturated. The supernatant is treated with potassium perfluorobutanesulfonate (67 mg).

Dilution with ether gives a solid which is washed with ether and dried in vacuo to give 93 mg of the title compound.

Example 59 (cis) -3- (C 2-Amino-1'-thiolyl) -1H-carboxy-1-methyl-1-ylethyloxy-imino (acetylamino) -4- (methoxycarboxylate) nyl) -2-oxo-1-azetidinesulfonic acid, potassium salt Slurry of (cis) -3- [2-amino-4-thiazolyl) -CO- (diphenylmethoxycarbonyl) -1-methylethoxyimino] acetyl-amino--4- (methoxycarbonyl) -2-oxo-1-15 azetidinesulfonic acid potassium salt in 0.4 ml of anisole, stirred at -12 ° C under argon and 0.9 ml of cold (-10 ° C) is added. ) trifluoroacetic acid. After 1.5 hours, 4 ml of ether and 2 ml of hexane are added and the resulting slurry is stirred for 15 minutes at -10 ° C and then for 20 minutes at ambient temperature. The solid is isolated by centrifugation and washed with ether. The pH of a suspension of this substance in 0.5 ml of cold water is adjusted to 6 with potassium hydroxide and then applied to a 30 ml HP-20AG column. Elution with water gives 30 mg of the title compound after evaporation (asotonithril is added and evaporated twice).

Analysis calculated for C 18 H 18 N 2 O 2 S 2 O 2 S: C, 12.19 H, 2.81; N, 12.98 Found: C, 29.08; H, 3.03; N, 12.19.

Example 60 (S) - (Trans) -3-Amino-4-ethynyl-2-oxo-1-azetidinesulfonic acid A) 2- (Trimethylsilyl) ethynylmagnesium bromide

In a flame-dried 50 ml flask maintained

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89 80271 under nitrogen pressure, 20 ml of dry tetrahydrofuran, 2.20 ml of trimethylsilylacetylene and 5.05 ml of a 3.06 M solution of methylmagnesium bromide in ether are added. The mixture is stirred for 140 minutes to give the title compound.

B) (S) - (trans) -4- [2- (Trimethylsilyl) ethynyl (-2-oxo-3-triphenylmethyl) amino] azide

To a flame-dried 250 ml three-necked flask is added 6.00 g of (S) - (cis) -4- (methylsulfonyl) -2-oxo-10β- (triphenylmethyl) aminoacetate. The flask is purged with nitrogen and then maintained under nitrogen pressure. After cooling the reaction mixture in a dry ice / isopropanol bath, 4.65 ml of a 30.06 M solution of methylmagnesium bromide in ether are added dropwise via hand syringe with vigorous stirring.

A solution of 2- (trimethylsilyl) ethynylmagnesium bromide prepared in Part A is added via a Teflon tube under nitrogen pressure (the flask containing the starting material is rinsed with 7 ml of tetrahydrofuran).

20 At the end of the addition, the cold bath is removed. After 45 minutes, a solution of 3.5 g of potassium bisulphate in 20 ml of water is added. Most of the tetrahydrofuran is removed on a rotary evaporator. The residue is transferred to a separatory funnel with ether and water. The aqueous layer 25 is separated and extracted twice with ether. The combined ether layers are washed once with saturated aqueous sodium chloride, dried over sodium sulfate and filtered. Removal of the solvent gives a foam which is chromatographed on a silica gel column. Elute with 2 liters of dichloromethane, 1 liter of 1% ether / dichloromethane, 2 liters of 2% ether / dichloromethane and 1.5 liters of 10% ether / dichloromethane (Frakt ίο 1 = 1000 ml; Fractions) 2 and 3 = 500 ml, and the final i ra k L io 4 = 250 ml) 1.30 g of the title compound are obtained in fractions 35 to 2-8 and 1.80 g of the corresponding trans isomer in fractions 90 80271 in fractions 12- 19. Fractions 9-11 contain 1.19 g of a mixture of cis and trans isomers.

C) (S) - (trans) -4-ethynyl-2-oxo-3 - [(triphenylmethyl) amino] azetidine 5 (S) - (trans) -4- [2- (trimethylsilyl) ethynyl] -2-oxo 3 - [(Triphenylmethyl) amino] azetidine (2.97 g) is dissolved in 30 ml of dichloromethane and 330 mg of tetrabutylammonium fluoride (containing 20-25% of water) are added.

After 20 minutes, the solvent is removed in vacuo. The residue is taken up in ethyl acetate and water. The organic layers are separated, washed once with water and once with saturated aqueous 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 give 2.35 g of the title compound as a powder (after drying under vacuum).

D) (S) - (trans) -3-Amino-U-ethynyl-2-oxo-1-azetidinesulfonic acid 20 (S) - (trans) -4-ethynyl-2-oxo-3- (E-triphenylmethyl) amino7azetidine (404 mg) and 560 mg of pyridine and sulfur trioxide complex are added to a 25 ml flask.

