WO1995023149A1 - Phenylalkylamino derivatives of condensed carbapenemes - Google Patents

Abstract

Compounds of general formula (I), salts and metabolically labile esters thereof; wherein R1 represents hydrogen or a hydroxyl protecting group; R2 represents hydrogen or a carboxyl protecting group; R3 represents hydrogen or a nitrogen protecting group; Z represents a methylene group optionally substituted by methyl; X represents a bond or a C1-2alkylene chain or an ethenylene chain; A represents a C2-4alkylene chain or A is a chain of 2 to 4 members one of which is an oxygen or sulphur atom or the group NH or a substituted derivative thereof and the other members are methylene groups; and R4 represents an optionally substituted phenyl group, a process for their preparation and their use as antibacterial agents.

Description

PHENYLALKYLA INO DERIVATIVES OF CONDENSED CARBAPENEMES .

This invention relates to phenylaikyiamino derivatives having antibacterial activity, to processes for their preparation, to compositions containing them and to their use in medicine.

European Patent Application publication No. 0 16953A2 describes 10-(1- hydroxyethyl)-11 -oxo-1 -azatricyclo[7.2.0.0³'⁸]undec-2-ene-2-carboxylic acid and certain 4 substituted derivatives thereof, including 4-amino, 4-alkylamino and 4- dialkylamino derivatives which have antibacterial activity.

European Patent Application publication No. 0422596A2 describes compound of the general formula

wherein R¹ is inter alia a 1-hydroxyethyl group, CO₂R² is a carboxy group which may optionally be esterified and ring B is a cyclic group which may be optionally substituted. Ring B may inter alia be a cyclohexane ring. The compounds have antibacterial activity. The specification specifically teaches compounds wherein B is an unsubsituted cyclohexane ring, but there is no teaching of specfic compounds wherein the cyclohexane ring is substituted.

European Patent Application No. 0507313A1 describes inter alia compounds of formula

wherein R¹ is inter alia an optionally substituted lower alkyl group, C0₂R² is a carboxy group which may be esterified, ring A is inter alia a cyclohexane ring and R^a is the group W^U³ [W³ is a bond, sulphur (which may be in the form of mono- or dioxide), oxygen, NH (which may be substituted) or a straight-chain or branched lower alkylene or alkenylene group which may be interrupted by sulphur (which may be in the form of mono- or dioxide), oxygen or NH (which may be substituted); U^a is carbamoyl, acyl which may be substituted, alkylammonium which may be substituted or a group of the formula

is a quaternized nitrogen-containing heterocyclic group which may be substituted and R^d is an alkyl group which may be substituted)] which compounds have antibacterial activity. Preferred compounds of this class are said to be 10-(1-hydroxyethyl)-11-oxo-1-azatricyclo [7.2.0.0³-⁸] undec-2-ene-2- carboxylic acid, derivatives wherein the group R^a is (CH₂)m K^a(CH₂)nU^a (wherein K^a is CH₂, 0, S or NH; m and n each is a whole number of 0 to 3; and U^a is an N-linked quaternary ammonium group.

We have now discovered that the introduction of certain N-aralkylamino groupings at the 4 position of 10-(1-hydroxyethyl)-11-oxo-1-azatricyclo [7.2.0.0³'⁸] undec-2-ene-2-carboxylic acid and structually similar compounds provides compounds with a particularly useful profile of activity as antibacterial agents.

According to the present invention, therefore we provide compounds of general formula (I)

salts and metaboiically labile esters thereof; wherein R., represents hydrogen or a hydroxyl protecting group; R₂ represents hydrogen or a carboxyl protecting group; R₃ represents hydrogen or a nitrogen protecting group;

Z represents a methylene group optionally substituted by methyl; X represents a bond or a C-^alkylene chain or an ethenylene chain; A represents a C₂^alkylene chain or A is a chain of 2 to 4 members one of which is an oxygen or sulphur atom or the group NH or a substituted derivative thereof and the other members are methylene groups; and R₄ represents an optionally substituted phenyl group.

In addition to the fixed stereochemical arrangement as defined in formula (I) the molecule contains a further asymmetric carbon atom at the 8-position, and another at the 4-position. It will be appreciated that all stereoisomers including mixtures thereof arising from these additional asymmetric centres, are within the scope of the compounds of formula (I).

The compounds of formula (I) are antibacterial agents and/or of use as intermediates for the preparation of other active compounds within the general formula (I). Compounds wherein R- represents a hydroxyl protecting group and/or wherein R₂ represents a carboxyl protecting group and/or wherein R₃ represents a nitrogen protecting group are in general intermediates for the preparation of other compounds of formula (I).

Salts of compounds of formula (I) include base addition salts, acid addition salts compounds and internal salts formed with the carboxylic acid grouping. Base addition salts for use in medicine are formed with bases that have a physiologically acceptable cation. Suitable cations include those of alkali metals (e.g. sodium or potassium), alkaline earth metals (e.g. calcium), amino acids (e.g. lysine and arginine) and organic bases (e.g. procaine, phenylbenzylamine, dibenzylethylenediamine, ethanolamine, diethanolamine, and N-methyl glucosamine). Acid addition salts may be formed with organic acids or inorganic acids. For use in medicine the salts will be those derived from physiologically acceptable acids e.g. acetic acid, succinic acid, hydrochloric acid, sulphuric acid and phosphoric acid.

Salts derived from bases wherein the cation is not physiologically acceptable or acids which are not physiologically acceptable may be useful as intermediates for the preparation and/or isolation of other compounds of the invention, and these salts also form part of the invention.

It will be appreciated that the compounds of formula (I) may be produced in vivo by metabolism of a suitable metabolically labile ester. Examples of suitable metabolically labile esters include acyloxyalkyl esters such as, acyloxymethyl or 1-acyloxyethyl e.g. pivaloyloxymethyl, 1-pivaloyloxyethyl, acetoxymethyl, 1- acetoxyethyl.l^l-methoxy-l-methy ethylcarbonyloxyethyl, 1- benzoyloxyethyl, isopropoxycarbonyloxymethyl, 1 -isopropoxycarbonyloxyethyl, cyclohexylcarbonyloxymethyl, 1-cyclohexylcarbonyloxyethyl ester, cyclohexyloxycarbonyloxymethyl, 1-cyclohexyloxycarbonyloxyethyl. 1-(4- tetrahydropyranyloxy)carbonyloxyethyl or 1-(4- tetrahydropyranyl)carbonyloxyethyl or 3-phthalidyl.

The compound of formula (I) salts thereof and metabolically labile esters thereof may form solvates (e.g. hydrates) and the invention includes all such solvates.

Particularly useful compounds of formula (I) for use in medicine as antibacterial agents are those in which the group R₁ , R₂ and R₃ represent a hydrogen atom and physiologically acceptable salts and metabolically labile esters thereof. These compounds exhibit antibacterial activity against a wide range of gram positive and gram negative, aerobic and anaerobic pathogenic microorganisms.

When R-_i is a hydroxy protecting group the group ORι is conveniently an ether or an acyloxy group. Examples of particularly suitable ethers include those in which R-_j is a hydrocarbylsilyl group such as trialkylsilyl, e.g. tri(Cι^alkyl)silyl such as trimethylsilyl or more especially t-butyldimethylsilyl. When the group OR-_j represents an acyloxy group then examples of suitable groups R-j includes alkanoyl e.g. acetyl or pivaloyl; alkenoyl e.g. allylcarbonyl; aroyl e.g. p- nitrobenzoyl; alkoxycarbonyl e.g. t-butoxycarbonyl; haloalkoxycarbonyl e.g. 2.2,2-trichloroethoxycarbonyl, or 1,1,1-trichloro-2-methyl-2- propoxycarbonyl; aralkyloxycarbonyl e.g. benzyloxycarbonyl or p- nitrobenzyloxycarbonyl; or alkenyloxycarbonyl e.g. allyloxycarbonyl.

When R₂ is a carboxyl protecting group examples of suitable groups R₂ include arylmethyl groups such as benzyl, p-nitrobenzyl, t-butylbenzyl or trityl, or alkenyl groups such as allyl or substituted allyl, t-butyl, haloalkyi e.g. trichloroethyl or tnalkylsilylalkyi e.g. trimethylsilylethyl. Preferred protecting groups R₂ include allyl or arylmethyl e.g. benzyl or p-t-butylbenzyl.

When R₃ is a nitrogen protecting group examples of suitable groups include optionally substituted allyloxycarbonyl, alkyloxycarbonyl or arylmethyloxycarbonyl.

Compounds wherein at least one of the groups R,_t R₂ and/or R₃ are a protecting group as defined above represent useful intermediates from the preparation of compounds of formula (I) having antibacterial activity.

When R is a subsituted phenyl group this phenyl group substituted by one to 3 substituents which may be the same or different and selected from halogen or amino, alkylamino, dialkylamino, acylamino, alkoxy, alkylenedioxy, alkyl, alkylthio, alkylsulphinyl, cyano, nitro, trifluoromethyl, trifluoromethoxy, CO₂R₅ or SO₂NR6R₇;

R₅ represents hydrogen, C^alkyl, C₂^alkenyl or C3^alkynyl; Re represents hydrogen, alkoxycarbonyl, allyloxycarbonyl or benzyloxycarbonyl; R₇ represents hydrogen, C^alkyl, C-^alkenyl or pheny C-^alkyl.

Unless otherwise specified the term alkyl as a group or part of a group refers to a straight or branched chain group e.g. methyl, ethyl, propyl or butyl.

The term acylamino refers to alkanoylamino, alkoxycarbonylamino, allyloxycarbonylamino or optionally substituted benzyloxycarbonylamino. The term alkylenedioxy refers to C1.3 alkenedioxy e.g. methylenedioxy or ethylenedioxy.

When A is a C₂-₄ alkylene chain this may be an ethylene, propylene or butylene chain.

When A is chain of 2 to 4 members and wherein one of the members is a group selected from oxygen sulphur or NH (or a substituent derivative thereof) examples of such chains include:

-OCH₂-. -OCH₂CH₂-, -CH₂-OCH₂-, -CH₂CH₂0-, -SCH₂CH₂-, -CH₂SCH₂- - CH₂CH₂S-, or -CH₂NH-CH₂-.

When a member of the chain A is a substituted 'NH' group examples of suitable substituents include C^alkyl e.g. methyl, benzyl, acetyl, C^alkoxycarbonyl, allyloxycarbonyl or an optionally substituent phenylmethyloxycarbonyl group.

The term halogen when used herein means fluorine, chlorine, bromine or iodine unless otherwise specified.

The general formula (I) as drawn includes at least 4 stereoisomers and mixtures thereof and these may be represented by the formulae (1a, 1b, 1c and 1d).

Id The solid wedge shaped bond indicates that the bond is above the plane of the paper. The broken bond indicates that the bond is below the plane of the paper.

The configuration shown for the carbon atom at the 8-position in formulae 1a and 1b is hereinafter referred to as the β configuration and in formulae 1c and 1d as the α configuration.

The configuration shown for the carbon at the 4 position in formulae 1b and 1d is hereinafter referred to as the α confirguation and in formulae 1 a and 1 c as the β configuration.

In general, in the specific compounds named below, the β-configuration at the

8-position corresponds to the S isomer and the β-configuration at the 4-position to the R isomer. The α configuration at the 8-position corresponds to the R isomer and the α-configuration at the 4-position corresponds to the S isomer.

The assignment of the R or S configuration at the 4- and 8- positions have been made according to the rules of Cahn. Ingold and Prelog, Experientia 1956, 12,

81.

Preferably the group R-i is a hydrogen atom

Conveniently the group R₂ is a hydrogen atom

Preferably the group R₃ is a hydrogen atom

A further preferred class of compounds are those wherein X is a bond.

Compounds of formula (I) wherein Z is methylene represent a further preferred class.

Compounds of formula (I) wherein A is a 2-4 alkylene chain or more particularly a propylene chain represent a further preferred class of compound according to the invention. For compounds of formula I wherein A is a C₂^alkylene chain, those in which the carbon atom at the 8- position is in the β configuration are preferred and within this class those compounds in which the carbon atom at the 4- position is in the α configuration are particularly preferred.

Examples of suitable R₄ groups include phenyl or a phenyl substituted by one to 2 groups selected from halogen e.g. fluorine, chlorine, bromine, alkoxy e.g. methoxy, methylenedioxy, hydroxy, nitro, trifluromethyl, alkylthio, e.g. thiomethyl, alkylsulphinyl e.g. methylsulphinyl, acylamino e.g. allyloxycarbonylamino, amino, C0₂R₅ (wherein R₅ is hydrogen or allyl) or SO₂NR₆R₇ (wherein R₇ is hydrogen, methyl, allyl or benzyl, and Re is hydrogen, allyloxycarbonyl or benzyloxycarbonyl).

A preferred class of compounds of formula (I) are those wherein R₄ is phenyl or phenyl substituted by one or 2 groups selected from halogen e.g. chlorine, fluorine or bromine, alkoxy e.g. methoxy, trifluoromethyl, carboxy, amino, nitro, allylaminosulphonyl, methylaminosulphonyl, benzylaminosulphonyl, methylenedioxy or alkylthio e.g. methylthio.

More preferably R4 represents phenyl or phenyl substituted by one or two groups selected from chlorine, fluorine, nitro, methylenedioxy or allylaminosulphonyl.

A preferred group of compounds of formula (I) are those wherein R-i, R₂ and R₃ represent hydrogen, X is a bond Z, is methylene, and A is a propylene group and physiologically acceptable salts or metabolically labile esters thereof.

Within this group particularly preferred compounds are those wherein R₄ is phenyl or phenyl substituted by one or two groups selected from halogen, alkoxy, trifluoromethyl, carboxy, amino, nitro, methylenedioxy, alkylthio or SO₂ NHR₇ wherein R₇ is methyl, allyl, or benzyl. More especially R-j represents phenyl, or phenyl substituted by one or 2 groups selected from chlorine, fluorine, nitro, methylenedioxy or allylaminosulphonyl.

