FI67853C - PROCEDURE FOR THE FRAMSTATION OF ANTIBACTERIAL 7-OXO-4-TIA-1-AZA-BICYCLO (3,2,0) HEPT-2-ENDERIVAT - Google Patents

Description

lv £ * · P '! ANNOUNCEMENT,

^ UTLÄGGNINGSSKRIFT O / ODO

^ (45) * '' ..... - ^ (51) Kv.lk. '/ Lnt.CI.4 C 07 D 499/00 FINLAND —FINLAND (21) Patent application - Patentansökning 793905 (22) Filing date— Ansökningsdag 13.12.79 (23) Start date - Giltighetsdag 1 ^ .12.79 (41) Become public - Blivit offentlig 19.06.80

National Board of Patents and Registration ..... .... .......

* _ / 44) Date of publication and date of publication. - 00

Patent and registration authorities Ansökan utlagd och ud.skriften publicerad 28.02.85 (32) (33) (31) Privilege claimed — Begärd priority 1 8.12.78 USA (US) 968663 True-Styrkt (71) Bristol-Myers Company, 345 Park Avenue, New York, New York 10022, USA (72) Marcel Menard, Candiac, PQ, Alain Martel, Delson, PQ, Canada (CA) (74) Oy Koister Ab (54) Method for new antibacterial 7 ~ oxo For the preparation of 4-thia-1-aza-bicyclo [3.2.0] hept - 2-ene derivatives - For the preparation of framstalin with avian antibacterial 7-oxo-4-thia-1-aza-bicyclo [3.2 .0 / hept-2 ender

The invention relates to a process for the preparation of novel 7-oxo-4-thia-1-aza-bicyclo [3.2.1] hept-2-ene derivatives of the formula I and their pharmaceutically acceptable salts, Y \ ^ Y ..... Ϊ> - < in which Z is hydrogen or an ester-forming group, X is methyl, phthalimidobutyl, CH 2 CCCH 2) = NOCH 3 or (CH 2) ^ O 2 ((JR) 2), where n is 2 or 3 and R is lower alkyl and Y is hydrogen, lower alkyl substituted by acetoxy or hydroxy, 26853 (CH3) 2NCH2, HCONHCH2, C6H5CH2CH (OH), CH3OCH, OCH CH (OH) or 2.2 -dimethyl-1,3-dioxolan-4-yl, provided that X is not methyl when Y and Z are each hydrogen, thus g compounds have a strong antibacterial effect.

The compounds of formula 1 are so-called penein j ohdannai sj.a. The formula of the Penern ring system is 4 // N ./'2 0 7 i \ and its systematic name is 7-oxo-4-t ia- 1-a t sa-b isyclo / T ,? Q / hept-2-ene. For simplicity, it is referred to as 2-pence and the numbering system used is as follows:

_6_______5 ^ S

// // // n N —3 0 4 \

The compounds of formula I may exist either as racemic mixtures (R, S form) or as separate dextrorotatory and levorotatory optical isomers (R and E forms). Preferred are compounds in which the configuration of the 5-carbon atom corresponds to the configuration of natural penicillin (5R- configuration) The substituents at the 5- and 6-positions of the 2,6-disubstituted penene compounds may be in the cis or trans position relative to each other. The isomers obtained when the 6-substituent of the penem has an asymmetric carbon atom are then identified as A, B, C and D isomers. The preferred isomer of this type of compound is isomer B. The promotion of the various optical and geometric isomers can be accomplished by standard separation and resolution methods well known to those skilled in the art.

Pharmaceutically acceptable salts include non-toxic carboxylic acid salts, e.g. non-toxic metal salts such as sodium, potassium, calcium, aluminum and magnesium salts, ammonium salt and salts of non-toxic amines such as trialkylamines, triethylamines, triethylamines (triethylamines), triethylamines. -yl-β-phenethylamine, 1-ephenamine, N, N'-dibenzylethylenediamine, N-alkylpiperidine and other amines used to form salts of penicillins and cephalosporins. In the presence of a basic group, the compounds may also form pharmaceutically acceptable acid addition salts, e.g. -inic acid, acetic acid, citric acid, oxalic acid, succinic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, ascorbic acid and malic acid. Compounds having an acidic group and a basic group may also exist in the form of an internal salt, i. in the form of an amphoteric ion. The preparation of the salts described above can be carried out according to the usual methods used for the formation of salts of β-lactam antibiotics such as penicillins and cephalosporins.

Suitable ester-forming groups are those commonly used in the β-lactam and peptide fields. Many such groups are known which have been used to protect the carboxyl group during subsequent chemical reactions and which, if desired, can be subsequently removed by conventional methods to obtain the free carboxylic acid. Known ester-forming groups include 2,2,2-trichloroethyl, tertiary alkyl of 4 to 6 carbon atoms, tertiary alkenyl of 6 to 7 carbon atoms, tertiary alkynyl of 5 to 7 carbon atoms, alkoxymethyl, alkanoylmethyl of 2 -7 carbon atoms, N-phthalimidomethyl, benzoylmethyl, halobenzoylmethyl, benzyl, p-nitrobenzyl, o-nitrobenzyl, benzhydryl, trityl, trimethylsilyl, triethylsilyl,? -Trimethylsilylethyl and the like.

The preferred ester-forming group is p-nitrobenzyl ester, which can be easily removed by catalytic hydrogenation. For the preparation of compounds having functional groups which are reduced under the above conditions, a preferred alternative is β-trimethylsilylethyl ester, which can be removed by treatment with fluoride ions. Easily removable ester-forming groups also include physiologically cleavable esters, i. esters known in the art of penicillin and cephalosporin chemistry to be easily cleaved in the body to the parent acid. Examples of such physiologically cleavable esters are indanyl, thalidyl, methoxymethyl, glycyloxymethyl, phenylglyloxymethyl, thienylglyloxymethyl or acyloxymethyl of the formula O-CH 2 C-Y 'wherein Y' is C 1 -C 4 alkyl or phenyl. Particularly preferred esters of this type are methoxymethyl, acetoxymethyl, pivaloyloxymethyl, phthalidyl and indanyl.

The compounds of formula I may exist in a variety of solvation states and the group of compounds thus includes both anhydrous and solvated forms (including hydrates).

Particularly preferred compounds of formula I are those wherein Y is X-hydroxyethyl.

Compounds of formula I and their pharmaceutically acceptable salts may be prepared by reacting a compound of formula 0- i J- -N P (Q)

o J

CO 2 Z 'wherein X and Y are as defined above, Z' is an ester-forming group and Q is phenyl, cyclized in an inert organic solvent at a temperature slightly above room temperature and the reflux temperature of the solvent, and optionally an ester-forming group the compound obtained is removed and, if desired, converted into a pharmaceutically acceptable salt.

The preparation of starting materials and final products can be described by the following reaction formulas: 6 67853

Reaction scheme, I (option 1) Y 0

^ .OAc H

I-XC-SNa Y-CH = CH-OAc CSI ^ 1 | _ ^ * N PH 7 · 5 '

0 X H

0 0 y II Y N

Y ^ y_ ^ SC-X _ ^ _ SC-X - ^ CHO I SOCl2

N \ co n ^ -N \ ^ 0H

O XH 2 o CO2 '/,' 0 9

v II γ_ II

V _rsc-x p0 -rsc-x Δ -1_> -> J N __ Cl base Λ-N \ ^ P03

O

co2z 'co2z'

V ^ Y

Π y -> yy> -x s? -N Deletion of the Z 'group ^ // ° co_z1 o Λ

2 CO2H

ö

II

Ac = CH3C- 0 = c6h5-7 67853

Reaction Scheme II (Option 2) _ p Y s oAc ^ Λ-C H -. C - S N a Y-CH = CH-OAc CSI v I _k—- -> J_ N ^ nrr ^ CT ^ h ^ -SAc _ \ _ / SAc - ^ CHO S0C12

J-N \ co_z '—N \ ^ 0H

O H 2 0 ^ CO 2 Z 'Y Y-s

\ -rSAc Ρ0Ί l-r SAC MA

- .. J> -a-> 1 „_ base m r * base i-Nv ^ 01 n - ^ - '3 o y * 0 C02Z' C02Z '

O

Y 0 v H

^ A- .SM II _ Y ~> v_ / SC-X Δ - ~ f x-c- Θ -f __> ^ H \ ^ P03 J n \ ^ P03 0 ^ J ^ co Z 'C02Z'

V ^ Y

i — r \ x. ^ .s I / - * · ΓΊ Removal of the group Vx ^ Z 'J // o \ sy ~ "—k CO Z' o x

2 CO H

O

Il _ ...

X-C- © = acyl uit Wave MA = heavy metal salt 8 67853

Reaction Scheme III (Option 3) ^ OAc., I 0 -, L S i -I a Y-CH = CH-OAc CSI ^ 3 v.

---> 1 - „J— N? H '5

O ^ H

Y_H1 — rsc03-> N — r sc03 -> I CHO SOC1

y N \ CO Z 'A— N \ PH

0 U2 / j 0 ^ CO ^ Z '

-j- "SC03 PO3 ^ - ^ scc) 3 MA

J-N Cl J-U ^ P0> 0 0 ^ CO2Z 'CO? Z' 0

υ ~ λ_ O II

prSH x - "- ©." Vrsc_x O ^ Ln ^ PO3 ^ 0J — N \ ^ p03 ^ CO22 'CO22'

Y

-rVx Υχ r ^ \ '// -—: -> Delete T \ _x yy N ~ - ^ y Z' group '//

O \ J M

CO Z 'O Λ co2h 9 67853

In Reaction Schemes I, the vinyl ester is converted to 4-acetoxy-2-azetidinone by a cycloaddition reaction with chlorosulfonyl isocyanate (CSI) followed by reduction with an organic reducing agent such as sodium sulfite. The CSI reaction is suitably carried out in a neutral organic solvent such as diethyl ether at a temperature of 0 ° C or below. The reduction step can be carried out in an aqueous or aqueous-organic reaction mixture at a temperature of 0 ° or below and a slightly basic pH. After the formation of U-acetoxy-2-azetidinone, the synthesis can be continued in three different ways. Alternatively, azetidinone is reacted with thiolic acid 0

II

X-C-SH or a salt thereof, in a suitable solvent (e.g. water or aqueous organic solvent). Displacement of the acetoxy group results in the incorporation of the desired 2-substituent into the azetidinone at this stage. This displacement reaction is preferably carried out at or below room temperature and with a slightly basic pH (^7.5). When Y Φ H, the cis and trans isomers of the resulting azetidinone are preferably separated at this stage of the process (e.g. by chromatography). Alternatives 2 and 3 convert 4-acetoxy-2-azetidinone to 4-acetylthio-2-azetidinone and 4-tritylthio-2-azetidinone, respectively, with nucleophilic displacement using thioacetic acid or triphenylmethylmercaptan (or its salt, respectively). , such as sodium salt).

H-thioazetidinone is then reacted with the glyoxyl O-slate ester HC-CO 2 Z ', where Z' is an ester-forming group such as p-nitrobenzyl or trimethylsilylethyl, or a reactive oxo derivative such as a hydrate in an inert organic solvent. (e.g. in benzene, toluene, xylene and the like solvent) and mainly at a higher than normal temperature (e.g. at 50 ° C - preferably at boiling temperature). When an ester hydrate is used, the water formed can be removed azeotropically or using a 10,678,83 molecular sieve. The hydroxy ester product is formed as a mixture of epimers and, if desired, can be purified by chromatography, or used directly in the next step.

The conversion of the hydroxy ester to the corresponding chloroester is carried out in a reaction with a chlorinating reagent, e.g. SOCl 2, POO 1, PCI or the like, in an inert organic solvent, e.g. , preferably in the presence of an aliphatic tertiary amine (e.g. triethylamine) or a heterocyclic tertiary amine (e.g. pyridine or collidine). The reaction is preferably carried out at a temperature between about -10 ° C and room temperature. The chloroester product is obtained as a mixture of epimers which, if desired, can be purified in the next step before use.

The phosphorane intermediate can be obtained by reacting a chloroester with triphenylphosphine in an inert organic solvent such as dimethylformamide, dimethyl sulfoxide, tetrahydrofuran, dimethoxyethane, dioxane or aliphatic, cycloaliphatic or aromatic hydrocarbon (e.g. in a solvent) in the presence of a base, preferably an organic tertiary amine such as triethylamine, pyridine or 2,6-lutidine. The reaction is preferably carried out at a temperature between room temperature and the boiling point of the solvent system.

In Option 1, the phosphorane intermediate is converted to the desired penem by cyclization in a thermally inert organic solvent at a temperature slightly above room temperature and the reflux temperature of the solvent system. The cyclization is preferably carried out at reflux temperature. Suitable inert organic solvents include aliphatic, cycloaliphatic or aromatic hydrocarbons (e.g. benzene, toluene, hexane, cyclohexaneL), 6785 3 halogenated hydrocarbons (e.g. methylene chloride, chloroform, chloroform, carbon tetrachloride), ethers (di-ethyl), diethyl ether, di-ethyl ), carboxylic acid amides (e.g. dimethylformamide), di-C 1 -C 6 alkyl sulfoxides (e.g. dimethyl sulfoxide) or C 1 -C 6 alkanol (e.g. methanol, ethanol, t-butanol), or mixtures thereof.

In options 2 and 3, phosphorane is converted to a heavy metal mercaptide of formula, s ---

J - i P (Q) M

0 3 L CO2z 'x

III

or

SHpCOOCH

'· R N \ ^ pO) 3

T

IIIa CO 2 Z 'wherein Q is phenyl, x is 1 or 2 and M is Cu (II), Pb (II) or Hg (II) when x is 2, or Ag (I) when x is 1. Mercaptide formation is performed by reacting phosphorane with a salt of Hg (II), Pb (II), Cu (II) or Ag (I) or (methoxycarbonyl) mercuric (II) acetate in a methanolic solvent and an organic or inorganic base such as aniline , in the presence of pyridine, collidine, 2,6-lutidine, alkali metal carbonate and the like base. The preferred base is pyridine. The reaction may be carried out at room temperature or, if desired, with moderate cooling or heating. The anion (A) of the heavy metal salt may be any anion which forms a salt soluble in the chosen solvent, e.g. The product is then reacted with an acylating agent which is 0

II

is capable of attaching a group X-C-, wherein X is the desired penem-2-subs-0 0

IT IT

substituent. The acylating agent (XC- +) may be an acid XC-OH or a reactive functional derivative thereof such as an acid halide (preferably an acid chloride), an acid azide, an acid anhydride, a mixed acid anhydride, an active ester, an active thioester, etc. The acylation is carried out in a neutral solvent. -hydrogenated hydrocarbon such as methylene chloride or ether such as dioxane, tetrahydrofuran or diethyl ether) and, when an acid derivative is used, an acid scavenger such as tri-lower alkylamine (e.g. triethylamine) or a tertiary organic base such as pyridine or collide. In the presence of 2,6-lutidine. When the free acid is used, the acylation is carried out in the presence of a suitable condensing agent, for example a carbodiimide such as N, N'-dicyclohexylcarbodiimide. Acylation of the mercaptide can be performed over a wide temperature range, but is preferably performed at -20 ° to + 2 ° C. The phosphorane obtained after acylation is cyclized as described above to give the desired penem ester.

Phosphorane formation via the mercaptide intermediate (Options 2 and 3) has been found to produce a much better product than that obtained using the more conventional Option 1.

Once the carboxyl-protected penem compound is formed, the ester group can be removed by conventional methods (e.g., by hydrolysis, hydrogenation, or photolysis). For example, removal of the p-nitrobenzyl ester can be accomplished by catalytic hydrogenation in the presence of a noble metal catalyst such as palladium or rhodium, including derivatives thereof such as oxides, hydroxides, or halides. The catalyst may be bound to a common support such as carbon or silicon. In this case, a non-reducing aqueous or non-aqueous inert solvent such as water, ethanol, methanol, ethyl acetate, tetrahydrofuran, diethyl ether or 13,678,83 dioxane is used. The hydrogenation can be carried out at atmospheric pressure or higher, and is preferably carried out at room temperature for about 1 to 3 hours, depending on the solvent and catalyst used. If an equivalent amount of a base such as an alkali metal or alkaline earth metal hydroxide or amine is used during the hydrogenation, the product can be recovered as the carboxylic acid salt. Removal of β-trimethylsilylethyl ester, another useful ester-forming group, is conveniently performed by treatment with a fluoride ion-producing compound. Other ester-forming groups can be removed in a similar manner by methods known to those skilled in the art.

Reaction Scheme IV (Options 1 and 2) -T "SC03 ^ SC03

I T _, I

0 Hl 0 ^ Nsb Y Ηλ SC0 „v-r base 1

Ck-N

O X B

deprotectionH ^ 14 67853

YvH] _ ^ - S C03 \ J — NN h— \

O H 0 XB

<fH0 0 CO Z 'X-C- Θ Φ v 0 Y-y ^ SC03 Yv ^ _- ^ sc-x

-OF. _OH X N

o 0 B

co2z Deletion of 'Z' group S0Clo

v 2 V

0

Y y II

"S -_„ SC0- -SC-X

^ 3 —f CO 2 Z 'PO 3 base

CHO

V I z,

\ [C02Z

V 0 Y ~ ^ _ ^ SC0T Y I!

-p 3 .SC-X

--OF. _ ^ P03

^ γΝ OH

C <V 'CO 2 Z' MA / base SOCl- ψ v 2 15 67853

O

CM Y 11

-r- "SM V_SC-X

0J-N \ ^ P03 jN ~ C1 O 0 C ° 2Z 'CO2z' 0 H ~ P0, XC- © base 0 * o Y ~ ^ ^ sc-x h_ sc-x QJN \ ^ P03 0 <^ - N \ ^ P03 C02Z 'co2z · V>' trv C02Z 'co ^, removal of the' Z 'group Removal of the Z * group v Ψ YVrVY Vrv iM IX> x

CO 2 H 0 CO 2 H

B = ring-type protecting group 16 6785 3

In Reaction Scheme IV, the Azetidinone Ring Type is protected with a conventional easily removable protecting group such as tri-organosilyl (e.g., trimethylsilyl or t-butyl idimotylsilyl), methoxymethyl, methoxyethoxymethyl, tetrahydropyranyl, or the like. The incorporation of the desired Y substituent at the 1-position of the azetidinone is then accomplished by reacting the appropriate electrophile with its N-snapped azetidinone in the presence of a strong base. Thereafter, the synthesis can be continued in two ways depending on the stage at which the azetidinone is deprotected.

According to Option 1, the N-protected intermediate is deprotected by conventional methods (e.g., hydrolysis with acid), followed by conversion to the 2,6-penem compound by esterification, chlorination of the hydroxy ester, conversion of the chloroester to phosphorane, conversion of the phosphorane to the heavy metal dimer, acylation of the mercaptide with XC - (+), cyclization of the thermally obtained phosphorane to give the 2,6-penem ester and removal of the ester group.

According to option 2, the N-protected azetidinone is converted to a heavy metal mercaptide, the mercaptide is acylated with group 0

II

The N-protecting group is deprotected, the deprotected azetidinone is reacted with a glyoxylate ester, chlorinated, the chloroester is reacted with phosphine to give phosphorane, the phosphorane is cyclized to give the penem ester and removed. an ester group to give a 2,6-penem compound.

In the preparation of 2-penem or 2,6-penem compounds according to the above methods, the free functional groups of the substituents X or Y which did not participate in the reaction can be temporarily protected in a manner known per se, such as e.g. free amino groups by acylation, tritylation or silylation, free hydroxyl groups, for example, by etherification or esterification, mercapto groups with ester ester, and free carboxyl or sulfonic groups, e.g. by esterification, including silylation. After the reaction has taken place, these groups can, if desired, be liberated, individually or together, in a manner known per se.

17 67853

The penem acid compounds of formula I may be converted into pharmaceutically acceptable salts by reacting the free acid with a stoichiometric amount of a suitable non-toxic acid or base in an inert solvent, followed by recovery of the desired salt, e.g. by freeze-drying or precipitation. The resulting mixtures of isomers can be divided into individual isomers by known methods. Mixtures of diastereomeric isomers, e.g., may be separated by fractional crystallization, using a chromatographic adsorption method (column or thin layer chromatography), or using other suitable separation methods. The racemates obtained can be divided into antipodes in the usual manner, e.g. by forming a mixture of diastereomeric salts with optically active salt-forming reagents, separating the diastereomeric salts and converting the salts into free compounds, or by fractional crystallization from optically active solvents.

The penem compounds of formula I and their pharmaceutically acceptable salts have been found to be effective broad-spectrum antibacterial agents useful in the treatment of infectious diseases caused by both gram-negative and gram-positive organisms.

Biological information

Representative compounds of formula I were subjected to experiments investigating their antibiotic activity in vitro against various microorganisms. Samples of said compounds, after dissolution in water and dilution with nutrient solution, showed the following minimum inhibitory concentrations (MICs) in mcg / ml against said microorganisms, determined by the tube dilution method after overnight incubation at 37 ° C.

Organism Compound (Example No.) 6785 3 18 MIC, micrograms / ml

Streptococcus pneumoniae 125 A9585

Streptococcus pyogenes A9604 '> 125

Staphylococcus aureus 1 A9537

Staph aureus +50% 2

Serum A9537

Staphylococcus aureus 32 A9606

Staphylococcus aureus 4 A15097

Streptococcus faecalis 125 A20688

Escherichia coli 8 A15119

Escherichia coli 2 A2 0 3 41-1

Klebsiella pneumoniae 125 A15130

Klebsiella species 4 A20458

Proteus mirabilis 8 A9900

Proteus mirabilis 63 A9716

Proteus morganii 63 A15153

Proteus rettgeri 32 A21203

Serratia marcescens 63 A20019

Enterobacter cloacae 16 A9659

Enterobacter cloacae 16 A 9 6 5 6

Pseudomonas aeruginosa 125

A9843A

Pseudomonas aeruginosa 125 A21213 19 67853 MIC, micrograms / ml

Organism Compound (Example No.)

Haemophilus influenzae A9833

Haemophilus influenzae A21S22

Bacteroides fragilis A20931

Bacteroides fragilis A20929 2

Streptococcus pneumoniae 1 A95 85

Streptococcus pyogenes 2 A9604

Staphylococcus aureus 8 A9537

Staph aureus +50%> 63

Serum A9537

Staphylococcus aureus 4 A9606

Staphylococcus aureus 8 A15097

Streptococcus faecalis 63 A20688

Escherichia coli 32 A15119

Escherichia coli 32 A20341-1

Klebsiella pneumoniae 63 A15 13 0

Klebsiella species> 125 A20468

Proteus mirabilis 63 A9900

Proteus vulgaris 63 A9716

Proteus morganii 63 A1S153

Providencia stuartii 32 A21205 20

Organism Compound (Example No.) _2 MIC, micrograms / ml 67853

Serratia marcescens 125 A20019

Enterobacter cloacae 125 A965 9

Enterobacter cloacae> 125 A9656

Pseudomonas aeruginosa> 125

A9834A

Pseudomonas aeruginosa> 125 A21213

Haemophilus influenzae A9833

Haemophilus influenzae A21522

Bacteroides fragilis A20931

Bacteroides fragilis A20329 7 4

Streptococcus pneumoniae .25 .016 A9585

Streptococcus pyogenes 8 .25 A 9 6 0 4

Staphylococcus aureus 8 · 03 A95 37

Staph aureus +50% 32 63

Serum A9537

Staphylococcus aureus 16 16 A9606

Staphylococcus aureus 63> 125 A15097

Streptococcus faecalis> 125 16 A20688

Escherichia coli 63 63 A15119

Escherichia coli> 125> 125 A20341-1

Klebsiella pneumoniae 125> 125 A15130 6785 3

Organism Compound (Example No.) 7 4_ MIC, micrograms / ml

Klebsiella species> 125> 125 A20468

Proteus mirabilis 63 32 A9900

Proteus vulgaris 125 A955 5

Proteus morganii 125 32 A15153

Proteus rettgeri 63 A21203

Serratia marcescens 63 16 A 2 0 019

Enterobacter cloacae 125> 125 A965 9

Enterobacter cloacae 125 125 A9656

Pseudomonas aeruginosa 125> 125

A9843A

Pseudomonas aeruginosa 125> 125 A 2 12 13

Haemophilus influenzae A9833

Haemophilus influenzae A21522

Bacteroides fragilis A20931

Bacteroides fragilis A20929

Pseudomonas aeruginosa A20599

Pseudomonas aeruginosa A992 5

Pseudomonas aeruginosa A20229

Proteus species A20543

Proteus mirabilis - 16 A9716

Providencia stuartii - 63 A21205 22 6 7 8 5 3 MIC, micrograms / ml

Organism Compound (Example No.) 5_

Streptococcus pneumoniae 2 A9585

Streptococcus pyogenes 16

A9 6 0H

Staphylococcus aureus 32 A 9 5 3 7

Staph aureus +50%> 63

Serum A9537

Staphylococcus aureus> 125 A9606

Staphylococcus aureus> 125

Alb 0 9 7

Streptococcus faecalis 125 A20688

Escherichia coli 63 A15119

Escherichia coli> 125 A20341-1

Klebsiella pneumoniae> 125 A15130

Klebsiella species> 125 A20468

Proteus mirabilis 63 A9900

Proteus vulgaris A9716

Proteus morganii 125 A15153

Proteus rettgeri 125 A21203

Serratia marcescens> 125 A20019

Enterobacter cloacae> 125 A9659

Enterobacter cloacae> 125 A9656

Pseudomonas aeruginosa 125

A9843A

Pseudomonas aeruginosa 125 A21213 MIC, micrograms / ml

Organism Compound (Example No.) 5_ 23 67853

Haemophilus influenzae A9833

Haemophilus influenzae A21522

Bacteroides fragilis A20931

Bacteroides fragilis A20929

Pseudomonas aeruginosa A20599

Pseudomonas aeruginosa A992 5

Pseudomonas aeruginosa A20229

Proteus species A2 0 5 4 3

Proteus mirabilis 63 A9716

Proteus mirabilis A95 55 6

Streptococcus pneumoniae 63 A95 85

Streptococcus pyogenes 125

A96 0U

Staphylococcus aureus 32 A95 37

Staph aureus +50% 32

Serum A9537

Staphylococcus aureus> 125 A9606

Staphylococcus aureus 63 A15097

Streptococcus faecalis 63 A20688

Escherichia coli 63 A15119

Escherichia coli 32 A20341-1 2i 678 5 3 MIC, micrograms / ml

Organism Compound (Example No.) 6

Klebsiella pneumoniae> 12 5 A1S130

Klebsiella species 63 A20468

Proteus mirabilis 125 A9900

Proteus vulgaris> 125 A9555

Proteus morganii 125 A1515 3

Proteus rettgeri 125 A21203

Serratia marcescens 125 A20019

Enterobacter cloacae 125 A9659

Enterobacter cloacae 32 A9656

Pseudomonas aeruginosa 125

A9843A

Pseudomonas aeruginosa 125 A21213

Haemophilus influenzae A9833

Haemophilus influenzae A21522

Bacteroides fragilis A20931

Bacteroides fragilis A20929 8 9

Streptococcus pneumoniae .06 .25 A9585

Streptococcus pyogenes .13 2 A9 6 0 4

Staphylococcus aureus 1 4 A95 37

Staph aureus +50% 16 63

Serum A9537 2S 67853 MIC, micrograms / ml

Organism Compound (Example No.) 8 9

Staphylococcus aureus 12 5 4 A9606

Staphylococcus aureus> 125 16 A15097

Streptococcus faecalis 125 125 A20688

Escherichia coli 16 4 A15119

Escherichia coli> 125 16 A20341-1

Klebsiella pneumoniae 125 32 A15130

Klebsiella species> 125 125 A20468

Proteus mirabilis 8 16 A9900

Proteus vulgaris 63 16 A9555

Proteus morganii 32 32 A15153

Proteus rettgeri 16 32 A21203

Serratia marcescens 63 32 A20019

Enterobacter cloacae 125 32 A 9 6 5 9

Enterobacter cloacae 63 63 A3 6 5 6

Pseudomonas aeruginosa 125 125

A9843A

Pseudomonas aeruginosa 125 125 A21213

Haemophilus influenzae A9833

Haemophilus influenzae A21522

Bacteroides fragilis A20931

Bacteroides fragilis A20929 26 6785 3 MIC, micrograms / ml

Organism Compound (Example No.) 10 11

Streptococcus pneumoniae 32 .25 A9585 32 .25

Streptococcus pyogenes 125 1 A9604> 125 1

Staphylococcus aureus A9537

Staph aureus +50% 125 2

Serum A9537> 63 8

Staphylococcus aureus A9606> 125 4

Staphylococcus aureus A15097> 125 8

Streptococcus faecalis> 125 63 A20688> 125 63

Escherichia coli> 125 16 A15119> 125 16

Escherichia coli> 125 16 A2 0341-1> 125 16

Klebsiella pneumoniae> 125 16 A15130> 125 16

Klebsiella species> 125 16 A20468> 125 16

Proteus mirabilis> 125 32 A9900> 125 16

Proteus vulgaris> 125 16 A21559> 125 16

Proteus morganii> 125 32 A15153> 125 16

Proteus rettgeri> 125 16 A21203> 125 16

Serratia marcescens> 125 16 A20019> 125 16

Enterobacter cloacae> 125 32 A9659> 125

Enterobacter cloacae> 125 16 A9656> 125

Pseudomonas aeruginosa> 125> 125 A9843A> 125 27 6 7 8 5 3 MIC, micrograms / ml

Organism Compound (Example No.) 10 11

Pseudomonas aeruginosa> 125> 125 A21213> 125

Haemophilus influenzae Δ9833

Haemophilus influenzae A21522

Bacteroides fragilis A20931

Bacteroides fragilis A20929 12

Streptococcus pneumoniae .03 A9585

Streptococcus pyogenes .06 A9604

Staphylococcus aureus .5 A9537

Staph aureus +50% 4

Serum A9537

Staphylococcus aureus 1 A9606

Staphylococcus aureus 2 A15097

Streptococcus faecalis 2 A20688

Escherichia coli 2 A15119

Escherichia coli 8 A2 0341-1

Klebsiella pneumoniae 8 A15130

Klebsiella species 63 A20468

Proteus mirabilis 2 A9900

Proteus vulgaris 4 A21559

Proteus morganii 8 A15153 28 6785 3 MIC, micrograms / ml

Organism Compound (Example No.) 12

Proteus rettgeri 2 A21203

Serratia marcescens 8 A20019

Enrerobacter cloacae 8 A9 6 5 9

Enterobacter cloacae 32 A9656

Pseudomonas aeruginosa 63

A 9 8 9 3 A

Pseudomonas aeruginosa A21213

Haemophilus influenzae A9833

Haemophilus influenzae A21522

Bacteroides fragilis A20931

Bacteroides fragilis A20929 14

Streptococcus pneumoniae. 9 A9585

Streptococcus pyogenes .5 A9604

Staphylococcus aureus 8 A9537

Staph aureus +50% 32

Serum A9537

Staphylococcus aureus 16 A9606

Staphylococcus aureus 32 A15097

Streptococcus faecalis> 63 A20688

Escherichia coli 32 A15119

Escherichia coli 32 A2 0 341-1 29 67853 MIC, micrograms / ml

Organism Compound (Example No.) 14

Klebsiella pneumoniae 63 AI 513 0 *

Klebsiella species 63 A2Q468

Proteus mirabilis 32 A9900

Proteus vulgaris 32 A21559

Proteus morganii 63 A15153

Proteus rettgeri 32 A21203

Serratia. marcescens 6 3 A 2 0 019

Enterobacter cloacae 63 A96S9

Enterobacter cloacae 63 A965 6

Pseudomonas aeruginosa> 63

A9 8 4 3A

Pseudomonas aeruginosa> 63 A21213

Haemophilus influenzae A983 3

Haemophilus influenzae A21522

Bacteroides fragilis A2 0 9 31

Bacteroides fragilis A20929 15 17 18

Streptococcus pneumoniae> 125 1 32 A95 85

Streptococcus pyogenes> 125 16 32 A9604

Staphylococcus aureus> 12 5 32 1.2 5 A9 5 3 7

Staph aureus +50%> 63> 63> 63

Serum A9537

Stahylococcus aureus> 125 32> 125 A960 6 30 6 7 8 5 3 MIC, micrograms / ml

Organism Compound (Example No.) 15 17 18

Staphylococcus aureus> 128> 125> 125 A15097

Streptococcus faecalis> 125> 128> 128 A20688

Escherichia coll> 125> 125> 125 A15119

Escherichia coli> 125> 125> 125 A20341-1

Klebsiella pneumoniae> 125> 125> 125 A15130

Klebsiella species> 125> 125> 125 A20468

Proteus mirabilis> 125> 125> 125 A9900

Proteus vulgaris> 125> 125> 125 A21559

Proteus morganii> 125> 125> 125 A15153

Proteus rettgeri> 125> 125> 125 A21203

Serratia marcescens> 125> 125> 125 A20019

Enterobacter cloacae> 125> 125> 125 A9559

Enterobacter cloacae> 125> 125> 125 A9 65 6

Pseudomonas aeruginosa> 125> 125> 125

A9 8 43A

Pseudomonas aeruginosa> 125> 125> 125 A21213

Haemophilus influenzae - - A9833

Haemophilus influnezae - - A21522

Bacteroides fragilis - - A20931

Bacteroides fragilis - - A20929

19 20 21 H

Streptococcus pneumoniae 63 16 4 32 A9585

Streptococcus pyogenes 63 32 4 32 A9604 31 67853 MIC, micrograms / ml

Organism Compound (Example No.) 2-9.20_ 21 22

Staphylococcus aureus> 63 63> 8 63 A9 537

Staph aureus + 50%> 32> 32> 32> 32

Serum A9537

Staphylococcus aureus> 63 63 63> 63 A9606

Staphylococcus aureus> 63 63> 63> 63 A15097

Streotococcus faecalis> 63 63> 63> 63 A20688

Escherichia coli> 63> 63 63 63 A15119

Escherichia coli> 63> 63 63 63 A20341-1

Klebsiella pneumoniae> 63> 63 63> 63 A15130

Klebisella species> 63> 63> 63> 63 A20468

Proteus mirabilis 63> 63> 63 63 A9900

Proteus vulgaris> 63> 63> 63> 63 A21559

Proteus morganii> 63> 63> 63> 63 A15153

Proteus rettgeri> 63 63 32 63 A21203

Serratia marcescens> 63> 63> 63 63 A20019

Enterobacter cloacae> 63> 63 63> 63 A9653

Enterobacter cloacae> 63> 63 63 63 A9656

Pseudomonas aeruginosa 63 63 63 63

A9843A

Pseudomonas aeruginosa 63 63 63 63 A21213

Haemophilus influenzae - - A9833

Haemophilus influenzae - - A21522

Bacteroides fragilis - - A20931

Bacteroides fragilis - - A2092 9 32 6785 3

The compound described in Example 11 was also tested in vivo in mice and its PD 2 (dose of compound, mg / kg required to give 50% protection to otherwise mice against lethal microorganism infection) was determined for the organism shown below.

S. aureus A9537

Compound S. aureus A 9537 Treatment- PDj-q infection organ (mg / kg / treatment) _nismi___

Example 11, Compound 6.6 x 10b 2 IM 7.7

The active compounds of formula I may be formulated into mixtures which contain, in addition to the active ingredient, a pharmaceutically acceptable carrier or diluent. The compounds can be administered both orally and parenterally. The pharmaceutical preparations may be in solid form, such as capsules, tablets or granules, or in liquid form, such as solutions, suspensions or emulsions. In the treatment of bacterial infections in humans, the active compounds may be administered orally or parenterally in amounts of about 5 to 200 mg / kg / day and preferably about 5 to 20 mg / kg / day in divided doses, e.g. three or four times a day. They are administered in dosage units containing, for example, 125, 250 or 500 mg of active ingredient in association with suitable physiologically acceptable carriers or diluents.

The following examples illustrate the invention. All temperatures are in degrees Celsius. For convenience, some abbreviations have been used in the examples. Of these abbreviations, the meaning of which is not obvious is defined below: 33 67 85 3 CSI chlorosulfonyl isocyanate pet.ether petroleum ether b.p. boiling point n.m.r. nuclear magnetic resonance h hour ether diethyl ether (unless otherwise stated)

Celite Johns-Manville Products Corporation’s buttermilk trademark psi pounds per square inch r.t. room temperature PNB p-nitrobenzyl m.p. melting point LAH lithium aluminum hydride n-BuLi n-butyllithium MIBK methyl isobutyl ketone

Et CgHj.-

Tr 'C'Wa

Me CH3-THF tetrahydrofuran

Ph phenyl DMF dimethylformamide TEA triethylamine PCBG p-nitrobenzylglyoxylate THP tetrahydropyranyl TFA trifluoroacetic acid HMPT (or hexamethylphosphoric triamide HMP A)

EtOAc ethyl acetate DMSO dimethyl sulfoxide

Ac CH3CO-

Ms CH 3 SO 2 EMAP 4-dimethylaminopyridine

Py pyridine EBA lithium diisopropylamide 34 67853

Example 1 2 '- (2'-Diethylphosphono-1'-ethyl) penein-3-carboxylic acid

Vo2h 9? (EtO) -P- (CH) CO CH -V (EtO) P- (CH.,) CO 2 H -► * Z / j 2 z 2 z 1_ 2

(EtO) P- (CH) COCl - ((EtO) - $ - CH) COSH

£ £ Δ 2 2 2 _3 4_

A mixture of compound 1 (11.2 g, 50 mmol) and 5-norm. NaOH (10 mL) was stirred and cooled (in an ice bath) for 15 minutes and at room temperature for 15 minutes. The mixture was extracted with ether and the extract was discarded. The aqueous solution was made acid-5-norm. HCl and extracted with CH 2 Cl 2 to give, after drying and evaporation of the solvent, 10.0 g (95%) of oil 2, nmr ef (CDCl 3), 9.1 (9H, m), 1.8-2.9 ( 9H, m), 1.2 (6H, t).

To a cooled solution (ice bath) of compound 2 (2.26 g, 10.76 mmol) was added dropwise oxalyl chloride (2.79 g, 1.88 mL, 21.5 mmol). The mixture was kept at room temperature for 6 h and then evaporated to dryness. (CO 2) 2 - residues were azeotroped with benzene to give 2.9 g (quantitative yield) of crude compound 3. IR δ C ° 1800; 1735 cm -1. NMR δ 9.3 (9H, m), λ max C 3.0-3.7 (2H, m), 2.0-3.0 (2H, m), 1.9 (6H This was treated with H 2 S / TEA according to the standard procedure to give 1.9 g (80%) of oil 9, which was estimated to be 8085 purity delta 80% .Nmr: <f 4.1 (4H, q ), 2.7-3.5 (2H, m), 1.7-2.5 (2H, m), 1.33 (6H, t).

? OAc 9 J-r-f (Eto) 2 ^ (CH2) 2 <Ss

Nh 1 N

5 i

To compound 4 (1.9 g, 8.4 mmol) was added 1 mol under N 2 protection. NaHCO 3 solution (10 mL) followed by the addition of compound 5 (0.813 g, 6.3 mmol) in Na 2 O (3 mL). The pH of the mixture was adjusted to 7-8 by adding more NaHCO 3. After standing for 4 h, the mixture was extracted with CHCl 3 to give, after drying and concentration, 1.05 g (56.4% of compound 5) of solid 6, m.p. 64-67 °, nmr λ 7.7 (NH), 5.3 (1H q), 4.2 (4H,) 3.8 (1H q) 3.5 (1H, q), 2.6- 3.2 (2H, m) 1.7-2.4 (2H, m), 1.3 (6H,) · s (CH)? (OEt) (CH) -1- (OEt)

rYY _, rfY

(/ -Nh CT

C02PNB

6 T_

A mixture of compound 6 (260 mg, 0.88 mmol) and p-nitrobenzylglyoxalate (198 mg, 0.88 mmol) was boiled in benzene (6 mL) under a Dean Stark apparatus for 16 h to give 453 mg of a heavy solvent after evaporation of the benzene. oil 7, nmr ^ * 8.3 (2H, d) 7.6 (2H, d) 5.3-5.7 (4H,) 4.9 (OH), 4.2 (4H,) 3.55 (1H g), 3.4 (1H, g), 2.5-3.2 (2H, m) 1.7-2.5 (2H, m) 1.3 (6H).

36 6 7 8 5 3 S- / <CH2I 2hott) 2 (CH2) 2t (OEt) 2

Π T - T

ο ^ -Ν · γ «co2pnb co2pnb 1 8

Crude compound 7 (504 mg, 0.88 mmol) was dissolved in 1 mol. pyridine in THF (0.9 mL). This was added dropwise, with stirring and cooling (ice bath) 1 mol. SOCl 2 in THF solution (0.9 mL) and the mixture was stirred cold for 15 min. and at room temperature for 40 min. Benzene (10 ml) was added and the solid was filtered off. The filtrate was concentrated in vacuo to give 463 mg (yield quantitative) of crude compound 8, nmrg 8.3 (2H, d), 7.6 (2H, d), 6.1 (1H, s), 5.7 (1H, m), 5.3 (2H, d), 4.2 (4H), 1.8-3.6 (6H, m) 1.3 (6H).

9? ς X (CH_) J- (OEt), JCH) - £ - (CEt)

RysY 2 2 2 _► P "Y

co2pnb co phb 8 9

To a solution of crude compound 8 (463 mg, 0.88 mmol) in THF (4 mL) was added triphenylphosphine (236 mg, 0.9 mmol) and 2,6-lutidine (96 mg, 0.9 mmol) and the mixture was allowed to stand at room temperature for 65 hours. It was then filtered, the filtrate was concentrated and the residual oil was chromatographed on a silica gel column eluting with a mixture of ethyl acetate and 2% ethanol to give 203 mg (30.6%) of oil 9 which solidified on standing, m.p. 12-12-12 ° C.

3 7 67853 <CH2> 2 ^ <0EC> 2 ^ (Ϊ [-ΥΎ -► —f V_ (CH2,;.>, OEtl2 J-Lr »3 </ - '" - f o y co pub

C02PNB

9 -

A solution of 9 (470 mg, 0.635 mmol) in toluene (30 mL) was boiled for 5 h, then concentrated and chromatographed on silica gel with ethyl acetate to give 167 mg (56%) of 10 as an oil, IR 1795, 1710 cm -1. nmr δ 8.3 (2H, d) 7.7 (2H, d) 5.7 (1H, m), 5.38 (2H, d) 4.1 (4H) 1.8-3.8 ( 6H, 1.35 (6H).