After purging the flask with nitrogen, 4.0 ml of dry pyridine are added and the mixture is heated to 80-85 ° C for 3 hours. The mixture is added to a vigorously stirred mixture of 4.0 ml of concentrated hydrochloric acid, 50 ml of water and 50 ml of ethyl acetate. The pH is adjusted to 3.15 with sodium bicarbonate. The aqueous layer is separated and extracted once with ethyl acetate. The combined organic layer is washed once with saturated aqueous sodium chloride, dried over sodium sulfate and filtered. Removal of the solvent in vacuo gives a foam which is taken up in 10 ml of dichloromethane. Formic acid (98%; 8 ml) is added and after 15 minutes the mixture is concentrated to 4 ml and 10 ml of dichloromethane are added to give a solid suspended in solution. Bullet 91 80271 is dated to give 100 ml of the title compound as a solid (apparent color defect on melting> 180 ° C).

Example 61 5? 3S- (30o (Z), 4- [7-3- [2- (2-Amino-4-thiazolyl) (methoxyimino) acetyl) amino] -N-ethynyl-O-oxo -1-acetidine sulfonic acid, potassium salt (Z) -2-amino (methoxyimino) -4-ethyl solacetic acid (100 mg), 85 mg of N-hydroxybenzotriazole-10 monohydrate and 113 mg of dicyclohexylcarbodi- the imide is placed in a 10 ml flask. The flask is purged with nitrogen and cooled in an ice-water bath. 0.6 ml of dimethylformamide are then added and the mixture is stirred for 10 minutes, during which time 0.6 ml of more dimethylformamide is added. Add (S) - (trans) -3-amino-4-ethynyl-2-oxo-1-azetidinesulfonic acid (95 mg; see Example 60) as a solid with 10 ml of dimethylformamide and 56 μl of triethylamine . The cold bath is removed and the mixture is stirred for 22 hours. Acetone-20 (3 mL) is added and any solids present are removed. . by filtration and washed with 4 ml of more acetone. All solvents are removed in vacuo and the residue is taken up in 5 ml of methanol and 162 mg of potassium perfluorobutanesulfonate are added and dissolved. After standing, the solid is precipitated and then isolated by centrifugation to give 68 mg of the title compound, m.p. > 230 ° C.

Example 62 E 3 S (R *]} - 3 - 2-amino - 4 - ylthio) - ZX 2 - furanyl - 3 0 methylene) Amino-2-oxo-1- Imidazolidinyl-1-carbonyl} '-amino] acetyl / amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

Following the procedure used in Example 4, but substituting (R) -2-amino- [E- [7- (3- (2-furanyl-methylene) amino] -2-oxo-1-imidazolidinyl) carbonyl) with amino-4-thiazoleacetic acid aminothiazole acetic acid gives the title compound, m.p. > 250 ° C.

Claims (8)

92 80271 A process for the preparation of novel therapeutically useful 1-sulfo-2-azetidinone derivatives of the general formula 1'H R3 10 N-1— C-CO-NH - --R „j, Γ n 3“ “H? N —k. o / N 2 1 1 L b | -N-SO 0 H 0 -R 13 wherein R 3 and are the same or different and represent hydrogen, C 1-4 alkyl or C 1-4 cycloalkyl or one of R 4 and R 4 is hydrogen and the other is alkynyl-1-yl, 3a R 3 is hydrogen, alkyl, cycloalkyl, alkylamino-carbonyl or alkyl substituted by one or more halogen, cyano, nitro, amino, mercapto, alkylthio, carboxyl or its sodium or potassium salt or its C 1-4 alkyl or benzyl ester, amido, alkoxycarbonyl, phenyl-methoxycarbonyl, diphenylmethoxycarbonyl, hydroxyalkoxyphosphinyl, dihydroxyalkoxyphosphinyl, hydroxy (phenylmethoxy) phosphinyl or a salt thereof, an azetidinone derivative having a hydrogen substituent at the 1-position: H r3 N -, - C-CO-NH --- Ru yUCF s tZl C-R13 0 ~ II 93 80271 or acylating the corresponding 2-at a cetidinone derivative having an amino substituent H r3 in the 3-position Process according to Claim 1, characterized in that a compound is prepared which has an alkyl which is optionally substituted by a carboxyl group or a C 1-4 alkyl or benzyl ester thereof. Process according to Claim 2, characterized in that a compound in which 15 is methyl is prepared. Process according to Claim 2, characterized in that a compound is prepared in which R 2 is carboxymethyl or 1-carboxy-1-methylethyl or a C 1-4 alkyl or benzyl ester thereof. Process according to one of Claims 1 to 4, characterized in that [3S- / 3Ct (Z), 4Λ77 "3 '(N-2-amino-4-thiazolyl) -1-carboxy-1-methyllethoxy) imine is prepared. (acetyl) amino) -4-methyl-2-oxo-1-azetidinesulfonic acid or a salt or ester thereof. 6. Intermediate 2-azetidinone derivatives of the general formula III'H R3 H 2 N 3 ij "^ 14 30 z ύ 4 μ 1T1, Ji-aN-SO, H 0... Where Rg and R ^ have the same meaning as in claim 1, and their salts.9 * 80271 5 H2N ~ 3 — Π-K4 jjj, 0 ^ -1 N-so3H Compound according to Claim 6, characterized in that the 2-position of the 2-azetidinone derivative has both a hydrogen and an alkyl substituent. Compound according to Claim 6, characterized in that the 4-position of the 2-azetidinone derivative has both a hydrogen and a methyl substituent. il 95 80271