Particularly preferred compounds of the invention include : (4S,8S,9R, 10S, 12R)-4-(4-nitrobenzylamino)-10-(1 -hydroxyethyl)-11 -oxo-1 -

3 8 azatricyclo-(7.2.0.0 ' )-undec-2-ene-2-carboxylic acid.

(4S,8S,9R, 10S, 12R)-4-(4-fluorobenzylamino)-10-(1 -hydroxyethyl)-11 -oxo-1 - 3 8 azatricyclo-(7.2.0.0 ' )-undec-2-ene-2-carboxylic acid.

(4S,8S,9R,10S,12R)-4-((4-(N-allyl)aminosulfonylbenzylamino)-10-(1- hydroxyethyl)-11 -oxo-1 -azatricyclo-(7.2.0.0 ^• )-undec-2-ene-2-carboxylic acid.

(4S,8S,9R, 10S, 12R)-4-(benzylamino)-10-(1 -hydroxyethyl)-11 -oxo-1 -azatricyclo- (7.2.0.0 ' )-undec-2-ene-2-carboxylic acid.

(4S,8S,9R, 10S, 12R)-4-(3,4-(methylendioxy)benzylamino)-10-(1 -hydroxyethyl)- 11-0X0-1- azatricyclo-(7.2.0.0 ^■ )-undec-2-ene-2-carboxylic acid.

(4S,8S,9R, 10S, 12R)-4-(3-chloro-4-fluorobenzylamino)-10-(1 -hydroxyethyl)-11 - Q oxo-1 -azatricyclo-(7.2.0.0 ^• )-undec-2-ene-2-carboxylic acid

Compounds according to the invention not only exhibit a broad spectrum of antibacterial activity against a wide range of pathogenic microorganisms but also have a very high resistance to all β-lactamases. Compounds of the invention are also relatively stable to renal dehydropeptidase.

Thus using a standard microtiter broth serial dilution test compounds of the invention have been found to exhibit useful levels of activity against a wide range of pathogenic microorganisims including strains of Staphylococcus aureus. Streptococcus faecalis. Escherichia col], Pseudomonas aeruαinosa.

Klebisiella. pneumoniae. Proteus microbilis Clostridium perfrinαens and

Bacteriodes fraαilis.

Compounds of the invention have also been found to exhibit a particularly advantageous profile of activity against gram negative micro-organisms.

The compounds of the invention may therefore be used for treating a variety of diseases caused by pathogenic bacteria in human beings and animals. Thus, according to another aspect of the present invention, we provide a compound of formula (I) or a physiologically acceptable salt thereof for use in the therapy or prophylaxis of systemic or topical bacterial infections in a human or animal subject.

According to a further aspect of the invention we provide the use of a compound of formula (I) or a physiologically acceptable salt thereof for the manufacture of a therapeutic agent for the treatment of systemic or topical bacterial infections in a human or animal body.

According to a yet further aspect of the invention we provide a method of treatment of the human or non-human animal body to combat bacterial infections which method comprises administering to the body an effective amount of a compound of formula (I) or a physiologically acceptable salt thereof.

While it is possible that, for use in therapy, a compound of the invention may be administered as the raw chemical it is preferable to present the active ingredient as a pharmaceutical formulation.

The compounds of the invention may be formulated for administration in any convenient way for use in human or veterinary medicine and the invention therefore includes within its scope pharmaceutical compositions comprising a compound of the invention adapted for use in human or veterinary medicine. Such compositions may be presented for use in conventional manner with the aid of one or more suitable carriers or excipients. The compositions of the invention include those in a form especially formulated for parenteral, oral, buccal, rectal, topical, implant, ophthalmic, nasal or genito-urinary use.

The compounds according to the invention may be formulated for use in human or veterinary medicine by injection (e.g. by intravenous bolus injection or infusion or via intramuscular, subcutaneous or intrathecal routes) and may be presented in unit dose form, in ampoules, or other unit-dose containers, or in multi-dose containers, if necessary with an added preservative. The compositions for injection may be in the form of suspensions, solutions, or emulsions, in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilising, solubilising and/or dispersing agents. Alternatively the active ingredient may be in sterile powder form for reconstitution with a suitable vehicle, e.g. sterile, pyrogen-free water, before use.

The compounds of the invention may also be presented for human or veterinary use in a form suitable for oral or buccal administration, for example in the form of solutions, gels, syrups, mouth washes or suspensions, or a dry powder for constitution with water or other suitable vehicle before use, optionally with flavouring and colouring agents. Solid compositions such as tablets, capsules, lozenges, pastilles, pills, boluses, powder, pastes, granules, bullets or premix preparations may also be used. Solid and liquid compositions for oral use may be prepared according to methods well known in the art. Such compositions may also contain one or more pharmaceutically acceptable carriers and excipients which may be in solid or liquid form.

The compounds of the invention may also be administered orally in veterinary medicine in the form of a liquid drench such as a solution, suspension or dispersion of the active ingredient together with a pharmaceutically acceptable carrier or excipient.

The compounds of the invention may also, for example, be formulated as suppositories e.g. containing conventional suppository bases for use in human or veterinary medicine or as pessaries e.g. containing conventional pessary bases.

The compounds according to the invention may be formulated for topical administration, for use in human and veterinary medicine, in the form of ointments, creams, gels, lotions, shampoos, powders, (including spray powders), pessaries, tampons, sprays, dips, aerosols, drops (e.g. eye ear or nose drops) or pour-ons. Aerosol sprays are conveniently delivered from pressurised packs, with the use of a suitable propellant, eg dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas.

For topical administration by inhalation the compounds according to the invention may be delivered for use in human or veterinary medicine via a nebuliser.

The pharmaceutical compositions for topical administration may also contain other active ingredients such as corticosteroids or antifungals as appropriate.

The compositions may contain from 0.01-99% of the active material. For topical administration, for example, the composition will generally contain from 0.01- 10%, more preferably 0.01-1% of the active material.

For systemic administration the daily dose as employed for adult human treatment will range from 5-100mg/kg body weight, preferably 10-60mg/kg body weight, which may be administered in 1 to 4 daily doses, for example, depending on the route of administration and the condition of the patient. When the composition comprises dosage units, each unit will preferably contain 200mg to 1g of active ingredient.

The duration of treatment will be dictated by the rate of response rather than by arbitrary numbers of days.

The compounds of formula (I) may be prepared by the cyclisation of a compound of formula (II)

(π) in which R₁₍ R₂, R₃, X, Z, A and R₄ have the meanings defined in formula (I) or is a group convertible thereto, and Y is an oxygen atom or a phosphine group, and if required or desired subjecting the resulting compound prior to or subsequent to any separation into its stereochemical isomers, to one or more of the following operations

(a) removal of one or more protecting groups.

(b) conversation of one compound of formula (I) into another compound of formula (I).

The cyclisation of a compound of formula (II) in which Y is oxygen is conveniently carried out by heating in the presence of an organic phosphite. The reaction is preferably carried out in a solvent or mixture of solvents at a temperature within the range 60-200°. Suitable solvents include hydrocarbons with an appropriate boiling point, for example aromatic hydrocarbons, such as toluene or xylene.

Suitable organic phosphites include acyclic and cyclic trialkylphosphites, triarylphosphites and mixed alkylarylphosphites. Particularly useful organic phosphites are the trialkylphosphites e.g. triethylphosphite or trimethylphosphite.

The cyclisation of a compound of formula (II) in which Y is a phosphine grouping is preferably carried out in a solvent at a temperature between 40- 200°C. Suitable solvents include hydrocarbons such as aromatic hydrocarbons, for example xylene or toluene, aliphatic hydrocarbons and halogenated hydrocarbons such as dichioromethane, chloroform and trichloroethane. Examples of suitable phosphine groups are triarylphosphines e.g. triphenyl phosphine or trialkylphospines e.g. th-t-butylphospine.

The hydroxyl, carboxyl and nitrogen protecting groups R_1f R₂ and R₃ may be removed by conventional procedures and in any order. More preferably however the hydroxyl protecting group R^ is removed prior to the removal of the carboxyl and or nitrogen protecting group. Such removal of the protecting groups is a further feature of the invention. The hydroxyl protecting groups may be removed by well known standard procedures such as those described in Protective Groups in Organic Chemistry, pages 46-119, Edited by J F W McOmie (Plenum Press, 1973). For example when R₁ is a t-butyldimethylsilyl group, this may be removed by treatment with tetrabutylammonium fluoride and acetic acid or by reaction with fluoride ions and a suitable phase transfer catalyst such as tetrabutylammonium bromide in the presence of acetic acid. A suitable solvence of fluoride ions included potassium fluoride or caesium fluoride. This process is conveniently carried out in a solvent such as tetrahydrofuran. Similarly when R₁ is a trichloroethoxycarbonyl group this may be removed by treatment with zinc and acetic acid.

The carboxyl protecting group R₂ may also be removed by standard processes such as those described in Protective Groups in Organic Chemistry, pages 192- 210, Edited by J F W McOmie (Plenum Press 1973). For example when R₂ represents an arylmethyl group this may be removed by conventional procedures using hydrogen and a metal catalyst e.g. palladium. When the group R₂ represents an allyl or substituted allyl group then this is preferably removed by treatment with an allyl acceptor in the presence of tetrakis(triphenylphosphine) palladium and optionally in the presence of triphenylphospine. Suitable allyl acceptors include sterically hindered amines such as tertbuylamine, cycNc secondary amines such as morpholine or thiomorpholine, tertiary amines such as triethylamine, aliphatic or cycloapliphatic β-dicarbonyl compounds such as acetylacetone, ethyl acetoacetate or dimedone, an alkanoic acids or alkali metal salts thereof such as acetic acid, propionic acid or 2-ethyl hexanoic acid or the potassium or sodium salt thereof, or 5,5-dimethyl-1,3-cyclohexadiene.

A particularly useful allyl acceptor is 5,5-dimethyl 1 ,3-cyclohexadiene.

The reaction is preferably carried out in an inert solvent such as an ether e.g. diethyl ether or tetrahydrofuran, an alkanol e.g. ethanol, an ester e.g. ethyl acetate or a halohydrocarbon e.g. methylene chloride, or mixtures thereof. The reaction is conveniently carried out in the temperature range 0°-40° more particularly at room temperature.

The nitrogen protecting group R₃ may be removed by conventional procedures. Thus for example if R₃ is an allyloxycarbonyl group this may be removed using the conditions described for removal of the carboxyl protecting group where R₂ is allyl.

Compounds of formula (I) may be converted into other compounds of formula (I). Thus compounds of the invention in which the group R₂ is a physiologically acceptable cation may be prepared from compounds of the invention in which R₂ is hydrogen by treatment with a suitable base. Conveniently the salt is formed in solution and then if required precipitated by the addition of a non- solvent e.g. a non polar aprotic solvent. Alternatively the sodium or potassium salt may be prepared by treating a solution of a compound of formula (I) in which R₂ represents a hydrogen atom with a solution of sodium or potassium 2- ethylhexanoate in a non-polar solvent such as diethyl ether.

Compounds wherein R₂ is a metabolically labile ester may be prepared from the corresponding compound wherein R₂ is a hydrogen atom or a cation by use of conventional esterification procedures.

Compounds of formula (I) wherein the group R₂ is a carboxyl protecting group and R₄ represents a phenyl group substituted by alkylsulphinyl may be prepared by oxidation of the corresponding compounds of formula (I) wherein R₄ is a phenyl group substituted by alkylthio. The oxidation is preferably carried out using a peracid e.g. peroxybenzoic acid such as m-chloroperoxybenzoic acid in an organic solvent such as a halogenated hydrocarbon e.g. methylene chloride. Preferably the reaction is carried out at a low temperature e.g. -78°C to -20°C.

Compounds of formula (I) wherein R₄ is phenyl substituted by amino may be prepared from the corresponding compound wherein R₄ is phenyl substituted by allyloxycarbonylamino by conventional means. Compounds wherein R is phenyl substituted by carboxy may be prepared from the corresponding allyl ester using conventional procedures.

Acid addition salts of compounds of formula (I) may be prepared by reaction with the appropriate acid, conveniently under non aqueous/conditions. Compounds of formula (II) in which Y = 0 may be prepared by treating a compound of formula (III) in which the group R.,, R₃, X, A, Z and R₄ have the meanings given above with an activated derivative of the acid (IV) in which R_2a is a carboxyl protecting group.

(IV)

(IH)

Suitable activated derivatives of the acid (IV) includes the corresponding acid halides e.g. acid chloride.

When the acid halide is used as the activated derivative of the acid (IV) then the reaction is preferably carried out in the presence of an acid acceptor such as a tertiary organic base for example pyridine or a trialkylamine in an aprotic solvent such as dichloromethane.

The compound of formula (II) in which Y is a phosphine group may be prepared by treating the intermediate (V) in which L is a leaving group such as a halogen e.g. chlorine.

(V)

with the corresponding phosphine e.g. triphenylphosine in the presence of a base. The reaction is conveniently carried out in a solvent such as dioxan in the presence of a tertiary organic base, e.g. 2,6 lutidine. The compounds of formula (II) are novel compounds and as such form a further aspect of the invention.

The compounds of formula (V) may be prepared from the corresponding hydroxy derivative (VI) by conventional means for converting hydroxyl groups into leaving groups.