J 1 R 2 V (CH 2) 2 - ^ - (OEt) 2 - +

CO2PNB CO2H

10 11

To a solution of 10 (59 mg, 0.126 mmol) in THF (3 mL) and ether (1 mL) was added NaHCO 3 (9 mg, 0.107 mmol), water (1 mL), and 10% Pd / Celite catalyst (60 mg) and the mixture were hydrogenated at 30 psi for 2 h. The product was isolated as usual to give 36 mg (86%) of compound 11 as an oil, IR (CHCl 3) 1798, 1730, 1710 cm -1. 9.0 (CO 2 H), 5.6- (1H, m), 4.4 (4H), 3.6 (1H, q), 3.15 (1H, q), 1.7-3.0 ( 4H, m), 1.3 (6H).

33 67853

Example 2 6-Acetoxymethyl-2-methylpenem-3-carboxylic acid

/ V. . S

Preparation of AcCT ^ -Γ \ co2h 1,3-diacetoxypropylene 1 (Ref. L.W. McTeer U.S. 2,866,813. CA 53 9063) CH2 = CH-CH0 Ac2 ° AcOCH2CH = CH-OAc + ch2 = ch-ch (oac) 2 cat. *

Catalyst preparation: A solution of boric acid (6.2 g) and oxalic acid (12.6 g) in water (HU ml) was evaporated to dryness to give a solid catalyst.

Method: Acrolein (140 g; 2.5 moles) was mixed with acetic anhydride (256 g, 2.5 moles) at room temperature. A 5 ml portion of this mixture was transferred to a 1 liter Erlenmeyer flask and treated with a few drops of a catalyst prepared by dissolving 1.0 g of solid catalyst in 5 ml of acetic anhydride. A vigorous exothermic reaction was initiated and the reaction mixture was maintained at 40-60 ° (controlled by cooling in an ice bath), and the remainder of the acroleic-acetic anhydride mixture was added to the flask in 10-15 mL portions, followed by the addition of a few drops of catalyst. To remove unreacted starting materials, the resulting mixture was distilled, followed by distillation of 1,1-diacetoxy.-propylene. Next, 51.6 g (13.06%) of product were obtained, b.p. 54-57 ° C / 1.2 mm.

NMR: δ f (ppm, CDCl 3), 7.4 (H 'd' J = 12) '5'3 ~ <H, m), 4.5 (2H, d, J = 7), 2.16 ( 3H, s), 2.05 (3H, s). IR: \) 1770, * c = o '1750, Ό 1680.

* c = o 67853

ΛΟ ^ Υ-Υ0 * 'a = o ^ 1- / AC

AcOCH CH = CH-OAc -9S1 J_> + i_v; H

CT (Τ '1 2a 2b

Procedure:

Chlorosulfonyl isocyanate (16.92 g; 0.12 mol) was added dropwise to cooled (ice-salt bath, -15 ° C) compound 1 (18.96 g; 0.12 mol). The pale yellow mixture was kept at 5 ° for 5 h, at which time it turned deep yellow.

This was diluted with ethyl acetate (20 ml), cooled to -30 ° C and added portionwise to a cooled (ice salt bath) mixture of water (3.4 ml), ice (17.0 g),

NaHCO 3 (0.3 g) and Na 2 SO 4 (3.4 g). The mixture obtained after the addition was stirred vigorously for 20 min. and to keep the pH between 7-8 a little more NaHCO 3 was added. The layers were separated and the aqueous layer was extracted with ethyl acetate (2 x 50 mL). The combined organic phase was dried (Na 2 CO 3: 1: 1). Filtered and evaporated to dryness to give 17.4 g of an oil. This was distilled under high vacuum (0.01-0.05 mm) from a hot air bath (temperature 55 ° -85 °) to remove compound 1. The non-distilled light brown oil was cooled, dissolved in ether and filtered through Celite to give, after evaporation to dryness, 5.28 g (22%) of a 1: 4 mixture of 2a and 2b (determined by nmr). as a colorless oil. NMR: 7.25 (H, NH), 6.0 (Q, 25H, d, J = 4.3), 5.8 (0.7, 5H, d, J = 15.), 4.5 (O , 5H, d, J = 6.5), 4.4 (1.5H, d, J = 4.5), 3.8 (0.25H, m), 3.5 (0.75H, m) , 2.13 (3H, s), 2.1 (3H, s). IR:)) c = Q 1780, 1740.

40 67853

OAC

Act / V-f CH 2 COSNa

__NH

^ She She AcS

2 AcO ^ N- / AcC / A- / ^ -1 ^ 'OAc

y_NH __NH J-NH

(Γ 0 3a 3b 3c

Procedural:

Sodium thioacetate was prepared by adding thioacetic acid (2.22 mL, 2.363 g) to a 1 mol solution. NaHCO 3 (31.0 mL) under nitrogen. This was added to a cooled (ice bath) solution of compound 2 (5.2 g; 25.9 mmol) in water (20 mL) and stirred for 4 h at room temperature. To make the reaction mixture homogeneous, a little acetone (20 mL) was added. The mixture was concentrated to remove acetone in vacuo and then extracted with methylene chloride. The extract was dried and evaporated to dryness to give 5.6 g of a mixture of isomers 3a and 3b (83.14%). The NMR spectrum of the crude oil showed that there was one trans and two cis compounds in the mixture. From this, a sample (550 mg) was chromatographed on silica gel (30 g, 10% H 2 O) and eluted with benzene-ether-methanol to give 200 mg of a 7: 1 mixture of 3a and 3b, followed by 150 mg of unknown cis (^ 5.55, d, J = 4.3) for which structure 3c was hypothetically proposed. NMR: 6.78 (H, NH), 5.52 (0.17H, d, J = 4.3), 5.18 (Q, 83H, d, J = 1.5), 4.37 (2H , d, J = 4.5), 3.45 (H, m), 2.35 (3H, s), 2.05 (3H, s). IR: C) 0 1765, 1740, 1600 cm -1.

41 67853

AccJ'N - [/ SAC ° 2H ^ O) ~ OH2C02C11 {0H * 2 / SAC

0 * -m ^ _N -CH-OH

o I

CO PNB 2 2 4

Procedural:

A mixture of crude compound 3 (2.17 g; 10 mmol) and p-nitrobenzylglyoxylate (2.5 g; 11 mmol) in benzene (200 mL) was boiled for 20 h under a Dean Stark trap, then concentrated in vacuo. to give 3.4 g of crude compound 4 as an oil. This was used as such without further purification. NMR: δ 7.5-8.5 (4H), 5.2-5.8 (4H), 3.4-5.1 (4H), 2-2.4 (6H). IR: ^ c_Q 1665, 1740, 1740, 1730, 1700. '

Aco / 'yYMc SOC12 Actf'S— | ^ Ac - N —C H - OH ^ H —0 H - C1

CO PNB 0 CO PNB

2 2 4 5

Procedure: To a cooled (ice bath) solution of crude compound 5 (3.3 g; 7.75 mmol) and pyridine (0.67 g; 8.5 mmol) in benzene (20 mL) was added dropwise thionyl chloride (1, 01 g; 8.5 mmol) in benzene (10 ml) and the mixture was stirred at the above temperature for 15 min. and at room temperature for 15 minutes. The benzene solution was decanted and the remaining semi-solid was washed three times with 15 ml portions of Benzene. The combined benzene solution was evaporated to dryness to give 1.90 g of crude compound 5 (55%). NMR: δ * 7.5 - 8.5 (4H), 6.12 and 6.2 (1H), 5.66 (1H, m), 5.4 (2H, d, J = 6), δ, 3-4 - (2H, m), 3.63 (H, m) 2.4 (3H, d), 2.1 (3H, s).

IR: 0 = 0 1765, 1740, 1730, 1700.

42 67853 yv Jjhc S Ac AC0 I-1 <Φ) 3Ρ ArO / X] - 1 HCl dioxane ^ —n- <p = p «J» 3

CO PNB CO PNB

2 'I 6

Procedure:

A mixture of crude compound 5 (1.90 g; 4.27 mmol), triphenylphosphine (1.572 g; 6 mmol) and 2,6-lutidine (0.542 g; 6 mmol) in dioxane (20 mL) was heated At 55 ° for 18 h. Cooled, filtered and evaporated to dryness to give 3.8 g of a crude dark oil.

This was chromatographed on silica gel to give 1.2 g (42%) of compound 6.

Actk '' 'W'SnC (AcikN— J-a ~ r ριφ, 3 * CO ^ NB Corpus ä 7

Procedural:

A solution of crude compound 6 (1.20 g; 1.79 mmol) in toluene (15 mL) was boiled for 5 h. It was cooled and evaporated to dryness to give an oil which was chromatographed on SiO 2 (30 g) and eluted. with benzene to give 0.4 g of compound 7 (57%). Anal. Calcd for C-, R, H, cN "O" S: 1 / Ib 2 7 C 52.04; H 4.11; N 7.14; Found: C, 51.77; H 4.08; N 7.30.

Separation of the cis and trans isomers was performed by careful chromatography on silica gel (60 g) eluting with benzene. cis isomer: ε (ppm, CDCl 3): 7.5-8.5 43 6785 3 (4H, aromatic), 5.67 (1H, d, J = 5, H-5), 5.28 (2H, AB quartet, benzyl), 4.33 (2H, d, AcOCH 2), 4.20 (1H, dt, H-6), 2.31 (3H, s, CH 3), 2.0 (3H, s, CH 3 CO ). δ p_Q 1770, 1740, 1730 cm-1, trans isomer: S (ppm, CDCl 3): 7.5-8.5 (4H, aromatic), 5.53 (1H, d, J = 2, H-5 ), 5.30 (2H, AB quartet, benzyl), 4.32 (2H, d, AcOCH 2), 4.27 (1H, dt, J = 5, J - 2, H-6), 2.31 (3H, s, CH 3), 2.0 (3H, s, CH 3 CO).) c = o 1770, 1740, 1730 cm -1.

^ rr> -ch3 \ θ2ΡΝΒ C02Na 7 i • Procedure:

To a solution of trans-7 (119 mg; 0.3 mmol) in ethyl acetate (15 mL) and water (7 mL) was added NaHCO 3 (25.2 mg, 0.3 mmol) and Pd / C. (110 mg) and this was hydrogenated at 30 psi for 4.5 h. The mixture was filtered and the layers were separated. The aqueous layer was washed with ether and freeze-dried to give 40 mg of solid compound 8 (48%).

c = 0 1765, 1740, 1600 cm -1, δ f (ppm, D 2 O): 5.52 (1H, H-5), 4.85 (2H, AcOCH 2), 4.0 (1H, H-6) , 2.65 (3H, CH 3), 2.40 (3H, CH 3 CO).

σ ΥΛ CO H

C02Na 2 8 11 44 6785 3

Example 3 2-Methylpenem-3-p-nitrobenzyl carboxylate (from a mercaptide intermediate)

. .SCOCH

/ 2CH "C0C1 —f '' '

I-f ---> 2 I

J— n pph. pyridine see 0 ^ 3 0 (

* min C0 „PNB

II C02PNB m 2

A solution of compound II (262 mg, 0.2 mmol), acetyl chloride (35 mg, 0.44 mmol) and 2 drops of pyridine in 10 mL of dichloromethane was stirred at 5 ° C for 1 h. The precipitated mercuric chloride was then filtered off and filtered off. the filtrate was washed successively with cold dilute hydrochloric acid, sodium hydroxide and finally brine. The organic solution was treated with hydrogen sulfide for 2 minutes at 5 ° C and stirring was continued at this temperature for another 10 minutes to precipitate the last traces of mercury salt. A little charcoal was added to the black mixture and the mixture was then filtered through a pad of Celite. Evaporation of the clear filtrate to dryness left 193 mg (80.7%) of compound III as a foam.

IR (CHCl 3) 1755 (-β-lactam) 1692 (SCOCH) 1620 (phenyl). '3 scccn —f'3 \ T. -> '/ CH3 J-n- ^ Fh Γ \ oy υ co2pnb

2 III C02 ™ 3 IV

Phosphorane III (75 mg, 0.126 mmol) was boiled in toluene (10 cm) for 2.5 hours under a nitrogen atmosphere. Evaporation of the solvent and purification of the residue gave a crystalline derivative (25 mg, 63%) whose physical and spectral values were in complete agreement with the values of the title product.

4 5 6785 3

Example 4 2- (4'-Phthalimido-1'-butyl) penem-3-carboxylic acid

C02H

° 0 °, 0 | / QT '\ ci TEA-H2S [pr \ SH

o O

1 2

To a solution of triethylamine hydrosulfide, previously prepared by bubbling 1 S gas through a solution of triethylamine (8.8 mL, 63.7 mmol) in methylene chloride (200 mL), was added dropwise at 0 ° C over 30 minutes. a solution of compound 1 (Gabriel Ber. 41, 2010) (10.55 g, 40.2 mmol) in methylene chloride (75 mL). The mixture was stirred at 0 ° C for 15 minutes and 2 hours at room temperature. The organic solution was diluted with methylene chloride (125 mL) and washed with 1N norm. HCl (2 x 15 mL), water (2 x 15 mL) and brine. Dry over MgSO 4 and evaporate the solvent to give 10.5 g (100%) of compound 2 as a white solid; mp. 93-94 ° C. n.m.r. (CDCl 3) J 7.5-8 (4H, m), 4.47 (1H, broad s), 3.5-3.9 (2H, m), 2.5-2.9 (2H, m ), 1.4 - 1.9 (4H, m). Anal. calc. for C 18 H 18 NO 2 S: C, 59.29; H, 4.97; N, 5.32; S, 1217. Found C, 58.92; H, 4.91; N, 5.42; S, 12.31.

Πί, '> - m 5 2 3 ***> 3> i

VwA [Qj

and V

J — NH 0 '> 0' o 4 46 6785 3

A suspension of compound 2 (3.04 g, 11.6 mol) in 1 mol. in sodium bicarbonate solution (11.6 ml) was stirred at room temperature under nitrogen for 15 minutes. Compound 3 (1.5 g, 11.6 mmol) was added and the resulting mixture was stirred at room temperature for 1.5 h. The reaction mixture was diluted with water and extracted with methylene chloride. The organic phase was dried and evaporated to dryness in vacuo to give 3.82 g of solid compound 4; mp. 95-96 ° C; I.R. (CHCl 3) 1775, 1710 cm -1, n.m.r. <f7.8 (4H, d, j = 2Hz), 7.05 (1H, broad s), 5.25 (1H, dd, J = 5Hz, J = 3Hz), 3.5-3.0 (2H, m), 1.5-2.0 (4H, m). Anal. calc. for C 16 H 16 N 2 O 4 S: C '57'62' H »4> 85; N> 8.43j S, 9.64. Found: C, 57.43; H, 4.82; N, 8.44; S, 9.71.

---- OP h2- 0> ° h- Λ-rl 'i rrsh "' VLyi ^ OH 0 C ° 2-2- <§) -N02 5

A benzene solution (30 mL) of Compound 4 (3.0 g, 9.04 mmol) and p-nitrobenzyl glyoxalate (2.22 g, 9.8 mmol) was boiled for 21 h under a Dean-Stark condenser filled with 3A. molecular sieve. Evaporation of the solvent gave 5.4 as an oil (100%). I.R. (fine) 3200-3600, 1770, 1710, 1525 cm -1, nmr (CDCl 3) 68.21 (2H, d, J = 9 Hz), 7.75 (4H, d, J = 2 Hz), 7, 52 (2H, d, J = 9 Hz), (1H, broad s), 5.32 (3H, 2s), 4.55 (1H, broad s), 3.5 to 3.7 (2H, m), 3.45 (1H, dd, Jgem = 15 Hz, = 5 Hz), 3.02, (1H, dd, Jgem = 15 Hz, Jtrdns = 3Hz), 2.4-2.9 (2H, m), 1.4 - 2.0 (4 H, m).

67853 47 C ° 2ch 2- {0y- ^ ° 2 CO2CH 2 ~ (2) _NO 2 5 £

Azetidinone glyoxalate 5 (4.9 g, 9.05 mmol) was treated with thionyl chloride (15 mL) at 0 ° C for 0.5 h and at room temperature for 1 h. Excess thionyl chloride was distilled off with benzene in vacuo to give compound 6 as a yellow syrup (5.0 g, 100%). n.m.r. (CDCl 3) δ 8.2 (2H, d, J = 9Hz), 7.72 (4H, broad s), 7.60 (2H, d, J = 9Hz), 6.1 (1H, broad s) , 5.50 - 5.85 (1H, m), 5.32 (2H, 2s), 3.4 - 4.0 (2H, m), 3.1 - 3.3 (1H, m), 2 Δ - 3.05 (1H, m), 2.50 - 2.85 (2H, m), 1.5 - 1.9 (4H, m).

CO 2 CH 2 - (O> -NO 2 6 - 7

A solution of compound 6 (21.6 g, 38.8 mmol) in tetrahydrofuran (85 mL, distilled over LAH) was treated with triphenylphosphine (10.2 g, 38.8 mmol) and 2,6-lutidine (5 .0 mL, 42.9 mmol) for 18 h at 40 ° C. The mixture was diluted with benzene-ether 1: 1 (30 mL), washed with water, 1-norm. HCl, saturated NaHCO 3, brine and dried over MgSO 4. Evaporation of the solvent gave a dark brown oil. It was passed through a silica gel (700 g) column (benzene-48 67853 ether mixture) to give 16.0 g (53%) of compound 7 as a thick oil. NMR (CDCl 3) d * 8.2 (2H, d, J = 9Hz), 7.8 (8H, d, J = 2Hz), 7.52 (16H, broad s), 5.2 (1H, broad s ) 4.78 (1H, 2s), 4.30 - 4.52 (1H, m), 3.5 - 3.8 (2H, m), 2.8 - 3.5 (2H, ra), 2 , 1-2.9 (4H, m), 1.5-1.9 (4H, m) $ 7

A solution of phosphorane 7 (5.0 g, 6.4 ramol) in toluene (35 ml) was boiled for 3 h. Evaporation of the solvent gave a residue which was passed through a silica gel (100 g) column. Elution with benzene followed by ether gave 600 mg of ester 8 as an oil, i.r. (fine 1790, 1710, 1520 cm -1 nmr (CDCl 3) no * 8.2 2 (2H, d, J = 9Hz), 7.8 2 (4H, d, J = 2Hz), 7.55 (2H, d , J = 9Hz), 5.69 (1H, dd, J = 4Hz, J - 2Hz), C 1 - SLX d 11 ώ 5.35 (2H, 2s), 4.12 (1H, dd, J = 16Hz, J = 4Hz), 3.50 μm cis (1H, dd, Jgem = 16Hz, Jtrans = 2Hz), 3.1-3.8 (2H, m), 2.5-3.0 (2H , m), 1.4 - 2.0 (4H, m).

49 67853 - '(1 °)

CO2CH2- (Oy ^ NO2 CO2H

8 i

A biphasic mixture of ester 8 (196 mg, 0.39 mmol) in ether (2 mL), tetrahydrofuran (4 mL) and sodium bicarbonate (32 mg, 0.39 mmol) in water (2 mL) was hydrogenated with 30% palladium / with diatomaceous earth (190 mg)

Parr1 in a shaker at 40 psi. After 4.5 hours, it was filtered through a pad of Celite and the pad was washed with water and tetrahydrofuran. The filtrate and washings were combined and the organic phase was separated. The aqueous solution was washed with ether, acidified to 1-norm. HCl (3 x 0.4 mL) and extracted (after each addition of acid) with ethyl acetate (4 x 2 mL). The organic extracts were washed with brine, dried over MgSO 4 and the solvent removed by evaporation to dryness to give the acid 9.67 mg (47%) as a yellow solid, i.r. (nujoli) 1775, 1705, 10690 cm -1, nmr (DMSO) δ 7.92 (4H, s), 5.71 (1H, dd, Jcis-4Hz, Jtrans = 2Hz), 3.90 (1H, dd , Jgem = 16 Hz,

Jcis = 4Hz) '3'47 (1H' dd 'Jgem = 16 Hz' Jtrans = 2Hz)> 3'3 '4'3 (3H, m), 2.7 - 3.05 2H, m), 1, Δ - 2.0 (4H, m).

50 67853

Example ^

Sodium 2- (acetonylmethyl-oxime) -penem-3-carboxylate XCO2Na .scocyl -? - cn ^ SCOCH2-C-CH3 - s' 2 3/3 _OH ^

0 ^ -0 J

I CO PM3 CO ^ PNB 2 1 ^

Ketal 1 (2.0 g, 4.54 mmol) was treated at 0 ° with 95% 3 TFA (20 cm) for 15 minutes. The mixture was diluted with brine and extracted with methylene chloride (4 x 30 cm). The methylene chloride extracts were washed with brine (3 times and brine, and dried over MgSO 4 (1.44 g, 80%)).

S (ppm, CDCl 3) 8.27 (2H, d, J = 9, Hm aromatic), 8.60 (2H, d, J = 9, Ho aromatic), 5.70-5.25 (m, CHO -PNB, HC-O, H-4, ° Cv'c = cr ^ A) **, 75 (1H, bs, OH), 3.76 (center of ABq,

H ^ ^ OH

CH2-C0), 3.47 (part of dd, J3_4 cig = 5, H-3), 3.05 (2H, 2dd,

Jgem = 15 'J3-4 trans = 3' H "3)> 1'30 '1'28 (1'67H' 2s ^ CHg), 1.98 (1.33 H, s, CH3) $ q = q (CHCl 3) 1780, 1755, and NQ 1525.

9 JSC0CHoJ; -CH

2SCOCH2CCH r-S * 3

1 * J-N DH

/ -N ΟΠ Ο γ '

I CO PNB

co2pnb 2 - 51 67853

A solution of ketone 2 (1.1 g, 3.63 mmol) in methylene chloride (50 cm -1) was treated at 0 ° under a nitrogen atmosphere with methoxylamine hydrochloride (334 mg, 1.1 eq.).

Triethylamine-3 (367 mg, 0.51 cm, 1 eq.) Was then added dropwise to the mixture. It was then stirred at room temperature for 18 h. The reaction mixture was diluted with methylene chloride, washed with water-brine (2 times), brine and dried over MgSO 4 (1.52 g, 98%).

^ (ppm, CDCl 3) 8.12 (2H, d, J = 8, H aromatic), 8.40 (2H, d, J = 8 H aromatic), 5.50-5.05 (4H, m, C ^ -PNB, H-4, HC-O), 3.80-3.60 (m, OCH 2, part of H-3 cis, part of OH), 3.55-270 (m, H- 3 part of cis, H-3 trans, part of CH 2 CO, OH), 1.97, 1.90, 1.88 (3H, 3s, CH 3) δ c = q (CHCl 3) 1770, 1750, 1690.

N-OCH

I) J h ~ -—OCH

SCOCH -C-CH II 3

/ 2 3 ^ SCOCH -C-CH

[SOCl2 - \ 2 3 cJ- γΟΗ pyFIdTIHi J__ C1 C02PHB co2? Hb

A cold (-15 ° C) solution of azetidinone 3 (1.52 g, 3 3.57 mmol) in THF (20 cm, distilled over LAH) was treated by the dropwise addition under a nitrogen atmosphere with pyrethane. dimx (325 mg, 0.332 cm, 4.10 mmol, 1.15 eq) and thionyl chloride (488 mg, 0.299 cm, 4.10 mmol, 1.15 eq). The mixture was stirred for 15 min. -15 ° C. The solid was filtered off and washed with benzene. The resulting solution was evaporated to dryness. The residue was dissolved in benzene and the solution was treated with charcoal (1.2 g, 76%).

ν (ppm, CDCl 3) 8.23 (2H, d, Hm aromatic), 7.80 (2H, d, Ho aromatic), 6.12, 6.08 (1H, 2s, HC-Cl), 5.75 - 5.55 (1H, m, H-4), 5.40, 5.30 (2H, 2s, CH 2 - PNB), 3.95 - 3.80 (3H, 3s, OCH 2), 3.80 - 2.95 (4H, m, 2H-3, CH 2 CO), 2.00 - 1.85 (3H, 4s, CH 2).

52 6785 30c = o (CHCl 3) 1790, 1765 (shoulder), 1700, V> NQ 1530.

H — OCH, —OCH,

II 3 II

scoch2-c-ch3 s scoch2-c-cm3 rT - JTI w, t / - "^ cl 0 Ύ '

\ CO PNB

CO PNB 2 i 1 2

A solution of chlorazetidinone 4 (1.2 g, 2.70 mmol) in THF (20 cm, distilled over LAH) was treated with triphenylphosphine (1.06 g, 4.05 mmol, 1.5 eq.) and 2,6-lutidine-3 nm (318 mg, 0.346 cm, 2.97 mmol, 1.1 eq.). The mixture was stirred for 4 days at room temperature under a nitrogen atmosphere. Diluted with ethyl acetate, washed with 2% aqueous HCl, Hail, 2% aqueous NaHCO 3, water and brine. The solution was then dried over MgSO 4 and the solvent was evaporated. The crude compound 5 was purified on silica gel (10 times the weight) on a column (ethyl acetate, 770 mg, 45%).

0 (CHCl 3) 1755, 1695, V 1630 - 1610, 152 5.

”U o im u 2

N-v-OCH

Il [wof’l scoch2-c-ch3 r-r3 \ il 4 5 I \ --1—> Γ,! > -cVC "CH3 j-ί toIueenl o * ~~

O [CO PNB

COjPNB

- 2

Phosphorane 5 (700 mg, 1.05 mmol) was boiled with toluene-ether-benzene). There was thus obtained compound 6 as a crystalline substance (251 mg, 62%, m.p. 116-125 °).

Evaporation of toluene gave a residue which was passed through a silica gel (1:15) column (4% 53 6 7 8 5 3

Anal. Calculated for C, „Ηη„ N., OrS:

J_ / _L / o O

C, 2.17; H, 4.38; Found: C, 51.15; H, 4.18; jj? 1Q 33> <f (ppm, CDCl 3) 7.70 (2H, d, Hm aromatic), 7., ^ (2H d, Ho aromatic), 5.00 (2H, s, CH 2 PNN), 4.85 (1H , 3.75-2.70 (7H, m, CH 3 O, CH 2, H-6), 1.77, 1.72, 1.65 (3H, s> CH}> ^ c = o (CHCl 3) 1787, 1742, 1705, yNO 1530. 3 UV (EtOH) λ max 318 (£ = 8420), 262 (¢ = 12.539).

- N — OCH „ς a — OCH1 —r \ ti 3 h2 (r \ ti

/> - CH -C-CH, - »I V-CH -C-CH

2 3 NaHC ° 3 CO PNB CO 2 2 2 6 7

A mixture of ester 6 (151 mg, 0.386 mmol) in 3 3 THF (20 cm), ether (40 cm) and NaHCO 3 (32 mg, 0.381 mmol) in water (20 cm) , was shaken in a Parr hydrogenator for 3 h at 35 psi using 30%

Pd / Celite catalyst (200 mg). The catalyst was filtered off and washed with water and ether. The resulting aqueous mixture was washed with 3 ethers (3 x 60 cm) and lyophilized (32 mg, 30%).

δ (ppm, DMSO) 5.50 (m, H-5), 3.75 (s, OCH 3), 0.77 (s, CH 3).

\] r_ (nujolitahdas) 1770, 1600, 1400.

<H 2 O) Araaks 300 (i - 2,800), 255 (£ = 2,400).

54 67853

Example 6 Γ y ^ <o - <, 2 0Λ-ν ^ 2 ^ co2h (_ ^ SAg - | ^ Svvjj ^ / Vvnch2ci 0Ργ3 + cicoch2ch2ci ‘J- * γ * 3

CO PNB CO PNB

2

I II III

A solution of I (1.1 g, 1.6 mmol) and II (0.16 mL, 1.6 mmol) in methylene chloride (30 mL) was cooled in an ice bath and 1 mol was added dropwise. methylene chloride solution of pyridine (1.7 mL, 1.7 mmol). The resulting reaction mixture was stirred at room temperature for 1 h and then filtered through Celite and washed with methylene chloride. The filtrate and washings were combined and washed successively with 1-norm. HCl (5 mL), water (5 mL), 1 mol. NaHCO 3 (5 mL) and brine, and then dried (MgSO 4) and evaporated to dryness in vacuo to give Compound III 900 mg (87%) as an amorphous solid. It was used without further purification in the next step. IR (CHCl 3) 1755, 1690 cm -1.

NMR (CDCl 3) δ 8.22 (2H, d, J = 9 Hz), 7.55 (15H, m), 6.72 (2H, d, J-9 Hz), 5.7 (1H, m ), 5.0 (2H, 2s), 3.55 (2H, 2s), 2.8 (4H, m).

t 55 6 7 8 5 3 rfSY ^ H2Cl, <> _ 0> P-- 0J-Μ \ ^ Ρφ3 C02PNB _—? o - <) 2 HI IV 0 ^ -ΝΎ ^ Φ3 CO PNB 2

V

A mixture of III (1.3 g, 2 mmol) and IV (0.65 mL, 3 mmol) was heated at 80 for 4 h. The reaction mixture was diluted with methylene chloride (10 mL) and washed with water (2 x 5 mL). . The organic layer was dried (MgSO 4) and evaporated to dryness in vacuo to give Compound V, 1.4 g (90%), as an amorphous solid. It was used without further purification in the next step.

IR (CHCl 3) 1755, 1690 cm -1 NMR (CDCl 3) δ 8.25 (2H, d, J = 9 Hz), 7.55 (15 H, m), 6.8 (2H, d, J = 9 Hz) ), 5.7 (1H, m), 5.1 (2H, 2s), 4.72 (2H, dq J = 12 Hz, J = 6 Hz), 2.6 (4H, m), 1.4 (6H, s), 1.28 (6H, s).

I O -> ΓΤ ’/ Μκ., 2

v CO PNB

2

VI

A solution of V (1.6 g, 2.06 mmol) in toluene (60 mL) was heated at reflux for 5 h.

The solvent was evaporated in vacuo and the residual oil was chromatographed on a silica gel column (30 g). Elution with benzene and then ether first removed the non-polar material and then eluted with ethyl acetate to give compound VI, 620 mg (62%) as a white solid, m.p. 83-4 ° C crystallized from ether.

IR (CHCl 3) 1790, 1710 cm -1 NMR: (CDCl 3) δ 8.2 (2H, d. J = 9 Hz), 7.6 (2H, dj J = 9 Hz) 7.5 (2H, s) 5.65 (1H, dd, Jtrans = 4 Hz,

Jcis = 2 Hz), 5.22 (2H, 2s), 4.75 (2H, dq J = 12 Hz, J = 6 Hz), 3.85 (1H, dd, J = 15 Hz, J. = 4 Hz), 3.5 (1H, da, J = 15 Hz, J · = 2 Hz), 2.8-3.3 (2H, m), 1.4 (6H, s), 1.28 (6H , s).

Llb I -► rr? (O - o <y-N 2

CO ΡΝΊ3 CO H

2 υ2

VI VII

To a solution of VI (200 mg, 0.4 mmol) in tetrahydrofuran (8 mL) and ether (4 mL) was added sodium bicarbonate (34 mg, 0.4 mmol), water (4 mL) and 30% Pd / Celite catalyst (200 mg) was then hydrogenated at 40 psi for 2 h. The mixture was filtered and the layers were separated. After washing with methylene chloride (2 x 5 mL), the aqueous phase was cooled with ice, acidified to 1-norm. HCl (1 mL), and extracted with chloroform (5 x 5 mL). The organic extracts were dried (MgSO 4) and evaporated to dryness in vacuo to give compound VII, 76 mg (52%) as an oil. IR (CHCl 3) 1790, 1710 cm -1 NMR (CDCl 3) δ 9.5 (1H, δ), 5.65 (1H, dd, J 1 = 4Hz, J = 2 Hz), 4.72 (2H, dq J = 12 Hz, J = 6 Hz), 4.2-5.1 (2H, m), 3.4-4.1 (2H, m), 2.7-3.4 (2H, m), 1 .35 (6H, s), 1.25 (6H, s).

57 67853

Example 7 «I rV ^ \ s 1 // F (OC H) J2 5 ^ 2 CO H 2 ^ SA9 ^ pf + C1C (CH) P (OC H) - ► / γ» 9 CO2PNB _Y '(CH2) 3 ^ (OC2H5.2 1 1

CO ^ FNB

2 To a cooled (ice bath) mixture of compound 1 (1.324 g, 2 mmol) and compound 2 (0.54 g, 2.2 mmol, crude) in Cl 2 Cl 2 (15 mL) was added dropwise 1 mol. pyridine / Cl 2 Cl 2 solution (2.2 mL, 2.2 mmol). The mixture was stirred at room temperature for 1 h and filtered through Celite. The filtrate was washed successively with 0.5 nomr. With HCl, IVO, 0.5 mol. NaHCO 3 and brine. Dried (MgSO 4) and filtered through Celite to give 0.9 g of an oil after evaporation to dryness, the oil was chromatographed on SiO 2 (10% H 2 O) and eluted with ethyl acetate to give 0.5 g of compound 3. (32.8 %). NMR δ (ppm, CDCl 3) 7.0-8.4 (m, 19H), 4.8-5.8 (3H, m), 4 *, 1 (4H, q), 3.3-4 , 2, (2H, m), 2.7 (2H, m), 1.9 (2H, m), 1.3 (6H, t).

58 6 7 8 5 3 (OEt) ΓΤ 1 -1- ‘0 ^ γΦ3

¢ 0 PNB ^ S

J Xs ^ NY (OEt) 2 CO PNB 2 4

Compound 3 (0.4 g, 0.52 mmol) was boiled in toluene (35 mL) for 4 h, and evaporated to dryness to give an oil of compounds 3, 4 and O 2 P = O * This was chromatographed on Pigs (10% H 2 O). ) and eluted with ethyl acetate to give 0.1 g of pure compound 4, followed by 0.15 g of compounds 3 and 4. NMRif (ppm, CDCl 3). 8.3 (2H, d), 7.67 (2H, d), 5.7 (H, q), 5.33 (2H, d), 4.2 (4H, q), 3.83 (H , q), 3.4 (H, q), 2.9 (2H, m), 1.9 (2H, m), 1.3 (6H, t), IR (fine) 1790 cm -1. kt Aarni) 1710 cm -1 (ester).

1 — rf ^ S'V / \ / ^ N "? (OEt) ___ CO PUB \ / \ 1 2 r \ y ^ ^ p (OEt) 2

J-N

4 O \ co2h 5

A mixture of compound 4 (0.1 g, 0.207 mmol), 30% Pd / Celite catalyst (0.1 g) and NaHCO 3 (17 mg, 0.207 mmol) in THF (10 mL), ether (5 mL) and water (5 mL) were hydrogenated for 2 h at an initial pressure of 40 psi. The mixture was filtered using Celite and the layers were separated. The basic aqueous layer was washed well with ethyl acetate and acidified to 1-norm. HCl. Extracted with CH 2 Cl 2 and dried (MgSO 4). The CH 2 Cl 2 solution was evaporated to dryness to give 48 mg of 5 59 67853 ° (66.5%). IK spectrum ./'17 90 (β-lactam) ^ 17Q0 (..Q_0Ii).

Example. 8 9 (0Me) 2 CC ^ Na ^ Cl 0 + (HeO) P - *

CCTPNE I

2 CO PNB

2 I 2 3

A mixture of compound 1 (1.07 g} 1> 66 mmol) and compound 2 (0.42 g, 3 mmol) in CH 2 Cl 2 (3 mL) was heated at 80 for 5 h. The crude oil was chromatographed on SiO 2: 11a (3% H 2 O) and eluted with ether, ether-ethyl acetate / mixture (1: 1) and ethyl acetate and 5% EtOH to give 1.0 g of compound 3 (82%). öpjy crystallized when in place, m.p. (from ether) 138-140 °.

NMR / (ppm, CDCl 3) 8.2 (2H, d) 7.0-8.0 (17H, m), 4.6-5.5 (3H, m), 3.8 (3H, s), 3.6 (3H, s), 1.5-3.5 (6H, m).

9 o .S. ^., Ρ (ΟΜβ), 9 1 -f'2 P (OMe) 2 f ^ 0 toluene. 1/7 0d — ΝγΡΦ, δ '"0i -> * γ CC ^ PNB co2pmb 3 4

Compound 3 (0.5 g, 0.69 mmol) was boiled in toluene (30 mL) for 4 h. It was evaporated to dryness, chromatographed on Pigs (3% H 2 O) and eluted with Et 2 O: EtOAc (1: 1) followed by with EtOAc-10% EtOH to give 0.18 g of compound 4 (58%). NMR δ (ppm, CDCl 3), 8.25 (2H, d), 7.6 δ ° 6785 δ (2Hj d), 5.65 (H, q), 5.3 (2H, d), 3.8 (3H, s) 3.6 (3H, s), 2.7-3.6 (2H, m), 1.5-2.5 (4H, m).

_ .S I (OMe) 2 I 30¾ Pd / rel ite ,, J, _NaHCO ^ CO PNB 2 9 CO Ma 5_

A mixture of compound 4 (50 mg, 0.112 mmol), NaHCO 3. (9.125 mg) and 30% Pd / Celite catalyst (50 mg) in THF (5 mL), Et 2 O (2.5 mL) and water (2.5 mL) were hydrogenated at an initial pressure of 40 psi ( 2 hours). The mixture was filtered using Celite and the layers were separated. The basic aqueous layer was washed well with ethyl acetate and lyophilized under high vacuum to give 28 mg of compound 5 (75.9%) (hygroscopic).

IR (KBr) 1770 cm -1 (γ 3 -actam), 1610 cm -1 (-C 0).

61 67853

Example 9 (1'R, 5R, 6R) and (1'S, 5S, 6S) 6- (1 * -hydroxy-1'-ethyl) -2-methylpenem-3-carboxylic acid (Isomer D) (describes the most preferred method To incorporate a 6-substituent in the middle of the synthesis)

OH

Λ -, - λ

CO ^ iia, K

A. Preparation of 4-Tritylthio-2-azetidinone Intermediates 1. 1- (Trimethylsilyl) -4-tritylthio-2-azetidinone fYsc * 3

A solution of 4-tritythio-2-azetidinone (345 mg, 1 mmol), 1,1,1,3,3,3-hexamethyldisilazane (80 mg, 0.5 mmol) and chlorotrimethylsilane (55 mg, 0 mmol) was added. .5 mmol) in dichloromethane (20 ml) was heated at reflux for 18 h. Evaporation of the reaction mixture to dryness left virtually pure title compound. Ε (ppm, CDCl 3): 7.32 (15H, m, aromatic), 4.22 (1H, dd, H-4), 2.57 (1H, dd, J = 4.1, J = 16, H-3), 2.22 (1H, dd, J = 2.2, J = 16, H-3), 0.3 (9H, s, CH 2).

62 6 7 8 5 3 2. 1- (t-butyldimethylsilyl) -4'-tritylthio-2-azetidinone / X-

I J

C (CH3) 3

Triethylamine (1.62 mL, 11.6 mmol) was added dropwise over 5 minutes to a cooled (0 °) and stirred solution of 4-tritylthio-2-azetidione (3.5 g, 10.1 mmol) and chlorine. t-butyldimethylsilane (1.68 g, 12.7 mmol) in DMF (35 mL). The reaction mixture was stirred at room temperature for 18 hours, diluted with water (250 mL) and ether (200 mL). The organic phase was washed with water (3 x 50 ml), dried and evaporated to dryness to leave an oil (4.33 g). Crystallization from pentane gave a total of 4.1 g (89%) of the title compound as a white solid, m.p. 113-4 °.

δ (ppm, CDCl 3): 7.45 (15H, m, aromatic), 4.2 (1H, dd, H-4), 2.63 (1H, dd, J = 4, J = 16, H-3 ), 2.13 (1H, dd, J = 2, J = 16, H-3), 1.0 (9H, s, t-Bu), 0.35 (6H, s, Me). tfG_0 1735 cm 1. Anal. calc. for C28H33NSO5: C, 73.15; H, 7.24; N, 3.05; S, 6.97%. Found: C, 73.27; H, 7.32; N, 2.97; S 6.94%.

67853 3. 1-Methoxymethyl-4-tritylthio-2-azetidinone

I —-► I

A— N J N.

O 2 H CH 2 OCH 3

A solution of 4-tritythio-2-azetidinone (1.38 g, 4.0 mmol) in THF (10 mL) was added to a well-stirred suspension of sodium hydride (200 mg of technical 50%, 4.1 nunol, washed). pentane) in THF (10 mL) kept at -15 °. Methanol (12 drops) was added and the mixture was stirred at 0-15 for 0.5 h. Methoxymethyl bromide (0.58 g, 4.6 mmol) was added and the mixture was stirred for 2 h, diluted with ether, washed with water and brine, dried and evaporated to dryness to leave an oil (1.72 g). Crystallization from pentane gave a white solid (1.41 g), m.p. 72-76 °, (*, ppm, CDCl 3): 7.3 (15H, m, aromatic), 4.4 (3H, m, NCl 2 O and H-4), 3.22 (3H, s, CH 2), 2.76 (2H, m, H-3).

4. 1- (methoxyethoxymethyl) -4-tritylthio-2-azetidinone. rf

To a suspension of tetrabutylammonium bromide (322 mg, 1 mmol) and potassium hydroxide (85%, 70 mg, 1.1 mmol) in dichloromethane (10 mL) cooled to 5 ° was added 4-tritylthio-2- azetidinone (345 mg, 1 mmol) and methoxyethoxymethyl chloride (187 mg, 1.5 mmol). The mixture was stirred at room temperature for 2 h, the solvent was evaporated off and the residue was partitioned between water and ethyl acetate. The dried organic phase was evaporated to dryness to leave a viscous oil (415 mg). Purification by column chromatography on silica gel, eluting with ether (5%) -dichloromethane, gave the title compound (206 mg, 48%) as an oil. , f (ppm, CDCl 3): 7.30 (15H, m, aromatic), 4.57 (2H, AB quartet, N-CH 2 O), 4.46 (1H, dd, H-4), 3 .50 (4H, s, OCH 2 CH 2 O), 3.30 (3H, s, CH 2), 2.75 (? H, m, H-3).

5. 1- (2'-Tetrahydropyranyl) -4-tritylthio-2-azetidinone 5β, 3N, 3'-n-butyllithium (1.6 mol, 1.6 ml, 2, 56 mmol) was added dropwise to a solution of 4-tritylthio-2-azetidinone (863 mg, 2.5 mmol) in THF maintained at -78 °.