Thus for example a compound of formula (V) in which L is a chlorine atom may be prepared by treating a compound of formula (VI) with thionyl chloride in an aprotic solvent such as dioxan or tetrahydrofuran and in the presence of a tertiary organic base e.g. 2,6-lutidine. Compounds of formula (VI) may be prepared from the reaction of a compound of formula (III) with glyoxylic ester (VII; CHOCO₂R_2a) preferably in the form of its hydrate or hemiacetal. The reaction is preferably carried out in an aprotic solvent such as toluene and in the presence of an activated molecular sieve. Compounds of formula (III) wherein R₃ is hydrogen may be prepared by reaction of the epoxide (VIII) wherein R₁ is a hydroxyl protecting group and R₈ is an alkyl group.

(VIII)

with the amine NH₂-ZXR . The reaction is preferably carried out in a solvent such as tetrahydrofuran or ethyl acetate and in the presence of a base such as potassium carbonate or triethylamine.

Compounds of formula (III) wherein R₃ is a nitrogen protecting group e.g. an alkoxycarbonyl, allyloxycarbonyl or benzyloxycarbonyl may be prepared by reacting the corresponding compound of formula (III) wherein R₃ is hydrogen with the appropriate chloroformate ester. This reaction is preferably carried out in a solvent such as tetrahydrofuran or ethyl acetate and in the presence of a base such as a tertiary amine e.g. triethylamine pyrridine or 2,6-lutidine.

The epoxides (VIII) are either known compounds or may be prepared by methods described for preparing the known compounds such as those described in EPA No. 0502488, or EPA No. 0586017.

In any of the formulae (I) to (VIII) shown above when there is an asymmetric carbon atom and no specific configuration is shown then the formula includes all possible configurations.

Specific stereoisomers of the compounds of formula (I) as defined in formulae 1a, 1b, 1c and 1d, essentially free of the other stereoisomers may be prepared by using the general processes described above starting with the appropriate stereoisomer of formula (II), (III) or (VIII) . The processes described above for preparing the compounds of formula (II) will in general give a mixture of stereoisomers.

The individual stereoisomers of the compounds of formula (II) may be prepared using the processes described above starting with the appropriate steroisomer of formula (lll) or (VIII).

In order that the invention may be more fully understood the following examples are given by way of illustration only.

In the Intermediates and Examples unless otherwise stated: All temperatures refer to °C. Solutions were dried over anhydrous sodium sulphate. All R_f values relate to thin layer chromatography (t.l.c.) on silica plates.

INTERMEDIATE 1 f3S.4R.6Ε.1^,S^-^ V1-ft-buWldimethylsilyloxy)ethvπ-^- 6^,-(1'-diethoxy phosphinyloxycvclohex-1 '-ene oxide)azetidin-2-one

INTERMEDIATE 2a

(3S.4R 3-ffR)-1-f -butyldimethylsilyloxy)ethvn-4-f(2'S.6'RV2'-N-allyloxycarbonyl

-N-(3.4-dichlorobenzylamino))-1'-oxocvclohex-6'-yl)azetidin-2-one.

To a solution of Intermediate 1 (2.75g) of ethyl acetate (25ml) with potassium carbonate (11.7g) at 0° under nitrogen, was added 3,4-dichlorobenzylamine (1.50ml) in ethyl acetate (6ml). The reaction mixture was stirred for 3 hours at 20°, then the ethyl acetate was decanted off and the residual solid was washed with ethyl acetate (10ml). The organic solution was washed with water (3x50ml) and brine (50ml), then dried and cooled to 0°. To the solution were added allyl chloroformate (2.4ml) and triethylamine (4.8ml) . The reaction mixture was stirred for 2 hours at 0°, then washed with a saturated aqueous solution of ammonium chloride (50ml), slightly acidic water (50ml), brine (2x50ml), dried and evaporated in vacuo. The residue was purified on silica gel (Rf=0.62 ethyl acetate/petroleum ether 3/2) to obtain the title compound as a pale yellow solid (670mg). IR vmax(nujol): 1763(C=0), 1718(C=0), 1703 (C=0). INTERMEDIATE 2b f3S.4R)-3-ffR)-1-ff-butyldimethylsilyloxy)ethyl)-4-ff2'S.6'R)-2'-N-allyloxycarbonyl -N-(4-bromobenzylamino)-1'-oxocvclohex-6'-vDazetidin-2-one.

To a solution of Intermediate 1 (2g) in ethyl acetate (20ml) with potassium carbonate (8.7g) at 0° under nitrogen , were added 4-bromobenzylamine hydrochloride (1.4g) and triethylamine (0.87ml). The reaction mixture was stirred for 3.5 hours at 10°, then further portions of 4-bromobenzylamine (0.35g) and triethylamine (0.22ml) were added. The reaction mixture was stirred for 3 hours at 10°, then the ethyl acetate was decanted off and the residual solid was washed with ethyl acetate (10ml). The organic solution was washed with water (2x50ml ) and brine (50ml), then dried and cooled to 0°. To the solution were added allyl chloroformate (1.2ml) and triethylamine (1.7ml). The reaction mixture was stirred for a further 1 hour at 20°, then further portions of allyl chloroformate (0.48ml) and triethylamine (0.84ml) were added. The reaction mixture was stirred for 1.5 hours at 20°, then washed with a saturated aqueous solution of ammonium chloride (50ml), slightly acidic water (50ml) brine(2x50ml), dried and evaporated ]n vacuo. The residue was purified on silica gel (Rf=0.74 ethyl acetate/ petroleum ether 7/3) to obtain the title compound as a white solid (91 Omg). IRυmax(CDCl3): 3414 (NH), 1763 (C=O), 1722 (C=O), 1697 (C=O).

INTERMEDIATE 2c (3S.4RV3-ffRV1-rf-butyldimethylsilyloxy.ethvn-4-ff2'S.6'R)-2'-N-allyloxycarbonyl

-N-f4-(trifluoromethyl.benzylamino) 1'-oxocvclohex-6'-yl)azetidin-2-one.

The title compound (530mg j was prepared as described for intermediate 2a, from Intermediate 1 (2.5g) and 4-(trifiuoromethyl)benzyl amine (1.5ml)

(Rf=0.62 ethyl acetate/petroleum ether 2/3). IR υmax(nujol): 3270-3190 (NH), 1761 (C=0), 1718 (C=0), 1705 (C=0).

INTERMEDIATE 2d f3S.4RV3- RV1-ff-butyldimethylsilyloxy>ethvn-4- 2'S.6'R)-2'-N-allyloxycarbonyl -N-f3.4-di-ff-butyldimethylsilyloxyVbenzylamino)-1'-oxocvclohex-6'-vhazetidin-2- one. To a solution of Intermediate 1 (1.64g) in ethyl acetate (15ml) with potassium carbonate (6.8g) at 0° under nitrogen, were added

3,4-di-(t-butyldimethylsilyloxy) benzylamine (2.02g) in ethyl acetate (6ml). The reaction mixture was stirred for 3 hours at 20°, then the ethyl acetate was decanted off and the residual solid was washed with ethyl acetate (10ml). The organic solution was washed with water (2x50ml ) and brine (50ml), then dried and cooled to 0°C. To the solutio'n, were added allyl chloroformate (0.85ml) and triethylamine (1.35ml). The reaction mixture was stirred for 1 hour at 20°, then further portions of allyl chloroformate (0.34ml) and triethylamine (0.45ml) were added. The reaction mixture was stirred for 0.30 hours at 20° then washed with a saturated aqueous solution of ammonium chloride (50ml), slightly acidic water (50ml), brine(2x50ml), dried and evaporated jn vacua The residue was purified on silica gel (Rf=0.57 ethyl acetate/petroleum ether 1:1 ) to obtain the title compound as a white solid (660mg). IR υmax(Nujol): 1763 (C=0), 1718 (C=0), 1701 (C=O).

INTERMEDIATE 2e fSS^RVS-qR I-^-butyldimethylsilyloxy^ethyl -f^'S.e'R^'-N-allyloxycarbonyl

-N-(4-(allyloxycarbonvπ-benzylamino)-1'-oxocvclohex-6'-vπazetidin-2-one.(Sche me 1)

To a solution of Intermediate 1 (2g) in ethyl acetate (20ml) with potassium carbonate (8.7g) at 0° under nitrogen, were added 4-(allyloxycarbonyl)benzylamine trifluoroacetate (1.39g) and triethylamine (1.2ml). The reaction mixture was stirred for 2.5 hours at 20°, then the ethyl acetate was decanted off and the residual solid was washed with ethyl acetate (10ml). The organic solution was washed with water (2x50ml ) and brine (50ml), then dried and cooled to 0°. To the solution were added allyl chloroformate (1.34ml) and 2,6-lutidine (2.2ml). The reaction mixture was stirred for 40 min. at 20°, then washed with a saturated aqueus solution of ammonium chloride (50ml), slightly acidic water (50ml), brine(2x50ml) dried and evaporated jn vacuo. The residue was purified on silica gel (Rf=0.78 ethyl acetate/petroleum ether 7:3) to obtain the title compound as a white foam (1.2g). I υmax(CDCl3): 1763 (C=0), 1718 (C=O), 1700 (C=0).

INTERMEDIATE 2f OS^RVS-ffRVI-ff-butyldimethylsilyloxy^ethyl -ff S.e'R^'-N-allyloxycarbonyl -N-(4-(allyloxycarbonylamino)benzylamino))-1'-oxocvclohex-6'-yl)azetidin-2-one. Using the process as described for Intermediate 2e the titie compound 1.98g was prepared as a white foam from Intermediate 1 (2.5g) and 4-aminobenzylamine (1.18ml) (Rf=0.75 ethyl acetate). IRυmax(CDCl3): 1761 (C=O), 1730 (C=0), 1693 (C=C)

INTERMEDIATE 2g (3S.4R)-3-f )-1-ff-butyldimethylsilyloxy)ethvn-4-ff2'S.6'R)-2'-N-allyloxycarbonyl -N-(4-nitrobenzylamino)-1'-oxocvclohex-6'-yl)azetidin-2-one.

To a solution of Intermediate 1 (2.5g) in ethyl acetate (25ml) with potassium carbonate (10.85g) at 0° under nitrogen, were added 4-nitrobenzylamine hydrochloride (1.97g) and triethylamine (1.45ml). The reaction mixture was stirred for 4 hours at 20°, then a further portion of triethylamine (0.36ml) was added. The reaction mixture was stirred for 1 hour at 20°, then the ethyl acetate was decanted off and the residual solid was washed with ethyl acetate (10ml). The organic solution was washed with water (2x50ml) and brine (50ml), then dried and cooled to 0°. To the solution were added allyl chloroformate (1.66ml) and 2,6-lutidine (2.74 ml). The reaction mixture was stirred for 1 hour at 0°, then washed with a saturated aqueous solution of ammonium chloride (50ml), slightly acidic water (50 ml), brine (2x50ml) dried and evaporated in vacuo. The residue was purified on silica gel (Rf= 0.58 ethyl acetate/petroleum ether 1/1) to obtain the titie compound as a white solid (1.00g). IRυmax(Nujol): 1761 (C=O), 1717-1705 (C=0).

INTERMEDIATE 2h f3S.4RV3-ffR)-1- -butyldimethylsilyloxy)ethvn^4-f(2'S.6'R)-2^,-N-allyloxycarbonyl -N-(4-fluorobenzylamino^-1'-oxocvclohex-6'-v azetidin-2-one. The titie compound was prepared as described for intermediate 2g, from Intermediate 1 (2g) and 4-fluorobenzylamine (0.95ml) as a white foam (1.3g) (Rf=0.52 ethyl acetate/petroleum ether 1/1). IR υmax(Nujol): 3100(NH) 1763 (C=0), 1718-1707 (C=0).

INTERMEDIATE 2i OS^RVS-ffRVI-ff-butyldimethylsilyloxy^ethyl -ff S.e'R^'-N-allyloxycarbonyl -N-(4-aminosulfonylbenzylaminoV1'-oxocvclohex-6'-vπazetidin-2-one. To a solution of Intermediate 1 (1.57g) in tetrahydofuran (15ml) at 0° under nitrogen, was added a solution of 4-(aminomethyl)benzensulfonamide (1.46g) and triethylamine (0.92ml) in tetrahydrofuran/water (1 :1) (15ml). The reaction mixture was stirred for 2.5 hours at 20° then washed with water (2x70ml), brine (70ml), dried and cooled to 0°. To the solution were added allyl chloroformate (1.05ml) and 2,6 lutidine (1.54ml). The reaction mixture was stirred for 1 hour at 0-20°, then washed with a saturated aqueous solution of ammonium chloride (70ml), slightly acidic water(70ml), brine (2x50ml), dried and evaporated jn vacuo. The residue was purified on silica gel (Rf=0.70 ethyl acetate) to obtain the title compound as a white foam (950mg). IRυmax(CDCl3): 1763 (C=0), 1724 (C=0), 1699 (C=0).

INTERMEDIATE 2i(a)

OS^R S-ffRVI-ff-butyldimethylsilyloxy^ethyl -f^'S.e'R^^'-N-allyloxycarbonyl -N-(4-(N-allyloxycarbonyl-N-allvπaminosulfonylbenzylamino)-1'-oxocvclohex-6'- vPazetidin-2-one. To a solution of 528mg of intermediate (2i) in 10 ml of ethyl acetate at 0°, was added 0.5ml of allylchloroformate and 1.3ml of triethylamine. The reaction solution was heated for 2 hours at 20° and for 2 hours at 40°, then it was washed with slightly acidic water (50ml), brine (2x50ml), dried and evaporated In vacuo. The residue was chromatographed on silica gel, using a mixture of ethyl acetate/petroleum ether (3/2), to afford 484mg of the title compound as a white foam (Rf=0.73 ethyl acetate/petroleum ether 4/1). IRυmax(CDCl3): 1763 (C=O), 1725 (C=0), 1699 (C=0).

INTERMEDIATE 2i(b) f3S.4RV3-ffRV1-ff-butyldimethylsilyloxy)ethvn-4-f(2'S.6'R)-2'-N-llyloxycarbonyl-

N-(4-fN-allyloxycarbonv)aminosulfonylbenzylamino)-1'-oxocvclohex-6'- l.azetidin-2-one.

To a solution of intermediate (2i) (84.5mg) in of ethyl acetate (1.5ml) at 0°, was added allylchloroformate (30μl) and triethylamine (100μl). The reaction solution was stirred for 1.30 hours at 20° , then it was washed with slightly acidic water (10ml), brine (2x1 Oml), dried and evaporated in vacuo. The residue was chromatographed on silica gel, using a mixture of ethyl acetate/petroleum ether (7/3), to afford 44.5mg of the title compound as a white foam (Rf=0.27 ethyl acetate/petroleum ether 4/1). IRυmax(CDCl3): 1759 (C=0), 1722 (C=0), 1699 (C=0).

INTERMEDIATE 2m

(3S.4R)-3-f(R)-1-ff-butyldimethylsilyloxy)ethvn-4-ff2^,S.6'RV2'-N-allyloxycarbonyl -N-f2-f4-bromophenvπethylamino))-1'-oxocvclohex-6'-vπazetidin-2-one. The title compound was prepared as described for intermediate 2d from

Intermediate 1 (2g) and 4-bromophenylethylamine (1.30ml) as a white solid ^* (1g). (Rf=0.78 ethyl acetate/petroleum ether 7/3). IR υmax(Nujol): 3143(NH), 1761 (C=0), 1718 (C=0), 1701 (C=C).

INTERMEDIATE 2n f3S.4RV3-ffR)-1-ff-butyldimethylsilyloxy)ethvn-4-ff2'S.6'R)-2'-N- benzyloxycarbonyl-N-(4-aminosulfonylbenzylamino)-1'-oxocvclohex-6'-vh azetidin-2-one.

The title compound was prepared as described for intermediate 2i, from Intermediate 1 (1.89g) and 4-(aminomethyl)benzensulfonamide (1.76g) as a white foam (1.1 g). (Rf=0.29 ethyl acetate/petroleum ether 7/3).

IR υmax(Nujol): 3248(NH), 1747 (C=O), 1720 (C=O).

INTERMEDIATE 2n(a) f3S.4R 3-ffRV1-ft-butyldimethylsilyloxy)ethvπ-4-ff2'S.6'RV2'-N- benzyloxy6arbonyl-N-(4-fN-benzyloxycarbonyl-N-benzyl)aminosulfonylbenzyl amino)-1 '-oxocvclohex-6'-yl)azetidin-2- one.

The title compound was prepared as described for intermediate 2i(a) from intermediate (2n) (1.00g) , benzylchloroformate (500μl) and triethylamine (800μl) as a white foam (599mg) .(Rf=0.78 ethyl acetate/petroleum ether 4/1 ).

INTERMEDIATE 2o

OS^RVS-ffRVI-rf-butyldimethylsilyloxy^ethyl -f^'S.e'R^'-N- allyloxycarbonyl-N-(4-fN-allyloxycarbonv-N-methvπaminosulfonylbenzylamino)- 1'-oxocvclohex-6'-yl.azetidin-2-one. To a solution of Intermediate (2i) (82.6mg) in ethyl acetate (1.5 ml) at 0°, was added methylchloroformate (20μl) and triethylamine (86μl). The reaction solution was stirred for 20 min. at 20° , then it was washed with amonium chloride (10ml), slightly acidic water (10ml), brine (2x10ml), dried and evaporated in vacuo. The residue was chromatographed on silica gel, using a mixture of ethyl acetate/petroleum ether (7/3), to afford 50.3mg of the title compound as a white foam (Rf=0.59 ethyl acetate/petroleum ether 7/3). IR υmaχ(Nujol): 3200 (NH), 1740 (C=0), 1624 (C=C).

INTERMEDIATE 2p

(3S.4R)-3-((R)-1-ff-butyldimethylsilyloxy)ethvn-4-ff2'S.6^,RV2'-N- benzyloxycarbonyl-N-(4-(methylthio)benzylamino))-1'-oxocvclohex-6'- yl)azetidin-2-one.

To a solution of Intermediate 1(2.0g) in ethyl acetate (20ml) with potassium carbonate (8.7g) at 0° under nitrogen, were added 4-methylthiobenzylamine (1.00g). The reaction mixture was stirred for 4 hours at 20°, then the ethyl acetate was decanted off and the residual solid was washed with ethyl acetate (10ml). The organic solution was washed with water (2x100ml) and brine (100ml), then dried and cooled to 0°. To the solution were added benzyl chloroformate (1.80ml) and 2,6-lutidine (1.95ml). The reaction mixture was stirred for 1.30 hours at 0°, then washed with a saturated aqueous solution of ammonium chloride (100ml), slightly acidic water (100ml), brine (2x100ml) dried and evaporated in vacuo. The residue was purified on silica gel (Rf= 0.73 ethyl acetate/petroleum ether 7/3) to obtain the titie compound as a white foam (500mg).

IR υmax(CDCl3): 3400 (NH), 1761 (C=O), 1718 (C=0), 1693 (C=0).

INTERMEDIATE 2q f3S.4R)-3-ffR»-1-ff-butyldimethylsilyloxy)ethvn-4-f(2'S.6'R)-2'-N- allyloxycarbonyl-N-(4-nitro-α-methylbenzylamino))-1'-oxocvclohex-6'-vπazetidin- 2-one.

To a solution of Intermediate 1 (2.5g) in tetrahydrofuran (15ml) at 0°C were added α-methyl-4-nitrobenzylamine hydrochloride (2.00g) and triethylamine (1.45ml) in water/tetrahydrofuran 2/1 (15ml) The reaction mixture was stirred for 4.30 hours at 20°, then was washed with water (2x150ml) and brine (150ml), then dried and evapored in vacuo. The crude was dissolved in dichloromethane (100ml) and cooled to 0°. To the solution were added allyl chloroformate (1.65ml) and 2,2,6,6-tetramethyl-piρeridine (3.50ml) and the reaction mixture was stirred for 20 hours at 20°. Further portions of allyl chloroformate (1.65ml) and 2,2,6,6-tetramethyl-piperidine (3.50ml) were added at 36h, 48h and 60h The reaction mixture was washed with slightly acidic water (3x200ml), brine (3x100ml) dried and evaporated in vacuo. The residue was purified on silica gel (Rf= 0.42 ethyl acetate/petroleum ether 3/2) to obtain the title compound as a white oil (680mg). IRυmax(Nujol): 3190 (NH), 1757 (C=O), 1724 (C=O), 1695 (C=C).

INTERMEDIATE 2r (3S.4RV3-ffR)-1-α-butyldimethylsilyloxy>ethyl -f(2'S.6'RV2'-N- allyloxycarbonyl-N-(2-nitrobenzylamino)-1'-oxocvclohex-6'-vπazetidin-2-one.

The title compound was prepared as described for intermediate 2p from

Intermediate 1 (2.28g), 2-nitrobenzylamine (1.36g) in ethyl acetate (5ml), allyl chloroformate (1.42ml) and 2,6 lutidine (2.1ml) as a white foam (1.60g).(Rf=0.56 ethyl acetate/petroleum ether 3/2).

IRυmax(CDCl3): 1763(C=O), 1722 (C=0), 1703 (C=O); 1528 (NO2); 1342

(NQ2).

INTERMEDIATE 2s OS^RVS-^RVI-ff-butyldimethylsilyloxy^ethyl -f^'S.e'R^'-N-allyloxycarbonyl benzylamino 1'-oxocvclohex-6'-vπazetidin-2-one. The title compound was prepared as described for intermediate 2p from Intermediate 1 (2.1 Og), benzylamine (950μl), allyl chloroformate (1.63ml), and 2,6 lutidine (2.30ml) as a white foam (1.42g). (Rf=0.60 ethyl acetate/petroleum ether 7/3).

IR υmax(Nujol): 1761 (C=0), 1699 (C=0).

INTERMEDIATE 2t f3S.4R)-3-f(R>-1-ft-butyldimethylsilyloxy)ethvn-4-ff2'S.6'R)-2'-N- allyloxycarbonylf4-chloro-2-nitro-benzylamino))-1'-oxocvclohex-6'-yl.azetidin-2- one.

The titie compound was prepared as described for intermediate 2p from Intermediate 1 (2.388g), 4-chloro-2-nitro-benzylamine (1.86g), allyl chloroformate (1.59ml), and 2,6 lutidine (2.33ml) as a yellow foam (1.274g).

(Rf=0.66 ethyl acetate/petroleum ether 3/2).

IR Vmax(Nujol): 1761 (C=0), 1720 (C=0), 1709 (C=0).

INTERMEDIATE 2u f3S.4R)-3-((R)-1-ff-butyldimethylsilyloxy)ethvn-4-ff2'S.6'R)-2'-N- allyloxycarbonylf3.4-(methylendioxy.benzylamino) 1'-oxocvclohex-6'- yl.azetidin-2-one.

The title compound was prepared as described for intermediate 2p from Intermediate 1 (2.605g), 3,4-(methylendioxy)benzyl amine (1.36ml), allyl chloroformate (1.73ml) and 2,6 lutidine (2.54ml) as a white solid (1.385g)

(Rf=0.63 ethyl acetate/petroleum ether 3/2).Mp=141.8-143.10.

IRυmax(Nujol): 1761 (C=0), 1720 (C=O), 1709 (C=0).

INTERMEDIATE 2v

(3S.4R)-3-ffR)-1-ff-butyldimethylsilyloxy)ethvn-4- 2'S.6'R)-2'-N- allyloxycarbonyl-N-f3-chloro-4-fluorobenzylamino)V1'-oxocvclohex-6'- vhazetidin-2-one.

To a solution of Intermediate 1 (2.127g) in ethyl acetate (20ml) at 0° under nitrogen was added potassium carbonate (9.23g) followed by 3-chloro-4- fluorobenzylamine (1.42g) in ethyl acetate (6ml) and the reaction mixture which was stirred for 4 hours at 20°. The ethyl acetate was decanted off and the residual solid was washed with ethyl acetate (10ml). The organic solution was washed with water (3x50ml) and brine (50ml), dried and cooled to 0°. To the solution were added allyl chloroformate (1.42ml) and 2,6-lutidine (2.1ml) . The reaction mixture was stirred for 1 hour at 0°, then washed with a saturated aqueous solution of ammonium chloride (50ml), slightly acidic water (50ml), brine (2x50ml), dried and evaporated in vacuo. The residue was purified on silica gel (Rp0.52 ethyl acetate/petroleum ether 3/2) to obtain the titie compound as a white foam (1 ,140g). IR vmax(nujol): 1763(C=0), 1720(C=O), 1697 (C=0).

INTERMEDIATE 2w f3S.4R)-3-ffR)-1-(f-butyldimethylsilyloxy)ethyl -^2'S.6'R)-2'-N- allyloxycarbonyl-N-(.4-t.methylthio)benzylamino))-1'-oxocvclohex-6'-yl)azetidin-2- one.

The title Compound (4.44g Rf=0.36 ethyl acetate/petroleum ether 1/1) was prepared as described for Intermediate 2a from Intermediate 1 (5.998g), and 4- (methylthio)benzylamine (3.84g). IR υmax(nujol): 3300 (NH), 1759 (C=0), 1718 (C=0), 1697 (C=0).

INTERMEDIATE 3a

Allyl(4S.8S.9R.10S.12R)-4-(N-allyloxycarbonyl-N-(3.4-dichlorobenzylamino 10-

3 8 f1-(f-butyldimethylsilyloxy)ethyl)-11 -oxo-1 -azatricvclo(7.2.0.0 ' )-undec-2-ene-2 -carboxylate.

A solution of triethylamine (1.18ml) and allyloxalylchloride (844mg) in dry xylene (4ml) at 0° under nitrogen, was stirred for 1 min., then a solution of intermediate (2a) (830mg) in dry xylene (8ml) was added. After 10 minutes the solution was washed with a 3% sodium hydrogen carbonate solution (50ml), water (3x50ml), brine (2x50ml), dried and concentrated to 20ml jn vacuo. The solution was heated at 80° and then was added 1.22ml of triethylphosphite. The solution was stirred at 80° for 20 hours and at reflux for 7 hours, then the solvent was removed in vacuo. The residue was chromatographed on silica gel, using a mixture of ethyl acetate/petroleum ether (1/4), to afford 432mg of the title compound as a yellow oil .(Rf=0.63 dichloromethane/diethyl ether 19/1. IRυmax(Film): 1778 (C=0), 1699 (C=O).

INTERMEDIATE 3b Allyl(4S.8S.9R.10S.12R)-4-fN-allyloxycarbonyl-N-f4-bromobenzylamino)-10-(1- (f-butyldimethylsilyloxy .ethyl)-11 -oxo-1 -azatricvclo(7.2.0.0 ' Vundec-2-ene-2- carboxylate.

A solution of triethylamine (1.26ml), allyloxalylchloride (897mg) and potassium carbonate (250mg) in dry dichloromethane (11ml) at 0°, under nitrogen, was stirred for 1 min., then a solution of intermediate (2b) (900mg) in dry dichloromethane (11 ml) was added. After 5 minutes the solution was washed with a 3% sodium hydrogen carbonate solution (50ml), water (3x50ml), brine (2x50ml), dried and evaporated in vacuo. The crude mixture was dissolved in dry xylene (50ml) and triethylphosphite (1.3ml) added, the solution heated at reflux for 6.5 hours, and then the solvent removed jn vacuo. The residue was chromatographed on silica gel, using a mixture of diethyl ether/dichloromethane (1/19), to afford of the title compound (280mg) as a yellow oil. (Rf=0.66 diethyl ether/dichloromethane). IRυmax(CDCl3): 1771(C=0), 1744(C=0), 1711(C=O),1700(C=O).