After stirring for 15 minutes, 2-chlorotetrahydropyran (560 mg, 4.7 mmol) was added and the reaction mixture was allowed to warm to room temperature over 1.5 hours. The reaction solution was diluted with ethyl acetate, washed with brine, dried and evaporated to dryness in vacuo to leave an oil (635 mg). Column chromatography on silica gel, eluting with dichloromethane-ether, gave an isomeric mixture of the title compounds with a small amount of starting material as an impurity. δ * (ppm, CDCl 3): 7.28 (15H, m, aromatic), 4.4 (H, dd, H-4), 2.9-2.2- (2H, m, H-3) , 4.1-3.2 and 2.2-0.7 (tetrahydropyranyl).

65 6785 3 B. Preparation of 3- (1'-hydroxy-11-ethyl) -1-methoxymethyl-4-tritylthio-2-azetidinones [3A, 3R, SC * 3 and the like. (- jn ^ a) (1'S, 3S, 4R and 11R, 3R, 4S) isomer (isomer C) in THF (5 ml) was prepared at -78 ° C from a solution of lithium diisopropylamide in n-butyllithium (1.6 mol). ., 1.0 mL, 1.6 mmol) and diisopropylamine (0.25 mL, 1.84 mmol). After 30 minutes, a solution of 1-methoxymethyl-4-tritylthio-2-azetidinone (491 mg, 1.42 mmol) in THF (6 mL) was added dropwise and the solution was stirred for 15 minutes. Acetaldehyde (3.0 ml) was added dropwise, after which, after 20 minutes, water (30 ml) was added. The mixture was acidified (to pH 3) with 2% HCl and extracted with ethyl acetate (5 x 20 mL). The combined organic phases were washed with brine, dried and evaporated to dryness to leave an oil which crystallized on trituration with ether: 440 mg, 80%, m.p. 188.5 to 9 ° C; 1 Hmr (CDCl 3) δ: 7.3- (15H, m, aromatic), 4.37 (2H, ABq, N-CH 2 O), 4.32 (1H, d, J = 2, H-4) , 3.17 (3H, s, OCH 3), 3.32-2.70 (2H, m, H-3 and H-5), and 1.12 ppm (3H, d, J = 7, CH 3); Anal. Calcd for C 18 H 18 NO 2 S: C 72.02, H 6.28, N 3.23, S 7.39; found: C 71.99, H 6.02, N 3.21, S 7.40%.

b) 1'S, 3S, 4R and 1'R, 3R, 4S) and (1'R, 3S, 4R and 1'S, 3R, 4S) (isomers C and B)

A solution of lithium diisopropylamide (0.482 mmol) in butyllithium (0.191 mL of a 2.52 M solution in hexane, 0.482 mmol) and diisopropylamine (0.067 mL, 0.482 mmol) is prepared in dry ether (3 mL) at -78 ° C. After 20 minutes, a solution of (4R and 4S) 1-methoxymethyl-4-tritylthio-2-azetidinone (0.171 g, 0.439 mmol) in a mixture of dry ether (1 mL) and dry THF (1 mL) was added dropwise. , and the resulting clear solution of 67853 66 was stirred at -78 ° C for 15 minutes. Tetrabutylammonium fluoride solution (0.96 ml of 0.5 M THF solution, 0.48 mmol) was then added. A precipitate formed as the mixture turned slightly pink. After 5 minutes, the reaction mixture at -78 ° C was quenched with freshly distilled acetaldehyde (0.2 mL, excess), and stirring was continued for another 15 minutes. Further work-up was carried out by adding the mixture to saturated ammonium chloride solution and extracting with ethyl acetate (2 x 25 ml). The combined organic phases were washed with brine and dried over anhydrous magnesium sulfate. Evaporation of the solvent in vacuo gave an oil (0.228 g) which was chromatographed on 10 grams of silica gel. A mixture of benzene and ethyl acetate (6: 4) gave 0.106 g (62% yield) of the substance and a mixture of two alcohol isomers, which were separated by chromatography on thick-layer plates (solvent mixture the same). The high Rf alcohol (0.033 g, 17%) was identical to the above isomer (isomer C): m.p. 188.5-189 ° C (ether-dichloromethane); a low Rf alcohol (0.030 g, 16%) isomer B '> was obtained as an oil which was difficult to crystallize from hexanes: m.p. 94-95 ° C. and (CHOC10). : 3600 (OH), 1760 cm -1 (C = O); 1 Hmr (CDCl 3) δ: 6.9-7.5 (15H, m, aromatic), 4.2 (2H, center of ABq, J- 11.5, CH 2 -O-CH 3), 4.28 (1H, d, J = 2.0, 4-H), 3.65 (1H, broad sextet center, H-1 '), 3.3 ( 1H, dd, 4 trans = 2'5 'J3 1' = 5.5, H3), 3.15 (3H, s, O-CH3), 1.55 (1H, broad s, OH-1 '), 1.05 (3H, d, J = 6.5, H-2 ') Anal Calcd for C 26 H 27 NO 3 S: C 72.02, H 6.28, N 3.23, S 7.39, found: C 71.77, H 6.36, N 3.15, S 7.43%.

67 67853 C. Preparation of trans-3-acetyl-1-methoxymethyl-4-tritylthio-2-azetidinone o, X usc4> 3 ___LDA CH3 Γ

| EtOAc - N

Lithium diisopropylamide was prepared under standard nitrogen protection at -78 ° C from diisopropylamine (0.34 mL, 2.4 mmol) and n-butyllithium (1.1 mL of a 2.2 M hexane solution, 2.4 mmol) in THF (3 mL). A solution of 1-methoxymethyl-4-tritylthio-2-azetidione (0.78 g, 2 mmol) in THF (3 mL) was added dropwise and after stirring for 20 min. At -78 ° C, ethyl acetate (0.53 g, 6 mmol) was added in one portion and stirring was continued for 0.75 h at -78 ° C. The reaction mixture was diluted with ether and washed with ammonium chloride solution, water and brine, dried and evaporated to dryness to give an oil (0.7 g). Purification was performed by chromatography on silica gel (20 g) and eluting increasingly with ether containing benzene. The appropriate fractions were evaporated to dryness to give the title compound as a colorless oil (0.32 g, 37%); 1 Hmr (CDCl 3) δ: 7.7-6.8 (15H, aromatic), 4.85 (1H, d, J = 2, H-4), 4.5 (2H, s, N-CH 2 - O), 3.9 (1H, d, J = 2, H-3), 3.22 (3H, s, CH 3) and 2.0 ppm (3H, s, CH 3); and Vmax: 1770, 1710 cm-1.

68 6785 3 D. Preparation of trans-3-acetyl-1- (t-butyldimethylsilyl) -4-tritylthio-2-azetidinone - »,

CT "Ν \ 5 / · Β" EtOAC O? -% si / t_EU

N (CH3) 2 Slv «cV2

Diisopropyllithium amide was prepared in the usual manner from diisopropylamine (0.18 mL, 1.24 mmol) and n-butyllithium (0.78 mL of a 1.6 M hexane solution, 1.24 mmol) in THF (8 mL). At -78 °, a solution of 1- (t-butyldimethylsilyl) -4-tritylthio-2-azetidinone (0.46 g, 1 mmol) in THF (8 mL) was added dropwise. After a stirring period of 5 minutes, ethyl acetate (1 ml) was added in one portion and the mixture was stirred at -78 ° C for 3 h. The mixture was acidified with cold hydrochloric acid (0.5-norm) to pH 6 and extracted with ethyl acetate (2 x 20 mL). The combined organic phases were dried and evaporated to dryness to give an oil (0.5 g) which was crystallized from pentane: a total of 200 mg, 40%; mp. 122-4 ° C; and V max: 1750 '1710 cm-1; m / z (CDCl 3) δ: 7-7.1 (15H, m, aromatic), 4.83 (1H, d, J = 2, H-4), 3.38 (1H, d, J = 2, H = 3), 1.80 (3H, s, CH 3), 0.92 (9H, s, Bu and 0.3 ppm (6H, s, OHg)).

E. Preparation of Trans-1- (t-butyldimethylsilyl) -3-formyl-4-triethylthio-2-azetidinone _ / scb '. Brain · ®] J — V * “' -Bu To a cooled (-78 ° C) solution of 4-isopropylamine (0.34 mL, 2.4 mmol) in tetrahydrofuran (5 mL) was added dropwise, under nitrogen, 1.5 mol. n-BuLi solution (1.6 mL, 2.4 mmol). After stirring for 30 minutes, a solution of 1- (t-butyldimethylsilyl) -1,4-tritylthio-2-azetidinone (1.0 g, 2.18 mmol) in tetrahydrofuran (5 mL) was added dropwise with stirring. continued for 30 minutes. Ethyl formate (0.8 mL, 9.9 mmol) was added and the cooled solution was stirred for 10 minutes. The reaction mixture was washed successively with cold 1N hydrochloric acid (5 ml), 1 mol. sodium bicarbonate (6 mL), water (10 mL) and brine. The organic layer was dried (H 2 SO 4), evaporated to dryness and crystallized from pentane to give 810 mg (76¾) of the formate as a white solid, m.p. 132-3 ° C; and (CDCl 3) max: 1760 '1715 cm -1; m / z (CDCl 3) δ f: 9.0 (1H, d, J = 1.2S Hz), 7.30 (15H, m), 4.7 (1H, d, J = 1.5Hz) and 3, Δ ppm (1H, t, J = 1.5 Hz).

Note: a) diisopropylamine was distilled over CaH and stored over KOH b) tetrahydrofuran was distilled over lithium aluminum hydride and stored on a 3A molecular sieve • c) ethyl formate was mixed with 1 CO 2 at room temperature and then distilled with P 2 O 2 *. n on top d) n-BuLi was titrated to 1-norm. hydrochloric acid.

F. Preparation of 1-tert-butyldimethylsilyl-D (S-d'-hydroxy-1'-ethyl) -4-tritylthio-2-azetidinones (4 isomers)

OH

—I '^ scb cA-YSC4

a) j -► 3 I

cA-NW </ - Vsi / tBu X (Me) 2 X <He) 2 n-Butyllithium (1.6 mol, 3.4 mL, 5.44 mmol) was added over 5 minutes to a solution of di- isopropylamine (0.847 mL, 6.23 mmol) in THF (30 mL) and maintained at -78 ° C. After 0.5 h, a solution of 1- (t-butyldimethylsilyl) -4-tritylthio-2-azetidinone (2.0 g, 6785370 4.4 mmol) in THF (20 mL) was added; After 15 minutes, acetaldehyde (10 mL) was added in one portion; after a further 15 minutes, water (100 ml) was added. The mixture was acidified (pH 5-6) with dilute hydrochloric acid and extracted with ethyl acetate (3 x 30 mL). The organic phases were washed with brine, dried and evaporated to dryness to leave an oil which was found by thin layer chromatography to be a mixture of four isomers (labeled A, B, C, D in descending order of polarity).

Crystallization of the oily residue in ethyl acetate-pentane gave isomers B and C as a white solid and isomers A and D remained in the mother liquors. Four pure compounds were obtained by preparative chromatography (Waters, 500) of the above solid and mother liquors. The relative amounts were: A, 17%; B, 32%; C, 39%; D, 12%.

When ether was used instead of THF in the above reaction and the reaction was quenched after 1 minute at -78 ° C, the relative amounts of A, B, C and D were: 12.9, 30.5 , 38.2 and 18.4%. When the reaction mixture was allowed to warm to 20 ° C over 2 hours before quenching, the relative amounts were: 13.4, 24.6, 44 and 18%. When one molar equivalent of dry magnesium bromide was added to the reaction mixture, the ratios changed to 19.2, 19.7, 30.1 and 31%.

Isomer A: This isomer exhibits cis stereochemistry at the C8-C4 position. It is a racemic mixture of the (1'S, 3R, 4R) and (1'R, 3S, 4S) enantiomers. Subsequently, the compounds obtained from Compound A are referred to as "Isomer A". They are a mixture of enantiomers and have the same configuration at C-p, C3 and Cl2. Compounds obtained from Compound A by a reaction which occurs with the inversion of the configuration are referred to as "Isomer D" if the inversion occurs at, and "Isomer C" when the inversion occurs at C 1; mp: 152-3 ° C; 1 Hmr (CDCl 3) δ: 8.06.8 (15H, m, aromatic), 4.30 (1H, d, J = 5.5, H-4), 3.78 (1H, m, H- 1 '), 3.10 (1H, dd, J = 5.5, J = 10, H-3), 1.22 (3H, d, J = 6.5, CH 2), 0.95 (9H, s, Bu), 0.27 (6H, 2s, CH 2). Anal. calc. for C30H3NO2Si: C 71.52, H 7.40, N 2.78, S 6.36%; found: C 71.28, H 7.41, N 2.48, S 6.19%.

67853

Isomer B: This isomer exhibits trans stereochemistry at the C3-C4 position. It is a racemic mixture of (1'R, 3S, 4R) and (1'S, 3R, 4S) enantiomers. Compounds having the same configuration at C 1-4 are referred to as "Isomer B"; and (CHCl 3): 1745 cm (C = O); mp

η O mä.KS

158-9 ° C; Hmr (CDCl 3) δ: 7.60-7.10 (15H, m, aromatic), 4.02 (1H, d, J = 0.8 H-4), 3.32 (1H, dd, J = 3.0, J = 0.8, H-3), 3.55-3.15 (1H, m, H-1 '), 0.88 (12H, CH 3, and t-Bu), 0.16 (6H, s, CH 3);

Isomer C: This isomer exhibits trans stereochemistry at the C8-C4 position. It is a racemate of (1'S, 3S, 4R) and (1'R, 3R, 4S) enantiomers. Compounds having the same configuration at C 1 -C 3 and are referred to as "Isomer C"; mp. 134-6 ° C; 1 Hmr (CDCl 3) J 1: 7.60-7.10 (15H, m, aromatic), 4.32 (1H, d, J = 1.8, H-4), 3.02 (1H, dd, J = 2.7, J = 1.8, H-3), 3.0-2.5 (1H, dq, J = 2.7, J = 6, H-11), 1.02 (3H, d , J = 6, CH 3), 0.95 (9H, s, t-Bu), 0.27 (6H, s, CH 3); and (CHCl 3)

Kate ·· 1735 1 <C = °) ·

Isomer D: This isomer exhibits cis stereochemistry at C 8 -C 10. It is a racemate of (1'R, 3R, 4R) and (1'S, 3S, 4S) enantiomers. Compounds having the same configuration at C 1 -C 3 and are referred to as "Isomer D": m.p. 171-2 ° C; Hmr (CDCl 3) δ: 7.80-6.90 (15H, m, aromatic), 4.70 (1H, d, J = 4.5, H-4), 3.02 (1H, dd, J = 4.5, J = 0.5, H-3), 2.39 (1H, dq, J = 0.5, J = 6.5, H-1 '), 1.0 (3H, d, J = 6.5, CH 3), 0.97 (9H, s, t-Bu), 0.32 (6H, s, CH 3). Anal. calc. for C 30 H 37 NO 2 SSi: C 71.52, H 7.40, N 2.78, S 6.36%; found: C 71.27, H 7.43, N 2.51, S 6.31%.

72 67 8 5 3

OH

JLj usc (!) 3 - _h5C ^ ^ -f 3 BH f b) I --- ► n ._Bu / -N - ^ / Bu 0 ^ - \ S / Bu

οχ V

N "®2" e2 2 trans isomer

A solution of trans-3-acetyl-1- (t-butyldimethylsilyl) - [4-tritylthio-2-azetidinone (1.0 g, 2 mmol) in THF (30 mL) was added dropwise under a nitrogen atmosphere. to a cooled (0 °) and stirred suspension of sodium borohydride (0.38 g, 10 mmol) in THF (120 mL). The ice bath was removed and the mixture was stirred at room temperature for 4 h. It was poured into ice-cold hydrochloric acid (1N, pH 6), stirred for 15 minutes and extracted with ether (3x). The combined ether extracts were dried and evaporated to dryness to give an oil (1.04 g) which crystallized from pentane to give the title compounds as a mixture of C and B isomers in a ratio of 70:30; mp. : 119-121 ° C; 84%.

O 0H

Ai_j ^ ch /.-N. -Nv 0 UsUCH) ° t (CI! 3> 5 \ -Bu3 2

Isomer B

A suspension of cuproiodide (4.78 g, 15 mmol) in ether (50 mL) was cooled to 0 ° C and 1.9 M was added under protection. methyllithium solution (26 mL, 50 mmol). The brown solution was stirred at 0 ° C for 10 minutes and then cooled to -60 ° C and trans-1-1 (t-butyldimethylsilyl) -3-formyl-4-tritylthio-2-azetidinone (2 , 43 g, 5.0 mmol) in a mixture of tetrahydrofuran (10 mL) and ether (40 mL). Stirring was continued for 3 h. The solution was warmed to -40 ° C and 1 mol was carefully added. ammonium chloride solution. The mixture was filtered using Celite and the organic 67853 phase was washed with 1 mol. ammonium chloride solution (3 x 5 mL) and then brine and dried over sodium sulfate. Filtration and evaporation to dryness gave the alcohol, isomer B, which was crystallized from warm pentane in 1.6 g (65%) yield; mp. 160-161 ° C; and (CHCl 3) ν max: 1730 cm -1;

1 Hmr (CDCl 3) δ: 7.32 (15H, m), H 05 (1H, s), 3.4 (1H, d, J = 3HZ, 3.2 5-3.55 (1H, m) , 1.6 (1H, s), 0.9 (12H, s) and 0.1 ppm (6H, s).

Note: a) tetrahydrofuran and ether were distilled over lithium aluminum hydride b) methyllithium was titrated to 1-norm. with hydrochloric acid c) copper (I) iodide was purified by continuous extraction with dry tetrahydrofuane in a Soxhlet extractor for 18 hours, followed by drying in a vacuum desiccator for 1 hour.

-Vf ** 3 _. cH ^ tisc * 3 0 Sx ^ tBu "tBu

Methylmagnesium iodide (0.1 mL, 0.1 mmol) was added dropwise to a cooled (0 ° C) and stirred solution of trans-1- (t-butyldimethylsilyl) -3-formyl-4-tritylthio-2-azetidinone ( 25 mg, 0.05 mmol) in THF (2 mL). The solution was stirred for 1.5 h at 0 ° C, poured into ammonium chloride solution, acidified with hydrochloric acid solution (1N) and extracted with ether. Drying and evaporation of the organic extracts left an oil with a starting material and a small amount of a mixture of the two trans-title compounds, with isomer B being the predominant component.

7 "6785 3 F. (1'S, 3S, 4R and 1'R, 3R, 4S) 1- (t-butyldimethylsilyl) -3- (1'-trimethylsilyloxy-1'-ethyl) -4-tritylthio-2-azetidinone (Isomer C) / OH OSi-

ΧγΑ _ ^ ApfA

s

A solution of (1'S, 3S, 4R and 1'R, 3R, 4S) 1- (t-butyldimethylsilyl) -3- (1'-hydroxy-1'-ethyl) -4-tritylthio-2-azetidinone (15 mg, 0.3 mmol) and azidotrimethylsilane (35 mg, 0.30 mmol) in dry THF (6 mL) were stirred at room temperature until the starting material disappeared (15 min). Purification of the reaction mixture by column chromatography (silica gel, CH 2 Cl 2) gave the title compound as a white solid (128 mg, 74%), m.p. 144-146 ° C. m / z (CDCl 3) δ: 7.10-7.60 (15H, m, aromatic), 4.30 (1H, d, J = 1.5, H-4), 2.25-2.89 (2H, m, H-3, H-1 '), 0.82-1.07 (12H, m, t-Bu, H-2'), 0.27 (6H, s, CH 3), -O , 10 (9H, s, -O-Si (CH3) 3; and <CHCl3W max: 1736 cm-1 (C = O).

G. (1'S, 3R, 4R and 1'R, 3S, 4S) 1 '- (t-butyldimethylsilyl) -3- (1'-methoxymethoxyether-1 * -ethyl) -4-tritylthio-2-azetidinone (Isomer A ) preparation och.och3 ° »1 AZl / sch f ~ * \ —N ..

J N O S i CT ^ SifMe). 2 t-Bu I 2 t-Bu n-butyllithium (about 12.5 ml of a 1.6 M solution of hexane, 20 mmol, just enough to give a permanent pink color) was added dropwise to a solution of (1'S, 3R, 4R and 1 'R, 3S, 4S) 1- (t-butyldimethylsilyl) -3- (1'-hydroxy-1'-ethyl) -4-tritylthio-2-azetidinone (Isomer A) (10.1 g, 20 mmol) THF (100 ml) and kept at -78 °.

After a stirring period of 15 minutes, a solution of bromomethoxymethyl ether (2 mL, 24 mmol) in THF (30 mL) was added dropwise. The mixture was stirred for 1 h at -78 ° C and 2 h at room temperature and poured into ammonium chloride solution (200 ml). Extraction with ethyl acetate (3 x 200 ml), washing with brine, drying over sodium sulphate and evaporation to dryness gave the crude title compound which was purified by chromatography on silica gel eluting increasingly with ethereal benzene mixtures (10.4 g, 95%). 1 Hmr (CDCl 3) δ · 7.1-7.5 (15H, m, aromatic), 4.47 (1H, d, H-4), 4.23 (2H, ABQ, J = 7.0). -CH 2 -O), 3.1-3.4 (2H, m, H-3 and H-1 '), 3.23 (3H, s, O-CH 3>, 1.37 (3H, d, J = 6.5, CH 2), 0.97 (9H, s, Bu) and 0.25 ppm (6H, 2s, CH 2).

H. (1'S, 3S, 4R and 1'R, 3R, 4S) 1- (t-butyldimethylsilyl) -3- (1'-formyloxy-1 * -ethyl) -4-tritylthio-2-azidinone (Isomer C) preparation OH <P0 ^ X __ ^ SC03 _j ^ SC <^ 3 V \

A solution of 1'S, 3S, 4R and 1'R, 3R, 4S) 1- (t-butyldimethylsilyl) -3- (1'-hydroxy-1 * -ethyl) -4-tritylthio-2-acetyl- dinone (isomer c) (50 mg, 0.1 mmol), p-bromobenzenesulfonyl chloride (100 mg, 0.4 mmol) and dimethylaminopyridine (24 mg, 0.2 mmol) in DMF (3 mL) were stirred at room temperature until the starting material had disappeared from view (0.5 h). The reaction mixture was then diluted with water and extracted with ether. The organic extracts were washed with brine, dried (MgSO 4) and evaporated to dryness. The title compound was purified by column chromatography. 1 Hmr (CDCl 3) δ: 7.80 (1H, s, CHO), 7.20-7.56 (15H, m, aromatic), 3.90-4.36 (1H, m, H-1 ' ), 4.07 (1H, d, J = 2, H-4), 3.22 (1H, broad s, H-3), 1.18 (3H, d, J = 6.5, H-2 '), 1.0 (9H, s, t-Bu), 0.31 (6H, s, di-CH 2).

76 6785 3 I. (1'R, 3S, 4R and 1'S, 3R, 4S) 1 '- (t-butyldimethylsilyl) -3-11-acetoxy-11-ethyl) -4-tritylthio-2-azetide- preparation of non (Isomer B) OH OAc J STr J, JiTr

Ac2 ° ^ Tl J — N \ /, pyridine J — N \ / S i — T- S i—— \ \

A solution of (1'R, 3S, 4S and 1'S, 3R, 4S) 1- (t-butyldimethylsilyl) -3- (1'-hydroxy-11-ethyl) -4-tritylthio-2-azetidinone ( 13.85 g, 27.5 mmol) in pyridine (75 ml) and acetic anhydride (50 ml) (prepared at 0 °) were stirred at room temperature for 40 h. The reagents were evaporated (the last residue was removed by azeotropic distillation with toluene 3 times). ), leaving an almost white solid. The crude derivative was crystallized from ether-petroleum ether to give the pure title compound (97.5%). 1 Hmr (CDCl 3 δ: 7.64-7.03 (15H, m, H aromatic), 4.60 (1H, m, J = 6, H-1 '), 3.92 (1H, d, J = 2, H-4), 3.55 (1H, dd, J = 2 ', J = 6, H-3), 1.79 (3H, s, CH 3 CO), 0.98 (3H, d, J = 6, CH 3), 0.88 (9H, s, t-butyl), 0.12 (6H, s, CH 2); ir (CHCl 3) ν max: 1775, 1740 cm -1 (C = O).

J. Preparation of 1- (t-butyldimethylsilyl) -3- (1'-paranitrobenzyldioxycarbonyl) -1 * -ethyl) -4-tritylthio-2-azetidinone (4 isomers)

\ tBu XtBU

"Isomer C" n-butyllithium (8.8 mL of a 1.6 M solution in hexane, 14 mmol, just enough to give a permanent pink color) was added dropwise to a solution of 77 6785 3 1- (t-butyldimethylsilyl) - A solution of 3- (1'-hydroxy-1'-ethyl) -4-trityl-lithio-2-azetidinone "Isomer C" (6.55 g, 13 mmol) in THF (70 mL) maintained at -78 ° C :in. After a stirring period of 15 minutes, a solution of paranitrile benzyl chloroformate (3.2 g, 14.8 mmol) in THF (30 mL) was added dropwise.

The mixture was stirred for 1 h at -78 ° C and poured into ammonium chloride solution (100 mL). Extraction with ethyl acetate (3 x 100 ml), washing with brine, drying and evaporation to dryness left 11 g of crude material. The pure title compound was obtained by chromatography on silica gel (220 g) and eluting increasingly with ethereal benzene mixtures; 93%, m.p. 118-119 ° C (ether); 1 Hmr (CDCl 3) δ: 8.35-7 (19H, m, aromatic), 5.12 (2H, s, benzyl), 4.08 (1H, d, J = 1.8, H-4), 4-3.5 (1H, dq, J = 6.5, J = 2, H-1 '), 3.10 (1H, dd, J = 2, J = 1.8, H-3), 1 , 2 (3H, d, J = 6.5, CH 3), 1.0 (9H, s, Bu) and 0.30 ppm (6H, 2s, CH 3); ir (CHCl 3) ν max: 1745 cm -1 (C = O); Anal. Calcd for C 39 H 18 + 2 N 2 O 8 SiS: C 66.83, H 6.20, N 4.10, S 4.69; , 90, H 6.26, N 4.11, S 4.59.

"Isomer B" Treatment of 1- (t-butyldimethylsilyl) -3- (1'-hydroxy-1'-ethyl- (4-triethylthio-2-azetidinone) "Isomer B" as described above gave pure 1- (t- butyldimethylsilyl) -3- (11-paranitrobenzyldioxycarbonyl-1'-ethyl) -4-tritylthio-2-azetidinone "Isomer B" as a foam, 95% 1 Hmr (CDCl 3) g: 8.32-6.90 (19H , m, aromatic), 5.1 (2H, s, benzyl), 4.65-4.20 (1H, m, H-1 '), 3.97 (1H, d, J = 1.5, H -4), 3.58 (1H, dd, J = 1.5, J = 5.8, H-3), 1.1 (3H, d, CH 3), 0.7 (9H, s; Bu and 0.2 ppm (6H, s, CH 3) and (membrane)

Vmax; 1755 '171.0 C = O · "Isomer A" Treatment of 1- (t-butyldimethylsilyl-3- (1'-hydroxy-1'-ethyl) -4-tritylthio-2-azetidinone "Isomer A as described above "Pure 1- (t-butyldimethylsilyl-3- (1'-paranitrobenzyldioxycarbonyl-1'-ethyl) -4-tritylthio-2-azetidinone" Isomer A "was obtained as an oil; 95%.

1 Hmr (CDCl 3) g: 8.3-6.7 (19H, m, aromatic), 4.95 (2H, ABq, benzyl), 4.53 (1H, p, J = 7.5, H-1 ' , 4.31 (1H, d, J = 6, H-4), 3.32 (1H, dd, J = 6, J = 7.5, H-3), 1.44 (3H, d, J = 6.5), 0.95 (9H, s, tBu) and 0.2 ppm (6H, 2s, CH 3).

7 8 67853 "Isomer D" In a similar manner, "Isomer D" of 1- (t-butyldimethylsilyl-3- (1'-hydroxy-1'-ethyl) -4-tritylthio-2-azetidinone gave pure 1- (t- butyldimethylsilyl) -3- (11-paranitrobenzyldioxycarbonyl-1'-ethyl) -4-tritylthio-2-azedidinone "Isomer D", 90% 1Hmr (CDCl3) 8.3-6.7 (19H, m, aromatic ), 5.20 (2H, ABq, benzyl), 4.72 (1H, d, J = 5, H-4), 3.50 (1H, dq, J = 6.5, H-1 '), 2.85 (1H, dd, J = 0.5, J = 5, H-3), 1.03 (3H, d, J = 6.5, CH 2), 1.0 (9H, s, t -Bu) and 0.35 ppm (6H, s, CH 3), mp 130-2 ° C, anal calcd C 66.83, H 6.20, N 4.10, s 4.70, found: C 66.56, H 6.28, N 3.96, S 4.89.

K. (1'S, 3S, 4 = and 1'R, 3R, 4S) 1- (t-butyldimethylsilyl) -3- (1'-methanesulfonyloxy-1''-ethyl) -4-tritylthio-2-azetidinone (Isomer C) Manufacture of OM s

I -—- ► ^ _N

(U ^ SilCH) ° UilCH3'2 \ -Bu t-Bu

A solution of (1'S, 3S, 4R and 1'R, 3R, 4S) -1- (t-butyldimethylsilyl) -3- (11-hydroxy-1'-ethyl) -4-tritylthio-2-azetidinone (Isomer C) (2.0 g, 4 mmol) in dichloromethane (80 mL) was treated at 5 ° C with methanesulfonyl chloride (0.99 g, 8.6 mmol) and triethylamine (0.87 g, 8.6 mmol). mmol).

After stirring at that temperature for 1 h under N 3, the solution was washed with brine, dried (MgSO 4) and evaporated to dryness. After crystallization from ether-petroleum ether, 1.9 g (81.9%) of mesylate were obtained, m.p. 120-122 ° C; 1 Hmr (CDCl 3) δ: 7.13-7.61 (15H, m, aromatic), 4.50 (1H, d, J = 2, H-4), 3.62 (1H, dq, J = 6 , 5, 2, H-1 '), 2.96 (1H, dd, J = 2, 2, H-3), 2.84 (3H, s, methanesulfonyl), 1.22 (3H, d, J = 6.5, H-2 '), 0.99 (9H, s, Si-t-Bu) and 0.30 ppm (6H, s, Si (CH 3) 2); and V max (CHCl 3): 1746 (C = O), 1343 and 1180 cm -1 (SO 2).

79 67853 L. (1'R, 3S, 9R and 1'S, 3R, 9S) 1- (t-butyldimethylsilyl) -3- (1 * -methanesulfonyloxy-1'-ethyl) -9-tritylthioazetidinone (Isomer B) manufacturing

MsO

oh A X, sc <t> ^ Λ-γ 3 J — N \ <° Nid- σ ^ Si-f- \ \

A solution of (1 * R, 3S, 9R and 1'S, 3R, 9S) 1- (t-butyldimethylsilyl) -3- (11-hydroxy-11-ethyl) -9-tritylthio-2-azetidinone (Isomer B) (5.03 g, 10 mmol), methanesulfonyl chloride (2.52 g, 22.0 mmol) and triethylamine (2.23 g, 22.0 mmol) in CH 2 Cl 2 (200 mL), stirred at 5 ° C for 1 h.

The solution was then washed with brine, dried (MgSO 4) and evaporated to dryness to leave a residue which crystallized as a white solid on trituration with ether (5.40 g, 93%); mp. 127-131 ° C. Hrnr (CDCl 3) δ: 7.20-7.63 (15H, m, aromatic), 9.51 (1H, dq, J = 5.0-6.2, H-1 '), 9, Δ (1H, d, J = 2.0, H-9), 3.60 (1H, dd, J = 5.0-2.0, H-3), 2.03 (3H, s, -CH 2 ), 1.01 (3H, d, J = 6.2, H-21), 0.90 (9H, s, t-Bu), 0.12 (6H, s, -CH 3); and (CHCl 3) ν max: 1745 cm -1 (C = O).

M. (1'S, 3S, 9R and 1'R, 3R, 9S) 3- (1 * -p-Bromobenzenesulfonyloxy-11-ethyl) -1- (t-butyldimethylsilyl) -9-tritylthio-2-azetidinone ( Preparation of Isomer C) oo <j> Br

Xa - \ 80 6785 3

Solution of (1'S, 3S, 1R and 1'R, 3R, 1S) 1- (t-butyldimethylsilyl) -3- (11-hydroxy-1'-ethyl) -4-tritylthio-2 -azedidinone (Isomer C) (2.5 g, 5 mmol) in dry THF (100 mL), cooled to -78 ° C and treated with 2.52 mol. with butyllithium / hexane solution (2.38 mL, 6 mmol). After 3-1 minutes, p-bromobenzenesulfonyl chloride (1.53 g, 6 mmol) dissolved in THF was added dropwise. The solution was stirred at -78 ° C for 3 h and then allowed to warm to room temperature. The solvent was then evaporated off and the desired product was purified by column chromatography (silica gel, CH 2 Cl 2) (3.36 g, 91.6%), m.p. 112-111 ° C; 1 Hmr (CDCl 3): 7.68 (1H, s, benzenesulfonyl), 7.28-7.60 (15H, m, aromatic), 1.59 (1H, d, J = 1.8, H1), 3 .68 (1H, dq, J = 6.2, H-1 '), 2.99 (1H, dd, J = 1.8, 2.0, H-3), 1.18 (3H, d, J = 6.2, H-2 '), 1.08 (9H, s, t-Bu), 0.10 and 0.38 (6H, 2S, -CH 2); and (CHCl 3) νmax: 1719 cm -1 1 (C = 0).

Preparation of N. (1'S, 3R, 1R and 11R, 3S, IS) -3- (1'-methoxymethyl-1'-ethyl) -1-tritylthio-2-azetidinone (Isomer A) OCH-10CH-,

och2och3 I

_. h-r · J_N Me (f-

<y \ '2 σ H

Si t-Bu

A cold (0 ° C) solution of HMPA-O (116 ml-13 ml) containing 1- (t-butyldimethylsilyl) -3- (1'-methoxymethyl-1'-ethyl) -1-tritylthio- Isomer A of 2-azetidinone (11 g, 20 mmol) was treated with sodium azide (2.7 g, 12 mmol). The cold bath was removed and the mixture was stirred for 30 minutes. It was then poured into cold water (1.3 L) and dried. The title compound was recrystallized from ethyl acetate-hexane (7.2 g, 83%) as a white solid, m.p. 173-171 ° C.

^ Hrnr (CDCl 3) δ '· 7.10-7. (15H, m, aromatic), 1.85 (2H, ABq, J = 7.1.0-CH 2 -O), 1.53 (1H, d, J = 5.2, H1), 1.12 ( 1H, s, NH), 81 67853 4.15 (1H, m, H-1 '), 3.5 (1H, m, H-3), 3.47 (3H, s, O-CH 2), 1 .5 (3H, d, J = 6, CH3). ir (KBr) max: 3400-3500 (N-H) and 1760 cm -1 (C = O).

0. Preparation of (1'S, 3S, 4R and 1'R, 3R, 4S) 3- (1'-methoxymethyloxy-1'-ethyl) -4-tritylthio-2-azetidinone (Isomer C) OCH OCH ° CH2OCH3 X .. . {_ ^ BrCH2OCH3 NaN, -f

Cold solution (dry ice-acetone bath) of (1'S, 3S, 4R and 1'R, 3R, 4S) 1- (t-butyldimethylsilyl) -3- (1 · -hydroxy-1'-ethyl) -4 -Tritylthio-2-azetidinone (5.03 g, 10 mmol) in THF (50 mL, distilled over LAH) was treated dropwise with 1.6 M. n-butyllithium in hexane (13.0 mL) until the pink color persisted. A solution of THF (20 mL) in bromomethyl methyl ether (1.49 g, 0.97 mL, 1.19 mmol) was added dropwise. The mixture was stirred at -78 ° C for 30 minutes and at 0 ° C for 3 h. It was poured into ice-cold ammonium chloride solution and extracted with ether. The ether extracts were combined, washed with water, dried (MgSO 4) and evaporated to dryness to give crude (1'S, 3S, 4R and 1'R, 3R, 4S) 1- (t-butyldimethylsilyl) -3- (1'-methoxymethyloxy). -1'-ethyl) -4-tritylthio-2-azetidinone (5.83 g, 100%) deprotected as described below:

A cold (ice bath) solution of the above derivative (5.83 g, 10 mmol) in HMPA-H 2 O (90 mL to 10 mL) was treated with sodium azide (1.365 g, 21 mmol). The cooling bath was removed and the mixture was stirred at room temperature for 2 hours. It was then slowly poured into ice-cold water (900 ml) and stirred for 30 minutes. The precipitate was collected by filtration and redissolved in methylene chloride. The solution was washed with water and brine and dried (MgSO 4) to give the title compound (3.0 g, 69%), m.p. 172-172.5 (from ethyl acetate-hexane); ir (CHCl 3) 82 6 7 8 5 3 \),: 3400 (N-H) and 1760 cm -1 (C = O); ^ Hmr (CDCl 3) δ '· 7.67-7.12 v max J -3.

(15H, m, H aromatic), 4.63 (2H, center of ABq, J = 6.0-CH 2 -O), 4.49 (1H, s, NH), 4.40 (1H, d, J = 3, H-4), 4.25-3.80 (1H, m, H-11), 3.35-3.15 and 3.26 (4H, s + m, CH 3 and H-3) and 1.30 ppm (3H, d, J = 6, CH 3).

P. Preparation of (1'R, 3S, 4R and 1'S, 3R, 4S) 3- (1'-formyloxy-11-ethyl) -4-tritylthio-2-azetidinone (Isomer B)

OSO φΒΓ OCHO

Α2- ^ 3 _ ^ Al_ ^ sc * 3 o '-' SA- J—

A solution of (1'S, 3S, 4R and 1'R, 3R, 4S) 3- (1'-p-bromobenzenesulfonyloxy-1'-ethyl) -1- (t-butyldimethylsilyl) -4-tritylthio 2-Azetidinone (Isomer C) in DMF (3 mL) was heated at 50 ° C for 48 h and then at 100 ° C for 4 h. The reaction mixture was then diluted with 11a and extracted with ether. The ether extracts were washed with brine, dried (MgSO 4) and evaporated to dryness. The title compound was obtained as white crystals after purification by column chromatography (silica gel, 5% CH 3 CN-CH 2 Cl 2) (2 mg, 4.8%), m.p. 131-132 ° C; 1 Hmr (CDCl 3) 8.07 (1H, s, CHO), 7.24-7.56 (15H, m, aromatics), 5.23 (1H, dq, J = 6.4, 7, H-1 '), 4.38 (1H, dm J = 2.4, H-4), 4.25 (1H, s, NH), 3.20 (1H, dd, J = 7, 2.4, H- 3), 1.43 (3H, d, J = 6.4, H-2 ') ;, and (CHCl 3)? Max = 3400 (NH), 1765 (C = O), 1725 cm -1 (C = 0).

83 6 7 8 5 3 Q. Preparation of (1'R, 3S, 4R and 1TS, 3R, 4S) 3- (1'-acetoxy-1'-ethyl) -4-tritylthio-2-azetidinone (Isomer B) OAc

OAc I

I STr) · .. / STr

^ _. TT

Ll ™ «· τ 0J — mh

^ ^ tBDMS

The pure derivative (1'R, 3S, 4R and 1'S, 3R, 4S) 1- (t-butyldimethylsilyl) -3- (11-acetoxy-11-ethyl) -4-tritylthio-2-azetidinone (5.77 g, 10.57 mmol) was dissolved in warm HMPT-water (60 mL, 10 mL). The solution was cooled to room temperature and NaN 3 (1.2 g) was added. Stirred for 45 minutes (the progress of the reaction was monitored by tlc) and slowly poured into stirred cold water (800 ml). The mixture was stirred for another 20 minutes. The crystalline material was collected and washed with water. It was redissolved in Cl 2 Cl 2, washed with water (twice) and brine and dried over MgSO 4. Evaporation of the solvent left a foam which crystallized from ether-petroleum ether (4.90 g, 96.5%, mp 143-144.5 ° C) ir (CH 2 Cl 2) + max 3395 (NH)> 1772> 1738 cm-1 (C = O). - '' Hinr (CDCl 3> S: 7.9-6.8 (15H, m, H aromatic), 5.12 (1H, center of dq, J = 6.5, 7.5, H-1 ') ), 4.33 (1H, d, J = 2.8, H-4), 4.20 (1H, bs, NH), 3.17 (1H, ddd, J3_lf = 7.5, J3_4 = 2, Δ, J 3_NH = 1, H-3), 2.1 (3H, s, CH 3 CO), 1.35 (3H, d, J-6.5, CH 2).

84 6785 3 R. Preparation of 3- (11-hydroxy-1'-ethyl) -4-tritylthio-2-azetidinone. (Mixture of four isomers)

OH? ÄC

_ SC4) ^ 0 -S 3 LDA CH -.- 1 -; - ► I -ph oho— * 3 r pyridim J r] J — n 3 Ck \ u ° 0 ^ i (CH3) 3 Q.i +

J \ - Si-Cl C

DMF -n (Et) <f · —N \ '. A solution of lithium diisopropylamide (0.74 mmol) in dry tetrahydrofuran (5 mL) was prepared from diisopropylamine (0.103 mL, 0.74 mmol) and BuLi at -78 ° C. (0.29 mL of a 2.52 M hexane solution). After the mixture was -78 ° C for 30 minutes, a solution of (R and S) 1-trimethylsilyl-4-tritylthio-2-azetidinone (0.292 g, 6.99 mmol) in dry tetrahydrofuran (2 mL) was added dropwise. . After 5 minutes, excess freshly distilled acetaldehyde (0.2 mL) was added in one portion. After 20 minutes at -78 ° C, tlc analysis showed complete disappearance of the starting materials and the reaction mixture was quenched by the addition of saturated ammonium chloride solution. Extraction with ethyl acetate (2 x 25 mL) followed by washing of the combined organic phases with saturated NH 4 Cl solution, brine and drying over anhydrous magnesium sulfate gave, after evaporation of the solvent, a yellow oil. Filtration of this oil on silica gel (10 g, eluting with CH 2 Cl 2 / EtOAc, 6: 4) gave a mixture of alcohols (0.215 g, 80%). This mixture (1Hmr) cannot be separated by hplc or tlc chromatography.

a: Acetylation

Acetylation of a sample of the mixture (0.065 g) with excess acetic anhydride (1.0 ml) and pyridine (1.4 ml) gave a mixture of acetates. HPLC analysis showed four components: a) 34.6%; b) 17.4%; c) 30.1%; d) 17.9%. Compound a) was identical to isomer B in a direct comparison, (hplc).