INTERMEDIATE 3c

Allyl(4S.8S.9R.10S.12R)-4-fN-allyloxycarbonyl-N-(4-ftrifluoromethvn benzylamino)-10-f 1 -( f-butyldimethylsilyloxy)ethvπ-11 -oxo-1 -azatricvclo

(7.2.0.0 ' )-undec-2-ene-2-carboxylate.

The title compound was prepared as described for intermediate 3a from intermediate (2c) (530mg), triethylamine (0.76ml) and allyloxalylchloride

(540mg) as a yellow oil (121.7mg) (Rf=0.46 dichloromethane/diethyl ether

19/1).

IRυmax(CDCl3): 1774 (C=O), 1713 (C=O), 1697 (C=0)

INTERMEDIATE 3d

Allyl(4S.8S.9R.10S.12R)-4-(N-allyloxycarbonyl-N-(3.4-di-(f-butyldimethylsilyloxy

)-benzylamino)-10-(1 -(f-butyldimethylsilyloxy)ethyl .-11 -oxo-1 -azatricvclo

(7.2.0.0^3,8)undec-2-ene-2-carboxylate.

The title compound was prepared as described for intermediate 3a from intermediate (2d) (4.87g), triethylamine (5.25ml) and allyloxalylchloride (3.70g) as a yellow oil (2.4g) (Rf=0.52 ethyl acetate/petroleum ether 1/9).

IR υmax(CDCl3): 1771 (C=O), 1713 (C=O), 1691 (C=0)

INTERMEDIATE 3e Allyl(4S.8S.9R.10S.12R -fN-allyloxycarbonyl-N-(4-(allyloxycarbonylbenzyl aminoVI 0-(1 -ff-butyldimethylsilyloxytethvπ-11 -oxo-1 -azatricvclo(7.2.0.0 '⁸.- undec-2-ene-2-carboxylate.

A solution of triethylamine (1.67ml) and allyloxalylchloride (1.20g) in dry xylene

(6ml) at 0°, under nitrogen, was stirred for 1', then a solution of intermediate (2e) (1.20g) in dry xylene/dry dichloromethane(3/1 ) (8ml) was added. After 10' minutes the solution was washed with a 3% sodium hydrogen carbonate solution (50ml), water (3x50ml), brine (2x50ml), dried and the dichloromethane residue was removed jn vacuo: after which was added triethylphosphite (1.70ml). The solution was then heated at 80°C for 6 hours and at reflux for 8 hours, then the solvent was removed jn vacuo. The residue was chromatographed on silica gel, using a mixture of ethyl acetate/petroleum ether (3/2), to afford the title compound (420mg) as a yellow oil (Rf=0.45 dichloromethane/diethyl ether 19/1). IRυmax(CDCl3): 1774 (C=0), 1715 (C=0).

INTERMEDIATE 3f

Allylf4S.8S.9R.10S.12R)-4-(N-allyloxycarbonyl-N-(4-(allyloxycarbonylamino) benzylamino .-10-(1 -(f-butyldimethylsilyloxy)ethvθ-l 1 -oxo-1 -azatricvclo

3 8

(7.2.0.0 ' Rιndec-2-ene-2-carboxylate. The titie compound was prepared as described for intermediate 3e from intermediate (2f) (1.00g), triethylamine (1.30ml) and allyloxalylchloride (970mg) as a yellow oil (274.3mg) (Rf=0.44 dichloromethane/diethyl ether 19/1). IRυmax(CDCl3): 1772 (C=0), 1720 (C=0), 1693 (C=O).

INTERMEDIATE 3g

Allyl(4S.8S.9R.10S.12R)-4-(N-allyloxycarbonyl-N-(4-nitrobenzylamino)-10-f1-(f-

3 8 butyldimethylsilyloxytethvD-11 -oxo-1 -azatricvclo(7.2.0.0 ' )-undec-2-ene-2- carboxylate.

The titie compound was prepared as described for intermediate 3e from intermediate (2g) (1.00g), triethylamine (1.48ml) and allyloxalylchloride (1.00g) as a yellow oil (672.3mg) (Rf=0.57 dichloromethane/diethyl ether 19/1).

IR υmax(CDCl3): 1776 (C=O), 1713 (C=0), 1524 (C=0).

INTERMEDIATE 3h Allyl(4S.8S.9R.10S.12R -(N-allyloxycarbonyl-N-(4-fluorobenzylamino V10-(1-(f

-butyldimethylsilyloxytethvn-11 -oxo-1 -azatricvclo(7.2.0.0 ' )-undec-2-ene-2- carboxylate.

The title compound was prepared as described for intermediate 3a from intermediate (2h) (1.88g), triethylamine (2.95ml)and allyloxalylchloride (2.10g) as a yellow oil (983.6mg) (Rf=0.48 dichloromethane/diethyl ether 19/1 ). IRυmax(Film): 1780(CO), 1715(C=0), 1699(C=0), 1600(C=C).

INTERMEDIATE 3i Allyl(4S.8S.9R.10S.12R -fN-allyloxycarbonyl-N-(4-(N-allyloxycarbonyl-N-allvn aminosulfonylbenzylamino .-10-(1 -(f-butyldimethylsilyloxytethyl .-11 -oxo-1 -

3 8 azatricvclo(7.2.0.0 ' Rιndec-2-ene-2-carboxylate.

The title compound was prepared as described for intrmediate 3a from intermediate 2i(a) (484mg), triethylamine (0.53ml) and allyloxalylchloride

(377mg) as a colourless oil (280.0mg) (Rf=0.66 dichloromethane/diethyl ether 19/1 )

IR υmax(CDCl3): 1774 (C=0), 1722 (C=0).

INTERMEDIATE 3m Allyl(4S.8S.9R.10S.12R)-4-(N-allyloxycarbonyl-N-(2-(4-bromophenvn ethylaminoV10-(1-(f-butyldimethylsilyloxy)ethvπ-11 -oxo-1 -azatricvclo(7.2.0.0 ' . -undec-2-ene-2-carboxylate.

A solution of triethylamine (1.37ml), allyloxalylchloride (970mg) and potassium carbonate (272mg) in dry xylene (13ml) at 0° under nitrogen, was stirred for 1 min. then a solution of of intermediate (2m) (1.00g) in dry xylene (13ml) was added. After 5 minutes the solution was washed with a 3% sodium hydrogen carbonate solution (50ml), water (3x50ml), brine (2x50ml), dried and concentrated to 20ml invacuo. Triethylphosphite was (1.4ml) added, the solution heated at reflux for 11 hours and then the solvent was removed in vacuo. The residue was chromatographed on silica gel, using a mixture of ethyl acetate/petroleum ether (1/3), to afford 429mg of the title compound as a yellow oil (Rf=0.73 ethyl acetate/petroleum ether 3/7). IR υmax(CDCl3): 1772 (C=0), 1713 (C=0), 1693 (C=0).

INTERMEDIATE 3n Benzyl(4S.8S.9R.10S.12R)-4-(N-benzyloxycarbonyl-N-(4-⁽N-benzyloxycarbonyl- N-benzvπaminosulfonylbenzylamino .-10-(1 -(t-butyldimethylsilyloxy)ethvπ-l 1 -ox

3 8

Q-1-azatricvclof7.2.0.0 ' Rιndec-2-ene-2-carboxylate.

The title compound was prepared as described for intermediate 3a from intermediate 2n(a) (599mg), triethylamine (584μl) and benzyloxalylchloride (557mg) as a colourless oil (391 mg) (Rf=0.59 dichloromethane/diethylether

19/1).

IR υmax(CDCl3): 1769 (C=0), 1722 (C=O).

INTERMEDIATE 3o

Allyl(4S.8S.9R.10S.12R)-4-fN-allyloxycarbonyl-N-(4-(N-allyloxycarbonyl-N- methvDaminosulfonylbenzylamino V10-(1 -(f-butyldirnethylsilyloχy)ethvθ-l 1 -oxo-1

3 8 -azatricvclo(7.2.0.0 ' )-undec-2-ene-2-carboxylate.

The title compound was prepared as described for intermediate 3a from intermediate (2o) (168mg), triethylamine (200μl) and allyloxalylchloride (145mg) as a yellow oil (40mg) (Rf=0.45 dichloromethane/diethylether 19/1 ).

INTERMEDIATE 3p Allyl(4S.8S.9R.10S.12R)-4-(N-benzyloxycarbonyl-N-(4-methyltio)benzylaminoV 10-M -(f-butyldimethylsilyloxytethvn-11 -oxo-1 -azatricvclo(7.2.0.0^3,8)-undec-2-en e-2-carboxylate.

The title compound was prepared as described for intermediate 3a from intermediate (2p) (2.22g), triethylamine (1.5ml) and benzyloxalylchloride (1.44g) as a yellow gum (779.8mg) (Rf=0.57 dichloromethane/diethylether 19/1). IR υ_max(CDCl3): 1772 (C=O), 1700-1691 (C=0).

INTERMEDIATE 3q

Allyl(4S.8S.9R.10S.12R)-4-(N-allyloxycarbonyl-N-(α-methyl-nitro)benzylamino)- 10-(1 -(f-butyldimethylsilyloxy)ethvn-l 1 -oxo-1 -azatricvclo(7.2.0.0³'⁸ϊ-undec-2- ene-2-carboxylate.

The title compound was prepared as described for intermediate 3a from intermediate (2q) (253mg), triethylamine (245μl) and allyloxalylchloride (156μl) as a colourless gum (33.8mg) (Rf=0.53 dichloromethane/diethylether 19/1). IR υmax(CDCl3): 1778 (C=0), 1715 (C=0), 1695 (C=0).

INTERMEDIATE 3r

Allylf4S.8S.9R.10S.12R -fN-allyloxycarbonyl-N-(2-nitrobenzylaminoV10-(1-(f- butyldimethylsilyloxy)ethvπ-11-OXQ-1 -azatricvclof7.2.0.0 ' )-undec-2-ene-2- carboxylate. The title compound was prepared as described for intermediate 3a from intermediate (2r) (1.60g), triethylamine (1.6ml) and allyloxalylchloride (990μl) as a yellow gum (526.3mg) (Rf=0.51 dichloromethane/diethylether 19/1). IR υmax( ilm): 1780 (C=0), 1717 (C=0), 1699 (C=0).

INTERMEDIATE 3s

Allyl(4S.8S.9R.10S.12R -(N-alMoxycarbonylbenzylamino 0-M-(f-

3 8 butyldimethylsilyloxy>ethyl)-11 -oxo-1 -azatricvclo(7.2.0.0 ' -undec-2-ene-2- carboxylate. The title compound was prepared as described for intermediate 3a from intermediate (2s) (1.42g), triethylamine (1.5ml) and allyloxalylchloride (980μl) as a colourless gum (546.9mg) (Rf=0.47 ethyl acetate/petroleum ether 1/4).

IR υmax(film): 1780 (C=O), 1699 (C=O).

INTERMEDIATE 3t

Allyl(4S.8S.9R.10S.12R)-4-fN-allyloxycarbonyl-N-(4-chloro-2-nitro-enzylamino)-

10-(1 -(f-butyldimethylsilyloxy.ethvn-11 -oxo-1 -azatricvclo(7.2.0.0³'⁸)-undec-2- ene-2-carboxylate.

The title compound was prepared as described for intermediate 3a from intermediate (2t) (1.27g), triethylamine (1.18ml) and allyloxalylchloride (750μl) as a yellow oil (441.7mg) (Rf=0.52 dichloromethane/diethylether 19/1).

IRυmax(CDCl3): 1776 (C=0), 1718 ^*C=O), 1699 (C=0), 1531 (C=C).

INTERMEDIATE 3u Allyl(4S.8S.9R.10S.12R -(N-allyloxycarbonyl-N-((3.4-methylendioxy)benzyl

3 8 aminoWI 0-(1 -(f-butyldimethylsilyloxy)ethyl .-11 -oxo-1 -azatricvclo(7.2.0.0 ' .- undec-2-ene-2-carboxylate.

The title compound was prepared as described for intermediate 3b from intermediate (2u) (1.36g), triethylamine (1.35ml) and allyloxalylchloride (1.10ml) as a yellow oil (573.3mg) (Rf=0.51 dichloromethane/diethylether 19/1). IR υmax(CDCl3): 1772 (C=0), 1715 (C=0), 1691 (C=0), 1651 (C=C).

INTERMEDIATE 3v Allyl(4S.8S.9R.10S.12R)-4-(N-allyloxycarbonyl-N-(3-chloro-4- fluorobenzylaminoM 0-(1 -(f-butyldimethylsilyloxy)ethvθ-l 1 -oxo-1 -azatricvclo- (7.2.0.0 ' undec-2-ene-2-carboxylate.

A solution of triethylamine (1.26ml) and allyloxalylchloride (0.80ml), in dry xylene (10ml) at 0° under nitrogen, was stirred for 1 min., then a solution of intermediate (2v) (1.104g) in dry xylene (12ml) was added. After 30 minutes the solution was washed with a 3% sodium hydrogen carbonate solution (100ml), water (3x100ml), brine (2x100ml), dried and concentrated to 40ml in vacuo. Triethylphosphite (2.36ml) was added to the reaction mixture which was then heated at 80° at 80° for 12.5 hours and at reflux for 10.5 hours, then the solvent was removed in vacuo. The residue was chromatographed on silica gel, using a mixture of ethyl acetate/petroleum ether (15/85), to afford of the title compound as a colourless oil (527.4mg, Rf=0.55 dichloromethane/diethyl ether 19/1). IRυmax( i-m): 1774 (C=O), 1715 (C=O), 1693 (C=0), 1501 (C=C).

INTERMEDIATE 3w

Allyl(4S.8S.9R.10S.12R)-4-(N-allyloxycarbonyl-N-(4-(methylthio)benzylaminoV 10-M-(f-butyldimethylsilyloxytethvD-11-oxo-1 -azatricvclo(7.2.0.0³-⁸.-undec-2- ene-2-carboxylate. The title compound was prepared as described for Intermediate 3a from intermediate (2w) (4.44g), triethylamine (4.40ml) and allyloxalylchloride (2.80ml) as a colourless oil (1.752g), (Rf=0.66 dichloromethane/diethyl ether 19/1). IRυmax(CDCl3): 1772 (C=O), 1713 (C=O), 1691 (C=O)

EXAMPLE 1a Allyl(4S.8S.9R.10S.12R -(N-allyloxycarbonyl-N-(3.4-dichlorobenzylaminoV10- Q

(1 -hvdroxyethvπ-11 -oxo-1 -azatricvclo(7.2.0.0 ' .-undec-2-ene-2-carboxylate. To a solution of of intermediate (3a) (432mg) in dry tetrahydrofuran (15 ml) under nitrogen at 0° was added glacial acetic acid (0.28ml) followed by of tetrabutylammonium fluoride trihydrate (925mg). The solution was stirred at 20° for 26 hours. The solution was then poured into ethyl acetate (25ml) and washed with a saturated solution of ammonium chloride (30ml), 3% sodium hydrogen carbonate solution (2x30ml), water (30ml) and brine (30ml), dried and evaporated jn vacuo. The residue was chromatographed on silica gel, using ethyl acetate as eluant, to give the title compound as a colourless oil (185mg) (Rf=0.41 ethyl acetate/petroleum ether 3/2).

IR υmax(CDCl3): 1776 (C=0), 1720 (C=0), 1700 (C=0), 1600 (C=C).

EXAMPLE 1b

Allyl(4S.8S.9R.10S.12R)-4-(N-allyloxycarbonyl-N-(4-bromobenzylamino)-10-(1-

3 8 hvdroxyethyl >-11 -oxo-1 -azatricvclo(7.2.0.0 ' .-undec-2-ene-2-carboxylate.

1ml of a solution of 0.4ml of glacial acetic acid and 4.8ml of tetrabutylammonium fluoride 1.1 M/tetrahydrofuran to (10ml) with tetrahydrofuran was added to intermediate (3b) (280mg) in dry tetrahydrofuran under nitrogen. The solution was stirred at 20°C for 4 hours, then a further portion of 0.3ml of solution of acetic acid/tetrabutylammonium fluoride was added and stirred at 20°C for 16 hours. The solution was then poured into 20ml of ethyl acetate and washed with a saturated solution of ammonium chloride (30ml) 3% sodium hydrogen carbonate solution (2x30ml), water (30ml) and brine (30ml), dried and evaporated in vacuo. The residue was chromatographed on silica gel, using ethyl acetate as eluant, to give 78.4mg of the title compound as a colourless oil.

(Rf=0.2 ethyl acetate/petroleum ether 1/1). IR υmaχ(CDCl3): 1776 (C=O), 1712 (C=O), 1696 (C=O).

EXAMPLE 1c

Allyl(4S.8S.9R.10S.12R)-4-(N-allyloxycarbonyl-N-(4-(trifluoromethyl.benzyl amino)-10-(1 -hvdroxyethyl V-11-oxo-1 -azatricvclo(7.2.0.0 ' )-undec-2-ene-2- carboxylate.

To a solution of intermediate (3c) (120mg) in distilled tetrahydrofuran (0.8ml) was added glacial acetic acid (0.15ml) followed by tetrabutylammonium fluoride trihydrate (790mg). The solution was stirred at 40° for 3 hours. The solution was then poured into ethyl acetate (15ml) and washed with a saturated solution of ammonium chloride (20ml) 3% sodium hydrogen carbonate solution (20ml), water (20ml) and brine (20ml), dried and evaporated jn vacuo. The residue was chromatographed on silica gel, using ethyl acetate/petroleum ether as eluant, to give the titie compound as a colourless oil (55.7mg) (Rf=0.25 ethyl acetate/petroleum ether 1/1). IR υmax(CDCl3): 1774 (C=0), 1713 (C=0), 1699 (C=0). EXAMPLE 1d

Allyl(4S.8S.9R.10S.12R)-4-(N-allyloxycarbonyl-N-(3.4-dihvdroxybenzylaminoV 10-(1-(f-butyldimethylsilyloxy ethvn-11 -oxo-1 -azatricvclo(7.2.0.0^3,8 undec-2- ene-2-carboxylate.

To a solution of of intermediate (3d) (1.80g) in distilled tetrahydrofuran (100ml) under nitrogen at 0° was added glacial acetic acid (0.60ml) followed by tetrabutylammonium fluoride trihydrate (2g). The solution was stirred at 20° for 40 min. The solution was then poured into of ethyl acetate (100 ml) and washed with a saturated solution of ammonium chloride (100ml) 3% sodium hydrogen carbonate solution (2x100ml), water (100ml) and brine (100ml), dried and evaporated jn vacuo. The residue was chromatographed on silica gel, using ethyl acetate/petroleum ether 3/2 as eluant, to give the title compound as a white foam (1.3g) (Rf=0.69 ethyl acetate/petroleum ether 1/1). IRυmax(CDCl3): 1772 (C=0), 1717 (C=0), 1695 (C=O).

EXAMPLE 1d(a) Allyl(4S.8S,9R.10S.12R)-4-(N-allyloxycarbonyl-N-(3.4-dihvdroxybenzylamino)-

3 8

10-(1-hvdroxyethvπ-11 -oxo-1 -azatricvclo(7.2.0.0 ' )-undec-2-ene-2-carboxylate The title compound was prepared as described in example 1 a from intermediate

(3d) (1.3g), glacial acetic acid (0.88ml) and tetrabutylammonium fluoride trihydrate (2.94g) as a white foam (310.7mg) (Rf=0.28 ethyl acetate/petroleum ether 4/1 ).

IR υmax(CDCl3): 3500-3400 (OH) 1776 (C=O), 1769 (C=O), 1720-1690 (C=O and C=C).

EXAMPLE 1e

Allyl(4S.8S.9R.10S.12R)- -(N-allyloxycarbonyl-N-(4-(allyloxycarbonvhbenzyl amino)-10-(1 -hvdroxyethvD-11 -oxo-1 -azatricvclo(7.2.0.0 ' )-undec-2-ene-2- carboxylate.

The title compound was prepared as described in example 1c from intermediate (3e) (420mg), glacial acetic acid (0.53ml) and tetrabutylammonium fluoride trihydrate (2.74g) as a white foam (168mg) (Rf=0.10 ethyl acetate/petroleum ether 1/1). IRυmax(CDCl3): 1774 (C=0), 1715 (C=0). EXAMPLE 1f

Allyl(4S.8S.9R.10S.12R)-4-(N-allyloxycarbonyl-N-(4-(allyloxycarbonylamino.

3 8 benzylamino)-10-(1 -hvdroxyethvD-11 -oxo-1 -azatricvclo(7.2.0.0 ' Rιndec-2-ene -2-carboxylate.

The title compound was prepared as described in example 1 c from intermediate

(3f) (274.3mg), glacial acetic acid (0.34ml) and tetrabutylammonium fluoride trihydrate (1.72g) as a white foam (103.7mg)(Rf=0.08 ethyl acetate/petroleum ether 1/1). IRυmax(CDCl3): 1772 (C=0), 1734 (C=0), 1718(C=0), 1690 (C=0).

EXAMPLE 1g

Allyl-f4S.8S.9R.10S.12R -(N-allyloxycarbony[-N-(4-nitrobenzylamino .-10-(1 -

3 8 hvdroxyethvπ-11 -oxo-1 -azatricvclo(7.2.0.0 ' )-undec-2-ene-2-carboxylate. The title compound was prepared as described in example 1 c from intermediate (3g) (590mg), glacial acetic acid (0.79ml) and tetrabutylammonium fluoride trihydrate (4.07g) as a yellow oil (233.8mg) (Rf=0.30 ethyl acetate/petroleum ether 1/1). IR υmax(KBr): 1776 (C=0), 1713 (C=O).

EXAMPLE 1h

Allyl(4S.8S.9R.10S.12R -(N-allyloxycarbonyl-N-(4-fluorobenzylamino 10-(1-

3 8 hvdroxyethylVI 1-OXQ-1 -azathcvclof7.2.0.0 ' )-undec-2-ene-2-carboxylate.

The title compound was prepared as described in example 1c from intermediate (3h) (108mg), glacial acetic acid (0.14ml) and tetrabutylammonium fluoride trihydrate (720mg) as a yellow oil (60.8mg) (Rf=0.30 ethyl acetate/petroleum ether 1/1).

IR υmax(CDCl3): 1774 (C=0), 1715 (C=0), 1693 (C=0), 1605 (C=C).

EXAMPLE 1i

Allyl(4S.8S.9R.10S.12R -(N-allyloxycarbonyl-N-(4-(N-aHyloxycarbonyl-N-allvn

-aminosulfonylbenzylaminoVI 0-(1 -hvdroxyethvn-11 -oxo-1 -azatricvclo(7.2.0.0^3,8

.-undec-2-ene-2-carboxylate.

The title compound was prepared as described in example 1c from intermediate (3i) (280mg), glacial acetic acid (0.28ml) and tetrabutylammonium fluoride trihydrate (1.46g) as a white foam (200mg) (Rf=0.10 ethyl acetate/petroleum ether 1/1).

IR υmax(CDCl3): 1776 (C=0), 1722 (C=0), 1710 (C=0).

EXAMPLE 1m

Allyl(4S.8S.9R.10S.12R)-4-(N-allyloxycarbonyl-N-(2-(4-bromophenvnethyl

3 8 amino .-10-(1 -hvdroxyethyl)-11 -oxo-1 -azatricvclo(7.2.0.0 ' )-undec-2-ene-2- carboxylate.

The title compound was prepared as described in example 1a from intermediate (3m) (429mg), glacial acetic acid (0.26ml) and tetrabutylammonium fluoride

1.1M/tetrahydrofuran (2.5ml) as a yellow oil (195.3mg) (Rf=0.27 ethyl acetate/petroleum ether 1/1).

IRυ_max(CDCl3): 1772 (C=0), 1713(C=0), 1693 (C=0).

EXAMPLE 1n

Benzyl(4S.8S.9R.10S.12R)-4-(N-benzyloxycarbonyl-N-(4-(benzyloxycarbonyl-N- (4-(N-benzyloxycarbonyl-N-benzyl.aminosulfonilbenzylamino).- 10-(1-

3 8 hvdroxyethvπ-11 -oxo-1 -azatricvclo(7.2.0.0 ' )-undec-2-ene-2-carboxylate.

The title compound was prepared as described in example 1c from intermediate (3n) (391 mg), glacial acetic acid (335μl) and tetrabutylammonium fluoride trihydrate (1.72g) as a white foam (150.1mg) (Rf=0.34 ethyl acetate/petroleum ether 7/3).

IRυ_max(CDCl3): 1774 (C=O), 1720 (C=0).

EXAMPLE 1o

Allyl(4S.8S.9R.10S.12R -(N-allyloxycarbonyl-N-(4-(N-allyloxycarbonyl-N- methvπ-aminosulfonylbenzylamino)-10-(1 -hvdroxyethylVI 1 -oxo-1 -azatricvclo

(7.2.0.0 ' .-undec-2-ene-2-carboxylate.

The title compound was prepared as described in example 1c from intermediate (3o) (38.4mg), glacial acetic acid (43μl) and tetrabutylammonium fluoride trihydrate (221 mg) as a colourlesss oil (15mg) (Rf=0.25 ethyl acetate/petroleum ether 7/3).

EXAMPLE 1p Benzyl(4S.8S.9R.10S.12R -(N-benzyloxycarbonyl-N-(4-(methylthioVbenzyl 3 8 aminoV-10-(1 -hvdroxyethvD-l 1 -oxo-1 -azatricvclo(7.2.0.0 ' )-undec-2-ene-2- carboxylate.

The title compound was prepared as described in example 1 c from intermediate (3p) (770mg), glacial acetic acid (900μl) and tetrabutylammonium fluoride trihydrate (4.65g) as a white foam (637.9mg) (Rf=0.17 ethyl acetate/petroleum ether 1/1). IR υmax(CDCl3): 3450-3150 (OH), 1776 (C=0), 1717 (C=0), 1699 (C=0).

EXAMPLE 1p(a) Benzyl(4S.8S.9R.10S.12R -fN-benzyloxycarbonyl-N-(4-(methylsulfoxide)-

3 8 benzylaminoVI 0-(1-hvdroxyethyl)-11 -oxo-1 -azatricvclo(7.2.0.0 ' )-undec-2-ene

-2-carboxylate.

To a solution of example (1p) (78.9mg) in distilled dichloromethane (5ml), at -

78° was added dropwise meta-chloro-perbenzoic acid (50%) (38.4mg) in dichloromethane (1.5ml). The solution was stirred at -78° for 1 hour, then a further portions of meta-chloro-perbenzoic acid (38.4mg; 50%) was added. The reaction mixture was stirred for 30 min. at -780C then washed with a sodium sulfite solution 3% (50ml), sodium hydrogen carbonate solution, 1 % (2x50ml), brine (50ml), dried and evaporated jn vacuo. to give 83.2mg of the title compound as a white gum (Rf=0.16 ethyl acetate).

IR υmax(Nujol): 1776 (C=O), 1717 (C=0), 1695 (C=O), 1086 (S=0), 1049

(S=0).

EXAMPLE 1q

Allyl(4S.8S.9R.10S.12R)-4-(N-allyloxycarbonyl-N-(4-nitro- -methylbenzyl

3 8 aminoVI 0-1 -hvdroxyethvB-11 -oxo-1 -azatricvclo(7.2.0.0 ' ϊ-undec-2-ene-2- carboxylate.