85 6 7 8 5 3 b: t-Butyldimethylsilyl derivatives

A mixture of alcohols (0.121 g, 0.34 mmol) was treated with t-butyl dimethylchlorosilane (0.117 g, 0.776 mmol) and triethylamine (0.10 mL, 7.14 mmol) in dry dimethylformamide (1 mL) for 36 h at room temperature. temperature. After dilution with ethyl acetate, the solution was washed with saturated ammonium chloride and dried over anhydrous magnesium sulfate. Evaporation to dryness gave an oil (0.716 g) with four components by HPLC, a = 3.7%; b = 60.6%; c = 31.1%; d = 4.6% (Identity of each not proven)

remarks:

Butyllithium and lithium hexamethyldisilazane were ineffective Order of increasing polarity 3

Acetylation of the product from 1-t-butyldimethylsilyl-4-tritylthio-2-azetidinone gave relative proportions: d = 29.5%; c = 24.1%; b = 33.8%; a = 12.6%.

Reaction of a mixture of alcohols from (R and S) 1- (t-butyldimethylsilyl) -4-tritylthio-2-azetidinone gave: a = 5.2%; b = 41.3%; c = 48%; d = 4.6%.

S. Preparation of (1'R, 3S, 4R and 1'S, 3R, 4S) 3- (1'-benzoxy-1'-ethyl) -4-tritylthio-2-azetidinone (Isomer B)

Ms0 <? <: 0φ sccb V "'- tx θ'-s, 0

A solution of di (1S, 3S, 4R and 1'R, 3R, 4S) 3- (1'-methenesulfonyloxy-1'-ethyl) -4-tritylthio-2-azetidinone (Isomer C) (035 mg, 2 mmol) and sodium benzoate (432 mg, 3 mmol) in 10% O-DMF (10 mL) were heated at 90 ° C for 7.5 h. The reaction mixture was then diluted with H 2 O and extracted with ethyl acetate. The organic extracts were washed with brine, dried (MgSO 4) and evaporated to dryness. Column chromatography (silica gel, 5% CH 3 CN-CH 2 Cl 2) of purified 67853 86 residue afforded the title compound as a white solid (108 mg, 23.2%), m.p. 158 ° C. 1 Hmr (CDCl 3) δ: 7.03-8.25 (20H, m, aromatic), 5.32 (1H, dq J = 6.19, H-11), 4.40 (1H, d , J = 2.5, HH), 4.30 (1H, s, NH), 3.40 (1H, dd, J = 9, 2.5, H-3), 1.50 (3H, d, J = 6.1, H-2 '); and (CHCl 3) max: 3400 (N-H), 1765 (C-O), 1715 cm -1 (C = O).

Preparation of 3- (1T-paranitrobenzyldioxycarbonyl-1'-ethyl) -4-tritylthio-2-azetidinone (4 isomers)

and PNB

"Isomer C" a) A solution of 1- (t-butyldimethylsilyl) -3- (1'-paranitrobenzyldioxycarbonyl-11-ethyl) -4-tritylthio-2-azetidinone "Isomer C" (1.3 g) in a mixture of TFA (5 mL), water (5 mL), dichloromethane (20 mL) and methanol (30 mL) was stirred for 2 days at room temperature. The solution was diluted with water and the aqueous phase was extracted with dichloromethane. The combined organic phases were washed with sodium hydrogen carbonate and water, dried and evaporated to dryness to leave an oil. Crystallization from ether gave the pure title compound (902 mg), m.p. 78-80 ° C; 1 Hmr (CDCl 3) 8.25-7 δ (19H, m, aromatic), 5.21 (2H, s, benzyl), 5.05 (1H, m, H-1 '), 4.40 (1H, s, NH), 4.27 (1H, d, J = 2.8, H-4), 3.37 (1H, dd, J = 5.3, 2.8, H-3) and 1.37 ppm (3H, d, J = 6.5, CH 2); ir (CHCl 3) max: 3390 (N-H), 1765 and 1745 (shoulder) (C = 0, and 1525 cm -1 (NO 2)).

b) Cold (Isomer C) of 1- (t-butyldimethylsilyl) -3- (1'-paranitrobenzyl-dioxycarboxyl-1'-ethyl) -4-tritylthio-2-azetidinone (9.11 g, 13.3 mmol) 0 ° C) HMPT-H 2 O (90 mL to 19 mL) solution was treated with sodium azide (1.82 g, 27.9 mmol). The cold bath was removed and the mixture was stirred for 30 minutes. It was then poured into water (1 L) and extracted 87 67853 with ether (5 x 200 mL). The ether fractions were combined and washed with water (5 x 200 mL), brine and dried over MgSO 4. Alternatively, since the title compound precipitated on dilution with water, it was filtered off and recrystallized from ether; 7.22 g, 89%, m.p. 78-80 ° C.

"Isomer B" 3- (1'-Paranitrobenzyldioxycarbonyl-1'-ethyl) -4-tritylthio-2-azetidinone "Isomer B" was prepared as described above for "Isomer C"; 92%, m.p. 155.5-156 ° C (from ether); 1 Hmr CCDCl 2 (δ): 8.25-6.80 (19H, m, aromatic), 5.20 (2H, s, benzyl), 4.95 (1H, m, H-1 '), 4.35 (1H, d, J = 2.9, H-4), 4.17 (1H, s, NH), 3.20- (1H, dd, J = 10.8, J = 2.9, H- 3) and 1.40 ppm (3H, d, J = 7.5, CH 3); ir (CHCl 3) νmax = 3480, 3390 (NH), 1772, 1750 (C = O), and 1525 cm -1 (NO 2). Anal. Calcd for? 32 ^ 28 ^ 2 ^ 6 ^ ^ 67.59, H 4.96, N 4.93, S 5.64, found: C 67.48, H 4.98, N 4.92, S 5.67.

"Isomer A" 3- (1'-Paranitrobenzyldioxycarbonyl-1'-ethyl) -4-tritylthio-2-azetidinone "Isomer A" was prepared as described above for "Isomer C"; mp. 205-206 ° C.

1 Hmr (CDCl 3) δ: 8.2-6.7 (19H, m, aromatic), 5.22 (2H, ABq, benzyl), 5.57-4.85 (1H, m, H-1 ' ), 4.65 (1H, NH), 4.50 (1H, d, J = 6.5, H-4), 3.65 (1H, dd, J = -6.5, 12, JN_R = 1 , H-3) and 1.52 ppm (3H, d, J = 7.5).

"Isomer D" 3- (1 * -paranitrobenzyldioxycarbonyl-1'-ethyl) -4-tritylthio-2-azetidinone "Isomer D" was prepared as described above for "Isomer C"; 1 Hmr (CDCl 3) δ: 8.15-6.70 (19H, m, aromatic), 5.23 (2H, ABq, benzyl), 5.20 (1H, m, H-1 '), 4, 75 (1H, NH), 4.52 (1H, d, J = 5.5, H-4), 3.42 (1H, J = 5.5, 3, H-3 and 1.5 ppm (3H , d, J = 6.5, CH 2) (J value for H-3 taken after D 2 O exchange).

88 67853 U. Preparation of (1'R, 3S, 4R and 1'S ^ R ^ S) 3- (1'-Methanesulfonyloxy-1'-ethyl) -4-tritylthio-2-azetidinone (Isomer B)

MsO MsO

V "3 _.

o 1-4- ^ \

A solution of (1'R, 3S, 4R and 1'S, 3R, 4S) 1- (t-butyldimethylsilyl) -3- (1'-methanesulfonyloxy-1'-ethyl) -4-tritylthio-2- azetidinone (Isomer B) (4.95 g, 8.5 mmol) and sodium azide (1.11 g, 17.0 mmol) in a mixture of 10% H 2 O-HMPA (50 mL) were stirred at room temperature for 30 minutes. The solution was then diluted with water (250 mL) and extracted with ether. The organic extracts were washed with brine, dried (MgSO 4) and evaporated to dryness. Crystallization of the residue (from ether-Petro-ether) gave the title compound (3.33 g, 83.8%); mp. 130-131 ° C. 1 Hmr (CDCl 3) δ: 7.20-7.62 (15H, m, aromatic), 4.97 (1H, dq, J = 6.4, 6.1, H1), 4.56 (1H, d, J = 2.8, H-4), 4.22 (1H, m, NH), 3.27 (1H, dd, J = 6.1, 2.8, H-3), 3.0 (3H, s, -CH 2), 1.63 (3H, d, J = 6.4, H-2f); ir (nujoli) Vmax: 3195 (n-H) '17 68 cm-1 (C = O).

V. Preparation of (1'S, 3S, 4R and 1'R, 3R, 4S) -3- (1'-Methanesulfonyloxy-1'-ethyl) -4-tritylthio-2-azetidinone (Isomer C) OMs OMs

Al_ ^ c * 3 t-Bu

A solution of (1'S, 3S, 4R and 1'R, 3R, 4S) -1- (t-butyldimethylsilyl) -3- (1'-methanesulfonyloxy-11-ethyl) -4-triethylthio-2 -azetidinone (Isomer C) (2.85 g; 4.9 mmol) in 10% aqueous HMPA (25 mL), treated with sodium 89 6785 3 azide (0.65 g, 10 mmol) and stirred for 25 h. At 0.5 ° C. Dilution of the solution with water (250 ml) caused the reaction product to crystallize apart. The crude mesylate was redissolved in dichloromethane, washed with brine, dried (MgSO 4) and evaporated to dryness. Trituration with ether gave the title compound as white crystals, m.p. 155-160 ° C; 1.80 g; 78.6%. ^ Hmr (C-DCl ^) <? ' 7.80, (UH, s, benzenesulfonyl), 7.30-7.65 (15H, m, aromatic), 5.13 (1H, dq, J = 6.5, 4.0, H-1 '), 4.50 (1H, d, J = 2.9, H-4), 4.40 (1H, s, N-H9, 3.40 (1H, dd, J = 4.0, 2, 9, H-3), 1.50 (3H, d, J = 6.5, H-21), and (CHCl 3)

Vmax: 3400 cm-1 (N_H) '1770 cm-1 (c = °) · W. (1'S, 3S, 4 = and 1'R, 3R, 4S) 3- (11-p-bromobenzenesulfonyloxy-1 * - Preparation of ethyl) -4-tritylthio-2-azetidinone (isomer C) OSO ΦΒΓ

03 ° 2φΒΓ Γ 2 JSC <K

χ.5 (Γφ 3 -J 3 \

A solution of (1'S, 3S, 4R and 1'R, 3R, 4S) 3- (1'-p-bromobenzenesulfonyloxy-1'-ethyl) -1- (t-butyldimethylsilyl) -4-tritylthio-2 -azetidinone (Isomer C) (1.4 μg, 2 mmol) and sodium benzoate (0.865 g, 6 mmol) in 10% H 2 O-HMPA (40 mL) were stirred at room temperature for 1 h. The solution was then diluted with Hao (100 mL) and extracted with ether. The ether extracts were washed with brine, dried (MgSO 4) and evaporated to dryness. The crude crystalline title compound was triturated with a small volume of ether and collected by filtration (0.92 g, 77%), m.p. 125-126 ° C.

1 Hmr (CDCl 3) δ: 7.80 (4H, s, benzenesulfonyl), 7.30-7.65 (15H, m, aromatic), 5.13 (1H, dq, J = 6.5, 4, 0, H-1 '), 4.50 (1H, d, J = 2.9, H-4), 4.40 (1H, s, NH), 3.40 (1H, dd, J = 4, 0.29, H-3), 1.50 (3H, d, J = 6.5, H-2 '); and (CHCl 3) O: 3400 cm -1

η o ψ ITlcLKS

(N-H), 1770 cm (C = O).

90 6 7 8 5 3 X. Preparation of (1'R, 3S, 4R and 1'S, 3R, 4S) 3- (1'-hydroxy-1'-ethyl) -4-tritylthio-2-azetidinone (Isomer B)

OH

oso Φβγ 1 ηςφ __. 3 O-vs 4 '* 0 s \ To a warm solution of (1fS, 3S, 4R and 1'R, 3R, 4S) 3- (1'-p-bromobenzenesulfonyloxy-1'-ethyl) -1- (t- butyldimethylsilyl) -4-tritylthio-2-azetidinone (Isomer C) in HMPA (5 mL) was added dropwise to 1 mL of H 2 O 2. The reaction mixture was kept at 90 ° C for 20 h, then diluted with ether and washed 4 times with brine. The organic solution was dried (MgSO 4), evaporated to dryness and the crude title compound was purified by column chromatography (silica gel, 15% CH 3 CN-Cl 12 Cl 2). Obtained as a white solid (122 mg, 44.5%), m.p. 187-189 ° C, which was found to be identical to a sample of the title compound prepared by another method.

Y. Preparation of 3- (1'-hydroxy-1'-ethyl) -4-tritylthio-2-azetidinone

Both isomers, (1'S, 3S, 4R and 1 * R, 3R, 4S) 3- (11-hydroxy-1'-ethyl) -4-tritylthio-2-azetidinone (Isomer C) and (11R, rS, -R and 1'S, 3R, 4S) -3- (1'-hydroxy-11-ethyl) -4-tritylthio-2-azetidinone (Isomer B) was prepared by the same method. For example, a solution of (1'S, 3S, 4R and 1'R, 3R, 4S) 1- (t-butyldimethylsilyl) -3- (1'-hydroxy-1'-ethyl) -4-tritylthio-2- azetidinone (Isomer C) (1.0 g, 2 mmol) and sodium benzoate (0.665 g, 6 mmol) in 10% H 2 O-DMF (40 mL) were stirred at room temperature for 18 h. The reaction mixture was then diluted with 0 and extracted with ether. The organic extracts were washed with brine, dried (MgSO 4) and evaporated to dryness. The crude title compound was crystallized from cold ether (0.47 g, 61%), m.p. 134-135 ° C. 1 Hmr. (CDCl 3) δ: 7.12-7.56 (15H, m, aromatic), 4.48 (1H, s, NH), 4.28 (1H, d, J = 2.8, H- 4), 2.94 (1H, dq, J = 6.5, 6.2, H-1 '), 3.06 (1H, dd, J = 6.2, 2.8, H-3), 2.18 (1H, s, -OH), 1.30 (3H, d, J = 6.5, H-2 '); and (CHCl 3) max: 3400 C * n-H), 1760 cm -1 (C = O).

In a similar manner, (1'R, 3S, 4R and 1'S, 3R, 4S) 1- (t-butyldimethylsilyl) -3- (1'-hydroxy-1'-ethyl) -4-tritylthio-2-azetidinone ( Isomer B), m.p. 190-192 ° C. 1 Hmr (CDCl 3) 7.10-7.55 (15H, m, aromatic), 4.45 (1H, d, J = 2.5, H-4), 4.28 (1H, s, NH), 4.10 (1H, dq, J = 6.4, 5.3, H-1 '), 3.08 (1H, dd, J = 5.3, 2.5, H-3), 1.50 (1H, s, -OH), 1.30 (3H, d, J = 6.4, H-21); ir (CHCl 3) νmax: 3400 (NH), 1760 cm -1 (C = Q) Z. (1'S, 3R, 4R and 1'R, 3S, 4S) 3- (11-methoxymethyl-1'-ethyl) Preparation of - (improved trobenzyl 2''-hydroxy-2n-acetate) -4-tritylthio-2-azetidinones (Isomer A) OCH_OCH OCH2OCH3 hn "· -. '55 * '„

Co ^ RRB

A mixture of 3- (11-methoxymethyl-1 * -ethyl) -4-tritylthio-2-azetidinone Isomer A (7.5 g, 17.3 mmol), paranitrobenzyl glyoxylate hydrate (4.7 g, 20.8 mmol) and toluene (300 mL) was heated at reflux for 1 h in a Dean-Stark apparatus filled with a 3 Å molecular sieve. The solution was cooled in ice and triethylamine (0.24 mL, 1.7 mmol) was added dropwise. The mixture was stirred for 1 h, washed with dilute hydrochloric acid, sodium bicarbonate and brine, dried and evaporated to dryness to give the title compound as a foam (10.5 g, 94%). 1 Hmr 67853 92 (CDCl 3) δ: 8.25-6.84 (19H, m, aromatic), 5.24 (2H, s, benzyl), 4.67-4.83 (3H, m, O-CH 2 and H-4), 4.34-4.55 (1H, m, H-2 "), 4.02 (1H, m, H-1 '), 3.54 (1H, m, H-3), 3.40 (3H, s, O-CH 2), 1.38 (3H, d, J = 6.5, CH 2), and (KBr) δ ks: 3360 (OH), 177 ° C -actam), 17 3 5 (ester C = 0) and 1605 cm -1 (aromatic).

AA. (1'S, 3S, 4R and 1'R, 3R, 4S) 3- (1'-methoxymethoxy-1'-ethyl) -1- (paranitrobenzyl 2 "-hydroxy-2" -acetate) -4-tritylthio- Preparation of 2-azetidinone (Isomer C)

OCH OCH OCH OCH

I 2 3 CHO I * t-Tr J. / STr CO PNB A. / ri —— 0tC ..

H o

CO ^ PNB

A solution of hydrogenated paranitrobenzyl glyoxylate (1.73 g, 7.11 mmol) was boiled in toluene (90 mL) using a Dean-Stark condenser filled with a 3 Å molecular sieve for 2 hours. To the boiling solution was added (1'S, 3S, 4R and 1'R, 3R, 4S) 3- (1'-methoxymethyloxy-1'-ethyl) -4-tritylthio-2-azetidinone (3.0 g, 6.93 mmol). and the mixture was boiled for an additional 2 h. The mixture was cooled to room temperature, triethylamine (70 mg, 97 γ1, 0.69 mmol) was added and stirred for 2 h. The reaction mixture was diluted with ether, washed with 1% aqueous HCl, water, 1%: with aqueous NaHCO 3, water and brine, dried (H 2 SO 4) and evaporated to dryness to give the title compound. (4.60 g, 100%); ir (CHCl 3) ν max: 3530-3100 (0-H), 1765, 1750 (C = O) and 1525 cm -1 (NO 2) and '' Hrnr (CHCl 3): 8.22, 8.18 (2H , 2d, J = 8, Hm aromatic), 7.67-7.0 (17H, m, H-aromatic), 5.3 (2H, bs, CH2-PNB), 5.30-5.02 (m , H-2 "), 4.89-4.52 (m, H1 'and O-H), 4.63, 4.59 (1H, 2d, J = 2, H-4), 4.33, 4.30 (center of ABq of 2H, 2, J = 7, J = 7, O-CH 2 -O), 4.1-3.67 (1H, m, H-1 '), 3 , 2 (1H, H-3), 3.1, 3.6 (3H, 2s, CH 2 -O), and 1.15 ppm (3H, d, J = 6.5, CH 3).

93 BB. (1'R, 3S, 4R and 1S, 3R, 4S) 3- (1'-acetoxy-1'-ethyl) -1-paranitrobenzyl-2 "-hydroxy-2" -acetate) -4-tritylthio-2- preparation of azetidinone 67853 OAc PAc J STr fH0 A ^ STr

y-T-f io2PNB, P

TEA 0 * ~ Ν-γ- CO PNB 2 "Isomer B"

A solution of hydrogenated p-nitrobenzyl glyoxylate (triturated with ether) (1.82 g, 30 mmol) was boiled in benzene through a Dean-Stark condenser filled with a 3 Å molecular sieve for 2 hours. To this was added azetidinone (1'R, 3S, 4R and 1'S, 3R, 4S) 3- (1'-acetoxy-1'-ethyl) -4-tritylthio-2-azetidinone (10.88 g, 25.2 mmol) and the mixture was boiled for another 1 h. The solution was cooled to room temperature and triethylamine (0.35 mL, 2.5 mmol) was added. It was then stirred for 2 h; the progress of the reaction was monitored by y, tie. Evaporation of the solvent gave a white foam in quantitative yields (100%, mixture of epimers). Alternatively, the solution may be washed with acid and base. and (CHP10) v). : 3520 (OH), 1775, 1745 cm-1 (C = (J); -j 3 2 * max 1 Hmr (CDCl 3): 8.2, 8.18 (2H, 2d, J = 8, Ho aromatic) , 7.80-6.90 (17H, m, H-aromatic), 5.28, 5.17 (2H, 24, CK 2 -PNB, 4.89 (0.67H, d, J = 7.2) , CH0), 4.80 (center of m, H-1 '), 4.38 (0.33 H, 2d, J = 8.8, CH0), 4.22 (D, 33H, d, J4_3 = 2.5, H-4), 4.09 (0.67H, d, J4_3 = 2.1, H-4), 3.65 (D, 67H, dd, 1 ^, = 5.8, J3_4 = 2.1, H-3), 3.47 (0.33H, dd, J, = 5.5, J3_4 = 2.5, H-3), 3.3 (0.33H, d, J = 8.8, OH), 3.23 (0.67H, d, J = 7.5, OH), 1.88, 1.86 (3H, 2s, CH 3 CD), 1.10, 1.06 (3H , 2d, J = 5.8, 6.3, CH 3).

CC. Preparation of 3- (1'-paranitrobenzyldioxycarbonyl-1'-ethyl) -1- (paranitrobenzyl 2 "-hydroxy-2" -acetate) -4-tritylthio-2-azetidinone (4 isomers) 94 67853

OCO.PNB OCO PNB

^

M y. m OH

</ ”Vh Ύ

C02PNB

"Isomer C"

A mixture of 3- (1'-paranitrobenzyldioxycarbonyl-11-ethyl) -4-tritylthio-2-azetidinone "Isomer C" (1.70 g, 0.3 mmol), paranitrobenzyl glyoxylate hydrate (815 mg, 3 .6 mmol) and toluene (50 mL) were heated at reflux for 7 days in a Dean-Stark apparatus filled with a 3 Å molecular sieve. The cooled solution was washed with dilute hydrochloric acid, sodium bicarbonate and brine, dried and evaporated to dryness to give the title compound (2.1 g) as an epimer mixture (carbon-2 "). Purification was carried out by chromatography on silica gel. More non-polar epimer at 2 ": 1 Hmr (CDCl 3) g: 8.25-6.80 (23H, m, aromatic), 5.30 and 3.12 (4H, 2s, benzyls), 4.65 (1H, d , J = 9, H-2 "), 4.45 (1H, d, J = 2.5, H-4), 4.45-4.10 (1H, m, H-11), 3.50 (1H, d, J = 9.2 "-OH), 3.28 (1H, dd, J = 2.5, J = 2.5, H-3) and 1.23 ppm (3H, d, J = 6.5, CH 2); and (CHCl 3) 3530-3200 (D-H), 1765, 1750 (C = O) and 1525 cm -1 (NO 9).

η 1

More polar isomer for C-2: Hmr (CDCl 3) δ: 8.25-6.85 (23H, m, aromatic), 5.25 and 5.08 (4H, 2s, benzyls), 5.05 (1H , d, J = 7, H-2 "), 4.35 (1H, d, J = 2.5, H-4), 4.40-4.05 (1H, m, H-1 '), 3.42 (1H, J = 7.2 "-OH), 3.33 (1H, dd, J = 2.5, 2.5, H-3), 1.23 (3H, d, J = 6); , 5, CHO); and (CHC10) 0. : 3520 3200 (0-H), 1755 (C = O) and 1525 cm (NO,).

"Isomer B"

A mixture of hydrogenated paranitrobenzyl glyoxylate (1.74 g, 7.66 mmol) and (1'R, 3S, 4R and 11S, 3R, 4S) -3- (1'-para-nitrobenzyldioxycarbonyl-1'-ethyl ) -4-Trityl io-2-azetidinone (3.63 g, 6.38 mmol) was boiled in toluene-67853 95 sa (70 mL) on a Dean-Stark condenser filled with 3A molecular sieves for 3 hours. The solution was cooled to room temperature and triethylamine (64.5 mg, 89 mL, 0.639 mmol) was added. It was then stirred for 4 h, diluted with ether and washed with 2% aqueous HCl, water, 2% aqueous NaHCO 3, water and brine. Dried and evaporated to dryness to give the pure title compound (5.02 g, 100%). 2 epimers were separated using a preparative silica gel plate. More non-polar epimer at 2 ": (CHCl 3): 3,500 (0-H), 1772, 1750 (C = O) 1525 cm -1 (NO 2); Hmr (CDCl 3) δ: 8.30-8.0 and 7.65-6.80, (23K, m, aromatic), 5.27 and 5.13 (4H, 2s, benzyls), 4.71 (1H, m, J-6.5, 6.5, H1 »), 4.28 (1H, d, J = 2.2, H-4), 4.23 (1H, d, J = 8.7, H-2"), 3.50 (1H, dd, J = 2.2, 6.5, H-3), 3.28 (1H, d, J = 8.7, 0-H) and 1.18 ppm (3H, d, J = 6.5, CH ^). More polar epimer: ir (CHCl 3) Vnjaks1 3480 <0-H) 1772, 1750 (C = O) and 152 5 cm -1 (NO 2); Hmr (CDCl 3) 8.35-6.90 (23H, m, aromatic), 5.15 (4H, benzyl), 4.72 (1H, d, J = 7.5, H-2 "0), 4.90-4.50 (1H, m, J = 6.5, 6.5, H-1 '), 4.10 (1H, d, J = 2, H-4), 3.68 (1H , dd, J = 2, 6.5, H-3), 3.82 (1H, d, J = 6.5, 0-H) and 1.15 ppm (3H, d, J = 6.5, CH 3).

"Isomer A" 3- (1 * -paranitrobenzyldioxycarbonyl-1'-ethyl) -4-tritylthio-2-azetidinone "Isomer A" was similarly obtained from 3- (11-paranitrobenzyldioxycarbonyl-1'-ethyl) -1- ( paranitrobenzyl 2 "-hydroxy-2" -acetate) -4-tritylthio-2-azetidinones "Isomer A". 1 Hmr (CDCl 3) δ: 8.3-6.7 (23H, m, aromatic), 5.17 (2H, benzyls), 5.0 (1H, m, H "1 ')> 4, 9 and 4, Δ (1H, 2d, J = 6, H-4, two epimers), 4.23 and 3.96 (1H, 2s, H-2 ", two epimers), 3.68 (1H, dd, J = 6 , 6, H-3 and 1.47 ppm (3H, 2d, J = 6.5, CH 2, two epimers).

"Isomer D" 3- (1'-paranitrobenzyldioxycarbonyl-1'-ethyl) -4-tritylthio-2-azetidinone "Isomer D" was similarly obtained from 3- (1'-paranitrobenzyldioxycarbonyl-1'-ethyl) -1 - (paranitrobenzyl 2 "-hydroxy-2" -acetate) A "Isomer D" mixture of -4-tritylthio-2-azetidinones. 1 Hmr (CDCl 3: 8.30-6.60 (23H, m, aromatic), 5.20 (4H, m, benzyls), 4.83 (1H, 2d, J = 5, H = 4), 5.50 -4.30 C2H, m, H1 'and H-2 "), 3.48 (1H, m, H-3), 3.15 (1K, m, O-H), 1.37 and 1.30 ppm (3H, 2d, CH 3>.

96 6 7 8 5 3 DD. (1'3,3S, 4R and 1 * R, 3R, 4S) 3- (1'-Methanesulfonyloxy-1 * -ethyl) -1- (paranitrobenzyl 2 "-hydroxy-2" -acetate) -4- Preparation of tritylthio-2-azetidinone (isomer C) (epimers at C ^ ")? t4s ροφ.

----- VY ··

CT \ H io2PNB

A solution of paranitrobenzyl glyoxylate hydrate (9.72 g, 42.6 mmol) in benzene (350 mL) was boiled for 2 h with azeotropic removal of water using a Dean-Stark trap. To that solution was added (1'S, 3S, 4R and 1'R, 3R, 4S) 3- (1'-methanesulfonyloxy-1'-ethyl) -4-tritylthio-2-azetidinone (16.62 g, 35 0.5 mmol) and boiling was maintained for a further 0.5 h. The reaction mixture was then cooled to room temperature, treated with triethylamine (0.5 μL; 3.5 mmol) and stirred for 3 hours to complete the reaction. Evaporation of the solvent left a white non-oily substance which was used as such in the next step. J'Hmr (CbCl2) 8.12 (2H, d, J = 9, Hm aromatic), 7.28 (17H, part of d,

Ho aromatic, trityl), 5.28 (2H, s, -CH2-PNB), 4.88 (0.5 H, s, H1 "), 4.62 Q., SH, m, H-2" and . H-4), 4.00 (2H, m, H-1 ', -OH), 3.15 (1H, m, H-3), 2.73 (3H, s, mesylate and 1.30 ppm ( 3 H, d, J = 6 Hz, H-2 '), and Jmax: 352 ° (0-H), 1775 (C = O) and 1765 cm-1 (C = O).

97 6 7 8 5 3 EE. (1'S, 3R, 4R and 1'R, 3S, 4S) 3- (1-methoxymethyl-1 * -ethyl) -1- (paranitrobenzyl 2 "-chloro-2" -acetate) -4-tritylthio-2- preparation of azetidinone (Isomer A) och2och3 ° ch2och3

ApYscE A_ ^ sca

CO PNB CO PNB

2 1

Pyridine (1.1 mL, 1M, 2 mmol) was added dropwise to a solution of 3- (11-methoxymethyl-11-ethyl) -1- (paranitrobenzyl-2 "-hydroxy-2" -acetate) -4 -tritylthio-2-azetidinone Isomer A (7 g, 10.9 mmol) in THF (350 mL) cooled to -15 ° C. Immediately thereafter, thionyl chloride (1.0 mL, 14.0 mmol) was added dropwise and the mixture was stirred at -15 ° for 0.5 h. The precipitate was removed by filtration and washed with benzene. The combined filtrates were evaporated to dryness, the residue dissolved in fresh benzene and the solution treated with activated charcoal, filtered and evaporated to dryness to leave the title compound as an oil (6.5 g, 90%);

Hmr (CDCl 3) δ: 6.65-8.35 (19H, m, aromatic), 5.24 (2H, s, benzyl), 3.43 (3H, s, OCH 2) and 1.42 ppm (3H, d, J = 6, CH 3).

67853 98 FF. (1'S, 3S, 4R and 1'R, 3R, 4S) 3-Cl'-ethoxymethyloxy-I'-ethyl) -1-paranitrobenzyl 2 "-chloro-2" -acetate) -4-specificthio-2-azetidinone ( Preparation of isomer C)

OCH OCH

OCH OCH I 2 3 STr X / STr ^ '"rY socl2 r f J —' N ^ -OH '(f"

I CCTPNB

C02PNB 2

A cold (ice-MeOH bath) solution of (1'S, 3S, 4R and 1 * R, 3R, 4S) 3- (1 * -methoxymethyloxy-1'-ethyl) -1-paranitrobenzyl 2 "-Hydroxy-2 '- acetate) -4-tritylthio-2-azetidinone (4.25 g, 6.62 mmol) in THF (60 mL, distilled over LAH) was treated by dropwise addition. pyridine (0.696 mL, 8.61 mmol) and thionyl chloride (0.530 mL, 8.61 mmol). The mixture was stirred for 30 minutes at -15 ° C. The precipitate was collected by filtration and washed with benzene. The THF-benzene solution was evaporated to dryness and the residue was redissolved in benzene. The resulting solution was treated with charcoal. Removal of charcoal on a Celite mattress followed by evaporation of benzene gave the title compound (4.86 g, 100%); ir (CHCl 3) δ k £ j: 1770 (C = O) and 152 5 cm -1 (NOg); 1 Hmr (CDCl 3) δ: 8.15, 8.12 (2H, 2d, H-aromatic), 7.70-7.00 (17H, m, H-aromatic), 5.62, 5-, 02 ( 1H, 2s, H-2 "), 5.27 (2H, s, CH2-PNB), 4.7 (1H, d, H-4), 4.7-3.7 (m, O-CH2- 0, H-1 '), 3.5-2.8 (m, H-3), 3.12, 3.08 (3H, 2s, O-CH 3), and 1.30-0.96 ppm ( 3H, m, CH 3).

GG. (1'R, 3S, 4R and 1'S, 3R, 4S) S-d'-acetoxy-1'-ethylD-1- (paranitrobenzyl 2 "-chloro-2" -acetate) -4-trityl Preparation of -acsedidinone 67853 OAc OAc j STr ^> Y'Tr soci> -> Tf C1 j-tly0H pyridimine 0 '# U'NyX *

1 CO PNB

CO2PNB 2 "Isomer B" (1'R, 3S, 4R and 1'S, 3R, 4S) 3- (1'-acetoxy-1'-ethyl) -1- (paranitrobenzyl-2 "-hydroxy-2" -acetate) A THF solution of -4-tritythio-2-azetidinone (10.88 grams of NH) (THF distilled over LAH) was treated at -15 ° C (ice-methanol bath) with pyridine (2.19 g) protected under a nitrogen atmosphere. , 2.24 mL, 27.7 mmol) and thionyl chloride (3.3 g, 2.02 mL, 27.7 mmol). The mixture was stirred for 20 min. -15 ° C. The salt was filtered off and washed with benzene. Evaporation of the solvent (THF + benzene) gave a residue which was dissolved in benzene (warm) and treated with charcoal. The suspension was filtered through a pad of Celite and the solvent was evaporated to leave a foam; ir (CH 2 Cl 2) νmax: 1780, 1740 cm -1 (C = O) 2 Hmr (CDCl 3: 8.17, 8.21 (2H, 2d, J = 8, Ho-aromatic), 7 .76-6.88 (17H, m, H-aromatic), 5.31, 5.16, 5.12, 4.73 (3H, 4s, CH2-PNB, CHCl3), 5.12-4.55 (1H, m, H-1 '), 4.35-4.25 (1H, m, H-4), 3.80-3.45 (1H, m, H-3), 1.90 (3H , s, CH 3 CO), 1.12 1.07 (3H, J = 6.5, CH 2).

100 67853 HH. 3- (1'-Paranitrobenzyldioxycarbonyl-1'-ethyl) -1- (paranitrobenzyl 2 "-chloro-2" -acetate) -4-triethylthio-2-azetidinones (mixture of epimers of C2 ")

0C0 PNB OCO PNB

av ** __

| CO PNB

CO PNB 2 2 "Isomer C11

Pyridine (58 mg, 0.73 mmol) was added dropwise to a solution of 3- (1'-paranitrobenzyldioxycarbonyl-1'-ethyl) -1- (paranitrobenzyl 2 "-hydroxy-2" -acetate) -3-trityl lithium "Isomer C" of 2-azetidinones (470 mg, 0.6 mmol; mixture of epimers from C-2) in THF (15 mL) cooled to -15 ° C. Immediately thereafter, thionyl chloride (86.5 mg, 0.73 mmol) was added dropwise and the mixture was stirred at -15 ° C for 0.5 h. The precipitate was removed by filtration and washed with benzene. The combined filtrates were evaporated to dryness, the residue was dissolved in fresh benzene and the filtrate was treated with activated charcoal, filtered and evaporated to dryness to leave the title compound as an oil; 530 mg; 100%.

1 Hmr (CDCl 3): 8.7-6.8 (23H, m, aromatic), 5.53 (1H, s, H-2 "), 5.30 and 5.17 (4H, 2s, benzyls), 4 , 52 (1H, d, J = 2, H-4), 4.20-3.70 (1H, m, H-1 '), 3.31 (1H, dd, H-3), 1.27 and 1.21 ppm (3H, 2d, J = 6.5), ir (CHCl 3) νmax: 1780, 1750 and 1525 cm -1 (NO 2).

"Isomer B" 3- (1-Paranobenzobutyldioxycarbonyl-1'-ethyl) -1-Paranitrobenzyl 2 "-chloro-2" -acetate) -4-Tritylthio-2-azetidinones "Isomer B" in C-2 "mixture of epimers) was prepared in quantitative yields as described above for" Isomer C ". 1 Hmr (CDCl 3) δ: 8.25-6.30 (23H, m, aromatic), 5.40-5.0 (4H, m , benzyls), 5.40-4.45 (1H, m, H-1 '), 4.82 and 4.57 (1H, 2s, H-2 "), 4.36 and 4.31 (1H, 2d, J = 2.5, H-4), 3.63 (1H, m, J = 2.5, J = 6.5, H-3), 1.25 and 1.18 ppm (3H, 67853 101 2d, J- 6.5, CH3); ir (CHCl 3) ψmax; 1780 »1750 (GRO), and 1525 cm-1 (NO2).

"Isomer A" "Isomer A" of 3- (1'-paranitrobenzyldioxycarbonyl-1'-ethyl) -1 '- (paranitrobenzyl 2 "-chloro-2" -acetate) -4-tritylthio-2-azetidinones 2 "mixture of epimers).

1 Hmr (CDCl 3) 8.30-6.80 (23H, m, aromatic), 5.45-4.80 (1H, m, H-1 '), 5.18 and 5.21 (4H, 2s , benzyls), 4.87 (1H, 2d, H-4), 4.22 and 3.87 (1H, 2s, H-2 "), 4.05-3.40 (1H, m, H-3 ), 1.57 and 1.50 ppm (3H, 2d, CH 2).

"Isomer D" "Isomer D" of 3- (1 "-paranitrobenzyldioxycarbonyl-1'-ethyl) -1 '- (paranitrobenzyl 2" -chloro-2 "-acetate) -4-tritylthio-2-azetidinones (C-2 "mixture of epimers).

1 Hmr (CDCl 3) δ: 8.30-6.70 (23H, m, aromatic), 5.32-5.10 (4H, m, benzyls), 5.48 and 5.30 (1H, 2s, H-2 "), 4.82 (1H, d, J = 5, H-4), 5.30-5.20 (1H, m, H-1 '), 3.15 (1H, m, H -3), 1.40 and 1.3 Opm (3H, 2d, J = 6.5, CH 3); and CHCl 3): 1780, 1750 (C = O) and 1525 cm -1 (NO 2).

II. (1'S, 3S, 4R and 1'R, 3R, 4S) 3- (1'-Methanesulfonyloxy-11-ethyl) -1- (paranitrobenzyl 2 "-chloro-2" -acetate) -4-tritylthio-2 -azetidinone (isomer C) (epimers of C2 ") OMs? Ms 50φ A ._ / SCt» 3 A- ^ 3 ÖJ — N OH (/ —Ν ^ Α

C02PNB CO 2 PNB

To a cold solution (5 ° C) of (1'S, 3S, 4R and 1'R, 3R, 4S) 3- (1'-methanesulfonyloxy-1'-ethyl) -1- (paranitrobenzyl 2 "-hydroxy-2" acetate) -4-tritylthio-2-azetidione (24.0 g, 35.5 mmol) in dry tetrahydrofuran (350 mL) was added dropwise to pyridine (3.65 g, 46.2 mmol) and thionyl chloride (5.5 mL). g, 46.2 mmol). After the mixture was stirred for 45 minutes, ether (100 ml) was added to precipitate the hydrochloride salt, 102 6785 3 which was filtered off. The filtrate was evaporated to dryness and the residue was redissolved in benzene (200 ml) and treated with charcoal. Evaporation of the solvent left an almost white foam which was used as such in the next step. 1 Hmr (CDCl 3) δ: 18 (2H, d, J = 9, Hm aromatic), 7.72 (17H, m, part of d, Ho aromatic, trityl), 5.57 and 5.12 (1H, s, H-2 "), 5.28 (2H, s, -CH 2 PNB), H, 73 (1H, 2d, H-4), 3.21 (1H, 2dq, H-3), 2 .78 (3H, 2s, mesylate and 1.21 ppm (3H, 2d, H-6H2; H-2 '); ir>) max 1779 cm-1 (C = O).

JJ. (1'S, 3R, 4R and 1'R, 3S, 4S) 3- (1'-methoxymethoxy-1'-ethyl) -1- (paranitrobenzyl 2 "-triphenylphosphoranylidene-2" -acetate) -4-tritylthio- 2-Azetidinone (Preparation of Isomer A> och2och3 khtKH)

Vs "3 _> Vf \

CC ^ PNB C02PNB

A mixture of S-d'-methoxymethoxy-1'-ethyl) -1- (paranitrobenzyl-2 "-chloro-2" -acetate) -4-tritylthio-2-azetidinone Isomer A (6.6 g, 10 mmol) ), triphenylphosphine (3.3 g, 12.5 mmol), 2,6-lutidine (1.3 mL, 11 mmol) and dioxane (140 mL) were heated at reflux for 2 days.

The solution was diluted with ether, washed with dilute acid (5% HCl), water, dilute sodium bicarbonate solution and brine, dried and evaporated to dryness. The residue was purified by chromatography on silica gel eluting with a mixture of 10% ether in benzene. Evaporation to dryness of the appropriate fractions left the title compound as a foam (1.4 g, 13.7%). IR (KBr) δ: 1750 (C = O) and

_ η IU3.XS

1660-1650 cm (C = 0, aromatic).

103 67853 KK. (1'S, 3S, 4R and 1'R, 3R, 4S) 3- (1'-methoxymethyloxy-1'-ethyl) -1- (paranitrobenzyl-4'-triphenylphosphoranylidene-2 "-acetate) -4-tritylthio- Preparation of 2-azetidinone (Isomer C) OCH2OCH, och2och3 \ 'sTr J STr p ^ 3' -Γ yt »'-'- y * · co2pnb

Dioxane (100 mL, distilled over LAH) solution of (1'S, 3S, 4R and 1'R, 3R, 4S) 3- (11-methoxymethyloxy-1'-ethyl) -1- (paranitrobenzyl-2 ‘’ chloro-2,1-acetate) -4-tritylthio-2-azetidinone (4.86 g, 6.62 mmol), triphenylphosphine (2.60 g, 9.93 mmol) and 2,6-lutidine ( 770 mg, 0.837 mL, 7.20 mmol) was heated at reflux for 4 h and kept in a hot bath (100 ° C) for 16 h. The mixture was diluted with ether, washed with 1% aqueous HCl, water, 10% NaHCO 3. aqueous solution, water and brine and dried (MgSO 4). The solution was evaporated to dryness and the residue was filtered through silica gel (65 g) on a column (5%, 10% and 20% ether in benzene) to give the title compound (2.8 g, 48%). IR (CHCl 3) max: 1795 (C = O), 1620 and 1605 (phosphorane) and 1515 cm -1 (NO 2) · LL. (1'R, 3S, 4R and 1'S, 3R, 4S) 3- (1'-Acetoxy-1'-ethyl) -1- (paranitrobenzyl-2'-triphenylphosphoranylidene-2'-acetate) -4-tritylthio-2-azetidinone (Isomer B) OAc OÄC I Crp) .... .STr V ^ "'rf J i Cl 2,6-lutidine Qj — Ν \ ^ ΡΦ3

0 CG PNB

CO PNB 2 2

Dioxane (100 ml, freshly distilled over LaH) solution containing crude (1'R, 3S, 4R and 1'S, 3R, 4S) 3- (1'-acetoxy-1'-ethyl) -67853.104 1 - (improved · Itobenzyl-2 "-chloro-2M-acetate) -4-tritylthio-2-azetidinone was treated with 2,6-lutidine (2.97 g, 3.23 mL, 27.72 mmol) and triphenylphosphine ( 9.91 g, 37.8 mmol) The mixture was boiled (oil bath 130 °) for 18 h, the solvent was evaporated off and the residue was redissolved in methylene chloride and the resulting solution was washed successively with dilute HCl, H 2 O, dilute NaHCO 3. Drying and evaporation of the solvent left the title compound as a solid which was triturated with ether and collected by filtration (14.6 g, 65.9%); IR (CH 2 Cl 2) max: 1750 (C = O) and 1620, 1610 cm -1 (phosphorane.i).

MM. 3- (1'-Paranitrobenzyldioxycarbonyl-1'-ethyl) -1- (paranitrobenzyl-2,1-triphenylphosphoranylidene-2 '' - acetate) -4-tritylthio-2-azetidinone

OC02PNB OCO..PMB

Α1γ · 5εφ3 _t J-N Cl, __ N ^ Ρφ Y o Y 3 co2pkb

Isomer B

A mixture of (1'R, 3S, 4R and 1'S, 3R, 4S) 3- (1'-paranitrobenzyldioxycarbonyl-1'-ethyl) -1- (paranitrobenzyl-2-chloro-2 "-acetate) - 4-Tritylthioazetidinone (Isomer B) (4.96 g, 6.22 mmol, mixture of C-2 "epimers), triphenylphosphine (2.47 g, 9.42 mmol) and 2,6-lutidine (740 mg, 0.80 mL, 6.91 mmol), boiled in dioxane (freshly distilled from LAH) for 30 h. The solution was diluted with ether and ethyl acetate, washed with 5% aqueous HCl, water, 10% aqueous NaHCO 3, water and brine, and dried (MgSO 4). Evaporation of the solvent gave a residue which was passed through silica gel (10 times the weight) on a column (10% ether in ether, ether, and ethyl acetate). The title compound was obtained as a crystalline solid (3.1 g, 49%), m.p. 189-190 ° C (from ether); IR (CHCl 3): (C = O), 1620, 1605 (phosphorane) and 1522 cm -1 (NO 2) · 105 6785 3

Isomer C

Isomer C of 3- (1'-paranitrobenzyldioxycarbonyl-1'-ethyl) -1- (paranitrobenzyl-2 "-triphenylphosphoranylidene-2" -acetate) -k-tritylthio-2-azetidinone was prepared as described above for isomer B. IR (CHCl 3): 1750 η -3 η max (C = O), 1610, 1620 (phosphorane) and 1520 cm -1 NO 2; H Hmr (CDCl 3) δ · 3.6 = 6.7 (H, aromatic ), 5.22 and k, 95 (benzyl), k, 70 (Hk), 2.6 (H3), 1.19 and 1.07 ppm (CH2).

Isomer D

A mixture of 3- (11-p-nitrobenzyldioxycarbonyl-1'-ethyl) -1- (p-nitrobenzyl-2 "-chloro-2" -acetate) -C-tritylthio-2-azetidinone isomer D (k , 598 g, k, k5 mmol; purity 77%, mixture of C-2 "epimers), triphenylphosphine (1.25 g, 5 mmol; Aldrich) and 2,6-lutidine (0.63 ml , 580 mg, 5.0 kM; anachemia) in dioxane (65 mL; distilled over LAH) was gently heated to reflux under N 2 H, followed by reaction with tlc (benzene: ether = 3: 1). cooled, diluted with ethyl acetate and washed successively with 0.1 N HCl, water, 2% NaHCO 3 and then brine, drying (Na 2 SO 4) and evaporating the solvents to give a dark colored oil which was purified by column chromatography (SiO 2). , 88 g, eluent 10-25% ether in benzene) to give 1.108 g (1.08 mmol, 2k yield, 3%) of the title compound as a yellowish foam: 1 Hmr (CDCl 3) tf: 1.08 (d, J = 6Hz, 1'-CH 2); IR (fine): 1745 cm -1 (s, C = O).

67853 106 NN. (1'S, 3S, 4R and 1'R, 3R, 4S) -3- (1'-methanesulfonyloxy-1'-ethyl) -1- (paranitrobenzyl-2,1-triphenylphosphoranidene-2 "-acetate) - Preparation of 4-tritythio-2-acetidinidinone (isomer C) oms? Ms A ^ _ / Cb. 3 ci * / "" γ * 1

X. COPNB

CO PNB 2

A solution of (1'S, 3S, 4R and 1'R, 3R, 4S) -3- (11-methanesulfonyloxy-1'-ethyl) -1- (paranitrobenzyl-2''-chloro-2 " acetate) -4-tritylthio-2-azetidinone (24.7 g, 35.5 mmol), triphenylphosphine (11.2 g, 42.7 mmol) and 2,6-lutidine (4.2 g, 39.1 mmol) in dry dioxane (350 ml), boiled under nitrogen for 19 h. The solvent was evaporated off and the crude product was redissolved in ethyl acetate and washed successively with dilute HCl, NaHCO 3 and brine. Purification was completed by chromatography on a silica gel column (cm Elution with 10% ether in dichloromethane (1.5 L) followed by ether (1.5 L) gave purified phosphorane: 12.36 g (40%).

1 Hmr (CDCl 3) 2.53 and 2.93 ppm (3H, 2s, mesylate); IR max: 1749 1620 cm-1 (c = O) · 00. (1'R, 3S, 4R and 1'S, 3R, 4S) -3- (11-hydroxy-1'-ethyl) -1- (paranitrobenzyl-2 Preparation of "-triphenylphosphoranylidene-2" -acetate) -4-tritylthio-2-azetidinone (Isomer B)

OH

OAc 1 ^ STr A-r5Tr - * n p *,

T CO PNB

C02PNB 2

Solution of phosphorane (1'R, 3S, 4R and 1'S, 3R, 4S) - (1'-acetoxy-1'-ethyl) -1- (paranitrobenzyl-2 "-triphenylphosphoranylidene-2 '' - acetate ) -4-tritylthio-2-azetidinone (4.43 g, 5.00 mmol) in methanol (10 mL) and THF (60 mL) was treated with 107 67853 in 1% aqueous NaOH (1 eq. , 200 mg

♦. X

In 20 ml H 2 O). The progress of the reaction was monitored by tlc. The mixture was diluted with ether-ethyl acetate and washed with HCl, aqueous NaHCO 3, H 2 O and brine. Evaporation of the solvent gave a residue which was crystallized from benzene-ether (3.7 g, 87.7%), m.p. 169.5 to 170.5 ° C. IR (CH 2 Cl 2) Found: 1745 (C = O) and 1620 cm -1 (phosphorane).

^ Heating the mixture increased the reaction.

PP. (1'S, 3R, 4R and 1'R, 3S, 4S) -Silver 3- (1'-methoxymethyl-1'-ethyl) -1- (paranitrobenzyl-2'r-triphenylphosphoranylidene-2 "-acetate) - Preparation of 2-azetidin-4-thiolate (Isomer A) och2och3 OCH2OCH3 i SAg Λ -> * __ / "Vt

O ^ p (^ 3 CO2PNB

Silver 3- (1'-methoxymethyl-1'-ethyl) -1- (paranitrobenzyl-2'-triphenylphosphoranylidene-2'-acetate) -3-tritylthio-2-azetidinone (Isomer A) was prepared elsewhere by paranitrobenz as described for isomer C of the -syldioxycarbonyl derivative. Yield 50%. IR (fine) λ max: 1745 cm -1 (C = O).

108 67853 QQ. (1'S, 3S, 4R and 1'R, 3R, 4S) -Silver-2- (1'-methoxyethyloxy-1'-ethyl) -1- (paranitrobenzyl-2 "-triphenyl-phosphoranylidene-2 "-acetate) -2-azetidinone-4-iolate (Isomer C) och2och3 OCH2 ° CH3 J SAg

T CO PNB

CO 2 NB 2 (1'S, 3S, 4R and 1'R, 3R, 4S) -3- (1'-methoxymethyloxy-1'-ethyl) -1- (paranitrobenzyl-2 "-triphenylphosphoranylidene-2" -acetate) - 4-Tritylthio-2-azetidinone (887 mg, 1.0 mmol) was first dissolved in hot (40 ° C) methanol (30 mL), treated with pyridine (103 mg, 0.105 mL, 1.3 mmol) and, after cooling, treated with 0.15 mol of silver nitrate. with methanol solution (8.7 mL, 1.3 mmol). The mixture was stirred for 1 h at 23 ° C, cooled (ice bath) and stirred for 20 minutes. The salt was filtered off and washed successively with cold methanol and ether (3 times, 671 mg, 87%). IR (CHCl 3) ν max: 171 + 5 (C = O), 1605 (phosphorane) and 1520 cm -1 (NO 2).

RR. Preparation of silver 3- (1'-paranitrobenzyldioxycarbonyl-1'-ethyl) -1- (paranitrobenzyl 2 "-triphenylphosphoranylidene-2" -acetate) -2-azetidinone-4-thiolate

0C02PNB OCO PNB

. of 9 *

CO PNB ¢ 0 PNB

2 2 "Isomer B" (1'R, S, R and 1'S, 3R, 4S) -3- (1'-paranitrobenzylcarbonyldioxy-1'-ethyl) -1- (paranitrobenzyl-2 "-triphenylphosphoranylidene) 2 "-acetate) -4-tritio-2-azetidinone (1.02 g, 1 mmol) was first dissolved in CH 2 Cl 2 (3 mL) and diluted with m.p. 109 67853 miles (55 ° C) in MeOH (20 mL). mL). The hot solution was first treated with pyridine (120 mL, 117 mg, 1.48 mmol) and hot (55 ° C) 0.15 mol. with a solution of silver nitrate in methane (8 mL, 1.2 mmol). The mixture was stirred at room temperature for 15 minutes, then at 0 ° C for 2 hours. It was then concentrated to a 10% solution on a rotary evaporator (no bath). The mercaptide was filtered off, washed twice with cold (-15 ° C) methanol and three times with ether. (917 mg, 100%). IR (nujcli) 1745 (C = O), 1600 (phosphorane) and 1517 cm-1 (no2).

"Isomer C11

Silver 3- (1 "-paranitrobenzyldioxycarbonyl-1'-ethyl) -1- (paranitrobenzyl-2" -triphenylphosphoranylidene-2 triphenylphosphoranylidene-2 "-acetate) -2-azetidinone-4-thiolate," Isomer C ", prepared as described above for "Isomer B": IR (nujol): 1745 (C = O) and 1600 cm -1 (phosphorane).

"Isomer D" 3- (1'-p-Nitrobenzylcarbonyldioxy-1 * -ethyl) -1- (p-nitrobenzyl-2 "-triphenylphosphoranylidene-2" -acetate) -4-tritylthio-2-azetidinone Isomer D (145 mg , 0.142 mmol) was prepared by first dissolving it in CH 2 Cl 2 (5 mL), removing CH 2 Cl 2 in 55-60 and adding hot MeOH (4 mL). To the above solution was added a hot solution of AgNO 2 in MeOH (0.15 mol, 1.14 mL, 0.17 mmol, 1.2 eq) followed by pyridine (14 μL, 0.17 mmol). , 1.2 eq.). Silver mercaptide began to precipitate immediately. The mixture was stirred at room temperature for 2 h and at 0 ° for 1 h. The mercaptide was collected by filtration and washed with ice-cold MeOH and ether to give 99 mg (0.11 mmol, 78%) of the title compound as a brownish solid; IR (nujol): 17 50 cm -1 (s, C = O).

no 67853 SS. (1'R, 3S, 4R and 1'S ^ R ^ SJ-silver-S-d'-hydroxy-1'-ethyl) -1- (paranitrobenzyl-2 "-triphenylphosphoranylidene-2" -acetate) -2- Preparation of azetidinone-4-thiolate (Isomer B) OH ™ _ / STr AgN03 -f

T CO_PNB

C02PNB 2

Solution * of (1'R, 3S, 4R and 1'S, 3R, 4S) -3- (11-hydroxy-1'-ethyl) -1- (paranitrobenzyl-2 "-triphenylphosphoranylidene-2" -acetate ) -4-tritylthio-2-azetidinone (1 g, 1.19 mmol) in MeOH (10 mL), treated with pyridine (124 μL, 121.3 mg, 1.53 mmol) and at 10 ° C. 15-M. with silver nitrate MeOH solution (15 mL, 2.25 mmol - or until no more precipitation of silver mercaptide occurred). The mixture was stirred for 1 h and concentrated on a rotary evaporator (no bath) to a mixture of about 10%. The solvent was filtered off. The filter cake was washed once with MeOH and 3 times with ether, and treated under high vacuum (954 mg, 100%). IR (nujol phase): 3500-3400 (0-H), 1752 (C = O), 1595 (phosphorane) and 1525 cm-1 (no2).

· ·

The crystalline material was first dissolved in Cl 2 Cl 2.

TT. (1'R, 3R, 4R and 11S, 3S, 4S) -4-acetylthio-3- (1'-p-nitrobenzyldioxycarbonyl-1'-ethyl) -1- (p-nitrobenzyl-2 " Preparation of triphenylphosphoranylidene-2 "-acetate) -2-azetidinone (Isomer D)

and PNB

OCO ^ PNB J · 2, A_ / SA? CH, COCl-Pyr d |

| C02PNB

CO PNB 2

To a stirred solution of silver 3- (1'-paranitrobenzyl-2 "-triphenylphosphoranylidene-2" -acetate) -2-acetethinone-4-thiolate (Isomer D) (85 mg, 0.095 mmol) Cl 2 in m 67853 (5 mL) of pyridine (30 μL to 0.37 mmol; Fisher) was added at 0-5 ° C to CH 2 Cl 2 (20 μL, 0.28 mmol) and the mixture was stirred at 0-5 ° C. At C for 30 minutes. The precipitate formed was filtered off and washed with CH 2 Cl 2. The filtrate and washings were combined, washed successively with brine, dilute HCl, saturated NaHCO 3 and then brine, dried (Na 2 SO 4) and evaporated to dryness to give 75 mg (0.091 mmol, 95% yield of crude compound) as a syrup: 1H (CDCl 3) δ: 2.33 (s, -SOCOCH 3); IR (fine) γ max: 1750 (β-lactam, ester), 1695 (thioester), 1520 and 1350 cm -1 (-NO 2).

UU. Preparation of (1'R, 5R, 6R and 1'S, 5S, 6S) -cis-p-nitrobenzyl-2-methyl-6- (1'-p-nitrobenzyldioxycarbonylmethyl-penem-3-carboxylate (Isomer D) oco2pnb oco pnb 11 _ '^ V- | ^ SA0 toluene-, JZn ^^ CH3

o ^ CO2PNB CC ^ PNB

A solution of the above acetylthioazetidinone (74 mg, 0.09 mmol) in toluene (30 mL) was heated at reflux for 7 h. After evaporation of the solvent, the residue was purified by hplc (SiO 2; eluent: benzene: ether = 3: 1) to give 24 mg (0.044 mmol, 49% yield) of the penem ester as a syrup. tNote: this oil could be crystallized from a mixture of THF-ether or CH 2 Cl 2-ether): 1 Hmr (CDCl 3) δ: 1.40 (3H, d, J = 6.5 Hz, 1'-CH 2), 2.38 (3H, s , 2-CHq), 4.07 (1H, dd, Jc c = 4Hz, Jc Ί = 9Ηζ, 6-H), 5.05-5.30- * 3 ojbb) l 5.34-5.59 ( 2H, AB type, 3-CO 2 CH 2 -Ar), 5.30 (2H, s, 1'-CO 2 -CH 2 -Ar), 5.1-5.6 (1H, m, 1'-H), 5, 68 (1H, d, J e = 4Hz, 5-H), 7.49-7.64-8.18-8.33 (4H, A2'B2 ', 1 aromatic Hs), 7.53-7 , 68-8.18-8.33 (4H, A2, B2 ', 3-aromatic Hs); ir (fine): 1780-lactam), 1750 (-CO 2), 1710 (ester), 1520 and 1350 cm -1 (-no 2).

112 6 7 8 5 3 VV. Preparation of (1'R, 5R, 6R and 1'S, SS, 6S) potassium and sodium 6- (1'-hydroxyethyl) -2-methylpenem-3-carboxylate (isomer D)

OH

0CO2PNB T s XL · '5, H, / M-Celite V «» 3 0J ^ N ^ "CH3 e = tteri-H2O 0" ^ f ^ p CO PNB 2

A solution of the above penem ester (24 mg, 0.044 mmol) in THF (5 mL) was stirred with ether, (10 mL), 1NO (5 mL), phosphate buffer (1.00 mL, 0.05 mL). mol., pH 7.00: Fisher) and 30% Pd-Celite catalyst (50 mg, Engelhard). This mixture was hydrogenated at 35 psi for 21.5 hours at room temperature. After removal of the catalyst (with Celite), the aqueous layer was separated, washed with ether and lyophilized to give 12 mg of a mixture of the title sodium and potassium salts as a white powder: 1Hmr (D 2 O · 1.23 (3H, d, J = 6Hz, 1'-CH ^), 2.27 (3H, s, 2-CH3), 3.85 (1H, dd, J5 6 = 4Hz, Jg ^ ΘΗζ, 6-H), 4.3 (1H, m, 1'-H ) and 5.65 ppm (1H, d, * J 6 = 4Hz, 1 sH); ir (nujol) O: 1755 (lactam) and 1570 cm -1 (-CO 2): uv ^ 20) 297 (£ 2300, calculated as K-salt), 258 (£ 1900, calculated as K-salt). This material was identical to the title compound prepared by aldol condensation of acetaldehyde using 2-methylpenem-3-carboxylic acid Dianione. (1Hmr, ir, uv).

67853 113

Example ^ (1'S, 5R, 6S and i'R, 5S, 6R) 6-1; -hydroxy-1 (ethyl) -2-methylpenem-3-carboxylic acid (Isomer C)

OH

CO H

OCO ^ NB OCO PNB OCO PNB OH

S * 9 ^ i_ / EAc -f- "5, —► i —► I> —c» o-J — 'Y », cr Ν'ν“ φ3 0 = s * -k- ~ ^

CO PNB CO PNB C02 ™ B C ° 2H

2 2

nrn dmb OCO.PNB OCO.PNB

OCO PNB OCO PNB t 2 | 2 ^ c »? Αγ_γ5Λ9 A ^ __ ^ SAc _ ^ SAC ^ λ-N s- N. <2-N ___ ys ^ N v cC- ^ sl <4. cr ^ '- ^ si <j- o ^ Si $ - *

OCO PNB OCC ^ PNB

XIfl / SAc A ^ sac

Ti - * · ——► Γ "v ^ 0" ο ^ - "Ύ · Ρ1

CO PNB CO ^ PNB

n't m. η ... 67853

Method A

1) (1'S, 3S, 4R and 1 * R, 3R, 4S) -4-acetylthio-3- (1'-paranitrobenzyldioxycarbonyl-1'-ethyl) -1- (paranitrobenzyl-2 "-triphenylphosphoranylidene) 2 "-acetate) -2-azetidinone (isomer C).

oco2pnb OCO-PNB 1 I ^, SAg ch3coci '"I j 0i ^ OPt3 ° ^

0 CO PNB

CO PNB 2

To a cold (ice-MeOH bath) solution of 1'S, 3S, 4R and 1'R, 3E, HS) -silver-3- (11-paranitrobenzyl-dioxycarbonyl-1'-ethyl) -1- (paranitrobenzyl-2 "-triphenylphosphoranylidene-2" -acetate) -2-azetidinone-4-thiolate (Isomer C) (1.14 g, 1.30 mmol) in Cl 2 Cl 2 (60 mL) was added pyridine (0.6 mL, 0.74 mmol) and by dropwise addition of acetyl chloride (236 mg, 0.213 mL, 3.00 mmol). The reaction mixture was stirred for 1 h at -15 ° C. The precipitate was filtered off and washed with ether. The filtrate was washed with 2% aqueous HCl, water, 2% aqueous NaHCO 3, water and brine and dried (MgSO 4). The residue obtained after evaporation of the solvent was triturated with ether (895 mg, 83.7%, m.p. 184-185 ° C with decomposition); IR (CHCl 3) tfmax; 1755, 1695 (C = O), 1620 and 1605 cm -1 (phosphorane). Anal. Calcd for C 18 H 18 N 2 O 2 SSi: C 61.38; H 4.42; N 5.11, S 3.90, found: C 61.26, H 4.49, N 4.88, S 4.26.

115 6785 3 2. (1'S, 5R, 6S and 1'R, 5S, 6R) -paranitrobenzyl 2-methyl-6- (1'-paranitrobenzyldioxycarbonyl-1'-ethyl) -penem-3-carboxylate (isomer C)

occym OCO, PNB

'Äv ·, .tQ- "·

C02PNB C02PNB

A solution of (1'S, 3S, 4R and 11R, 3R, US) -4-acetylthio-3-C1T-paranitrobenzyldioxycarbonyl-1f-ethyl) -1- (paranitrobenzyl-2 "-triphenylphosphoranylidene-2" -acetate) -2-azetidinone (Isomer C) (855 mg, 1.04 mmol) in toluene (60 mL) was heated at reflux for 4.5 h. After concentrating the solution, the residue was passed through a silica gel (10 g) column (1% ether in benzene). to give the pure title compound (393 mg, 69.6%), m.p. 157-158 ° C (from CHCl 3 -ether); ir (CHCl 3) νmax: 1785, 1745, 1710 (C = O) and 1525 cm -1 (NO 2); 1 Hmr (CDCl 3) δ: 8.30-7.2 (8H, m, H-aromatic) δ .46 (1H, d, J = 1.8, H-5), 5.40-5.0 (5H, m, z CH2-PNB and H-1 '), 3.95 UH, dd, J = 1.8, J = 5.4, H-6), 2.35 (3H, s, CH 3) and 1.43 ppm (3H, d, J = 5.4, CH 3); C 53.04, H 3.89, N 7.73, found C 52.76, H 3.86, N 7.69.

3) (1'S, 5R, 6S and 1'R, 5S, 6R) -6- (1'-hydroxy-1'-ethyl) -2-methyl-penem-3-carboxylic acid (Isomer C)

OH

OCO PNB J

\: o2pnb c02H

A mixture of (1'S, 5R, 6S and 1'R, 5S, 6R) -paranitrobenzyl 2-methyl-6- (1'-paranitrobenzyldioxycarbonyl-1'-ethyl) -penem-3-carboxylate (206 mg, 0.379 mmol), 116 67853 from THF-ether-H 2 O (30 mL, 40 mL, 20 mL), 0.05 mol. pH 7 buffer solution (7.64 mL, 0.38 2 mmol) and 30%. Pd-Celite catalyst (500 mg) was hydrogenated in 42 psi H 2 on a Parr shaker for 16 hours. The catalyst was filtered off and washed with water. The aqueous phase was washed with ether (3 times), acidified by portionwise addition of cold 1% aqueous HCl until pH 2.5 and extracted with ethyl acetate (15 x 20 mL) between each HCl addition. The ethyl acetate extracts were combined and washed with brine (3 x 30 mL). Evaporation of the solvent and trituration of the residue with ether gave the title compound (57 mg, 65.6%); ir (KBr): 3580-3300 (0-H), 1755 and 1660 cm-1 (C = O); uv (EtOH) ν max 311 (£ 6538), 262 ((3672); m / z (DHSO-d 6) 5.57 (1H, d, J = 1.7, H-5), 4.02 (1H, m, H-11), 3.75 (1H, dd, J = 1.7, J = 3.5, H-6), 2.23 (3H, s, CH 3) and 1.23 ppm (3H, d, CHgj.

Method B

1) Silver (1'S, 3S, 4R and 1'R, 3R, 4S) -1- (t-butyldimethylsilyl) -3- (11-paranitrobenzyldioxycarbonyl-11-ethyl) -2-azetidinone-4-thiolate ( isomer C)

OCO PNB 1 2 SAO

'ns "3 -►' Ti»

\ tBu ^ BU

Isomer C (1 g, 143 mmol) of 1 '- (t-butyldimethylsilyl) -3- (1'-paranitrobenzyldioxycarbonyl-1'-ethyl) -4-tritylthio-2-azetidinone was dissolved in hot (40 ° C) methanol with stirring. (12 ml). A solution of silver nitrate (0.59 g) in methanol (12 ml) was added followed by pyridine (0.13 ml). The mixture was stirred vigorously for 1 h at room temperature and 2 h at 0 °. The solid silver mercaptide was collected by filtration and washed with ether, 352 mg (4G%).

IR max: 1735 cm-1 (C = O) · 117 6785 3 2) (1'S, 3S, 4R and 1'R, 3R, 4S) 4-acetylthio-1- (t-butyldimethylsilyl-3- ( 1'-paranitrobenzyldioxycarbonyl-1'-ethyl) - '2-azetidinone (isomer C) oco2pnb oco2pnb Λ-Υ5'9 \ o 1 tBu ^ tBu

To a solution of isomer C of silver 1- (t-butyldimethylsilyl) -3- (1'-paranitrobenzyldioxycarbonyl-1'-ethyl) -2-azetidinone-4-thiolate (880 mg) in dichloromethane (40 mL) and stirred at 0 ° C, pyridine (0.57 ml) was added followed by acetyl chloride (0.49 ml) dropwise. The mixture was stirred for 0.5 h at 0 °, the solids removed by filtration and the filtrates diluted with ether, washed with aqueous hydrochloric acid (2%), water, sodium bicarbonate (2%) and brine, dried and evaporated to dryness to leave the title compound as an oil ( 610 mg).

1 Hmr (CDCl 3) δ: 8.2 and 7.48 (4H, 2d, aromatic), 5.40 (1H, d, J = 2.2, H-4), 5.2 (2H, s, benzyl ), 5.3-4.9 (1H, m, H-11), 3.42 (1H, dd, J = 2, H-3), 2.32 (3H, s, CH 2), 1.40 (311, d, J = 6.5, CH 2), 0.95 (9H, s, t-Bu) and 0.2 ppm (6H, CH 2) · 3) (1'S, 3S, 4R and 11R, 3R , 4S) -4-acetylthio-3- (1'-paranitrobenzyldioxycarbonyl-1'-ethyl) -2-azetidinone (Isomer C)

OCO, PNB OC02PNB

XO- __ ^ tBu

Isomer C (1.4 g) of the above S-acetyl-Nt-butylmethylsilyl-azetidinone derivative was dissolved in a mixture of TFA (0.5 ml), water (0.5 ml), methanol (3 ml) and dichloromethane (2 ml) and stirred at room temperature for 48 hours. The solution was diluted with water (100 mL) and extracted with dichloromethane (4 x 20 mL). The combined organic extracts were washed 118,678,53

Sodium bicarbonate (2%) and brine, dried and evaporated to dryness to leave the crude title compound as an oil. Purification was performed by chromatography on silica gel (30 g) eluting with 5% ether in benzene; (650 mg). Crystallization from benzene gave a white solid. 1 Hmr (CDCl 3) cp: 8.15 and 7.45 (4H, 2d, aromatic), 6.18 (1H, NH), 5.19 (2H, s, benzyl), 5.05 (2H, m, H -4 and H-1 '), 3.35 (1H, dd, J = 2.5, 4.5, H-3), 2.34 (3H, s, CH 2) and 1.42 ppm (3H, d, J = 6.5, CH 3); and 175 °> 1698 cm -1 (C = O).

4) (1'S, 3S, 4R and 1'R, 3R, 4S) -4-acetylthio-3- (1'-paranitrobenzyldioxycarbonyl-11-ethyl) -1- (paranitrobenzyl-2M-hydroxy-2 " acetate) -2-azetidinones (epimers of C-1 ") (isomer C)

OCOjPNB

ΟΟΟ, ΡΝΒ I SAC

- * X- 0 ίθ2Ρΐ®

A mixture of isomer C of 4-acetylthio-3- (1'-paranitrobenzyldioxycarbonyl-1'-ethyl) -2-azetidinone (750 mg), paranitrobenzyl glyoxylate hydrate (525 mg) and benzene (50 ml), heated by boiling for 3 days using a Dean-Stark apparatus filled with a 3A molecular sieve. A second portion of glyoxylate (52 mg) was added and boiling was continued for another 2 days. The mixture was diluted with ether, washed with hydrochloric acid (2%), water, sodium hydrogen carbonate (2%) and water, dried and evaporated to dryness to leave an oily residue (975 mg). Chromatography on silica gel, eluting with benzene-ether (85:15) gave pure title compounds. Hrnr (CDCl 3) rf: 8.2 5-6.75 (8H, m, aromatic), 5.30 and 5.12 (4H, 2s, benzyls), 5.05-4.70 (1H, H-2 "), 4.45-4.35 (1H, 2d, H-4), 4.50-4.10 (1H, m, H-1 '), 3.30 (1H, m, H -2 and 1.25 ppm (3H, 2d, CH 2).

119 67853 5) (1'S, 3S, rR and 1'R, 3R, 4S) -4-acetylthio-3- (1'-paranitrobenzyldioxycarbonyl-1'-ethyl) -1- (paranitrobenzyl-2 "- triphenylphosphoranylidene-2 "-acetate) -2-acetidininone (Isomer C)

OCO PNB QCO „PNB

OCO PNB γ- 2

A _, - ssc son ,. A-fSAC ^, "rfSAC

^., 0 "" o ^ H-vcl

L An dkto CO PNB

CO2PNB co2pnb 2 4-Acetylthio-3- (11-paranitrobenzyl-dicycarbonyl-1'-ethyl) -1- (paranitrobenzyl-2 "-hydroxy-2" -acetate) -2-azetidinone Isomer C (577 mg, 1 mmol) was dissolved in dry THF (10 mL) and pyridine (95 mg, 1.2 mmol) was added to the solution. The solution was cooled to 0 ° and thionyl chloride (143 mg, 1.2 mmol) was added slowly. The mixture was stirred for 30 min.

At 0 °, diluted with a small amount of ether and insoluble salts were removed by filtration and washed with ether. The combined filtrates were evaporated to dryness to give a crude mixture of epimers of the C-2 "chloro compound. It was dissolved in THF (20 mL), triphenylphosphine (314 mg, 1.2 mmol) and 2,6-lutidine (129 mg, 1, 2 mmol) and the solution was stirred at 45 ° C for 4 days The solids were removed by filtration, washed with benzene and the combined filtrates evaporated to dryness to leave an oil with spectral properties and tlc behavior identical to a sample of the title compound prepared by acylation. the corresponding silver thiolate.

The desired penem product is obtained by cyclization as described in Example 9.

6785 3 120

Example 11 (11R, 5R, 6S and 1'S, 5S, bR) -6- (1'-Hydroxy-X'-ethyl) -2-methylpenem-3-carboxylic acid (Isomer B)

OH

inQ-.

VO H 2

Method A

OH CH.,

oco2pnb oco.pnb oco0pnb i V

Λ-Γ5'1 '^ r-rsV-cH -' Lf ^ ^ ΖΓρφ - * IX ^ 3 ~ 3 o ^ y

O "f 3 \ θ p„ E CO2H

CO PNB CO PNB 2 2 2

Method B

CH0 OH \ 3 OH OH PH I \ Λ-γ5Α9 DTMS / TEA '' f ^ V-CH. _TjT / - ρφ3 2, * cci J-Ι Ρφ3 Λ-y Λ-y

T pyridine [co.pnb rr> H

CO 2 FNB 3) H 3 O + CO 2 Pfre 2 * -

Method A

1) (1'R, 3S, 4R and 1'S, 3R, 4S) -4-acetylthio-3- (1'-paranitroxy-1'-ylcarbonylcarbonyl-1'-ethyl) -1- (paranitrobenzyl) nlt thiphenylphosphoranylidene-2 "-acetate) -2-acetidino. ^ (isomer B)

OCO.PNB OC02PNB

J 2 SAg J SAc / · - «- S AcCl - SV” CO PNB C02PN8 121 67853

A solution of (1'R, 3S, 4R and 1'S, 3F, 4S) -silver-3- (1'-paranitrobenzyldioxycarbonyl-1'-ethyl) -! '-paranitrobenzyl 211-triphenylphosphoranylidene-2 "-acetate) -2-acetidinone 4-thiolate (Isomer B) (917 mg, 1.03 mmol) in CH 2 Cl 2 ( 20 mL), treated at -15 ° C (ice-MeOH bath) with pyridine (242 μL, 247 mg, 3.13 mol) and added dropwise to acetyl chloride (142 μL, 157 mg, 2.0 mmol). The mixture was stirred for 15 minutes at -15 ° C and the solid was filtered off and washed with ether The organic solution was washed with 2% aqueous HCl, water, 2% aqueous NaHCO 3, water and brine and dried over MgSO 4. The residue obtained after evaporation of the solvent was crystallized from ether (710 mg, 80%, m.p. 18 3-18 5 ° C; IR (CHCl 3) tfmax: 1755, 1695 (C = O), 1620, 1605 (phosphorane) and 1625 cm -1). (N02).

2) (1'R, 5R, 6S and 1'S, 5R, 6R) -paranitrobenzyl 2-methyl-6- (1'-paranitrobenzyldioxycarbonyl-11-ethyl) penem-3-carboxylate (isomer B)

OCO PNB fO / NB

J ____ / W \ ph », —i— · Pt"

0 yh to PNB

CO PNB 2 2

A solution of (1'R, 3S, 4R and 1'S, 3R, 4S) -4-acetylthio-3- (1'-paranitrobenzyldioxycarbonyl-1'-ethyl) -1-paranirrobenzyl-2 '-triphenylphosphoranylidene-2 "-acetate) -2-azetidinone (550 mg, 0.791 mmol) was boiled in toluene for 7 h. The concentrated solution obtained by evaporation of the solvent was passed through a silica gel column (10 times the weight of silica gel) to give the title compound (0.5% ether / benzene). - * 2% ether in benzene) as a white solid, 329 mg, c 77%, mp 134-13 5 ° C, (CH 2 Cl 2 -ether), ir (CHCl 3): 178 5, 1745, 1705 (C = O) and 152 5 cm -1 (NO 2); 1 Hmr (CDCl 3): 8.20 (2, d,

Ho aromatic), 7.60 (2H, d, Hm aromatic), 5.55 (1H, d, J = 1.5, Hs), 5.5-4.75 (5H, m, 2CH2 "PNB, H -1 '), 3.86 (1H, dd, • 122 67853 J = 7.8, J-1.5, H-6), 2.38 (3H, s, CH 3 and 1.50 ppm (3H, d = J = 6.3, CH 3) Anal. Calcd for (-H 2 H 2 N 3 N 3 (+ 10 H 2 O 3 O 4> H 3.89, N 7.73, s 5.90; found: C 53, 05, H 3.98, N 7.63, S 6.02.

3) (1'R, 5R, 6S and 1'S, 5S, 6R) 6- (1'-hydroxy-1 * -ethyl) -2-methyl-penem-3-carboxylic acid (Isomer B)

?B

OCO.PNB J

__3-- »" rf VCH3 p> -

CO PNB

A mixture of (1'R, 5R, 6S and 1'S, 5S, 6R) -paranitrobenzyl 2-methyl-6- (1'-paranitrobenzyldioxycarbonyl-1'-ethyl) penem-3-carboxylate (Isomer B ) (65 mg, 0.12 mmol), 0.05 mol. The pH buffer solution (1.06 eq.), H 2 O-THF-ether mixture (10 mL, 10 mL, 25 mL) was shaken on a Parr hydrogenator for 16 h at 50 psi H 2 using 30% Pd / Celite catalyst (200 mg). The catalyst was filtered off and washed with a small volume of water. The aqueous layer was washed with ether (3 times), acidified by portionwise addition of cold 1% aqueous HCl, extracted with ethyl acetate between each addition of HCl, and saturated with brine and extracted thoroughly with ethyl acetate. The ethyl acetate extracts were combined, washed with brine (5 times) and dried (MgSO 4). Evaporation of the solvent gave a solid residue which was triturated with methylene chloride (19.9 mg, 71%). and (nujoli) $ m: 3500 (0-H), 178 δ, 1672 cm -1 (C = O); uv (EtOH) Amax: 260 (£ 3450) (309 <£ 6400); 1 H-NMR (DMSO d 6) tf: 5.54 (1H, d, J = 1.5, H-5), 3.88 (1H, m, H-1 '), 4.2-3.5 ( 2H, bs, O-H), 3.65 (1H, dd, J = 6.5, J = 1.5, H-6), 2.28 (3H, s, CH 2) and 1.15 ppm ( 3H, d, J = 6, CH 3).

123 67853

Method B

1) (1'R, 3S, 4R and 1'S, 3R, 4S) -4-acetylthio-3- (1,1-trimethylsilyloxy-1'-ethyl) -1- (paranitrobenzyl-2 "-triphenylphosphoranylidene) 2 "-acetate) -2-acetiaminone (isomer B)

OH 0TMS

_f8 ** 3 TMSCl AcCl -f ^ SAC

3 'TEA' C5H5N 0 ^ “^ γΡΦ3

I CO PNB

C02PNB 2

Suspension of (1'R, 3S, 4R and 1'S, 3R, 4S) -silver-3- (1'-hydroxy-1'-ethyl) -1- (paranitrobenzyl-1 "-triphenyl-phosphoranylidene-2" acetate) -2-azetidinone-4-thiolate (505 mg, 0.715 mmol) in THF (25 mL) was cooled to -15 ° C (ice-MeOH bath), triethylamine (289 mg, 398 μL) was added dropwise. 2.86 mmol), trimethylchlorosilane (310 mg, 362, 2.85 mmol) and finally imidazole (50 mg, 0.734 mmol) were stirred for 3 h at -15 ° C and room temperature for 16 h (IR of the sample showed that it had no hydroxyl group). The mixture was cooled to -15 ° C, diluted with CH 2 Cl 2 (20 mL), treated with pyridine (226 mg, 231 μL, 2.86 mmol) and acetyl chloride (168 mg, 152 μL, 2.14 mmol). , stirred for 0.5 h, diluted with ether, washed with dilute HCl, water, 5% aqueous NaHCO 3, water and brine and dried. The solvent was removed on a rotary evaporator and the residue was purified by filtration through a silica gel column (1:10, 3% 10% ether in benzene) to give the title compound (360 mg, 84.2%) which included a small amount of desilylated derivative (30 mg, 7%). .8%). IR (liquid film) \:,: 1750, 1790 (C = 0), η me ie · 1620 (phosphorane) and 1518 cm ”(Γ ^) · 124 6 7 8 5 3 2) (l'R, 3S, 4R and 1'S, 3R, 4S) -4-acetylthio-3- (1'-hydroxy-1'-ethyl) -1- (paranitrobenzyl) -2M-triphenylphosphoranylidene-2 "-acetate) -2-azetidinone ( isomer B)

OH

? ™ S λ ^ SAc I SAC H cP '' 'j - ("ΓΊ —3--' n j> 4>, J — N Ρ (Ν 3

CO PNB

iO PNB 2 2

A solution of (1'R, 3S, 4R and 11S, 3R, 4S) -4-acetylthio-3-11-trimethylsilyloxy-1'-ethyl) -1- (paranitrobenzyl-2 "-triphenylphosphoranylidene-2" - acetate) -2-azetidinone (360 mg, 0.504 mmol) was treated with TFA (3 drops) and stirred at room temperature for 18 h. The mixture was diluted with ethyl acetate, washed with water, dilute aqueous NaHCO 3, water and brine and dried (MgSO 4). ). Evaporation of the solvent gave the title compound (334 mg, 100%); IR (CHCl 3) ν max: 1755, 1690 (C = O), 1620, 1605 (phosphorane) and 1520 cm -1 (NO 2).

3) (1'R, 5R, 6S and 1'S, 5S, 6R) -paranitrobenzyl 2-methyl-6- (1'-hydroxy-1'-ethyl) -penem-3-carboxylate (isomer B) oh oh J * / SAc JV · -

ΡΗΒ2ΡΗΒ C02PNB

A solution of (1'R, 3S, 4R and 1'S, 3R, 4S) -4-acetylthio-3- (1'-hydroxy-1'-ethyl) -1- (paranitrobenzyl-2 "-triphenyl- phosphoranylidene-2 "-acetate) -2-azetidinone (410 mg, 0.638 mmol) in toluene (40 mL) was boiled for 7 h. Toluene was partially distilled off. The residue was passed through silica gel (1:10) on a column (3%, 4%, and 5% ether in benzene) to give the title compound (151 mg, 65%) as a white solid, m.p. 161 to 161.5 ° C; ir (CDCl 3) V v: ma 125 67853 3600, 3500-3400 (OH), 1780, 1608 (c = 0) and 1525 era * 1 (NOj); 1 HHmr (CDCl 3) cT: 3.2 ° (2H, d, J-7, Ko aromatic), 7.6 ° (2H, d aromatic), 5.57 (1H, d, J = 2, H-5 ), 5.29 (2K, center of ABq, J = 15, CH2-PNB), 4.2 (1H, dq, J = 7, J = 6, H-1 '), 3.67 UH, dd , J = 7, J = 2, H-6), 2.33 (3H, s, CH 2) and 1.33 ppm (3H, d, J = 6, CH 2);

Anal. calc. for C 10 H 19 O 2 S: C 52.74, H 4.43, N 7.69,

Xb lb l b S 8.80; Found: C 52.67, H 4.41, N 7.71, s 6.96.

4) (1'R, 5R, 6S and 1'S, 5S, 6R) -6- (11-hydroxy-1'-ethyl) -2-methyl-penem-3-carboxylic acid (Isomer B)

OH

0H J c Ύτ>. 'ΧΟ-

0 'CO H

CO PNB 2 2

A mixture of (1 ', 5R, 6S and 1'S, 5S, 6R) -paranitrobenzyl-6- (11-hydroxy-11-ethyl) -2-methylpenem-3-carboxylate (89 mg, 0.244 mmol) ), THF-t 1 O-ether mixture (15 mL, 10 mL, 30 mL), 0.05 mol. The pH 7 buffer solution (5.06 mL, 0.253 mmol) and 30% Pd / Celite catalyst (250 mg) were shaken on a Parr hydrogenator for 3.5 h at 45 psi H 2. Further work-up as described above gave the title compound (32 mg, 57%).