The title compound was prepared as described in example 1 c from intermediate (3q) (33mg), glacial acetic acid (43μl) and tetrabutylammonium fluoride trihydrate (220mg) as a colourless gum (10.4mg) (Rf=0.36 ethyl acetate/petroleum ether 3/2).

IR υmax(CDCl3): 1776 (C=0), 1695 (C=0).

EXAMPLE 1r

Allyl(4S.8S.9R.10S.12R -(N-allyloxycarbonyl-N-(2-nitrobenzylaminoV10-(1- hvdroxyethvn-11 -oxo-1 -azatricvclo(7.2.0.0 ' )-undec-2-ene-2-carboxylate. The title compound was prepared as described in example 1 c from intermediate (3r) (526.3mg), glacial acetic acid (700μl) and tetrabutylammonium fluoride trihydrate (3.63mg) as a yellow gum (261 mg) (Rf=0.29 ethyl acetate/petroleum ether 1/1). IRυ_max(CDCl3): 1776 (C=0), 1715 (C=0), 1699 (C=0).

EXAMPLE 1s

Allyl(4S.8S.9R.10S.12R -(N-allyloxycarbonylbenzylamino)-10-(1 -hvdroxyethyl

3 8

H 1 -oxo-1 -azatricvclo(7.2.0.0 ' )-undec-2-ene-2-carboxylate. The title compound was prepared as described in example 1 c from intermediate

(3s) (546.3mg), glacial acetic acid (790μl) and tetrabutylammonium fluoride trihydrate (3.48g) as a colourless oil (254.6mg) (Rf=0.19 ethyl acetate/petroleum ether 2/3).

IR υmax(CDCl3): 1774 (C=0), 1711 (C=0), 1691 (C=0).

EXAMPLE 1t

Allyl(4S.8S.9R.10S.12R -(N-allyloxycarbonyl(4-chloro-2-nitrobenzylamino .-

3 8 10-(1 -hvdroxyethvπ-11 -oxo-1 -azatricvclo(7.2.0.0 ' )-undec-2-ene-2-carboxylate

The title compound was prepared as described in example 1c from intermediate (3t) (413.2mg), glacial acetic acid (530μl) and tetrabutylammonium fluoride trihydrate (2.32g) as a colourless oil (161.9mg) (Rf=0.48 ethyl acetate/petroleum ether 1/1).

IR υmax(CDCl3): 1776 (C=0), 1710 (C=0).

EXAMPLE 1u

Allyl(4S.8S.9R.10S.12R)-4-(N-allyloxycarbonyl(3.4-(methylendioxy)benzyl

3 8 amino)-10-(1 -hvdroxyethvn-11 -oxo-1 -azatricvclo(7.2.0.0 ' .-undec-2-ene-2- carboxylate.

The title compound was prepared as described in example 1c from intermediate (3u) (505.7mg), glacial acetic acid (680μl) and tetrabutylammonium fluoride trihydrate (2.99g) as a colourless oil (220.3mg) (Rf=0.20 ethyl acetate/petroleum ether 1/1).

IRυmax(Film): 3400 (OH), 1776(C=O), 1699(C=0).

EXAMPLE 1v Allyl(4S.8S.9R.10S.12R)-4-(N-allyloxycarbonyl-N-(3-chloro-4-

3 8 fluorobenzylaminoV10-(1-hvdroxyethyl)-11 -oxo-1 -azatricvclo(7.2.0.0 ' )-undec-

2-ene-2-carboxylate

To a solution of Intermediate (3v) (527.4mg) in distilled tetrahydrofuran (3.5ml) was added glacial acetic acid (0.7ml) followed by tetrabutylammonium fluoride trihydrate (3.2g) . The solution was stirred at 40° for 2.5 hours. The solution was then poured into of ethyl acetate (100ml) and washed with a saturated solution of ammonium chloride (100ml) 3% sodium hydrogen carbonate solution (100ml), water (100ml) and brine (100ml), dried and evaporated in vacuo. The residue was chromatographed on silica gel, using ethyl acetate/petroleum ether7/3 as eluant, to give the title compound

(271.2mg) as a white foam (Rf=0.17 ethyl acetate/petroleum ether 1/1).

IR υmax(CDCl3): 1774 (C=0), 1717 (C=0), 1695 (C=0).

EXAMPLE 1w

Allyl(4S.8S.9R.10S.12R)-4-(N-allyloxycarbonyl-N-(4-(methylthio)benzylamino)-

3 8 10-(1 -hvdroxyethylVI 1 -oxo-1 -azatricvclo(7.2.0.0 ' )-undec-2-ene-2-carboxylate

The title compound was prepared as described in Example 1v from intermediate (3w) (1.75g), glacial acetic acid (2.23ml), and tetrabutylammonium fluoride trihydrate (9.80g) as a colourless gum (640mg) (Rf=0.15 ethyl acetate/petroleum ether 2/3).

IRυmax(CDCl3): 1772 (C=O), 1715 (C=O), 1693 (C=O).

EXAMPLE 2a (4S.8S.9R.10S.12R)-4-(3.4-dichlorobenzylamino)-10-(1 -hvdroxyethvn-11 -oxo-1 - azatricvclo-(7.2.0.0 3 ' 8 Vundec-2-ene-2-carboxylic acid.

To a solution of example (1a) (185mg) in dry tetrahydrofuran (3ml) at 20° under nitrogen, was added 5,5-dimethyl-1,3-cyclohexandione (190mg). After 5 minutes a solution of tetrakis(triphenylphosphine)palladium (155mg) in dry tetrahydrofuran (3ml) was added to the above solution. After 50 minutes, ethyl acetate (30ml) was added and the product was extracted with water (3x20ml). The aqueous solutions were combined and washed with ethyl acetate (3x40ml) and diethyl ether (2x40ml). The organic solvent remaining was removed in vacuo.The aqueous solution was freeze-dried to obtain title compound as a white solid (45.8mg). HPLC (column= Lichrosphere RP-C18, flow=1 ml/min., eluant=acetonitrile

/buffer (pH=7.4) 25/75): Rt=7.5 min, 90% purity.

IRυmax(Nujol): 3346 (OH), 3194(NH), 1759 (C=0).

EXAMPLE2b

(4S.8S.9R.10S.12R -(4-bromobenzylaminoV10-f 1 -hvdroxyethvn-11 -oxo-1 -

3 8 azatricvclo-(7.2.0.0 )undec-2-ene-2-carboxylic acid. The title compound was prepared as described in example 2a from of example (1 b) (105mg), 5,5-dimethyl-1 ,3-cyclohexadiene (108mg) and tetrakis(thphenylphosphine)palladium (109.8mg) as a white solid (22.8mg). HPLC (column= Lichrosphere RP-C18, flow=1 ml/min., eluant=acetonitrile /buffer (pH=7.4) 20/80): Rt=9.0 min, 80% purity. IR υmax(Nujol): 3360 (OH+NH), 1772 (C=0), 1593 (C=C).

EXAMPLE 2c

(4S.8S.9R.10S.12R)-4-(4-(trifluoromethylbenzylamino)-10-(1 -hvdroxyethyl .-11 -

3 8 oxo-1 -azatricvclo-(7.2.0.0 ' undec-2-ene-2-carboxylic acid.

The titie compound was prepared as described in example 2a from example

(1c) (50mg), 5,5-dimethyl-1 ,3-cyclohexadiene (52mg) and tetrakis(tripheriylphosphine)palladium (42mg) as a white solid (15.2mg).

HPLC (column= Lichrosphere RP-C18, flow=1 ml/min., eluant=acetonitrile

/buffer (pH=7.4) 25/75): Rt=6.0 min, 70% purity.

IR υmax(Nujol): 3335 (OH), 3182 (NH), 1761 (C=0), 1593 (C=C).

EXAMPLE 2d

(4S.8S.9R.10S.12R)-4-(3.4-(dihvdroxybenzylamino)-10-(1 -hvdroxyethvn-11 - oxo-1 -azatricvclo-(7.2.0.0 3 ' 8 -undec-2-ene-2-carboxylic acid.

The title compound was prepared as described in example 2a from example

(1d) (310m)g, 5,5-dimethyl-1,3-cyclohexadiene (340mg) and tetrakis(triphenylphosphine)palladium (280mg) as a white solid (214.5mg).

HPLC (column= Novapac C18 S4, flow=1 ml/min., eluant=acetonitrile /buffer

(pH=7.4) 1/9): Rt=1.3 min, 80% purity.

IR υmax(Nujol): 3422 (OH), 3173 (NH), 1751 (C=0).

EXAMPLE 2e (4S.8S.9R.10S.12R)-4-(4-carboxybenzylamino )-10-(1 -hydroxyethylV11 -oxo-1 -

3 8 azatricvclo-(7.2.0.0 ' .-undec-2-ene-2-carboxylic acid. The title compound was prepared as described in example 2a from example (1e) (84.6mg), 5,5-dimethyl-1,3-cyclohexadiene (126mg) and tetrakis(triphenylphosphine)palladium (104mg) as a white solid (34.7mg).

HPLC (column= Lichrosphere C18 S5, flow=1 ml/min., eluant=acetonitrile /buffer (pH=7.4) 15/85): Rt=4.78 min, 60% purity. IRυmax(Nujol): 3348 (OH), 3180 (NH), 1761 (C=0).

EXAMPLE 2f

(4S.8S.9R.10S.12R)-4-(4-aminobenzylamino V10-(1 -hydroxyethvIM 1 -oxo-1 -

3 8 azat cvclo-(7.2.0.0 ' )-undec-2-ene-2-carboxylic acid.

The title compound was prepared as described in example 2a from example

(1f) (103mg), 5,5-dimethyl-1 ,3-cyclohexadiene (149mg) and tetrakis(triphenylphosphine)palladium (123mg) as a white solid (59.5mg).

HPLC (column= Lichrosphere RP-18, flow=1 ml/min., eluant=acetonitrile /buffer

(pH=7.4) 7/93): Rt=7.00 min, 80% purity.

IRυmax(Nujol): 3344 (OH), 3211 (NH), 1755 (C=O), 1610 (C=C).

EXAMPLE 2g

(4S.8S.9R.10S.12R -(4-nitrobenzylamino)-10-M -hvdroxyethvn-11 -oxo-1 -

3 8 azatricvclo-(7.2.0.0 ' Rιndec-2-ene-2-carboxylic acid.

The titie compound was prepared as described in example 2a from of example

(1 g) (233mg), 5,5-dimethyl-1 ,3-cyclohexadiene (248mg) and tetrakis(triphenylphosphine)palladium (203mg) as a white solid (59.9mg).

HPLC (column= Lichrosphere RP-18, flow=1 ml/min., eluant=acetonitrile /buffer

(pH=7.4) 22 78): Rt=4.00 min, 80% purity.

IRυmax(Nujol): 3500(OH.NH), 1778(C=O), 1717 (C=O).

EXAMPLE2h

(4S.8S.9R.10S.12R)-4-(4-fluorobenzylamino)-10-(1 -hvdroxyethyl .-11 -oxo-1 - azatricvclo-(7.2.0.0 ' )-undec-2-ene-2-carboxylic acid.

The title compound was prepared as described in example 2a from example

(1h) (60mg) 5,5-dimethyl-1,3-cyclohexadiene (68mg) and tetrakis(triphenylphosphine)palladium (55mg) as a white solid (37.9mg). HPLC (column= Lichrosphere RP-C-18, flow=1 ml/min., eluant=acetonitrile

/buffer (pH=7.4) 15/85): Rt=7.00 min, 75 purity.

IR υmax(Nujol): 3346-3182 (OH.NH), 1763 (0=0), 1610 (C=C).

EXAMPLE 2i

(4S.8S.9R.10S.12R)-4-((4-(N-allvnaminosulfonylbenzylamino)-10-(1-

3 8 hvdroxyethyl )-11 -oxo-1 -azatricvclo-(7.2.0.0 ' ϊ-undec-2-ene-2-carboxylic acid.

The titie compound was prepared as described in example 2a from of example

(1 i) (130mg) 5,5-dimethyl-1 ,3-cyclohexadiene (213mg) and tetrakis(triphenylphosphine)palladium (180mg) (after purification by HPLC) as a white solid (5.9mg).

HPLC (column= Lichrosphere C18 S5, flow=1 ml/min., eluant=acetonitrile /buffer

(pH=7.4) 22/78): Rt=4.5 min, 90 purity.

IRυmax(Nujol): 1755 (C=O), 1587 (C=C).

EXAMPLE 21

(4S.8S.9R.10S.12R -(3.4-dimethoxybenzylamino)-10-f1-hvdroxyethvn-11-oxo-

3 8 1 -azatricvclo-(7.2.0.0 ' )-undec-2-ene-2-carboxylic acid.