Example 12 (1'S, 5R, 6R and 1'R, 5S, 6S) -6- (11-hydroxy-1'-ethyl) -2-methylpenem-3-carboxylic acid (Isomer A) f ,,

Xco2h 126 6 7 8 5 3 1) (1'S, 3R, 4R and 1,1'R, 3S, 4S) -4-acetylthio-3- (1,1-methoxymethoxy-11-ethyl) -1- (paranitrobenzyl-2 "- triphenylphosphoranylidene-2" -acetate) -2-azetidinone (isomer A) OCH, OCH3 f "20Cl% Ac I —f 'ί ^ ν' * 3

1 CO PNE

co2pnb 2 4-Acetylthio-3- (1'-methoxymethoxy-1'-ethyl) -1- (para-nitrobenzyl) -2 "-triphenylphosphoranylidene-2" -acetate) -2-azetidinone Isomer A was prepared as described elsewhere for isomer C of the paranitrobenzyldioxycarbonyl derivative in 85% yield. IR (fine): 1750 and 1690 cm-1 (C = O).

2) (1'S, 3R, 4S and 1,1R, 3S, 4S) -4-acetylthio-3- (1,1-hydroxy-1,1-ethyl) -1- (paranitrobenzyl-2 "-triphenylphosphoranylidene-2" acetate) -2-azetidinone (Isomer A)

OH

? CH2OCH3 I cAc £ _> ____ 'Vf

U T CO.PNB

6o2PNB 2 4-Acetylthio-3- (1'-methoxymethoxy-1'-ethylD-1-industrial nitrobenzyl-2 "-triphenylphosphoranylidene-2 ', - acetate) -2-acetidinone Isomer A (500 mg, 0.68 mmol) was added to a cooled solution (0 ° C) of trifluoroacetic acid (50 ml) and water (10 ml) and stirred for 15 minutes on ice and for 3 h at room temperature The reaction mixture was evaporated to dryness, dichloromethane was added and the solution was washed with sodium hydrogen carbonate. , water and brine, dried and evaporated to dryness to give the title compound (450 mg, 96%): IR (fine): 3400 (OH), 1745 and 1690 cm-1 (C = O).

67853 3) (I'SjSRjBR and 1'R, 5S, 6S) -paranitrobenzyl 6- (1'-hydroxy-1'-ethyl) -2-methylpenem-3-carboxylate (Isomer A).

ALRIGHT

CH JL

V ”- Tu-

0 ^ “ΝΎ ^ Φ3 Vo2PNB

£ θ PNB

Prepared as described for the isomer C of the paranitrobenzyldioxycarbonyl derivative in H5% yield; 1 Hmr (CDCl 3) δ: 7.93 (UH, ABq, aromatic), 5.68 (1H, d, J = 4.0, H-5), 5.33 (2H, ABq, benzyl), 4 , 3 (1H, m, H-1f), 3.8 (1H, dd, J = 4.0, H-6), 2.41 (3H, s, CH 2), 2.31 (1H, s, OH), and 1.42 ppm (3H, d, J = 6, CH 2); ir (CHCl 3) ν max: 3100-3600 (OH), 1780 and 1710 cm -1 (C = O).

4) (1'S, 5R, 6R and 1'R, 5S, 6S) -6- (1'-hydroxy-1'-ethyl) -2-methyllipenem-3-carboxylic acid (Isomer A)

oh ° H

'tcQ-1 ·· - · tcQ- ”·

CO2PNB CO2H

A mixture of paranitrobenzyl 6- (11-hydroxy-1'-ethyl) -2-methyl-penem-3-carboxylate isomer A (82 mg, 0.2 mmol), Pd / Celite catalyst (30% , 400 mg), THF (10 mL), ether (25 mL), water (10 mL) and buffer (0.05 mol, pH = 7,

Fisher® SO-B-I08) (4 mL) was hydrogenated for 4 hours on a Parr shaker at an initial hydrogen pressure of 45 psi. The catalyst was removed by filtration through Celite and washed with water. The filtrates were washed with ether and the aqueous layer was acidified with cold hydrogen chloride solution (0.25 mol) and extracted with ethyl acetate (5 x 10 mL). The combined organic extracts were washed with brine, dried and evaporated to dryness. The foamy solid 12s 67853 was triturated with ether to give a white solid (20 mg, 14%). IR (nu j ol) '0: 3500 (OH), 17 85 and 1665 cm -1 (C = O) j uc (EtOH) / \: 301 (E .. 592 2), 260 (t' '- / bU).

ITtclKS ·

Example 13 (1'S5R, 6S and 1 'R, 5S, 6R) - β-Trimethylsilyl ether 6- (1'-acetoxy-1'-ethyl) -2-methylpenem-3-carboxylate (Isomer C) OAc

I H H

Lryne co 2 - [Si- 3- (1'-hydroxy-11-ethyl) -1- (β-trimethylsilylethyl 2 "-triphenylphosphoranylidene-2" -acetate) -trityl-tao-2-azetidinone

ST- 0H

—F 11 LDA. Me 'y-f STr h3 2) <: h3cho CO2 CO2

To a stirred solution of diisopropylamine (185 mg, 1.1 β4 mmol) in tetrahydrofuran (5 mL) at -78 ° C was added n-butyllithium (1.3 mL, 2.0 mmol). After 5 minutes, a solution of 1- (p-trimethylsilylethyl 2'-triphenylphosphoranylidene-2'-acetate) -4-tritylthio-2-azetidinone (1.27 g, 1.67 mmol) was added dropwise with stirring over 20 minutes. ) in tetrahydrofuran (15 ml). After 2 minutes, freshly distilled acetaldehyde (1 ml) was added and the solution was stirred for 5 minutes. Hydrochloric acid (12.6 mL, 0.3 mol) was added and the mixture was allowed to warm to 23 ° C. Water and ethyl acetate (20 mL each) were added, shaken, and separated. The organic phase was washed with water and saturated sodium chloride solution (20 ml each), dried and the solvent evaporated in vacuo to give 1.37 g of crude product. The product was adsorbed from methylene chloride onto silica gel (7 g) and applied (dry) to a column containing 28 g of silica gel. The column was eluted with ether (100 ml) and then with a 1: 1 mixture of ether / ethyl acetate (50 ml> 20 ml of the fractions from the column were discarded. The remainder were combined and the solvent 67853 was evaporated in vacuo to give 1.03 g of product. To a column containing 50 g of silica gel (wet), the column was eluted with ether (C b 8 0 ml) and then with ethyl acetate (200 ml). The latter fractions were combined (predominantly low Rf spot in tlc) and the solvent evaporated in vacuo to give the partially purified title compound, 440 mg (33%); IR V (H) and 1750 cm-lactam and ester); ^ Hmr (CHCl3) ^: the separation is too weak to indicate other than aromatic and trimethylsilyl on the basis of the peaks.

Silver 3- (11-hydroxy-1'-ethyl) -1H-trimethylsilyl ethyl 2-triphenylphosphoranylidene-2 "-acetate) -2-acetidinone-4-thiolate

OH OH

MaA-rSTr agnoypyridine ^ MeA-J * '*

J-N J> Ph J-N? PhT

co2 co2

A solution of silver nitrate (425 mg, 2.5 mmol), pyridine (79 mg, 1.0 mmol) and water (10 mL) was added to a solution of the above compound (403 mg, 0.50 mmol). in ether (10 ml). The mixture was stirred vigorously for 1 h. The precipitate was collected by filtration and washed with water and ether to give the title mercaptide 267 mg (80%). IR δ: 340 (J (OH) and 1750 cm-1 (Jb-lactam and ester).

67853 130 u'-acetylthio-3- (1'-acetoxy-1'-ethyl) -1 - ([9-trimethylsilylethyl-2 "-triphenylphosphoranylidene-2" -acetate) -2-azetidinone oh ° Ac JL. ^ Ag .... 1 sac {AcCl / pyridimx Me '"' η-pc ° 2 co2

A solution of acetyl chloride (70 mg, 0.88 mmol) in methylene chloride (1 mL) was added dropwise to a solution of the above silver mercaptide (267 mg, 0.40 mmol) and pyridine (70 mg, 0.88 mmol) in methylene chloride (1 mL). 5 ml) at 0 ° C. The mixture was stirred at 0 ° C for 1.5 h and then at 23 ° C for 15 minutes. The precipitate was filtered off and the solution was washed with Q.1 mol. with hydrochloric acid and 0.1 mol. sodium bicarbonate solution (10 mils each). The solvent was evaporated in vacuo to give the title compound 153 mg (59%); ir Vjuagk * 31 + 50 (OH), 17 50 (β-lactar and ester) and 1690 cm-1 (thioester); 1 Hmr (CDCl 3) δ: 7.5-8.2 (m, 15H, Ph), 5.85 (br, 1H, H-4), 3.0-5.0 (unresolved, 4H, OCH, OCH 2) , H-3), 2.0-2.6 (3 singlets; 6H, OAc, She), 0.9-1.7 (m, 5H, CH 2, CH 2 Si) and 0.20 ppm (s, 9H, SiMe3).

(1'S, 5R, 6S and 1'R, 5S, 6R) -N-trimethylsilylethyl 6- (1 * -acetoxy-1'-ethyl) -2-methylpenem-3-carboxylate (Isomer C) OAc

9Ac I H H

0PN ^ PPh3 Δ / -H

A solution of the above phosphorane (150 mg, 0.23 mmol) in toluene (15 mL) was heated at reflux for 2 h.

The solution was mixed with 1 g of silica gel and the solvent was evaporated off in vacuo. The silica gel was applied to a 4 g portion of silica gel on a column (dry) and eluted with ether. The first 131 67853 5 ml fraction (one spot with a high Rf value on a tlc plate) gave, by evaporation of the solvent, 65 mg (76%) of the title compound as a waxy solid. IR0,: 1790 β-lactam), 1740 (ε-Y max.

teri) and 1700 cm (OAc); 1 H (CDCl 3) δ: (d, J = 2Hz, 1H, H-5), 5.4 (m, 1H, H-1 '), 4.3 (m, 2H, OCH 2), 3.90 (q, J = 2Hz, 4Hz, 4Hz, 1H, H-7), 2.37 (s, 3H, 2-CH), 2.11 (s, 3H, OAc). 1.42 (d, J = 6.5 Hz, 3H, 2'-CH 2), 1.1 (m, 2H, CH 2 Si) and 0.05 ppm (s, 9H, SiMe 2). The product was found to be a single isomer.

Example 14

6-formamidomethyl-2-methylpenem-3-carboxylic acid, sodium and potassium salts 0 g? S

HCN \ ._ CH

H La // 3 trans-1- (t-butyldimethylsilyl) -3-methanesulfonyl-oxymethyl-4-tritylthio-2-azetidinone 0H uu 5 HH sccj> HH sci H * j _ MsO I Γ ° '^ Si ) 9 0 \ -Bu ^ t_Bu

A solution of trans-1- (t-butyldimethylsilyl) -3-hydroxymethyl-4-tritylthio-2-azetidinone (8.9 g, 16.36 mmol) in dichloromethane (50 mL) was treated with dichloromethane at 5 ° C. with methanesulfonyl chloride (1.4 mL, 18 mmol) and triethylamine (2.5 mL, 18 mmol). Stirring was continued for 1 h under protection. The solution was then washed successively with cold 1-norm. with hydrochloric acid, 1 mol. sodium hydrogen carbonate and brine, dried (MgSO 4) and evaporated to dryness in vacuo. The residue (mixture of hydroxy and mesylate compound) was treated a second time in the same manner as above to give mesylate (90 g, 97%) as an armorphic solid. It was used as such without further purification in the next step. An analytical sample was recrystallized from methylene chloride, m.p. 167-168 ° C; IR (fine) νmax: 1775 cm -1 (CDCl3) cT: 7.3 (15H, m), 4.4 (1H, d, J = 2Hz), 3.9 (1H, dd, J = 8Hz, 4Hz), 3.2 (2H, bs), 2.8 (3H, s), 0.95 (9H, s) and 0.3 ppm (6H, s).

67853 132 Trans-3-Hiethanesulfonyloxymethyl-4-tritylthio-2-acetyldinone and trans-3-azidomethyl-4-tritylthio-2-acetylxone HHT> SC: ^ s' "· A Ssc ^ -i MSck f 3__ , MSO ^ | _--> "3] \ i (CH3'2 t-Bu

A solution of trans-1- (t-butyldimethylsilyl) -3-methanesulfonylmethyl-4-tritylthio-2-azetidinone (21.0 g, 37.0 mmol) in HMPA (90 mL) was cooled in an ice bath and treated with sodium azide ( 2.7 g, 41.2 mmol) in IVOs (10 mL). The reaction mixture was stirred at room temperature for 1 h, diluted with ethyl acetate, washed with H 2 O (5 x 100 mL), dried (MgSO 4) and evaporated to dryness in vacuo (4.

where. Trans-3-methanesulfonyloxymethyl-4-trityl-thio-2-azetidinone was diluted with HMPA (90 mL), treated with sodium azide (2.7 g, 41.2 mmol) at room temperature in 1 H (10 mL), heated at 60 ° C for 2 h and triturated with cold water. The crude azide was diluted with benzene-ether (5: 1) and washed with water (5 x 20 mL). Evaporation of the solvent followed by crystallization from ether gave 18.0 g (77%) of the azide as a white solid. An analytical sample was recrystallized from CH 2 Cl 2 / ether, m.p. 174-175 ° C; Anal. Calcd for O 2 O 2 • 2 H 2 O 3: C, 68.97; H 5.03; N 13.99; Found: C, 68.78; H 5.00; N 14.16; ir (nujoli) ν max: 2100 λ max max 17 1765 cm -1: Hmr (CDCl 3) δ: 7.35 (15H, m), 4.75 (1H, bs), 4.4 (1H , d J = 2Hz), and 3.1-3.7 ppm (3H, m).

133 6785 3

Trans-3-aminomethyl-4-trityl-spore-2-acetinone

H H H H

"S'ff 3 _____,« / '' "f-f 3 cr n’h cy nh

To a solution of trans-3-azidomethyl-4-tritylthio-2-azetidinone (10.0 g, ?.5 mmol) in dry methanol (500 mL) was added ammonium chloride (19.0 g) and zinc powder (1.0 g). ) and the suspension was stirred at room temperature for 5 h. The reaction mixture was filtered and evaporated to dryness. The residue was partitioned between l-norm. parts soluble in hydrochloric acid and benzene. The aqueous layer was basified with 1 mol. sodium hydrogen carbonate and extracted with methylene chloride. The extracts were washed with brine, dried (MgSO 4) and evaporated to dryness in vacuo. The crude amine was crystallized from ether, IM, .05 g (79%); mp. 139-9 ° C, Anal. Calcd for C 10 H 14 Cl 2 Cl 2: C, 70.56; H 5.73; N 7.08; Found: C, 70.68; H, 5.9 M-; N 7.27; ir (CHCl 3) tfmax: 3400 and 1760 cm -1; 1 Hmr (CDCl 3) δ: 7.35 (15H, m) 5.15 (1H, m), 4.3 (1H, bs), 2.7-3 , Δ (3H, m) and 1.3 ppm (2H, m).

Trans-3-phthalimidomethyl-4-tritylthio-2-azetidinone o

H H H H H

------ * ^

o XH O

A solution of trans-3-aminomethyl-4-tritylthio-2-azetidinone (13.9 g, 37.2 mmol) and N-carbethoxyphthalimide (8.3 g, 37.9 mmol) in benzene (200 mL). heated at reflux for 15 h. The solvent was evaporated off in vacuo and the residue was crystallized from ether to give 17.4 g (93%) of the title compound; mp. 172-I73 ° C; Anal. Calcd for C 31 H 24 N 2 O 3 S: C 73> 78> H 4.79; N 5.55; Found: C, 73.92; H 4.87; N 5.49; ir (CHCl 3) Vmax ·· 1770 and 1715 cm -1; Himr (CDCl 3) δ: 7.8 (4H, m), 7.3 (15H, m), 4.45 (1H, d, J = 2Hz), 3.3-4.1 (3H, m) and 3.3-4.5 ppm (1 H, m).

Trans-3-phthalimidomethyl-1- (paranitrobenzyl-2R-hydroxy-2'-acetate) -4-tritylthio-2-azetidinone -

o / - »„ 0 </ “Y

CO PNB

A mixture of trans-3-phthalimidomethyl-4-tritylthio-2-azetidinone (17.4 g, 34.52 mmol), paranitrobenzyl glyoxylate hydrate (9.4 g, 41.4 mmol) and triethylamine (4.8 mL, 34.5 mmol) in tetrahydrofuran (250 mL) was stirred at room temperature for 20 h. The reaction mixture was evaporated to dryness in vacuo and the residue was treated with charcoal in benzene. Evaporation of the solvent gave crude hydroxy glyoxylate (25 g, quantitative) as an amorphous solid. It was used without further purification in the next step; ir (CHCl 3) νmax: 1770 and 1715 cm -1; 1 Hmr (CDCl 3) δ: 8.1 (2H, d, J = 9Hz), 7.55 (3H, d, J = 9Ha), 7.3 ( 19 H, m), 5.0-5.4 (2H, bs), 4.3-5.0 (2H, m) and 2.8-3.8 ppm (4H, m).

135 67853

Trans-3-phthalimidomethyl-1- (paranitrobenzyl-2 1-chloro-2 1 -acetate) -4-tritylthio-2-azetidinone CO PNB 2 2 To a cooled solution (ice bath, 0 ° C) containing trans -3-Phthalo-iridomethyl-1- (paranitrobenzyl-2H-hydroxy-2'-acetate) -4-tritylthio-2-azetidinone (25 g, 35 mmol) in tetrahydrofuran (150 mL) was added dropwise 1 -Mol. a solution of thionyl chloride in tetrahydrofuran (46 mL, 45 mmol) followed by 1 M. tetrahydrofuran solution of pyridine (46 mL, 45 mmol). The reaction mixture was stirred at room temperature for 20 minutes, diluted with petroleum ether (50 mL) and filtered through a pad of Celite / charcoal. The solvent was evaporated in vacuo to give chloroazetidinone (26 g, quantitative) as an amorphous solid. It was used without further purification in the next step, ir (CHCl 3): 17 7 5 and 1720 cm -1. 1 Hmr (CDCl 3) δ: 8.12 (2H, d, J = 9Hz), 7.60 (2H, d, J = 9Hz), 7.3 & 19H, m), 5.25 (2H, m), 4 , 7-5.4 (1H, m), 4.55 (1H, bs) and 3.3-4.0 ppm (3H, m).

67853 136

Trans-3-phthalimidomethyl-1- (paranitrobenzyl-2-triphenylphosphoranylidene-2'-acetate) -4-tritylthio-2-azetidinone (§rW-r ____ © Hih

CO PNB CO PNB

2 2

A mixture of trans-3-phthalimidomethyl-1- (paranitrobenzyl-2'-chloro-2'-acetate) -4-tritylthio-2-azetidione (26 g, 35.5 mmol), triphenylphosphine (10 , 25 g, 39.1 mmol) and 2,6-lutidine (4.6 ml, 39.1 mmol) in dioxane (200 ml) were heated at 100 ° C for 20 h. The reaction mixture was filtered through Celite and evaporated to dryness. The residue was chromatographed on a silica gel column (350 g) eluting with benzene- * benzene / ether (1: 1) to give phosphorane (21 g, 62%) as a white solid, ir (CHCl 3) +: g: 1750 and 1710 cm -1. 1 Hmr (CDCl 3) δ: 7.4 (38H, m), 4.8-5.4 (3H, m), 4.6 (2H, m) and 3.7 ppm (1H bs).

Trans-3-phthalamidomethyl-1- (paranitrobenzyl-21-triphenylphosphoranylidene-21-acetate) -4-tritylthio-2-acetidininone 0 CTCOJI bf 3

CO ^ NB

A cooled (ice bath) suspension of trans-3-phthalimidomethyl-1- (paranitrobenzyl-2'-triphenylphosphoranylidene-2-acetate) -4-tritylthio-2-azetidinone (18.2 g, 18.83 mmol) in tetrahydrofuran (30 mL), water (30 mL) and acetone (30 mL), treated dropwise with sodium sulfide (4.97 g, 20.7 mmol) in acetone / water 1: 1 (30 mL), and heated to reflux. h. The reaction mixture was diluted with water, acidified to 1-norm. hydrochloric acid and extracted with diclocrime-67853 137 methane. The organic extracts were washed with brine and evaporated to dryness in vacuo to give 17.1 g (8.8%) of the title compound as an amorphous pale yellow solid. It was used without further purification in the next step.

ir (fine) ^. : 3150-3600, 1750 and 1700 cm -1 Hmr maxs (CDCl 3) rf: 7.4 (38H, m) and 3.3-5.5 ppm (8H, m).

Trans-3-phthaloisoimidomethyl-1- (paranitrobenzyl 2'-triphenylphosphoranylidene-2'-acetate) -4-tritylthio-2-azetidinone (o) S> 'M. ? hsc *.

---- *

CO PUB do PUB

2 2

A solution of trans-3-phthalimidomethyl-1- (paranitrobenzyl-2'-triphenylphosphoranylidene-2'-acetate) -4-tritylthio-2-azetidinone (17.1 g, 17.54 mmol) in dichloroxymethane (125 mL), treated by the dropwise addition of N, N'-dicyclohexylcarbodiimide (3.62 g, 17.54 mmol) in dichloromethane (30 mL) at room temperature. The solution was filtered through Celite and evaporated to dryness to give the title compound (18.23 g, quantitative) as an oil. It was used without further purification in the next step, ir (fine) \). : 2110, _-i n vmax '1755 3a 1710 cm; Hmr (CDCl 3) δ: 7.5 (38H, m), 4.6-5.3 (4H, m) and 3.9 ppm (2H, bs).

13 8 67853

Trans-3-aminomethyl-1- (paranitrobenzyl-2'-triphenylphosphoranylidene-21-acetate) -4-tritylthio-2-azetidinone (1) 1. ri 'Π ° = 7.r / = N 1 -' * J-N ^ Ρφ

oy CO PHB

CO PUB 2 2

A solution of trans-3-phthaloisoimidomethyl) -1- (paranitrobenzyl 2 * -triphenylphosphoranylidene-2'-acetate) -4-tritylthio-2-azetidinone (5.9 g, 6.16 mmol) in tetrahydrofuran (40 mL), cooled to -20 ° C, was treated dropwise with hydrazine (0.2 mL, 6.16 mmol) under N-shielding and stirring was continued for 30 min. The reaction mixture was acidified to 1-norm. hydrochloric acid and washed with ether; the aqueous phase was basified with 1 mol. sodium hydrogen carbonate and extracted with methylene chloride. The organic extracts were washed with brine, dried (MgSO 4) and evaporated to dryness. The residue was purified on a silica gel column (60 g) eluting with ether -> ethyl acetate to give 3.38 g (66%) of aminophosphorane as an amorphous solid, ir (CHCl 3) ν max: 1730 »1710 cm -1; 1 Hmr (CDCl 3) δ: 6.5-8.1 (34H, m), 3.8-5.3 (6H, m) and 0.9-1.9 ppm (2H, m).

Trans-3-formamidomethyl-1- (paranitrobenzyl-2'-triphenylphosphoranylidene-2'-acetate) -4-tritylthio-2-azetidinone

I CO PUB

CO PUB 2 To a cooled (ice bath) solution of trans-3- (aminomethyl) -1- (paranitrobenzyl 2'-triphenylphosphoranylidene-2'-acetate) -4-tritylthio-2-azetidinone (5.0 g, 6.04 mmol) in dichloromethane (50 mL) was added dropwise a solution of acetic formic anhydride (600 mg, 6.8 mmol) in dichloromethane (5 mL) under N 2 protection, followed by a solution of 67853 139 containing triethylamine. (1 mL, 7 mmol) in dichloromethane (2 mL). Stirring was continued for 30 minutes. The solution was washed successively with 1-norm. with hydrochloric acid, water, 1 mol. sodium bicarbonate and saline. The organic layer was dried (MgSO 4), evaporated to dryness and the residue chromatographed on a silica gel column (50 g). Elution with ether-ethyl acetate gave 2.0 g (39%) of formamide. as an amorphous solid, ir (CHCl 3) tfmax: 1740, 1685 and 1620 cm -1; 1 Hmr (CDCl 3) δ: 6.6-8.2 (35H, m), and 2.5-5.3 ppm (7H , m).

Trans-silver-3- (formamidometyyli-1- (para-2'-triphenylphosphoranylidene-2'-acetate) -2-azetidinone-4-thiolate

M II O 4 H

• vtr '' __ vt-r · J — N, Ρώ -N *. ., Ρφ, O * 3 c 3 CC ^ PMB ^ o2pmb

A solution of trans-3-formamidomethyl-1- (paranitrobenzyl 2'-triphenylphosphoranylidene-2'-acetate) -4-tritylthio-2-azetidinone (550 mg, 0.64 mmol) in dichloromethane (10 ml) was evaporated to dryness and diluted with hot methanol (20 mL). The solution was stirred at 60 ° C and treated with a preheated (60 ° C) solution of 0.15 mol. silver nitrate in methanol (5.7 mL, 0.86 mmol), followed by a solution of a 1.5 M solution of pyridine in methanol (0.57 mL, 0.86 mmol). The cream solution was stirred at room temperature for 30 minutes, then in an ice bath for 2 h. The solid was filtered off, washed with cold methanol and ether, and dried to give 300 mg (65%) of the silver salt as a beige solid. It was used without further purification in the next step.

140 67853

Trans-4-acetylthio-3-formamidometyyli-L- (paranitrobentsyy-yl-2'-triphenylphosphoranylidene-2'-acetate) -2-azetidinone

HC.N --► H

H I. ___ N Ρφ O ^ 1- -γ-3 CO2PNB co ^ pnb To a cooled solution (ice bath) of trans-silver 3-formamidomethyl-1- (paranitrobenzyl-2'-triphenylphosphoranylidene-2'-acetate) -2-azetidinone -4-Thiolate (800 mg, 1.11 mmol) in dichloromethane (10 mL) was added dropwise under 1 N mol. dichloromethane solution of acetyl chloride (1.33 mL, 1.33 mmol) followed by 1-mol. dichloromethane solution of pyridine (1.33 mL, 1.33 mmol). The solution was stirred in a cold bath for 1 h, and then filtered through Celite. The filtrate was washed successively with 1-norm. with hydrochloric acid, water, 1 mol. sodium bicarbonate and brine and the organic layer was dried (MgSO 4) and evaporated to dryness. The residue was purified on a silica gel column (5.0 g) and eluted with ethyl acetate- * 10% methanol in ethyl acetate to give 450 mg (62%) of the title compound; ir (CHCl 3) 0: 1755, 1685 and η -] O * Illci X s 1620 cm -1; Hmr '(CDCl 3) cf: 8.18 (2H, d, J = 9Hz), 7.0-8.0 (20H, m), 6.75 (2H, d, J = 9 Hz), 6.3 (1H, m), 5.5 (1H, m), 5.2 (2H, bs), 4.9 (1H, bs), 3.6 (1H, m), 3.0 (1H, m) and 2.2 ppm (3H, 4 s).

141 67853

Paranitrobenzyl 6-formamidomethyl-2-methylpenem-3-carboxylate

«H H n H

X · "" ·· Ps \

T CO PNB

CO PMB 2

A solution of trans-4-acetylthio-3-formamidomethyl · -1- (paranitrobenzyl-21-triphenylphosphoranylidene-2 * -acetate) -2-azetidinone (450 mg, 0.686 mmol) in toluene (10 mL) was heated at reflux for 12 hours. . Evaporation to dryness and purification on a silica gel column eluting with ether → 10% methanol in ether gave 100 mg (39%) of the penern compound as an amorphous solid, ir (CHCl 3). : 1780 11 3 Ymax and 1690 cm -1; Hmr (CDCl 3) δ: 8.2 (2H, d, J = 9Hz), 8.2 (1H, s), 7.6 (2H, d, J = 9Hz) , 6.9 (1H, m), 5.55 (1H, s), 5.35 (2H, 2s), 3.3-4.1 (3H, m) and 2.33 ppm (3H, s) .

6-formamidomethyl-2-methylpenem-3-carboxylic acid, sodium and potassium salts

CV ™ Vo2Qa © a „dP

A mixture of paranitrobenzyl 6-formamidomethyl-2-methylpenem-3-carboxylate (80 mg, 0.21 mmol), palladium / Celite catalyst (30%, 100 mg), tetrahydrofuran (10 mL), ether (25 mL ) and a 0.05 mol buffer solution pH 7 (4.46 mL, 0223 mmol) were hydrogenated for 3 h on a Parr shaker at an initial hydrogen pressure of 45 psi. The catalyst was removed by filtration through Celite and washed with water. The filtrate and washings were combined and the phases were separated. The aqueous phase was washed with ether (3 x 15 mL) and freeze-dried. The crude solid was purified by hplc to give 18 mg of a mixture of sodium and potassium salts.

142 67853 uv H20) ^ kB ’. 299 (£ 4933), 259 (£ 4094); and (new) V max: 3100-3650 3a 1755 cm -1; 1 Hmr (D 2 O) 8.15 (1H, s), 5.53 (1H, d, J = 1.4Hz), 4.0 (1H, m), 3.74 (2H, d, J = 5Hz), 3.25-4.25 (1H, m) and 2.27 ppm (3H, s).

Example 15 (1'S, 5R, 6S and 1 + R, 5S, 6R) -6- (11-hydroxy-1'-propyl) -2-methylpenem-3-carboxylic acid, sodium salt (Isomer C)

Et, 1H

-ΐο ~.

\ Xy: a

Trans-1-t-butyldimethylsilyl-3-propionyl-4-tritylthio-2-azetidinones o - fSTr

+ LDA + CH CU C-OCH - ► J

i 3 2 3 J— N

Ό ^ U.. ^ SiMe 1 ^ 2 -f- 2

Procedure: nBuLi (37.50 mL, 1.6 M / hexane solution, 60 mmol) was added dropwise to a cooled (dry ice-acetone bath) and stirred solution of diisopropylamine (8.50 mL, 60 mmol) in dry THF (200 mL). The mixture was stirred cold and 1-t-butyldimethylsilyl-4-tritylthio-2 * -azetidinone (22.9 g, 50 mmol) in dry THF (100 mL) was added. After 15 minutes, methyl propionate (40 mL, excess) was added and the reaction mixture was kept at -78 ° C for 4 h. The cooling bath was then removed and the internal temperature was allowed to rise to 0 ° C (40 min). The mixture was poured into ice-HCl (pH 66) and extracted with ether. The layers were separated and the aqueous layer was extracted with ether. The combined ether solution was washed with water and brine and dried (Na 2 SO 4). Evaporators 143 6785 3 were dried in vacuo to give an oil in quantitative yields. This contained a mixture of starting material and the title compound.

It was used as such and purified in the next step. IR (fine) λ max: 1710 (_ £ _) 1750 cm-1 (β-lactam).

1-t-butyldimethylsilyl-3- (1'-hydroxy-1'-propyl) -4-tritylthio-2-azetidinones

OH

^ A_vSTr ^ .wSIr

+ NaBH4 -V I

J-N. rA— N.

0 ^ SiMe 0 ^ siMe 2 2

Procedural:

A solution of 1-t-butyldimethylsilyl-3-propionyl-4-tritylthio-2-azetidinone (26 g, 50 mmol) and sodium borohydride (7.6 g, 200 mmol) in THF (400 mL). stirred at room temperature for 18 h. It was poured into ice-HCl (1N) (pH 6) and extracted with ether. The acidic phase was extracted several times with ether and the combined ether solution was washed with brine, dried (Na 2 SO 4) and evaporated to dryness to give 25.0 g of an amorphous solid. This crude product was chromatographed on Pigs (ACT. 1, 400 g) and first eluted with CH 2 Cl 2 to give 10.8 g of 1-t-butyldimethylsilyl-4-tritylthio-2-azetidinone. Elution with 20% ether in CH 2 Cl 2 gave 10.3 g of the title compound as a mixture of two isomeric trans alcohols. This was separated by hplc (Water Associates, System 500) and using a mixture of 10% ethyl acetate in CH 2 Cl 2 as eluent. Isomer C, white solid, 3.8 g; mp. (petroleum ether) 134-136 ° C. '' 'Hmr (CDCl 3) δ: 7.1-7.8 (15H, m, STr), 4.35 (H, d), 3.1 (H, dd), 2.5 (H, m ), 0.7-1.7 (5H, s) and 0.25 ppm (6H, s). Anal. Calcd for C 12 H 12 NO 2 Si: C 71.91, H 7.59, N 2.71; received; C 71.51, H 7.60, N 2.96. Isomer B, white solid, 5.4 g; mp (pentane-petroleum ether) 97-99 ° C. 1 Hmr (CDCl 3) δ: 7.1-7.8 (15H, m, STr), 4.15 (H, d), 3.4 (H, dd), 3.2 14 4 - 67853 ( H, m), 0.7-1.7 (5H, m), 0.85 (9H, s) and 0.1 ppm (6H, s).

9 7-9 9 ° C. 1 Hmr (CDCl 3): 7.1-7.8 (15H, m, STr), 4.15 (H, d), 3.4 (H, dd), 3.2 (H, m), 7-1.7 (5H, m), 0.85 (9H, s) and 0.1 ppm (6H, s) The total yield of these two alcohols (based on the starting material recovered) was 67.5%.

(1'S, 3S, 4R and '1'R, 3R, 4S) 1-t-butyldimethylsilyl-3- (1'-paranitrobenzyldioxycarbonyl-1'-propyl) -4-tritylthio-2-azetidinone (Isomer C)

? H OCO PNB

_ ^ STr. 2 ^ * STr Ί j + C1CO2PNB n-BuLi '· <p J— N * j ® SiMe2 0 ^ SiMe ^

Procedure To a cooled (dry ice-acetone bath) solution of (1'S, 3S, 4R and 1'R, 3R, 4S) tert-butyldomethylsilyl-3- (1'-hydroxy-1'-propyl) -4-tritylthio-2- Azetidinone (Isomer C) (3.1 g, 6 mmol) in dry THF (20 mL) was added dropwise a 1.6 mol solution under N 2 protection. n-BuLi / hexane (4.88 mL, 7.8 mmol) with stirring at -78 ° C for 25 minutes. Paranitrobenzyl chloroformate (1.56 g, 7.2 mmol) in dry THF (10 mL) was then added dropwise and the resulting mixture was stirred at -78 ° C for 4 h. Diluted with ether and washed with NH 4 Cl solution and brine. The organic phase was dried (Na 2 SO 4) and evaporated to dryness to give 4.2 g of the title compound (quantitative yield). 1 HHmr (CDCl 3): 8.2 (2H, d), 7.0-7.7 (17H, m), 5.13 (2H, s), 4, C 5 (H, d), 3.75 (H , dt), 3.25 (dd), 0.55-1.8 (5H, m), 0.9 (9H, s) and 0.25 ppm (6H, d).

67853 145 (1'S, 3S, 4R and 1'R, 3R, 4S) -3- (1'-Paranitrobenzyldioxycarbonyl-1-propyl) -4-tritylthio-2-acetidinone (Isomer C) oco2pnb oco2pnb vJ - .. ^ STr I KMPT NcLN '^ ^^

c - (10% H O; 30> H

SiMe 2 n

Procedure: To a cooled (ice bath) solution of (1'S, 3S, 4R and 1'R, 3R, 4S) -lt-butyldimethylsilyl-3- (1'-paranitrobenzyldioxycarbonyl-1'-propyl) -4-tritylthio- 2-Azetidinone (Isomer C) (4.2 g, 6 mmol) in HMPT (40 mL) with 10% H 2 O was added sodium azide. (0.78 g, 12 mmol). The mixture was stirred at room temperature for 1 h. Diluted with water (100 mL) and extracted with benzene / petroleum ether (1: 1) (4 x 15 mL). The organic phase was washed several times with water (6 x 30 mL) and brine. Dried (Na 2 SO 14) and evaporated to dryness to give 3.5 g of a solid (quantitative yield). It was treated with pentane and filtered to give 3.4 g of a pale yellow solid; mp. 84-86 ° C; 1 Hmr (CDCl 3) δ: 8.2 (2H, d), 7-7.7 (17H, n), 5.2 2H, s), 4.95 (H, dt), 4.4 (NH) , 4.25 (H, d), 3.4 (H, dd), 1.7 (2H, m) and 0.95 ppm (3H, t).

67853 146 (1'S, 3S, 4R and 1f R, 3R, 4S) -3- (11-paranitrobenzyldioxycarbonyl-1'-propyl) -1- (paranitrobenzyl-2 "-hydroxy-2" -acetate) -4- tritylthio-2-azetidinone (isomer C)

OCO PNB

,, | Jj + PNB glyoxylate ^ 7 * N ^ jch

H. CO PNB

Procedural:

To a solution of (1'S, 3S, 4R and 1'RjSRjHSi-S-d'-paranitrobenzyldioxycarbonyl-1'-propyl) -4-tritylthio-2-azetidinone (Isomer C) (3.2 g, 5 .5 mmol) and paranitrobenzyl glyoxylate hydrate (1.362 g, 6 mmol) in dry THF (50 mL) were added a catalytic amount of TEA (H drop) and Na 2 SO 4 + (to absorb the formed H 2 O). The resulting mixture was stirred at room temperature for 6 h. It was filtered and evaporated to dryness to give 4.35 g of an amorphous solid (in quantitative yields). 1 Hmr (CDCl 3) 8.25 (4H, dd), 7-7.9 (19H, m), 5.28 (2H, s), 5.1 (2H, s), 4.8 (H, d), 4.4 (H, dd), 4.1 (H, dt), 3.4 (H, m), 1.1-1.8 (2H, m) and 0.8 ppm (3H, t).

(1'S, 3S, 4R and 1'R, 3R, 4S) -3- (1'-paranitrobenzyldioxycarbonyl-1'-propyl) -1- (paranitrobenzyl-2 "-chloro-2" -acetate) -4- tritylthio-2-azetidinone (isomer C)

OCO PNB OCOjPNB

d ....., _ dTr \ A. (__> st · '

Py / THF I

cr N \ ^ ° H soci2 / thf * ¢ / —Nyd C ° 2pNB X2? nb

Procedure To a cooled (ice-salt bath) solution of the above glyoxylate (4.35 g, 5.5 mmol) in dry THF (30 mL) was added 1 M. py / THF solution (7 mL, 7 mmol) and then 1-mol was added dropwise. SOCl 2 / THF solution (7 mL, 7 mmol). The resulting mixture was stirred at the above temperature for 1 h. The mixture was diluted with benzene (30 mL), stirred cold (ice-water bath) for 30 min, and filtered using Celite charcoal. The filtrate was evaporated to dryness to give 3.8 g of an amorphous solid (85.3%). 1 Hmr (CDCl 3) δ: 8.15 (4H, d), 6.75-7.7 (19H, m), 5.65 (H, s), 5.2 (2H, s), 5.1 (2H, s), 4.5 (H, m), 3.85 (H, m), 3.4 (H, m), 1.25-2.0 (2H, m) and 0 .9 ppm (3H, t).

(1'S, 3S, 4R and 1'R, 3R, 4S) -3- (1f-paranitrobenzyldioxycarbonyl-1'-propyl) -1- (paranitrobenzyl-2 "-triphenylphosphoranylidene-2" -acetate) -4 -tritylthio-2-azetidinone (Isomer C)

OCO PNB OC02PNB

xy ^ STr ^ oJ_l Cl lutidine * 0J — I "^ ** 3 C02PNB co2pnb

Procedural:

To a solution of the above chlorine compound (3.7 g, 4.568 mmol) in dioxane (35 mL) was added 03P (1.197 g, 5 mmol) and lutidine (0.54 g, 5 mmol).

The mixture was heated in an oil bath at 100 ° C for 3 days. It was cooled, diluted with ether and washed successively with 1-norm. with riCl, 1 mol. NaHCO 3 and brine. Dried (Na 2 SO 4) and filtered through Velite charcoal. The filtrate was evaporated to dryness to give 3.6 g of an oil. This was chromatographed

SiO 2 (120 g) and eluted with benzene, benzene ether to give 1.45 g of the title compound as an amorphous solid (31%); IR (fine) ν max: 1750 cm -1 (broad).

14 8 67853

(1'S, 3S, 4R and 1'R, 3R, 4S) -4-Acetylthio-3- (1,1-paranixrobenzyldioxycarbonyl-1'-propyl) -1- (paranitrobercryl-2 "-triphenylphosphoranylidene-2 :, -acetate i) -2-acetidone (isomer C) qco PNB

0C0 PNB QCO PNB I 2 SAc

1 py / H °° H ~ "0 ^^ ρφ3 py / MeOH

I T 6o PNB

C02? NB CO PNB 2

Procedure:

To a hot solution (60 ° C) of the above phosphorane (1.4 g, 1.35 mmol) in MeOH (40 mL) was added with stirring a hot solution of AgNO 3 (0.3 g, 1.76 mmol). )

In MeOH (10 mL) followed by pyridine (0.107 g, 0.11 mL, 1.76 mmol). Silver mercaptide began to precipitate immediately. The mixture was stirred at room temperature for 15 minutes and at 0 ° C for 2 h. It was filtered and the solid was washed well with cold MeOH and ether; 1.2 g, (quantitative yield); mp. 113-115 ° C with decomposition; IR (nujol) ν max: 1740-1760 cm-1 (broad). This solid was used as such. To a cooled (ice bath) solution of the above mercaptide (1.2 g, 1.35 mmol) in Cl 2 Cl 2 (15 mL) was added acetyl chloride (0.118 g, 0.107 mL, 1.5 mmol) Cl 2 Cl 2. (2 mL) followed by pyridine (0.119 g, 0.122 mL, 1.5 mmol) in CH 2 Cl 2 (2 mL). The mixture was stirred at 0 ° C for 30 minutes. It was filtered through Celite to remove the silver salt and the filtrate was washed successively with HCl (0.5 M), H 2 O, NaHCO 3 (0.5 M) and brine. The CR 2 Cl 2 solution was dried (MgSO 4) and evaporated to dryness to give 0.94 g of the oxycone compound as an amorphous solid (83.4%). IR (fine) \). : 17 50 cm_J · (wide).

vmax 14 s 67 8 5 3 (1'S, 5R, 6S and 11R, 5S, 6R) -paranitrobenzyl 6- (1'-paranitrobenzyldioxycarbonyl-1'-propyl) -2-methylpenem-3-carboxylate (isomer C) oco2pnb QC02pnb

J-M

CO PNB

co2pnb 2

Procedure:

A solution of the above phosphorane (0.4 g, 1.077 mmol) in toluene (35 mL) was heated to reflux and 5 mL of toluene was distilled off. The yellow solution was boiled for 7.5 h. It was evaporated to dryness to give 0.75 g of a thick oil. This was chromatographed on SiO 2 (ACT 1.3 g) and eluted with benzene and benzene-ether to give the title compound as a solid, 0.32 g (53.4%); mp. (from pentane) 150-162 ° C; 1 Hmr (CDCl 3) δ: 7.3-8.4 (8H, m, aromatic), 5.4 (H, d), 5.3 (4H, benzyls, m), 5.0 (H, dt), 4.0 (H, dd), 2.35 (6H, s), 0.8 (2H, dq) and 1.0 ppm (3H, t).

(1'S, 5R, 6S and 1'R, 5S, 6R) -6- (1'-hydroxy-1'-propyl) -2-methylpenem-3-carboxylic acid (Isomer C), sodium salt 0CO2 pnb TV CH, ypd-Celite, '“'" rf5} - CH3 ^ Fosf aattipus- N _ + kur Χ °° 2

Procedural:

A mixture of the above ester (48 mg, 0.086 mmol) and 30% Pd / Celite catalyst (100 mg) in THF (10 mL), Et 2 O (20 mL), H 2 O (10 mL) ) and phosphate buffer (pH 7, 2 mL) were hydrogenated for 23 h at an initial pressure of 50 psi. Filtered through Celite and separated the layers. The organic layer was washed with H 2 O 67853

15G

(2 x 5 mL) and the combined aqueous layer was washed with ethyl acetate (2 x 10 mL). The aqueous layer was then freeze-dried to give the oxycone compound as a white salt, 30 mg; IR (KBr) γ max: (β-lactam), and 1600-1650 cm -1 (broad, -CO 2); uv ^^^ g5 2 & s (£ 1105) and 305 (£ 1244).

Example 16 6-Dimethylaminomethyl-2-methyl-penem-3-carboxylic acid 2h 1- (t-butyldimethylsilyl) -3-dimethylaminomethyl-4-tritylthio-2-azetidinone (cis and trans).

J1 j seΦ ch3v / sA

Ψ2 Me2NH -f 3 0J-1 ^ s / e2 NaBH3CN 'CH3 o ^ -N \ s / e2.

^ t-Bu ^ t-Bu

To a solution of dimethylamine (18.5 ml of a 2N methanol solution) (36.9 mmol) in methanol (80 ml) was added a solution of hydrogen chloride in methanol (2.5 ml of a 5N methanol solution), followed by trans. -1- (t-butyldimethylsilyl) -3-formyl-4-tritylthio-2-azetidinone (3.0 g, 6.16 mmol) and sodium cyanoborohydride (0.27 g, 4.31 mmol). The mixture was stirred at room temperature for 3.5 h, poured into glacial acid mixture (pH = 2) and basified with sodium hydroxide (1N NaOH, pH = 9). The mixture was extracted with ether and the ether phase was washed with brine, dried and evaporated to dryness to give the title compound as a crude oil (3.0 g).

151 67853 cis- and trans-3-dimethylaminomethyl-4-tritylthio-2-azetidinone

He> sa, 3 Ρ5 3 Μ, Ν3 «'[" Γ J Jj Ί ^ Γ ~ * ** 0 ^ Si Ή-Βυ

A solution of the above crude compound (3.0 g, 6 mmol) in hexamethylphosphoric triamide (HMPT, 16 mL) in water (10%) was cooled (5 °) and treated with sodium azide (0.78 g, 12 mmol). The mixture was stirred at room temperature for 1.5 h, poured into ice water and extracted with ether (5 x 30 mL). The organic phases were extracted with hydrochloric acid (1N) and the acidic extracts were washed well with ether to remove HMPT. The acidic phase was basified (1-norm, MaOH) and extracted with dichloromethane. The organic layer was washed with brine, dried and evaporated to dryness to give the title compounds as an amorphous white solid (1.5 g, total 62.5%). The mixture of isomers was separated with Waters Prep 500, eluting with methanol (5%), ammonia (0.2%), ethyl acetate (95%). Trans isomer: 1.0 g, m.p. 129-131 ° C (pentane); (ppm, CDCl 3): 6.8-7.8 (15H, m, aromatic), 4.5 (1H, NH), 4.28 (1H, d, J = 2.5, H-1!), 3.35 (1H, m, H-3), 2.75-2.1 (2H, m, H-1 '), 2.3 (6H, s, CH 2). Cis-isomer: 0.5 g, m.p. 132-133 ° C (ether pentane); (ppm, CDCl 3): 7.7-6.7 (15H, m, aromatic), * +, 72 (1H, NH), 4.5 (1H, d, J = 5.3, H-4), 3.5 (1H, m, H-3), 2.85-2.35 (2H, m, H-1 '), 2.31 (6H, s, CH 2 ). The cis: trans ratio can be varied by changing the conditions.

cis- and trans-6-dimethylaminomethyl-2-methylpenem-3-carboxylic acid

The title compound was prepared from cis- and trans-3-dimethylaminomethyl-4-tritylthio-2-azetidinone by the method of Example 15. (ppm, CDCl 3): 5.5 (1H ', d, J = 1.3), 3.7 (1H, dt, J = 1.3, J = 8), 2.8 (2H, d, J = 8), 2.35 (6H, s), 2.3 (3H, s).

152 p · .7 67853

Example 1 / (1'R, 5R, 6S and 1'S, 5S, 6R) -6- (11-hydroxy-11-propyl) -2-methyl-penem-3-carboxylic acid, sodium and potassium salts (Isomer B)

HQ _.H

Xco2h 1 f R, 3S, 4R and 1'S, 3R, 4S) 1-t'-butyldimethylsilyl-3- (1'-formyloxy-1'-propyl) -4-tritylthio-2-azeidinone (Isomer B)

OH HCO 2 H, Et 3 N OCHO

. STr DMAJ? , Ac 2 O 1) J-N CH Cl 2 O 2 SiMe 2 2 0 SiMe 2 2 4-Dimethylaminopyridine (DMAP) was prepared according to the method described by a) H.C. Brown et al., Org. Synth. Collect. Butter. 5, 977 (1973) and b) Helmet Vorbruggen et al., Angew.

Chem. Int. Ed., 17, 569, (1978).

Procedure: To a cooled (0 ° C) solution of (1'R, 35.4R and 1'S, 3R, 4-S) -lt-butyldimethylsilyl-3- (1'-hydroxy-1'-propyl). li) -4-Tritylthio-2-azetidinone (Isomer B) (3.612 g, 7 mol) in CH 2 Cl 2 (50 mL), Et 2 N (4.48 mL, 35 mmol) was added. , HCO 2 H (0.63 mL, 16.8 mmol) and DMAP (0.854 g, 7 mmol) were added dropwise followed by acetic acid hydride (7.14 g, 70 mmol). The clear yellow solution was stirred at -40 ° C and the mixture was milky. It was poured into a mixture of ice-1-norm. HCl (pH 6) and the layers were separated. The CH 2 Cl 2 solution was washed with 1 M mol. NaHCO 3 and brine. It was dried (Na 2 SO 4) and evaporated to dryness to give 3.8 g of a solid residue. This was treated with pentane and filtered to give 3.7 g of a white solid (96.8%); mp. 12-127 ° C; ir (fine) O, 1720 ° (H-C-) 0 and 1750 cm-3 (β-lactam); Hmr (CDCl3): 7.1 (H, s, HC-) 6.7-7.7 (15H, m), 4.8 (H, m), 4.05 (H, d, J = 1.5), 3.7 (H, m, 153 67853 J = 15 5, J = 7), 1.4 (2H, m), 0.95 (9H, s), 0.8 (3H , t) and 0.1 ppm (6H, s); Anal. Calcd for C 18 H 18 NO 8 SSi: C, 70.42; H 7.20; M 2.57; Found: C, 70.2; H 7.33; II 2.73.

(1'R, 3S, 4R and 1'S, 3R, 4S) -3- (1'-formyloxy-1'-propyl) -4-tritylthio-2-azetidinone (Isomer B)

OCHO OCHO

-JSTt \ A_ ^ STr HMPT]

+ NaN -> I

^ SiMe 3 «<> (Τ 'NX

2 ^ H

Procedure: To a cooled solution (ice bath) of (3.7 g, 6.77 mol) in HMPT (40 mL) with 10% H 2 O was added NaN (0.91 g, 14 mmol). The mixture was stirred at room temperature for 1.5 hours. It was poured into ice water (200 ml) and extracted with ether (4 x 40 ml). The ether solution was diluted with petroleum ether and washed thoroughly with water and brine to remove HMPT. It was dried (^ 250 ^) and evaporated to dryness to give 2.92 g of a thick colorless oil (in quantitative yields). Hmr (CDCl 3) δ: 8.1 (H, h_q_ 7.1-7.7 (15H, m, -STr), 5.23 (H, m, J = 7), 4.38 (H, d, J = 2.5), 4.3 (H, -NH), 3.35 (H, dd, J = 2.5, J-7), 1.75 (2H, m) and 1.0 ppm ( 3H, t).

15 4 67853 (11R, 3S, 4R and 1'SjSR ^ SJ-S-d'-formyloxy-1'-ethyl) -1- (paranitrobenzyl-2 "-hydroxy-2" -acetate) -4- tritylthio-2-azetidinone (isomer B)

OCHO OCHO

_> STr 1 _ ^ STr

P jb glyoxylate

^ - \ H Et ^ N / THF V cr \ ^ 0H

CO_PNB

2

Procedural:

A mixture of 3- (1'-formyloxy-1'-propyl) -4-tritylthio-2-azetidinone (Isomer B), (2.9 g, 6.77 mmol), PNB glyoxylate (1, 59 g, 7 mmol), Et 2 N (5 drops) and Na 2 SO 4 (dry, 5.0 g) in THF (50 mL) were stirred at room temperature for 18 h. Filtered and evaporated to dryness to give an amorphous solid. in quantitative yields (4.33 g); 1 Hmr (CDCl 3) δ: 8.2 (2H, d), 7.1-7.8 (18H, m), 5.2 (2H, d), 4.9 (H, m), 4.65 and 4.3 (H, 4.65 (1/2 H, s)) 4.3 (1/2 H, s) /, 4.2-4.3 (H, d, 1/2 H at 4.2 , 1/2 H at 4 ·, 3), 3.65 (H, m), 1.4 (2H, m) and 0.8 ppm (3H, t).

(1'R, 3S, 4R and 1S, 3R, 4S) -3- (1,1-formyloxy-1'-propyl) -1- (paranitrobenzyl-2 "-chloro-2" -acetate) -4- tritylthio-2-azetidinone (isomer B)

OCHO OCHO

/ 1 .. ^ STr \ ^ ·· -—r »STr I-1 Py / THF j gj ΝγΟΗ SOCl2 / THF O ^ N \ ^ - C1

CO PNB CO PNB

2 2

Procedure: To a cooled solution (ice-salt bath) of the above glyoxylate (4.3 g, 6.77 mmol) and 1-mol. A solution of py / THF (8 mL, 8 mmol) in dry THF (30 mL) was added dropwise 1-mol. SO 2 Cl 2 / py solution (8 mL, 8 mmol). The resulting solid mixture was stirred at the above temperature for one hour. Dilute with benzene (30 mL) and stirring was continued for 67853 155. The mixture was filtered using Celite charcoal and the filtrate was evaporated to dryness to give 4.1 g of an amorphous solid (92%). ir (fine) J: 1720 0 0 K Ks (H-C-), 1750 (-C-PNB) and 1780 cm -1 (β-lactam); 1 Hmr (CDCl 2) δ: 8.25 (2H, d), 7.8 (H, s, "" 7-7.75 (17H, m), 5.25 HC-), (2H, d), 5.0 (H, m), 4.6 (H, s), 4.4 (H, d), 3.7 (H, m), 1.6 (2H, m) and 0.9 ppm ( 3H, t).

(1'R, 3S, 4R and 1'S, 3R, 4S) -3- (1'-formyloxy-1'-propyl) -1- (paranitrobenzyl-2 "-triphenylphosphoranylidene-2" -acetate) -4-tritylthio -2-azetidinone (isomer B)

OCHO OCHO

/ STr. _ ^ STr || 03P dioxane || J — n ci \ 7! ! * —N ρφ O νγ 'lutidine ° Vs ^ 3

CO PNB CO PNB

2 2

Procedural:

A mixture of the above chlorine compound (4.0 g, 6.07 mmol), OgP (1.834 g, 7 mmol) and lutidine (0.749 g, 7 mmol) in dioxane (40 mL) was heated to 100 ° C: s (oil bath) 2 days. It was cooled, diluted with ether and washed successively with cold 1N HCl solution, 1M. NaHCO 3 and brine. The organic solution was dried (NagSO 4) and filtered using Celite charcoal.

The solution was evaporated to dryness to give an oil which was chromatographed on SiO 2 (Act. 1, 200 g) and eluted with benzene and benzene-ether to give 2.60 g of the title compound as an amorphous solid (48.45%). ; IR (fine) tmax: 1720 "* max (H-C-O-), and 1750-1760 cm-1 (-CO2-riB and -actam).

67853 156 (1'R, 3S, 4R and 1'S, 3R, 4S) -4-acetylthio-3- (1'-formyloxy-1'-propyl) -1- (paranitrobenzyl-2 "-triphenylphosphoranylidene- 2 "-acetate) -2-azetidinone (isomer B)

PCH0 QCHC OCHO

STr _S 'SA9 _ λ SAc' oflyp * 3 * C02PNB co2pnb cc2pnb. Procedure: Warm solution (60 ° C) with 0.15 mol. A solution of AgNOg-CH 2 OH (8.7 mL, 1.3 mmol) was added to a mixture of the above phosphorane (0.88 g, 1 mmol) and pyridine (0.103 g, 1.3 mmol) in MeOH: (5 mL) heated to 60 ° C. The mixture was stirred at room temperature for 15 min. and at 0 ° C for 2 h.

It was filtered and washed with cold MeOH to give 0.53 g of silver mercaptide as a yellow solid (71%) which was used as such. To a cooled (ice bath) mixture of the above mercaptide (0.53 g, 0.71 mmol) and pyridine (0.079 g, 1 mmol) in Cl 2 Cl 2 (10 mL) was added dropwise CH 2 Cl 2 (0.079 g). , 1 mmol) in Cl 2 Cl 2 (5 mL). After stirring at 0 ° C for 1 h, the mixture was filtered. The filtrate was washed well with cold 0.5 mol. With HCl solution, 0.5 mol. NaHCO 3 and brine. Dried (Na 2 SO 4) and evaporated to dryness to give 0.1 g of an oil, (63%); Ir (fine) ^,: 1700-1760 ”* 1 0 iriciKs cm (broad ,, and fb lactam).

-C '67853 157 (1'Κ, 35, Μ-Κ and 1'S, 3R, 4S) - and acetylthio-3- (1'-hydroxy-1'-propyl) -1- (paranitrobenzyl-2 "-triphenylphosphoranyl- di-2 "acetate) -2-azetidinone (Isomer B)

OCHO pH

nA_ ^ SA0 Jw5 * 1 I HCl / MeOH_ ^ j

f I

CG PNB CO PNB

2 2

Procedural:

The above formate (1.0 g, 1.45 mmol) in THF (10 mL) was treated with HCl / MeOH (10 mL, prepared from 2 mL of concentrated HCl and diluted with MeOH at room temperature). To a volume of 24 ml). The mixture was kept at room temperature for 0.5 h. It was basified with 1 M NaHCO 3, extracted with ethyl acetate, washed with brine and dried (£30%). Evaporated to dryness to give 0.9 g of crude title compound. This was chromatographed on Pigs and eluted with ether and ether-ethyl acetate (1: 1) to give 0.6 g of pure title compound as an amorphous solid (62.5%); 1 Hmr (CDCl 3) δ: 82.5 (2H, d), 7.3-δ, Ι (17H, m, aromatic), 5.6 (H, m), 5.2 (2H), 4.9 (H), 4.4 (H, m), 2.3 (3H, SAc), 1.5 (2H, m) and 0.9 ppm (3H, t).

67853 158 (1'R, 5R, 6S and 1'S, 5S, 6R) -paranitrobenzyl 6 - (. 1'-hydroxy-1,1-propyl) -2-methylpenera-3-carboxylate (isomer B) OH oh onY, 3 - *

T CO.PNB

CO PNB 2 2

Procedural:

The above phosphorane (0.2 g, C, 3 mmol) was heated in toluene (45 mL) at reflux and 5 mL of toluene was distilled off. The resulting solution was boiled for 6 h. It was cooled and evaporated to dryness to give 0.2 g of an oil. This was chromatographed on Pigs and eluted with ether to give 0.1 g of the title compound as a white solid; (87%); mp. (from pentane) 133-135 ° C; 1 Hmr (CDCl 3) 8.3 (2H, d), 7.6 (2H, d), 5.6 (H, d), 5.35 (2H, d), 4.15 (H, m) , 3.8 (H, m), 2.4 (3H, s, CH 2), 2.2 (H, OH), 1.7 (2H, m) and 1.05 ppm 3H, t).

(1'R, 5R, 6S and 1'S, 5S, 6R) -6- (1'-hydroxy-1'-propyl) -2-methylpenem-3-carboxylic acid (Isomer B), K and Na salts a mixture of

OH? H

- .Ei>; ·· 'CO PNB CO (Ua + K) 2 1

Procedural:

A mixture of the above ester (0.07 g, 0.185 mmol), 30% Pd / Celite catalyst (150 mg) and a buffer solution (pH 7, 4 mL) in THF (15 mL), Et 2 O (25 mL) and deionized water (15 mL) was hydrogenated for 4 h at an initial pressure of 48 psi. Filtered using Celite and the layers were separated. The aqueous layer was washed with ethyl acetate and then lyophilized to give 91 mg of a solid; ir (KBr Vma] <s: 1780 (β-lactam) and 1650 cm -1 (broad, -CO 2 "); uv H 2 O 2 max: 255 (£ 983) and 300 (£ 1092).

67853 159

Example 18 (1'R, 5R, 6S and i'S, 5S, 6R) -6- (1'-hydroxy-2'-phenylethyl) -2-methylpenem-3-carboxylic acid (Isomer B)

H. yPH

3

TO H

Trans-1- (t-butyldimethylsilylD-S-phenylacetyl-4-ylthio-thio-4-azetidinone r- / iTr

+ LDA + <J> CH CO Et -I

σ 2 0 ^ —N- | iMe2

A solution of t-butyldimethylsilyl-4-tritylthio-2-azetidinone (18.32 g, 40 mmol) in dry THF (100 mL) was added dropwise to a cooled (-78 ° C) LDA solution / prepared under protection. At -78 ° C by dropwise addition of 1.6-mol, n-BuLi (101.25 mL, 162 mmol) to a solution of diisopropylamine (22.95 mL, 162 mmol) in dry THF (150 ml) and stirred at -78 ° C for 30 minutes. The mixture was stirred at -78 ° C for 30 minutes and ethyl phenyl acetate (15.66 g, 15.12 mL, 93.6 mmol) in dry THF (50 mL) was added and the reaction mixture was stirred at -78 ° C for 2 h.

It was poured into a mixture of ice and 1N HCl (pH 5-6) and extracted several times with ether. The ether solution was washed with brine and dried (Na 2 SO 4). It was evaporated to dryness to give 33.7 g of a crude solid. This was dissolved in ether (10 ml) and triturated with pentane (200 ml). The solid was filtered off and washed several times with pentane to give 18.3 g of a white solid (79.6%), m.p. 141-143 °. 1 Hmr (CDCl 3: 7.0-7.6 (20H, m), 4.8 (H, d), 3.7 (H, d), 3.53 (H, s), 3.43 (H , s), 1.5 (9H, s) and 0.3 ppm (6H, s).

160 6785 3 1- (t-butyldimethylsilyl-3- (1'-hydroxy-2'-phenylethyl) -4-tritylthio-2-azetidinone (2 trans diastereomers)

OH

n </ - Ν ^

A mixture of trans-1- (t-butyldimethylsilyl) -3-phenylacetyl-4-tritylthio-2-azetidinone (28.8 g, 50 mmol) and NaBH 4 (0.5 g, 0.25 mmol) in THF (200 ml), stirred at room temperature for 18 h, the mixture was poured into a mixture of ice and 1N HCl and extracted with CH 2 Cl 2, washed with brine and dried (Na 2 SO 4). to give an amorphous solid (27.7 g) A portion of the solid (23.0 g) was chromatographed on SiO 2 and eluted with hexane / ether to give an off-white solid (14.4 g) which was found to be a 1: 1 mixture of (1'R, 3S, 4R and 1'S, 3R, 4S) and (1'S, 3S, 4R and 1 * R, 3R, 5S) isomers. 1 Hmr (CDCl 2) 7-7.7 (20H, m), 4.37 (1 / 2H, d), 4.18 (1 / 2H, d), 3.3-3.8 (H, m), 3.45 ( 1 / 2H, dd), 3.1 (1 / 2H, dd) # 2.7 (2H, m), 0.87 (9H, d) and 0.25 ppm (6H, s).

1- (t-butyldimethylsilyl) -3- (1'-formyloxy-21-phenylethyl) -4-tritylthio-2-azetidinone oh O (H. Ocho); ^ STr V * STr Γ i -, "rf. '" Rf 0 ^ N _ | _iMe2 0 ^ NSiMe2

isomer B isomer C

To a cooled solution (-40 ° C) of a mixture of the above alcohols (14.4 g, 24.9 mmol) in CH 2 Cl 2 (250 mL) was added EtgN (15.93 mL, 125 mmol), HClOH. (2.24 mL, 59.75 mmol) and DMAP (3.04 g, 24.9 mmol). After stirring for 5 minutes, acetic anhydride (2.35 mL, 249 mmol) was added dropwise. The clear solution was stirred at -40 ° C for 15 minutes, whereupon it turned into a white, cloudy mixture. It was kept at -40 ° C for another 45 minutes, (total time 1 h). It was poured into ice and 1-norm. HCl, and the layers were separated. The CH 2 Cl 2 solution was washed well with cold 1-norm. With HCl, IVO, 1-mol. NaHCO 3 and brine. Dried (MgSO 4) and evaporated to dryness to give 14.0 g of an amorphous solid. This was separated by hplc (Water Associates, System 500) to give "Isomer B" 6.0 g, m.p. 172-173 ° C and "Isomer C" 6.0 g, m.p. 188-189 ° C.

Total yield of pure compound, i.e. 12 g (73.2%).

Isomer C: 1 H m (CDCl 3) 6.8-7.7 (21H, m), 5.05 (H, dt), 4.05 (CH, d) 3.65 and 3.75 (H, two doublets ), 2.7-2.9 (2H, s), 0.88 (9H, s) and C, 2 ppm (6H, s). Isomer B: 1 Hmr (CDCl 3) ci ': 7.75 (H, s), 6.9-75, (20H, m), 4.3 (H, dt), 3.95 (H, d), 3, 37 (H, dd), 2.95 (H, s), s), 2.85 (H, s), 0.9 (9H, s) and 0.2 ppm (6H, s).

3- (11-Formyloxy-2 * -phenylethyl) -4-tritylthio-2-azetidinone (1'R, 3S, 4R and 1'S, 3R, 4S enantiomers)

OCHO

^ OCHO STR 0 <_fSTl

Si «e2 N ^ h

Isomer B

To a cooled solution (ice bath) of the above formate (5.9 g, 9.375 mmol) in HMPT with 10% water (50 mL) was added NaN® (1.3 g, 20 mmol).

O

The mixture was stirred at room temperature for 1.5 hours. It was poured into ice water (300 ml) and extracted with ether (3 x 100 ml). The ether solution was washed well with water and brine. It was dried (Na 2 SO 4) and evaporated to dryness to give a solid residue. This was treated with petroleum ether and eluted with 67853 162 to give 4.4 g of a white solid (92%), m.p. 169-171 ° C. Analysis calculated for C 75.43; H 5.51; N 2.84; Found: C, 75.04; H 5.64; N 2.78.

1 Hmr (CDCl 3): 7.9 (H, s), 7.1-7.6 (20H, m), 5.4 (H, m), 4.6 (H, NH), 4.2 (H , d), 3.3 (H, dd), 3.15 (H, s) and 3.0 (H, s).

3- (1'-Formyloxy-2'-phenylethyl) -1- (paranitrobenzyl-2 "-hydroxy-2n-acetate) -4-tritylthio-2-azetidinone C1'R, 3S, 4R- and 11S, 3R, 4S enantiomers) H ·> ° CHO »° CHO STr φΛ / ^ STr φ \> <_ ^ STr

'"I f -► I

. M-N 2 - OH

Ύ

CO ^ NB

A suspension of PNB glyoxylate (2.37 g, 10.16 mmol) in dry benzene (100 mL) was boiled under a Dean Stark apparatus (filled with a 3A molecular sieve) for 2 hours. The above N-H compound (4.2 g, 8.537 mmol) was then added and boiling was continued for another hour. After cooling to room temperature, Et 2 N (0.12 mL, 0.85 mmol) was added and the mixture was stirred at room temperature for 1.5 h.

It was evaporated to dryness to give the title compound in quantitative yield as a mixture of two alcohol isomers.

"1 Hmr (CDCl 3) 2: 8.0-83, (2H, two doublets), 7.5 and 7.6 (H, two singlets), 7.0-7.4 (20H, m), 5, 25 (2H, d), 4.9 (H, OH), 4.25 and 4.35 (H, two doublets), 3.5-4.5 (H, m, broad), 3.1-3 .3 (H, m) and 2.9 ppm (2H, m).

67853 163 3- (1'-Formyloxy-2'-phenylethyl) -1- (paranitrobenzyl-2 "-chloro-2" -acetate) -4-tritylthio-2-azetidinone (1'R, 3S, 4R- and 1'S , 3R, 4S-enantiomer)

H OCHO H., .OCHO

_ ^ »STr ^> 5Tr J— Νγ-0Η d ^ NY" cl C02PNB co2pnb

To a cold (ice-salt bath) solution of the above glyoxylate (6.0 g, 8.537 mmol) in dry THF (30 mL) was added 1 mol of pyridine. THF solution (10 mL, 10 mmol) and then 1-mol was added dropwise. thionyl chloride in THF (10 mL, 10 mmol). After one hour at the above temperature, the mixture was diluted with benzene (30 ml) and stirring was continued cold for 30 minutes. The mixture was filtered through a pad of Celite charcoal and evaporated to dryness to give 6.0 g of an amorphous solid (98%): "Hrnr (CDCl 3) +: 8.2 (2H, m), 7-7.7 (23H , m), 5.8 (H, s), 5.25 (2H, s), 4.35 (H, d), 3.5-4.0 (H, m), 3.3 (H, m) and 2.9 ppm (2H, d).

3- (1'-formyloxy-2'-phenylethyl) -1-paranitrobenzyl-2 "-triphenylphosphoranylidene-2" -acetate) -4-tritylthio-2-azetidinone (1'R, 3S, 4R- and IS, 3R, 4S-enantiomer).

H. .OCHO STr .V ^ 0010 STr ♦ o <1_> ^ <. s I ~ J-fj Ρφ oi_N Cl ° 3

C02PNB C02PNB

J

A mixture of the above chlorine compound (6.0 g, 8.333 mmol), 03P (2.489 g, 0.5 mmol) and lutidine (1.0165 g, 1.1 mL, 9.5 mmol) was present. in dioxane (50 mL) was heated at 110 ° C (pour temperature) for 18 h. The mixture was cooled and filtered through Celite. The filtrate was diluted with ethyl acetate and washed with cold 1N HCl, H 2 O, 16 4 67853 l-mol. NaHCO 3 and brine. It was dried (Na 2 SO 4) and evaporated to dryness to give 3.0 g of crude product.

This was chromatographed on SiO 2. and eluted with ether / hexane (1: 1) and ether to give 4.0 g of the title compound. Sp. (needles from ether) 235-237 ° C (decomposed), (01%); IR (membrane) λ max: 1720, 1750 cm-1.

4-Acetylthio-3- (1'-formyloxy-2'-phenylethyl) -1- (para-nitrobenzyl-2,1-triphenylphosphoranylidene-2 "-acetate) -2-azetidinone (1'R, 3S , 4R- and 1'S, 3R, 4S-enantiomers) H 0CH0 H .. OCHO Φ H '"X ^ 0CH0> SAc _ / Ίτ __ / SAg __ —f da -y' Ρφ Cr-

0 + ~ A V * 3 (/ “" ν '3 T

Ιθ2ΡΝΒ t02 ™ B C ° 2PNB

To a boiling solution of the above phosphorane (3.6 g, 3.8 mmol) and pyridine (0.33 g, 4.2 mmol) in CH 2 Cl 2 (30 mL) and MeOH (30 mL) was added by dropwise addition of 0.15 mol. AgNO 2 / MeOH solution (28 mL, 4.2 mmol). The mixture was stirred at room temperature for 2.15 h. It was concentrated to low volume (1 mL), cooled and filtered to give the silver mercaptide as a yellow solid (2.3 g, 77%). To a mixture of this mercaptide and pyridine (0.277 g, 3.5 mmol) in ice-cold Cl 2 Cl 2 (20 mL) was added dropwise a solution of CH 2 Cl 2 (0.27 g, 3.5 mmol) in CH 2 Cl 2 (15 mL). The mixture was stirred at room temperature for 3 h, filtered through Celite and the filtrate was washed with cold 1 N HCl, H 2 O, 1 M NaHCO 3 and brine, dried (MgSO 4) and evaporated to dryness to give 1. 0 g 67853 165 amorphous solid (83.8%). 1 Hmr (CDCl 3) δ: 8.2 (2H, d), 7.0-8.0 (23H, m), 4.5-5.7 (4H, m), 2.6-3.3 (3H, m), and 2.3 ppm (2H, d, SAc).

4-acetylthio-3- (1'-hydroxy-2'-phenylethyl) -1- (paranitrobenzyl-2 "-triphenylphosphoranylidene-2" -acetate) "2-azetidinone (1'R, 3S, 4R- and 1'S, 3R, 4S-enantiomers and) H oh H „,> ° CHO φ SAc <> <_> SAc _s [----- ρφ Λ-N ρφ3 3 Τ CO ΡΝΒ CO ΡΝΒ 2 2

A solution of the above phosphorane (1.8 g, 2.416 mmol) in THF (10 mL) was treated with a 1-norm solution. HCl / MeOH (10 mL) and the mixture was stirred at room temperature for 4 h. It was concentrated to remove methanol, diluted with cold water, basified with 1 M mol. NaHCO 3 and extracted with CHCl 3. The CHCl 3 solution was dried (MgSO 4) and evaporated to dryness to give 1.65 g of an amorphous solid.

This was chromatographed on Pigs and eluted with ether / ethyl acetate to give 1.30 g of the title compound (75%). 1 Hmr (CDCl 3) δ: 8.2 (2H, d), 6.7-8.0 (22H, m), 4.0-6.0 (5H, m), 2.5-3.5 ( 3H, mm) and 2.2 ppm (3H, SAc).

166 67853

Paranitrobenzyl 6- (1'-hydroxy-2'-phenylethyl) -2-methylpenem-3-carboxylate (i, 5R, 6S and 1'S, 5S, 6R enantiomers) H. * ° h H.> 0H> 7 ς rC *. - · or 3 'co2pnb co2pnb

A solution of the above phosphopran (1.2 g, 1.67 mmol) in toluene (80 mL) was heated to reflux (10 mL was distilled off to remove moisture present and the low boiling solvent) for 6 hours. Evaporated to dryness and the crude product was chromatographed on SiO 2. The title compound was obtained by eluting the column with ether to give 0.65 g of an amorphous solid (89%). 1 Hmr (CDCl 3) δ: 8.2 (2H, d), 7.6 (2H, d), 5.4 (H, d), 5.2-5.4 (2H, d), 4, 0-4.5 (H, m), 3.7-4.0 (H, dd), 3.0 (2H, d), and 2.3 ppm (3H, s).

6- (1'-Hyaroxy-21-phenylethyl) -2-methylpenem-3-carboxylic acid (1'R, 5R, 6S and 11S, 5S, 6R enantiomers)

H. OH

H ... + ° H

NC02PNB 1

A mixture of paranitrobenzyl ester (0.33 g, 0.75 mmol), 0.05 mol. buffer solution (pH 7, 17.4 ml), THF (30 ml),

Et 2 O (30 mL), distilled H 2 O (60 mL), and 30% Pd / Celite catalyst (0.69 g) were hydrogenated for 24 h at an initial pressure of 50 psi. The mixture was filtered through Celite and the organic layer was washed with water. The combined aqueous layer was washed several times with ethyl acetate and lyophilized for 18 h to give the title compound as a yellow solid salt. This was treated with 167 67853 small amounts of water, acidified with cold 1-norm. HCl and extracted well with CHCl 3. The CHCl 3 solution was dried (MgSO 4) and evaporated to dryness to give a solid residue. This was treated with ether and filtered to give 30 mg of a white solid (13.2%), m.p. 165-167 ° C; IR (nujol) tmax: 3580 (OH, sharp), 1660 and 1760 cm-1; uv (MeOH) -max: 310 (£ 5490) and 254 (£ 4880).

Example 13 (4'R, 5R, 6S and 4'S, 5S, 6R) -6- (2 ', 21-dimethyl-11,3'-dioxolan-4'-yl) -2-methylpenem-3-carboxylic acid (isomer C)

Nco h A. Manufacture of compounds Ö Ö, ίzf ’· Γί x / X / * ·.

(4'R, 3S, 4R and 4'S, 3R, 4S) and (4'S, 3S, 4R and 4'R, 3R, 4S) 1- (t-butyldimethylsilyl) -3- (2 ', 2'-dimethyl- 1 ', 3'-dioxolan-4'-yl) -4-tritylthio-2-azetidinone ("Isomer C" and "Isomer B") 67853 168

Ethyl O- (2-methoxy-2-propyl) glycolate.

^ 0Et POCl3 iHG

To a solution of ethyl glycolate (16 μg, 0.150 mol; freshly distilled) and 2-methoxypropene (16.4 g, 0.216 mol, 2 μl 95% pure) in CH 2 Cl 2 (150 mL) was added 0-5 phosphorus oxychloride (3 drops, 35 mg, 0.23 mmol) and the mixture was stirred at 0-5 ° for 15 minutes and at room temperature for 1.5 h. This was then quenched with pyridine (30 drops), stirred for 45 minutes and the solvent was evaporated off. . The residue was diluted with pentane (150 mL) and dried over K 2 CO 3. After filtration, the solvent was evaporated to give 27.83 g (0.158 mol, 100%; purity 94.9%) of the title compound as a colorless oil: 1 Hmr (CCl 4) g: 1.25 (3H, t, J = 7Hz -CH 2 CH 3). ), 1.28 (6H, s, Me 2), 3.12 (3H, s, -OCH 3), 3.88 (2H, s, -OCH 2 CO-), 4.10 (2H, q, J = 7Hz, -CH2CH3); ir (fine) \ / k: 1760 and 1735 cm’’1 (ester).

67853 169 1) J. Meinwald et al., Tet. Lett., 4327 (1973) 2) M.S. Newman and M.C. Vander Zwan, J. Org. Chem., '38, 2910 (1973).

(3S, 4R and 3R, 4S) -1- (t-butyldimethylsilyl) -3- (11-keto-2'- (2 "-methoxy-2" -isopropyloxy) -1'-ethyl) -4-trityl- thio-2-azetidinone - ^ STr 1) LDA / THF ^) _f'STr /> C OMe / "v <

To a stirred solution of diisopropylamine (18.5 mL, 0.134 mol) in THF (400 mL; freshly distilled from LAH) at -78 ° C was added n-butyllithium (1.6 M in hexane, 90 ml, 0.144 mol) under the protection of the N 2 atmosphere. After 30 minutes, a solution of 1- (t-butyldimethylsilyl) -4-tritylthio-2-azetidinone (50.0 g, 0.109 mol) in THF (100 mL) was added dropwise over 10 minutes and the mixture was stirred for 5 minutes. To this pink solution was added ethyl O- (2-methoxy-2-propyl) glycolate (23.94 g, 0.136 mol) and the mixture was stirred for 1 h. After removal to a dry ice bath, saturated NH 4 Cl solution (200 mL) was added and followed by brine (100 mL). The aqueous phase was extracted with Et 2 O (3 x 100 mL). The combined organic extracts were washed with brine, dried (Na 2 SO 4) and evaporated to dryness to give 60.95 g (0.103 mol; crude product yield 94.6%) of the title compound as a crude orange oil. This raw material was used in the next reaction. A pure sample was obtained by column chromatography (SiO-, eluent: 2% 67853 170

Et 2 O in benzene); 1 Hmr (CDCl 3) δ: 0.30 (6H, s, Si-CH 2), 0.95 (9H, s, t-Bu), 1.12 (3H, s, CH 2)> 1, Δ (3H, s, CH 2), 3.15 (3H, s, OCH 2), 3.57 (1H, AB's A, J = 17Hz), 3.77 (1H, d, O g Cm J = 1.6Hz, H-3), 3.97 (1H, AB's 3, J = 17Hz), 4.83 (1H, d, J = 1.6Hz, H-4), 7.1-7, Δ (15H, m, aromatic Hs); ir (fine) ^ max; 1750> 1725> 1710 cm -1 (C = O); tic, Rf 0.53 (benzene; Et 2 O = 4: 1), Rf 0.61 (hexane: EtOAc ethyl acetate = 2: 1).

(3S4R and 3R, 4S) -1- (t-butyldimethylsilyl) -3- (1'-hydroxy-2'-methoxyisopropyloxyethyl) -4-tritylthio-2-azetidinone (mixture of epimers of C-1 ') _ / STr NaBH4. -ySTr / x κ y

Solution of crude 1- (t-butyldimethylsilyl) -3- (1'-keto-2 '- (2' -methoxy-2 "-isopropyloxy) -1'-ethyl) -4-triethylthio-2- azetidinone (50.95 g, 0.103 mol) in THF (100 ml) was diluted with absolute ethanol (350 ml) and to this solution was added haBH 4 (4.88 g, 0.156 mol) at 0 ° C. at 2h and quenched by the slow addition of 67853 171 brine (280 mL) The mixture was extracted with Et 2 O 3 (3 x 150 mL) and the extracts were washed with brine, dried (Na 2 SO 4) and evaporated to dryness to give a yellow residue which was redissolved in CH 2 Cl 2. This was re-dried (Na 2 SO 4) and evaporated to dryness to give 57.1 g (0.0966 mol, crude yield 93.8%) of the title compound as a crude yellow foam; 1 Hmr (CDCl 3) &lt; : 0.17 (s, SiCH 2), 0.80, 0.87 (2s, Si-tBu), 1.22, 1.25 (2s, CH 2), 3.03 (s, OCH 2), 4, 32 (d, J = 2Hz, H-4), 7.0-7.7 (m, aromatic Hs); ir (fine) ^ max 31 + 60 1745 (C = O), 1595 (aromatic): Rf 0.47 and 0.42 (hexanes: EtOAc ethyl acetate = 2: 1).

This crude material was used without purification in the next step.

(4R, 3S, 4R and 4'S, 3R, 4S) and 4'S, 3S, 4R and 4R, 3R, 4S) -1- (t-butyldimethylsilyl) -3- (2 ', 2', 2'-dimethyl- 1 ', 3f-dioxolan-4S-yl) -4-tritylthio-2-azetidinone (Isomer C and Isomer JD_ _ -r-S5 ™ P-T5 ° H'H2 ° uMe I, v xOMe J— N ^ / o ^ vsi / X * ^ / ^

A solution of (3S, 4R and 4R, 4S) -1- (t-butyldimethylsilyl) -3- (11-hydroxy-2'-methoxyisopropyloxyethyl) -4-tritylthio-2-azetidinone (Cl * mixture of diastereomers) (57.1 g, 0.0966 mol; crude) in CH 2 Cl 2 (500 ml) was treated at room temperature with p-toluenesulfonic acid monohydrate (200 mg) and 2,2-dimethoxypropane (20 ml) and then stirred for 1 hour. The mixture was washed with saturated NaHCO 3 and then brine, dried (Na 2 SO 4) and evaporated to dryness to give 49.64 g (0.088 mol, crude yield 91.9%) of a mixture of the title compounds (Isomer B and Isomer C) as a yellowish foam. This was purified. By HPLC (Waters 500 Silicagel; eluent hexane: ethyl acetate = 9: 1) and crystallization to give 14.28 g (25.5 mmol, 26.4%) of the title compound (Isomer C) as white crystals, mp 146 -147 ° C (pentane); 1 Hmr (CCl 4) g: 0.27 (6H, s, Si-CH 2), 0.95 (9H, s, Si-tBu), 1.15 (6H, S, di-Me), 2.5-2.9 (1H, m, H-4 '), 2.97 172 δ 7853 t, J = 1.8 Hz, H-3), 3.25-3.9 (2H, m, H-5 »), 4.27 (1H, d, J = 1.8Hz, H-4) ), 7.1-7.6 (15H, m, aromatic Hs); ir (nujol) ^ ma.kS: 1 z (C = O) and 1595 cm -1 (aromatic); Rf 0.45 (hexanes: AtOAc ethyl acetate = 4: 1) and 14.50 g (25.9 mmol, 25.9% yield) of the title compound (Isomer B) as white crystals: m.p. 144-145 ° C (Et 2 O-pentane); 1 Hmr (CCl 2) f: 0.02 (6H, s, SiMe), 0.8 33 (9H, s, Si-tBu), 1.13, 1.18 (6H, 2s, diMe), 2 Δ 5-2.8 (1H, m, H-4 '), 3.3-4.1 (2H, m, H-5'), 3.48 (1H, dd, J 34 = 1.5 Hz, J3_4 = 5.0Hz, H-3), 3.93 (1H, d, J4_3 = 1.5Hz, H-4), 7.1-7.7 (15H, m, aromatic Hs); ): 1350 (C = O) and 1595 cm -1 (aromatic); Rf 0.37 (hexanes: EtOAc ethyl acetate ^:!). Analysis calculated for c33H41NO3SSi: 2.50, S 5.73, obtained (isomer C): C 70.23, H 7.30, N 2.41, S 5.53, and (Isomer B): C 70.52, H 7.31 N 2.40, S 5.05.

B. Preparation of Penem (Isomer C) (4'R, 3S, 4R and 4'S, 3R, 4S) -3- (2 ', 2'-dimethyl-1', 3'-dioxolan-4'-yl) -4-Tritylthio-2-azetidinone (Isomer C)

to -V-Q

_ ^ STr Ρλ, _> STr pT NaN3_ „'| —f rJ N / HMPT-H-0 j — N ^ u ^ Si, 2 0 Mi /

To a stirred solution of (4'R, 3S, 4R- and 4'S, 3R, 4S) -1- (t-butyl-dimethylsilyl) -3- (2 ', 2'-dimethyl-1', 3'-dioxo -lan-4'-yl) -4-tritylthio-2-azetidinone (Isomer C), (14.3 g, 25.6 mmol) in hexamethylphosphoric triamide (230.4 mL) was added slowly (over 20 minutes) at 0-5 ° In C, a solution of sodium azide (2.50 g, 38.4 mmol; 1.5 eq.) In Hau (25.6 mL). The mixture was stirred at room temperature for 2 L and poured into cold water (2.5 L). The resulting white precipitate was collected, washed with Et 2 O and dried to give 11.26 g (25.3 mmol, crude yield 98.8%) of the title compound as a white solid. The pure material was obtained by crystallization from 67853 173 CH O mixture; mp. 192-193 ° C (dec.); 1 Hmr (CDCl 3) δ S: 1.33, 1.37 (6H, 2s, di-Me), 3.27 (1H, t, J = 3Hz, H-3), 3.8-9.4 ( 3K, m, H-4 'and H-5'), 4.40 (1H, d, J = 3Hz, H-4), 4.47 (1H, br, NH, exchanged) and 7.1-7 .7 ppm (15H, m, aromatic Hs); ir (nujol) Vmax: 3220 (NH), 1760 (C = O) and 1950 cm -1 (aromatic); Rf 0.31 (hexanes: EtOAc ethyl acetate = 3: 2).

(4'R, 3S, 4R and 4'S, 3R, 4S) -3- [2 ', 2'-dimethyl-1', 3'-dioxolan-41-yl) -1- (p- nitrobenzyl 2 "-hydroxy-2" -acetate) -4-tritylthio-2-azetidinone (mixture of epimers of C-2 ") (Isomer C) .STr CHO

qJ— NH 4 t ^ N / THF --N

CO ^ PNB

A suspension of p-nitrobenzyl glyoxylate hydrate (6.57 g, 28.95 mmol; 1.15 eq.) In benzene (500 mL) was heated at reflux using a Dean-Stark trap for 2 hours. Evaporation of the solvent gave p-nitrobenzylglyoxylate as an oil. A mixture of this oil and (4R, 3S, 4P and 4fS, 3R, 4S) -3- (2 ', 2 * -dimethyl-1', 31-dioxolan-4'-yl) -trityl-lithio-2-azetidinone (Isomer C) (11.2 g, 25.2 mmol) in THI (350 mL, distilled over LAH) was treated with triethylamine (289 mg, 2.86 mmol) at room temperature under N 2. protected for 18 h (overnight). After evaporation of the solvent, the residue, diluted with CH 2 Cl 2 (200 ml), was washed successively with 1N saline. HCl (2.9 mL), saturated NaHCO 3 and brine, dried (Na 2 SO 4) and evaporated to dryness after addition of Et 2 O (30 mL) to give 17.2 g (26.3 mmol, crude yield 100 purity 95.8 -i>) of the title compound as a white foam; Rf 0.40 and 0.30 (benzene: Et 2 O = 3: 2). Each isomer was separated by hplc (SiO 2 9 as eluent benzene: Et 2 O = 3.2) and purified by crystallization from CH 2 Cl 2 -Et 2 O. Isomer I: Rf 0.40 (benzene: Et 2 O = 3: 2), m.p. . 153-154 ° C; 1 Hmr (CDCl 3) δ: 1.20 (6H, s, di-Me), 3.1 (2H, m, H-3 and OH), 3.5-4.2 (3H, m, H-4 67853 174 and H-51), 4.55 (1H, d, J = 2Hz, H-4), 5.12 QH, br, H-2 "), 5.30 (2H, s, QCH 2 Ar) and 7.1-8.3 ppm (19H, m, aromatic Hs); ir (nujol) Umajjs: 3370 (OH), 1775 (β-lactam) and 1745 cm-1 (ester); Analysis calculated for C36H34N2O8S : C 66.04, H 5.23, N 4.28, found: C 65.85, H 5.64, N 4.11.

Isomer II: Rf 0.30 (benzene: Et 2 O = 3: 2); mp. 164-165 ° C; 1 Hmr (CDCl 3) tf: 1.17 (6H, s, di-Me), 3.2 (2H, m, H-3 and OH), 3.4-4.0 (3H, m, H-4 ' and H-5 '), 4.57 (1H, d, J = 2Hz, H-4), 5.23 (1H, br, -2 "), 5.27 (2H, s, -OCH 2 Ar), and 7.1-8.3 ppm (19H, m, aromatic Hs); ir (nujol): 3340 (OH), 1765-lactam) and 1740 cm-1 (ester) Analysis calculated for C 36 H 34 N 2 O 8 S: C 66'04ί H 5'23 »N 2 + 28; s 4.90; found: C 66.01; H 5.34; N 4.28; S 4.75.

4'R, 3S, 4R and 3'S, 3R, 4S) -3- (2 ', 2'-methylmethyl-11,3'-dioxolan-4'-yl) -1- (p-nitrobenzyl-2 "chlorine -2 "-acetate) -4-tritylthio-2-azetidinone (mixture of epimers of C-2") (Isomer C) ^ 3 ·, _SOCl2-Pyr_ ^ - |> STr J tl m thf 0 ^ -

I CO PNB

C02PNB 2

To a stirred solution of (3fR, 3S, 4R 3a 4'S, 3R, 4S) -3- (2 ', 2'-dimethyl-1 *, 3'-dioxolan-41-yl) -1- (p-nitrobenzyl- 2 "-hydroxy-2" -acetate) -4-tritylthio-2-azetidinone (Isomer C) (17.13 g, 25.07 mmol; mixture of epimers from C-2 ") in THF (250 mL) was added. At -15 ° C under N 2 protection under pyridine (2.84 mL, 35.1 mmol ·) followed immediately by thionyl chloride (2.20 mL, 30.1 mmol; Anachemia). minutes at -15 ° and then the white precipitate was filtered off.After washing with benzene, the filtrates and washings were combined and evaporated to dryness.Dissolved in benzene (250 ml) the residue was treated with activated charcoal, filtered and evaporated to dryness to give 17.94 g 67853 175 ( 26.65 mmol, crude yield 100%; purity 94.1%) of the crude title compound as a white foam: Rf 0.76 (benzene:

Et 2 O = 3: 2); 1 Hmr (CDCl 3) δ: 1.20 (6H, s, diMe), 3.17 (1H, m, H-3), 3.4-3.9 (3H, m, H-41 and H-Sr) , 4.67, 4.72 (1H, 2d, J = 2.5 Hz, H-4), 5.30 (2H, s, OCH 2 Ar), 5.83 (s, H-2. ') and 7.1-8.3 ppm (19 H, m, aromatic H 5); ir (fine) Vmax: 1770 cm (β-lactam and ester). This material was used without purification in the next step.

(4'R, 3S, 4R and 4 * S, 3R, 4S) -3 - (', 21-dimethyl-11,3'-dioxolan-41-yl-1- (p-nitrobenzyl-2 "-triphenylphosphoranylidene) 2 "-acetate) -4-tritylthio-2-azetidinone (isomer C) ~ t ° i pv Joi i-iL Cl dioxane IJ ρφ o <r 3

C02PNB C02PNB

A mixture of 4'R, 3S, 4R and 4'S, 3R, 4S) -3- (2 *, 2'-dimethyl-1 ', 3'-dioxolan-4-yl) -1- (p-nitrobenzyl -2 ,, -chloro-2 "-acetate) -4-tritylthio-2-azetidinone (Isomer C) (17.87 g, 25.0 mmol; purity 94.1% of a mixture of C-2" epimers) , triphenylphosphine (7.27 g, 27.5 mmol) and 2,6-lutidine (3.19 mL, 27.5 mmol) in dioxane (350 mL; distilled over LAH) were heated at reflux under N 2 for 40 h. Evaporation of the solvent in vacuo gave 29.5 g of a dark oil which was purified by column chromatography (SiO 2 330 g; eluent .20-50% Et 2 O in benzene) to give 10.5 g of a yellowish solid. This solid was rinsed with Et 2 O to give 7.49 g (8.33 mmol, 33.3% yield) of the title compound as slightly yellow crystals; 1 Hmr (CDCl 3) δ: 1.07 (s, di-Me) and 7.1-8.2 ppm (m, aromatic H 5); and (nujoli) Vmax: 1760 cm -1 (C = O). An analytical sample was obtained by crystallization from CH 2 Cl 2 -Et 2 O: m.p. 231-232 ° C; Analysis calculated for C 51 H 47 N 2 O 2 PS: C 72.14; H 5.27; N 3.12; S 3.57; 1.6 67853 Found: C 72.18; H 5.43; N 2.98; S 3.41;

Rf 0.17 (benzene: Et 2 O = 1: 1).

(4R, 3S, 4R and 4'o, 3R, 4S) -Silver-3- (2 ', 2'-dimethyl-11,3,3-dioxolan-4'-yl) -1- (p-nitrobenzyl) 2 "-triphenyl] phosphoranylidene-2" -acetate) -2-azetidinone-4-thiolate (Isomer C) 7 ^ tQ · Vr- ^ STr Α, ΝΟ, -Pyr I n * MeOH J- N prj, < / -N ^ Ph Ύ 3 ίο PNB C02rm 2

Solution of (4'R, 3S, 4R and 4'S, 3R, 4S) -3- (1,2'-dimethyl-1 ', 3'-dioxolan-4-yl) -1- (p-nitrobenzyl) -2 '* - triphenylphosphoranylidene-2'-acetate) -4-tritylthio-2-azetidinone (Isomer C) (319 mg, 0.355 mmol) in CH 2 Cl 2 (10 mL) was evaporated to dryness to give an oily residue which redissolved in hot methanol (8 mL; 60 °). To this solution was added a hot MeOH solution of AgNO 2 (0.15 mol, 4, C mL, 0.60 mmol) at 60 ° and then pyridine (29 μL, 0.36 mmol). The mixture was stirred at room temperature for 5 h and at 0 ° C for 1 h. The precipitate was collected and washed with ice-cold methanol and then cold Et 2 O to give 255 mg (0.334 mmol, yield 94, i%) of the title compound as a brownish solid; And tnujoli)

Ws: 1750 cm-1 (s = c = 0) · 41R, 3S, 4R and 4'S, 3R, 4S) -3- (2 ', 2'-dimethyl-1', 3'-dioxolan-4 ' -yl) -1- (p-nitrobenzyl-2 "-triphenylphosphoranylidene-2" -acetate) -4-acetylthio-2-azetidinone (Isomer C) N ΡΦ,

0 ^ ‘V 3 ^ J

ro PNB

C07PN3 2

To a solution of (4'R, 3S, 4R and 41S, 3R, 4S) -silver-3-67853,177 (2 ', 2'-dimethyl-1,3'-dioxolan-4'-yl) -1 - (p-Nitrobenzyl <'' triphenylphosphoranylidene-2 "acetate) -2-azetidinone-4-thio [*] (Isomer C) (25 * + mg, 0.333 mmol) in CH 2 Cl 2 (15 and pyridine (100 μL, 1.24 mmol; 3.72 eq.) Acetyl chloride (71 μL, 1.0 mmol; 3.0 eq.) was added at 0-5 ° C. stirred at 0-5 ° C for 40 minutes After the precipitate was filtered through Celite, the filtrate was washed successively with brine containing 1N HCl (1.25 mL), saturated NaHCO 3 then brine, dried ( Na 2 SO 4) and evaporated to dryness to give 200 mg of an oil which was crystallized from Et 2 O to give 155 mg (0.222 mmol, 66.7% yield) of the title compound as white crystals: 1 Hmr (CDCl 3) 1.23 (s, di -Me), 2.20, 2.33 (2s, -SAc) and 7.2-8.3 ppm (m, aromatic Hs): ir Cnujoli) t / max · 17 50 (Ji-lactam and ester) and 1690 cm -1 (thioester) the sample was obtained by crystallization from CH 2 Cl 2 -Et 2 O: m.p. 177-178 ° C; Analysis calculated for C37H35N2O8PS: C, 63.60; H 5.05; N 4.01; S 4.59; Found: C, 63.34; H 5.32; N 3.83; S 4.31.

Rf 0.62 (ethyl acetate).

(4'R, 5R, 6S and 4'S, 4S, 6R) -p-Nitrobenzyl-6- (2 ', 2'-dimethyl-1', 3'-dioxolan-4'-yl) -2-methylpenem-3 -carboxylate (Isomer C) Ö- ^ SAC toluene-.

oJZi ρφ3 * '

J CO.PNR

co2pnb 1

A suspension of (4'R, 3S, 4R and 4'S, 3R, 4S) -3- (2 *, 2r-dimethyl-1 ', 3'-dioxolan-4'-yl) -1- (p-nitrobenzyl -2 "-triphenylphosphoranylidene-2" -acetate) -4-acetylthio-2-azetidinone (Isomer C) (443 mg, 0.634 mmol) in toluene (70 ml) was heated at reflux under N 2 for 6 h. Evaporation of the solvent gave a white solid which was purified by column chromatography (SiO 2 10 g; eluent 10% Et 2 O in benzene) to give 247 mg (0.587 mmol, 67853,178 yield 92.7%) of the title compound as a white solid; 1 Hmr (CDCl 3) δ: 1.42 (6H, s, di-Me), 2.38 (3H, s, 2-CH 2), 3.8-4.5 (4H, m, H-6, H- 4 »and H-5 '), 5.02-5.25-5.33-5.57 (2H, AB type, -OCH 2 Ar), 5.57 (1H, d, J = 1.8 Hz, H -5) and 7.52-7.67-8.12-8.27 ppm (4H, 7 ^ ^ 2 ^ aromatic Hs); ir (nujoli) (max: 1760-lactam) and 1700 cm-1 (ester). An analytical sample was obtained by crystallization from CH 2 Cl 2 -Et 2 O: m.p. 167-168 ° C; uv (EtOH)? max: 265 (* · 14 »000) 3a 314 m / u (£ 10,000);

Analysis calculated for C 18 H 18 N 2 O 2 S: C, 54.28; H 4.79; N 6.66; S 7.63, found: C 54.15; H 4.78; N 6.54; S 7.64;

Rf 0.62 (benzene-Et 2 O = 1: 1).

(4'R, 5R, 6S) and 4'S, 5S, 6R) -6- (21,2'-dimethyl-1 ', 3'-dioxolan-41-yl) -2-methylpenem-3-carboxylic acid (Isomer C ) / <! / Deg.

_,> sv H / Pd-C V - "" ·, -r-> S \ on> CH>

\ o PNB CO H

2 ^

A solution of (4 * R, 5R, 6S and 4 * S, 5S, 6R) -nitrobenzyl-6- (2 ', 2'-dimethyl-1', 3'-dioxolan-4-yl) -2- methyl penem-3-carboxylate (Isomer C) (195 mg, 0.464 mmol) in THF (20 mL) was stirred in Et 2 O (20 mL), H 2 O (20 mL), NaHCO 3 (39 mg, 0.46 mmol). mmol) and 10% Pd-C catalyst (200 mg, Engelhard). This mixture was hydrogenated at 35 psi for 4 h at room temperature. After removal of the catalyst (using Celite) the aqueous layer was washed with ethyl acetate (2 times), saturated with NaCl, acidified to 1-norm. HCl (0.47 mL) and immediately extracted with ethyl acetate (3 x 20 mL). The brine washed extracts were dried (Na 2 SO 4) and evaporated to dryness to give 94 mg of a yellowish solid which was rinsed with pentane to give 89 mg (0.31 mmol, 179 6785 3% yield 67%) of the title compound as a yellowish solid: m.p. 132-133 ° C; Rf 0.60 (acetone: HOAc = 5: 0.7); 1 Hmr (CDCl 3) δ: 1.37, 1.43 (6H, 2s, di-Me), 2.36 (3H; s, 2-CH 3>, 3.9-4.6 (4H, m, H -6, H-41 and H-5 ') and 5.59 ppm (1H, d, J = 1.7 Hz, H-5), ir (nujol) δ: 1760 (β-lactam) and 1660 cm-1. 1 (CO 2 H); uv (EtOH) /? Max: 309 (£ 630 °) and 263 m -1 (£ 3800).

Example 20 (4'S, 5R, 6S and 4'R, 5S, 6R <) -6- (2 ', 21-dimethyl-N, 31-dioxolan-4'-yl) -2-methylpenem-3-carboxylic acid (Isomer B) „Π>“ · co2h (4'S, 3S, 4R and 4'R, 3R, 4SÖ> -3- (2 ·, 2'-dimethyl-1 *, 3'-dioxolan-4'-yl) -4 -tritylthio-2-azetidinone (Isomer B) ivrr V "

HMPT-H ^ O

/> <

The title compound was prepared as described in Example 51 "Isomer C for (4'S, 3S, 4R and 4'R, 3R, 4S) -1- (t-butyldimethylsilyl) -3- (2 ', 21-dimethyl-1' From 3'-dioxolan-3'-yl) -4-tritylthio-2-azetidinone (Isomer B) (14.4 g, 25.8 mmol), yield 10.8 g, 24.3 mmol, 94.1% mp 155 ° C (CH 2 Cl 2 -Et 2 O) Rf 0.24 (hexanes: ethyl acetate = 2: 1);

Hmr (CDCl 3) 1.37, 1.40 (6H, 2s, di-Me), 3.23 (1H, dd, J 3-4 = 2.5

Hz, J3_4, = 5Hz, H-3), 3.7-4.5 (4H, m, H-4 », H-5», NH), 4.50 (1H, d, J = 2.5Hz , H-4) and 7.1-7.6 ppm (15H, m, aromatic Hs); ir (nujoli) 3170 (NH) and 1745 cm-1 (C = O);

Analysis calculated for C 27 H 27 NO 3 S: 180 67853 C 72.78; H 6.11; N 3.14; S 7.20; Found: C, 72.16; H 6.11; N 3.14; S 7.17.

(4'S, 3S, 4R and 4'R, 3R, 4S1 -3- (2 ', 2'-dimethyl-1', 3 * -dioxolan-4'-yl) -1- (p-nitrobenzyl-2 "- hydroxy-2 "-acetate) -4-tritylthio-2-azetidinone (mixture of epimers of C-2") (Isomer B)

cA STr VHO ofS

^ "'" 1-f co2pmb - [^ STr 0 ^ ~ N \ h "Et ^ N / THF> qJ-Ν, ^ Η

C02PNB

The title compound was prepared as described in Example 151 for "Isomer C" (4'S, 3S, 4R and 4R, 3R, 4S) -3- (2 ', 2'-dimethyl-1', 3'-dioxolane- 4'-yl) -4-tritylthio-2-azetidirone (Isomer B) (10.8 g, 24.3 mmol); yield 15.8 g, 24.1 mmol, 99.3%); yellowish foam; Rf 0.29 and 0.2 2 (benzene: ethyl acetate = 1: 1); 1 Hmr (CDCl 3) δ: 1.28, 1.34 (2s, di-Me), 3.4-4.4 (m, H-3, H-4,, H-5 ', H-2 ", 0H), 4.39, 4.53 (2d, J = 2Hz, H-4), 5.15, 5.25 (2s, OCH 2 Ar) and 7.1-8.3 ppm (m, aromatic Hs ); (fine) ^ ^ g ^ 3440 (br, OH), 1760 (C = O), 1520, 1350 cm -1 (NO 2).

(4'S, 3S, 4R and 4'R, 3R, 4S) -3- (2 ', 2'-dimethyl-1', 3'-dioxolan-4'-yl) -1- (p-nitrobenzyl-2 " -Chloro-2 "-acetate) -4-Tritylthio-2-azetidinone (mixture of C-2" epimers) (Isomer B)

Vk._SSTr SOCl, -pyr - (^ STr

Ql-i OH ^ '

T CO PNB

COPND 2 2

The title compound was prepared as described in Example 51 "Isomer C for (4'S, 3S, 4R and 4'R, 3R, 4S) -3-18186785 3 (2 ', 2'-dimethyl-1', 3 '- dioxolan-4'-yl) -1- (p-nitrobenzyl-2R-hydroxy-2 "-acetate) -4-tritylthio-2-azetidinone (Isomer B) (14.9 g, 22.8 mmol; mixture of C-2 "epimers); yield 14.1 g, 20.9 mmol, 91.9%; yellowish foam; Rf 0.52 (benzene: Et 2 O = 3: 2); 1 Hmr (CDCl 3) *:

1.30, 1.38 (6H, 2s, di-Me), 3.4-4.5 (4H, m, H-3, H-4,, H-5,) S

4.57 (1H, d, J = 3 Hz, H-4), 5.13 (s, H-2 "), 5.27 (s, OCH 2 Ar; and 7.1-8.3 ppm (19H, m , aromatic Hs); ir (fine): 17 8 0 cm -1 (γ4-lactam, ester).

(41S, 3S, 4R and 4'R, 3R, 4S) -3- (2 ', 2'-dimethyl-1', 3'-dioxolol-41-yl) -1- (p-nitrobenzyl-2 " triphenylphosphoranylidene-2 "-acetate) -4-tritylthio-2-azetidinone (Isomer B) dioxane 0

V C ° 2? NB

CO PNB 2

The title compound was prepared as described in Example 51 for "Isomer C" (4'S, 3S, 4R and 3'R, 3R, 4S) -3- (2 ', 2'-dimethyl-11,3'-dioxolan-4 '-yl) -1- (p-nitrobenzyl-2 "-chloro-2" -acetate) -4-tritylthio-2-azetidinone (Isomer B) (14.0 g, 20.8 mmol; C-2 " mixture of epimers), yield 4.64 g, 5.16 mmol, 24.8%, mp 190-195 ° C (dec.) (CH 2 Cl 2 -Et 2 O), 1 Hmr (CDCl 3) δ 1.12, 1.20, 1.27, 1.35 (4s, di-Me) and 7.0-8.1 ppm (m, aromatic Hs); ir (CH 2 Cl 2) νmax: 17 50 cm-lactam ester) Analysis calculated for C H 2 N 2 O 2 PS: C 72.14, H 5.27, N 3.12, S 3.37, found: C 71.90, H 5.57, N 3.07, S 3.55;

Rf 0.21 (benzene: Et 2 O = 1: 1).

182 6 7 8 5 3 (4'S, 3S, 4R and 4'R, 3S, 4R) -Silver-3- (2 ', 2'-dimethyl-1', 3'-dioxolan-4'-yl) -1 - (p-Nitrobenzyl-2 "-triphenylphosphoranylidene-2" -acetate) -2-azetidinone-4-thiolate (Isomer B) ' ρφ3 «eon ^ ο · ^" γρ * = C02PNB co2pnb

The title compound was prepared as described in Example 51 "Isomer C for (4'S, 3S, 4R and 4'R, 3S, 4R) -3- (2 ', 2'-dimethyl-1', 3f-dioxolan-4 '-yl) -1- (p-nitrobenzyl-2 "-triphenylphosphoranylidene-2" -acetate) -4-tritylthio-2-azetidinone (Isomer B) (1.00 g, 1.12 mmol); yield 580 mg, 0.760 mmol, 67.8%, mp 129-135 ° C (dec), λ IR (nujol) λ max: 1745 cm -1 lactam, ester.

(4'S, 3S, 4R and 4'R, 3R, 4S) -3- (21,2 * -dimethyl-1 ', 31-dioxolan-4'-yl) -1- (p-nitrobenzyl-2 "-triphenylphosphoranylidene -2 "-acetate) -4-acetylthio-2-azetidinone (Isomer B) SH9, O -., - (SAc oi-'VPh" Vh "

CO PNB CO PNB

2 *

The title compound was prepared as described in Example 51 for "Isomer C" (4'S, 3S, 4R and 4'R, 3R, 4S) -silver-3- (2 ', 21-dimethyl-1', 3'-dioxolane). -4'-yl) -1- (p-nitrobenzyl-2 "-triphenylphosphoranylidene-2" -acetate) -2-acetidinin-4-thiolate (Isomer B) (2.46 g, 3.22 mmol) ); yield after purification by column chromatography 67853 183 CSiO2 32 g, eluent 10% to 50% ethyl acetate in CH2Cl2 = 1: 1); ^ Hmr (CDCl 3) $ ·. 1.23, 1.27, 1.30 (3s, di-Me), 2.22, 2.33 (2s, SAc) and 7.3-8.3 ppm (m, aromatic Hs); ir (fine) Vmax 175 (β-lactam, ester) and 1695 cm-1 (thioester).

(4'S, 5R, 6S and 4TR, 5R, 6S and 4'R, 5S, 6R) -p-nitrobenzyl-6- (2 *, 2'-dimethyl-1 ', 3'-dioxolan-4'-yl) -2-methylpenem-3-carboxylate (Isomer B)

& 3 CC ^ PNB

co2pnb

The title compound was prepared as described in Example 51 for "Isomer C" (4'S, 3S, 4R and 4'R, 3R, 4S) -3- (2 ', 2'-dimethyl-1 *, 3 * -dioxolane- 4'-yl) -1- (p-nitrobenzyl-2 "-triphenylphosphoranylidene-2" -acetate) -4-acetylthio-2-azetidinone (Isomer B) (200 mg, 0.286 mmol); yield 64 mg, 0.15 mmol, 53%; mp. 151-152 ° C (CH 2 Cl 2 / Et 2 O); Rf 0.67 (Benzene: Et 2 O = 1: 1); - 1 Hmr (CDCl 3) δ: 1.29, 1.38 (6H, 2s, di-Me), 2.30 (3H, s, 2-CH 3), 3.6-4.4 (4H, m , H-6, H-4 ', H-5'), 5.00-5.18-5.28-5.46 (4H, ABq, -OCH 2 Ar), 5.47 (1H, J = 1, 5Hz, H-5) and 7.42-7.55-8.05-8.15 ppm (4H, A 9'B9t, aromatic Hs); _ 1 LL e -j ir (fine): 1785 cm (β-lactam) and 1710 cm x (ester); uv (EtOH) Δmax * 266 13,300) and 314 myu (£ 9,700);

Analysis calculated for C 19 H 20 N 2 O 7 S: C, 54.28; H 4.79; N 6.66; S 7.63; Found: C, 54.00; H 4.75; N 6.68; S 7.61.

184 67853 (4'S, 5R, 6S and 4'R, 5S, 6R) -6- (2 ', 2'-dimethyl-1', 3'-dioxolan-4'-yl) -2-methylpenem-3-carboxylic acid (isomer B> 0 -., - NC0 PNB 2

The title compound was prepared as described in Example 51 for "Isomer C" (4'S, 5R, 6S and 4'R, 5S, 6R) -p-nitrobenzyl-6- (2 *, 2 * -dimethyl-1 ', 3 (-dioxolan-4'-yl) -2-methylpenem-3-carboxylate (Isomer B) (79 mg, 0.19 mmol); yield after recrystallization from Cl 2 Cl 2 pentane mixture 9 mg, 0.03 2 mmol, 17%; Rf 0.54 (acetone: HOAc - 5: 0.5); 1 Hmr (CDCl 3) δ: 1.35, 1.44 (6H, 2s, di-He), 2.37 (3H, s, 2-CH 2), 3.6-4.5 (4H, m, H -6, H-4 ', H-5') and 5.56 ppm (1H, brs, H-5); ir (fine) v / ma ^ s: 1785 cm-2 (γ9-lactar); uv (EtOH)? ma] <s' · 307 (t 4300) and 262 m / u (£ 3700).

Example 21 (1'R, 5R, 6S and 1'S, 5S, 6R) -6- (1,1-hydroxy-2'-methoxymethoxy-2'-ethyl) -2-ethyl) -2-methylpenem-3- carboxylic acid (isomer C)

QH

- "CCH.

2: (1'R, 3S, 4R and 1'S, 3R, 4S) -S'-N'-dihydroxyethyl-1- (p-nitrobenzyl-2 "-triphenylphosphoranylidene-2" -acetate) - 4-Acetylthio-2-azetidinone (Isomer C)?

CO PNB CO ^ PNB

2 ^ 185 6 7 8 5 3

A solution of (4'R, 3S, 4R and 4'S, 3R, 4S) -3- (2T, 2'-dimethyl-1 ', 3'-dioxolan-4'-yl) -1- (p-nitrobenzyl -2 "-triphenylphosphoranylidene-2" -acetate) -4-acetylthio-2-acetidine (Isomer C) (472 mg, 0.676 mmol) in trifluoroacetic acid (1.0 mL) and IVO (0.1 mL) was administered be at room temperature for 30 minutes. The mixture was added dropwise to a cold solution of NaHCO 3 (2.5 g) in H 2 O (50 mL) and extracted with CH 2 Cl 2 (3 x 20 mL). The extracts were washed with saturated NaHCO 3 and then brine, dried (Na 2 SO 4) and evaporated to dryness to give 458 mg (0.695 mmol, crude yield 100%; purity 97.3%) of the crude title compound as a yellowish foam; 1 Hmr (CDCl 3): 2.20, 2.32 (2s, SAc) and 7.2-8.3 ppm (aromatic Hs); ir (fine) · 3420 (OH), 174 δ lactam, ester) and 1690 cm -1 (thioester), Rf 0.16 (ethyl acetate).

(1'R, 3S, 4R and 1'S, 3R, 4S) -3- (1'-hydroxy-21-methoxymethoxy-1'-ethyl) -1- (p-nitrobenzyl-2 "-triphenylphosphoranylidene-2" acetate) -4-acetylthio-2-azetidinone (Isomer C)

9H 0H

HO: SAc ru rv ^ SAc 1 BrCHjOCHj-dimethylaniline |

J-N Ρφ CH Cl J-N

3 2 2 3

CO ^ PNB CO ^ PNE

To a solution of (1'R, 3S, 4R and 1'S, 3R, 4S) 3- (1 ', 2'-dihydroxyethyl) -1- (p-nitrobenzyl-2 "-triphenylphosphoranylidene-2" - acetate) -4-acetylthio-2-azetidinone (Isomer C) (291 mg, 0.430 mmol; purity 97.3%) and bromomethyl methyl ether (55.2 mg, 0.442 mmol; 4 drops) in CH 2 Cl 2 (8 ml), N, N'-dimethylaniline (58.8 mg, 0.483 mmol; 5 drops) was added at 0 ° C and the mixture was stirred at room temperature for 20 h. Additional bromomethyl methyl ether (2 drops) and N, N'-dimethylaniline ( 2 drops), and the mixture was stirred for another 4 h.

67853 186 The mixture diluted with CH 2 Cl 2 was washed successively with 1-norm. HCl, saturated NaHCO 3 and brine, dried (Na 2 O 2) and evaporated to dryness. The crude residue was purified by hplc (SiO 2, eluent ethyl acetate) to give the title compound as an oil (31 mg, 0.186 mmol; 42.2% yield);

Rf 0.2 * 1 (ethyl acetate; Hmmr (CDCl3) 2.20, 2.32 (2s, SAc), 3.30 (s, OCH2), 4.52 (s, -OCH2O-) and 7.4- 8.3 ppm (m, aromatic Hs); ir (fine),: 3420 (OH), 1755 (br, β-lactam 1 Iuq.KS / and ester) and 1690 cm-thio ester).

(1'R, 5R, 6S and 1'S, 5S, 6R) -p-Nitrobenzyl 6- (1'-hydroxy-2'-methoxymethoxy-2'-ethyl) -2-methylpenem-3-carboxylate

9H

f toluene _ t J— N 3 QJ — Δ Co2pnb

C02PNB

A solution of (1'R, 3S, 4R and 1'S, 3R, 4S) -3- (1'-hydroxy-21-methoxymethoxy-1'-ethyl) -1- (p-nitrobenzyl-2 "-triphenyl- phosphoranylidene-2 "-acetate) -4-acetylthio-2-azetidinone (Isomer C) (167 mg, 0.238 mmol) in toluene (30 mL) was heated at reflux under N 2 for 8 h. Evaporation of the solvent in vacuo gave an oily residue which was purified by hplc (SiO 2, eluent ethyl acetate) to give 68 my (0.16 mmol, 67% yield) of the title compound as an oil;

Rf 0.61 (ethyl acetate), 0.15 (benzene: Et 2 O = 1: 1); 1 Hmr (CDCl 3) δ: 2.38 (3H, s, 2-CH 2), 3.35 (1H, br, OH), 3.40 (3H, s, OCH 3), 3.6-3.8 ( 2H, m, H-2 '), 3.90 (1H, dd, J5 = 2Hz, J6 = 4Hz, H-6), 4.18 (1H, m, H-1'), 4.67 (2H , s, · -OCH 2 O-), 5.03-5.27-5.38-5.62 O 2, ABq, OCH 2 Ar), 5.65 (1H, d, J = 2Hz, HS) and 7.55- 7.70-8.15-8.30 ppm (4H, Ag'B 2, aromatic Hs); ir (fine) V,: 3450 (OH), 1785 (β-lactam), 1710 (ester) and 1520 cm (NO2); u.v (EtOH): 266 (£ 13,000) and 313 m / u (£ 9,100); Anal Calcd for C 18 H 20 N 2 O 2: C 50.94, H 4.75, N 6.60; Found: C 51.13, H 4.77, N 6.36.

67853 187 (1'R, 5R, 6S and 1'S, 5S, 6R) -6- (1'-hydroxy-21-methoxymethoxy-2'-ethyl) -2-methylpenem-3-carboxylic acid (Isomer C)%

pH

^ xqr · ^ -¾1 2

A solution of (1 * R, 5R, 6S and 1'S, 5S, 6R) -p-nitrobenzyl-6- (1'-hydroxy-2'-methoxymethoxy-2'-ethyl) -2-methylpenem-3 carboxylate (Isomer C) (51 mg, 0.12 mmol) in THF (10 mL) was stirred in Et 2 O (10 mL), Et 2 O (10 mL), NaHCO 3 (10 mg, 0.12 mmol). ) and 10% Pd-C catalyst (50 mg; Engelhard). It was hydrogenated at room temperature at 32 psi for 3 h. After the catalyst was filtered off through Celite, the separated aqueous layer was washed with Et 2 O (3 times) and saturated with NaCl. The aqueous phase, which was acidified at 0 ° C to 0.1 norm. HCl (1.2 mL), immediately extracted with ethyl acetate (3 x 15 mL). The extracts were washed with brine, dried (Na 2 SO 4) and evaporated to dryness to give 22 mg of a yellowish solid which was rinsed with a small amount of Et 2 O to give 20 mg (0.069 mmol; 58% yield) of the title compound as a slightly yellow solid; 1 Hmr (DMSO-d 6 S: 2.28 (3H, s, 2-CH 3), 3.27 (3H, s, OCH 3), 3.49 (2H, d, J = 6.2Hz, 2'-H) , 3.87 (1H, dd, J6_5 = 1.7 Hz, ^ 6_1 »= 3.3Ηζ, 6-H), 4.58 (2H, s, -OCH2O-) and 5.55 ppm (1H, d, J = 1.7 Hz, 5-H); ir (KBr) λmax: 3410 (OH), 1755 (β-lactam) and 1655 cm-2 (CO2H); uv (EtOH) [max '308 (£ 6800) and 262 m / u (£ 4200), mp 137-8 ° C (dec.).

67853 188

Example 22 (1'S, 5R, 6S and 1'R, 5S, 6R) -6- (1,1-hydroxy-2'-methoxymethoxy-2R-ethyl) -2-methylpenem-3-carboxylic acid (Isomer B)

OH

Ojo> ^

CO H

(1'S, 3S, 4R and 1'R, 3R, 4S) -3- (1 ', 2'-dihydroxyethyl) (p-nitrobenzyl 2'-triphenylphosphoranylidene-2' 'acetyl) -4- acetylthio-2-azetidin.none (Isomer B), ° h THF-Η, ο _ “^ ΓΤ" 0 rfJ-A-YP * j "0 co2pnb co2pNb

The title compound was prepared as described in Example 53 for "Isomer C" (4'S, 3S, 4R, and h'o 3r) 9c5R> Hd) -3- (2f, 2'-dimethyl-1 ', 3'- dioxolan-4'-yl) -1- (p-nitrool, +.

ea_C3yy 2 "-triphenylphosphoranylidene-2" -acetate) -4-acetyls from azetidinone (Isomer B) (1.03 g, 1.47 mmol); blockade 970 mg, 1.47 mmol, 100%; yellowish foamy material (1 H, CDCl 3): 2.20, 2.32 (2s, -SAc) and 7.3-8.2 ppm. (Blj 'matical Hs); ir (fine) ν max: 3410 (OH), 1750 (β-ester) and 1690 cm -1 (thioester): Rf 0.16 (EtOAc).

189 67853 (1'S, 3S, 4R and 1'R, 3R, 4S) -3- (1'-hydroxy-2'-methoxymethoxy-1'-ethyl) -1- (p-nitrobenzyl-2 "-triphenylphosphoranylidene -2 "-acetate) -H-acetylthio-2-azetidinone (Isomer B)

OH ° H

- _ ^ SAc / V'OV '^^ C

j BrCH OCH / dimethylaniline j j ο ^ -Ν-γ »· ΡΒ ch2ci2 cr ^ -γ C02PNB"

The title compound was prepared as described in Example 53 for "Isomer C" (1 * S, 3S, 4R and 1'R, 3R, HS) -3- (1 ', 2'-dihydroxyethyl) -1- (p- nitrobenzyl 2 "-triphenylphosphoranylidene-2" -acetate) -4-acetylthio-2-azetidinone (Isomer B) (485 mg, 0.736 mmol); yield 205 mg, 0.292 mol, 39.6%; oil; ν r (CDCl 3) ε: 2.22, 2.33 (2s, SAc), 3.32 (s, OCH 3),> 4.57 (S 2 O 2 F 3 O-) and 7.2-8.3 ppm (aromatic H 5) ir (fine) N 2 O 2 · 3 + + 20 (OH), 1755 (γ-lactam, ester) and 1690 (thioester) R f 0.32 (EtOAc).

(1'S, 5R, 6S and 1'R, 5S, 6R) -p-Nitrobenzyl 6- (1'-hydroxy-1'-methoxymethoxy-2'-ethyl) -2-methylpenem-3-carboxylate

^ oj "SAc 0 ^ KjH

”Γ * Ρφ -» I T ^ 'V-CH

o * 3 Δ cy— co2pnb lo.pnb

The title compound was prepared as described in Example 53 for "Isomer C" (1'S, 3S, 4R and 1'R, 3R, 4S) -3- (1'-hydroxy-2'-methoxymethoxy-1'-ethyl) -1- (p-nitrobenzyl 2 "-triphenylphosphoranylidene-2" -acetate) -N-acetylthio-2-azetidinone (Isomer B) (205 mg, 0.292 mmol) and hydroquinone (10 mg, C, 09 mmol); yield 38 mg, 0.090 mmol, 31%; 152-154 ° C; Rf 0.23 (benzene: Et 2 O = 1: 1); 1 Hmr (CDCl 3) δ: 67853 190 2.37 (3H, s, 2-CH 3), 3.40 (3H, s, OCH 3>, 3.4-3.9 (3H, m, H-6, H -2 "), 4.15 (1H, m, H-1 '), 4.67 (2H, s, -OCH 2 O-), 5.10-5.27-5.39-5.56 (2H, ABq, -OCH 2 Ar), 5.67 (1H, d, J = 1.5 Hz, H-5) and 5.55-5.16-8.15-8.27 ppm (4H, A2'B2? aromatic H3); ir (CH2Cl2 suspension)? max: 3370 (OH), 1785 (γ-lactam) and 1700 cm-1 (ester); uv (THF-EtOH = 1: 1) max: 265 (£ 10400) and 314 (£ 7800).

(1'S, 5R, 6S and 1'R, 5S, 6R) -6- (1'-hydroxy-2'-methoxymethoxy-2'-ethyl) -2-methylpenem-3-carboxylic acid <isomer B)

OH OH

^ ° 2PNB. C02H

The title compound was prepared as described in Example 53 for "Isomer C" (1'S, 5R, 6S and 1'R, 5S, 6R) -p-nitrobenzyl-6- (1'-hydroxy-2'-methoxymethoxy-2'-ethyl ) -2-methylpenem-3-carboxylate (Isomer B) (36 mg, 0.085 mmol); yield 7.5 mg, 0.026 mmol; 30%; yellowish crystals; 1 Hmr (CDCl 3) δ: 2.36 (3H, s, 2-CH 2), 3.39 (3H, s, OCH 9), 3.6-3.9 (3H, m, H-6, H-2 ' ), 4.15 (1H, m, H-1 '), 4.66 (2H, s, OCH 2 O) and 5.67 ppm (1H, d, J = 1.4 Hz, H-5); ir (CH 2 Cl 2) Vmax: 1785 γ-lactam) and 1675 cm -1 (CO 2 H); uv (EtOH) Amax: 308 2900) 3a 263 m / u 2900).

Claims (1)

A process for the preparation of antibacterial 7-oxo-4-thia-1-aza-bicyclo [3.2.1] hept-2-ene derivatives and their pharmaceutically acceptable salts, Y "ί-rsv_ • / / x J- (I) ° wherein Z is hydrogen or an ester-forming group, X is methyl, phthalimidobutyl, CH 2 C (CH 3) = NOCH 3 or (CH 2) n PO (O) 2 where n is 2 or 3 and R is lower alkyl, and Y is hydrogen, lower alkyl substituted with acetoxy or hydroxy, (CH3) 2NCH2, HCONHCH2, C6H5CH2CH (OH), CH3OCH2OCH2CH (OH), or 2,2-dimethyl-1,3-dioxolan-4- yyl, provided that X is not methyl when Y and Z are each hydrogen, characterized in that the compound of formula 0 v "YS - CX Ί! JN P (Q), 0 ^ 3 co2z * where X and Y have the same meaning as above, Z 'is an ester-forming group and Q is phenyl, is cyclized in an inert organic solvent at a temperature slightly above room temperature and the reflux temperature of the solvent , and if desired, the ester-forming group is removed and, if desired, the resulting compound is converted into a pharmaceutically acceptable salt.