To a solution of Intermediate 1 (1.1g) in ethyl acetate (10ml) with potassium carbonate (3.0g) at 10°C under nitrogen, was added 3,4-dimethoxybenzylamine (0.35ml). The reaction mixture was stirred for 1 hour at 10°C, then cooled to 0°C at which point were added allyl chloroformate (0.5ml) and triethylamine (0.6ml) . The reaction mixture was stirred for 1 hour at 20°C, then washed with a saturated aqueous solution of ammonium chloride (30ml), slightly acidic water (30ml), brine (2x30ml), dried and evaporated jn vacuo. The residue was purified on silica gel (eluant diethyl ether) to obtain the (3S.4R)-3-((R)-1-(t-butyldimethylsilyloxy)ethyl -((2'S.6'R.-2'-N- allyloxycarbonyl-N-(3.4- dimethoxybenzylamino))-1'-oxocvclohex-6'-vn azetidin-2-one (360mg; intermediate (21)). A solution of of triethylamine (0.31ml) and allyloxalylchloride (0.18 ml) in dry dichloromethane (7ml) at 0°, under nitrogen, was stirred for 3 min., then a solution of 300mg of intermediate (21) in dry dichloromethane (2ml) was added to the above. After 30 minutes, at 20°, the solution was washed with 3% sodium hydrogen carbonate solution (50ml), water (3x50ml), brine (2x50ml), dried and evaporated in vacuo. The residue was dissolved in xylene (10ml) and 5eq. of t ethylphosphite was added. The solution was refluxed for 24 hours, then the solvent was removed jn vacuo. The residue was chromatographed on silica gel, using a mixture of diethyl ether/petroleum ether (1/1), to afford of

Allyl(4S.8S.9R.10S.12R)-4-fN-allyloxycarbonyl-N-(3.4-dimethoxy benzyl amino )-10-(1 -(t-butyldimethylsilyloxytethyl )-11 -oxo-1 -azatricvclo(7.2.0.0^3, . undec-2-ene-2-carboxylate (40.3mg). This was dissolved in dry tetrahydrofuran (5ml) and, under nitrogen at 0° was added glacial acetic acid (0.015ml) followed by 0.19ml of tetrabutylammonium fluoride 1 M/tetrahydrofuran. The solution was stirred at 20° for 21 hours. The solution was then poured into 10ml of ethyl acetate and washed with a saturated solution of ammonium chloride (20ml) 3% sodium hydrogen carbonate solution (2x20ml), water (20ml) and brine (20ml), dried and evaporated jn vacuo. The residue was chromatographed on silica gel (diethyl ether/ethyl acetate 4/1 ) to give 11.4mg of

Allyl(4S.8S.9R.10S.12R -(N-allyloxycarbonyl- N-(3.4-di

3 8 methoxybenzylamino 10-(1-hvdroxyethyl)-11 -oxo-1 -azatricvclo(7.2.0.0 ' ) undec-2-ene-2-carboxylate This was dissolved in dry tetrahydrofuran (1 ml) at

20° under nitrogen , 5,5-dimethyl-1 ,3-cyclohexandione (12mg) was added. A solution of tetrakis(triphenylphosphine)palladium (12mg) in dry tetrahydrofuran

(1ml) was added to the above solution after 5 minutes. After 60 minutes, ethyl acetate (5ml) was added and the product was extracted with water (3x2ml). The aqueous solutions were combined and washed with ethyl acetate (3x5ml) and diethyl ether (2x5ml). The organic solvent remaining was removed jn vacuo.The aqueous solution was freeze-dried to obtain the title compound as a white solid (3.6mg). HPLC (column= Lichrosphere RP-C18, flow=1 ml/min., eluant=acetonitrile /buffer (pH=7.4) 9/1): Rt=9.0 min, 88% purity. IRυmax(Nujol): 1757 (C=0), 1583 (C=C).

EXAMPLE 2m (4S.8S.9R.10S.12R -((4-bromophenvDethylamino V-10-(1 -hvdroxyethvπ-11 -oxo -1 -azatricvclo-(7.2.0.0 ' undec-2-ene-2-carboxylic acid. The title compound was prepared as described in example 2a from example 1 m (156.4mg) 5,5-dimethyl-1 ,3-cyclohexadiene (155.2mg) and tetrakis(triphenylphosphine)palladium (126.2mg) (after purification by HPLC) as a white solid (1.1 mg), HPLC (column= Lichrosphere 018 S5, flow=1 ml/min., eluant=acetonitrile /buffer

(pH=7.4) 25/75): Rt=6.00 min, 85 purity.

IR υmax(Nujol): 3333-3188 (OH.NH), 1752 (C=0), 1636-1603 (C=C).

EXAMPLE 2n

(4S.8S.9R.10S.12R)-4-((4-(N-benzvnaminosulfonylbenzylamino)-10-M -hvdroxy

3 8 ethyl)-11 -oxo-1 -azatricvclo-(7.2.0.0 ' .-undec-2-ene-2-carboxylic acid.

To a solution of of example (1n) (66.4mg) in 9 ml of H2θ/isoprpyl alcohol/ethyl acetate 1/1/1 at 20°C under nitrogen, was added of glacial acetic acid (4.5μl) , 16mg of Pd/C 10% and H2 . After 45 minutes, the Pd was filtered and washed with water (10ml) and ethyl acetate (10ml). The aqueous solutions were washed with ethyl acetate (10ml), diethyl ether (2x1 Oml) and concentred jn vacuo. The aqueous solution was freeze-dried to obtain the title compound as a white solid (13.3mg). HPLC (column= Lichrosphere RP-C18, flow=1 ml/min., eluant= acetonithle/buffer (pH=7.4) 25/75): Rt=5.0 min, >80% purity. IR υmax(Nujol): 3184 (OH), 1752 (0=0).

EXAMPLE 2o (4S.8S.9R.1 OS.12R)-4-((4-(N-methyl.aminosulfonylbenzylaminoV10-(1 -hvdroxy

3 8 ethvO-11 -oxo-1 -azatricvclo-(7.2.0.0 ' .-undec-2-ene-2-carboxylic acid.

The title compound was prepared as described in example 2a from example

(1 o) (15mg) 5,5-dimethyl-1 ,3-cyclohexadiene (19mg) and tetrakis(triphenylphosphine)palladium (16mg) as a white solid (5mg). HPLC (column= Lichrosphere RP-C18, flow=1 ml/min., eluant=acetonithle

/buffer (pH=7.0) from 0/100 to 40/60): Rt=14.0 min, 50% purity.

IRυmax(Nujol): 3427 (COOH), 3190 (NH), 1750 (0=0).

EXAMPLE 2q (4S.8S.9R.10S.12R -((4-nitro α-methylbenzylaminoV10-(1 -hvdroxyethvn-11 - oxo-1- azatricvclo-(7.2.0.0 ' .-undec-2-ene-2-carboxylic acid. The title compound was prepared as described in example 2a from of example (1q) (15mg) 5,5-dimethyl-1,3-cyclohexadiene (9.8mg) and tetrakis(triphenylphosphine)palladium (6.4mg) as a white solid (5.5mg). HPLC (column= Lichrosphere RP-C18, flow=1 ml/min., eluant=acetonitrile /buffer (pH=7.4) from 15/85 to 40/60): Rt=13.5 min, 50% purity. IR υmax(Nujol): 3184 (NH), 1755 (C=O), 1599 (C=C).

EXAMPLE 2r

(4S.8S.9R.10S.12RΪ-4-(2-nitrobenzylaminoV10-(1 -hvdroxyethvn-11 -oxo-1 - 3 8 azatri cvclo-(7.2.0.0 ' -undec-2-ene-2-carboxylic acid.

To a solution of of example (1 r) (71.7mg) in dry tetrahydrofuran (2ml) at 20° under nitrogen, was added 5,5-dimethyl-1 ,3-cyclohexandione (57.3mg) and Pd(PPh3)2θl2 (28.6mg). After 2 hours at 200, a further portion of Pd(PPh3)2d2 (0.1eq.) was added but the reaction did not work. Then, after 45 minutes, Pd(PPh3)4 (47.5mg) was added, and the product was extracted with water (3x20ml). After 45 minutes, ethyl acetate (70ml) was added and the product was extracted with water (10ml). The aqueous solution was washed with ethyl acetate (2x1 OmI) and diethyl ether (2x10ml), then the remaining organic solvent was removed jn vacuo and the aqueous solutions was freeze-dried to obtain title compound as a white solid (12.6mg). The initial ethyl acetate solution was back extracted with water (50ml), and the above procedure was repeated once more to give the title compound as a white solid (17.6mg). HPLC (column= Lichrosphere RP-C18, flow=1 ml/min., eluant=acetonithle /buffer (pH=7.4) from 15/85 to 40/60): Rt=10.81 min, 80% purity. IRυmax(Nujol): 3437 (NH), 1757 (C=0), 1582 (C=O).

EXAMPLE 2s (4S.8S.9R.10S.12R)-4-(benzylaminoV10-(1 -hvdroxyethyl V11 -oxo-1 -azatricvclo-

(7.2.0.0³'⁸)-undec-2-ene-2-carboxylic acid.

The title compound was prepared as described in example 2a from example

(1s) (127.3mg) 5,5-dimethyl-1,3-cyclohexadiene (82mg) and tetrakis(triphenylphosphine)palladium (15.3mg) as a white solid (76.3mg) HPLC (column= Lichrosphere RP-C18, flow=1 ml/min., eluant=acetonitrile

/buffer (pH=7.4) from 15/85 to 60/40): Rt=4.86 min, >90% purity.

IR υmax(Nujol): 3437 (NH), 1761 (0=0), 1593 (C=C).

EXAMPLE 2t (4S.8S.9R.10S.12R -(4-chloro-2-nitro-benzylaminoH 0-(1 -hvdroxyethyl 3 8 11 -oxo-1 -azatri cvclo-(7.2.0.0 ' Vundec-2-ene-2-carboxylic acid.

The titie compound was prepared as described in example 2a from example (1t)

(88.3mg) 5,5-dimethyl-1,3-cyclohexadiene (48.8mg) and tetrakis(triphenylphosphine)palladium (9.1 mg) as a white solid (23.5mg). HPLC (column= Lichrosphere RP-C18, flow=1 ml/min., eluant=acetonitrile

/buffer (pH=7.4) from 25 75 to 60/40): Rt=5.31 min, >80% purity.

IRυmax(Nujol): 3430(OH), 3180(NH), 1780(C=0).

EXAMPLE2u (4S.8S.9R,10S.12R -(3.4-(methylendioxy)benzylamino)-10-(1-hvdroxyethyl)-

38 11-OXO-1-azatricvclo-(7.2.0.0 ' )-undec-2-ene-2-carboxylicacid.

The title compound was prepared as described in example 2a from example

(1u) (136.8mg) 5,5-dimethyl-1,3-cyclohexadiene (80.7mg) and tetrakis(triphenylphosphine)palladium (15mg) as a white solid (25.8mg). HPLC (column= Lichrosphere RP-C18, flow=1 ml/min., eluant=acetonitrile

/buffer (pH=7.4) from 15/85 to 60/40): Rt=5.84 min, 80% purity.

IRυmax(Nujol): 3408 (NH), 1747 (C=O).

EXAMPLE 2v (4S.8S.9R.10S.12R -(3-chloro-4-fluorobenzylamino)-10-(1 -hvdroxyethyl )-11 -

3 8 oxo-1 -azatricvclo-(7.2.0.0 ' )-undec-2-ene-2-carboxylic acid

To a solution of Example (1v) (226.8mg) in dry tetrahydrofuran (3.5ml) at 20° under nitrogen, was added 5,5-dimethyl-1 ,3-cyclohexandiene (132mg). After 5 minutes a solution of of tetrakis(triphenylphosphine)palladium (28.4mg) in dry tetrahydrofuran (1ml) was added to the above solution. After 45 minutes, of ethyl acetate (30ml) was added and the product was extracted with water (30ml). The aqueous solution was washed with ethyl acetate (2x40ml) and diethyl ether (2x40ml). The solvent was removed in vacuo to obtain to give the title compound as a white solid (65mg). HPLC (column= Lichrosphere RP-C18, flow=1 ml/min., eluant=acetonitrile /buffer (pH=7.4) 1/3-3/2): Rt=4.03 min, 85% purity. IR υmax(Nujol): 3454 (OH), 3362 (NH), 1759 (C=O), 1616 (C=C).

EXAMPLE 2w (4S.8S.9R.10S.12R -(4-(methylthio)benzylamino)-10-(1-hvdroxyethyl.-11-oxo-

3 8

1 -azatricvclo-(7.2.0.0 )undec-2-ene-2-carboxylic acid. The title compound was prepared as described in Example 2v. from Example (1w), (98.1mg), 5,5-dimethyl-1,3-cyclohexadiene (57mg) and tetrakis(triphenylphosphine)palladium (10.7mg) as a white solid (25mg).

HPLC (column= Lichrosphere RP-C18, flow=1 ml/min., eluant=acetonitrile

/buffer (pH=7.4) 9/41): Rt=7.57 min, 85% purity.

IR υmax(Nujol): 3458 (OH), 3186 (NH), 1738 (0=0), 1582 (C=C).

Pharmacy Example

Dry Powder for Injection active ingredient 538mg per vial.

Fill sterile vials with the sterile active ingredient. Purge the vial head space with sterile nitrogen; close the vials using rubber plugs and metal oversells (applied by crimping). The product may be constitute by dissolving in Water for Injection (10ml) or other suitable sterile vehicle for injection shortly before administration.

The antibacterial activity of the compounds of the invention may be readily determined using conventional test procedures. For example the antibacterial activity of the compounds of the invention was determined using a standard mictoriter broth serial dilution test. In this test the broth was incubated with approximately 10⁵ colony forming units of the test organism and incubated at 35° for 18 hours in the presence of test compound. Results obtained using the rest procedure are given in the table below and are expressed as minimum inhibitory concentrations (MIC) in micrograms/ml. Example No's

Organism 2g 2h 2i 2s 2q 2t

S Aureus 663E 0.25 0.25 0.25 0.25 0.12 0.12

E Coli TEMI 0.06 0.5 0.12 . 0.25 0.25 0.12

E Cloacae 0.5 2 1 0.5 2 1

C Perfringens 0.03 0.06 0.03 0.01 0.03 0.6

The compounds of the invention are essentially non toxic at therapeutically useful doses. For example no adverse effects were observed when compounds of the invention were administered to mice or rats at therapeutically useful dose levels.

Claims

Claims

(I) A compounds of general formula (I)

σ)

salts and metabolically labile esters thereof; wherein R₁ represents hydrogen or a hydroxyl protecting group; R₂ represents hydrogen or a carboxyl protecting group;

R₃ represents hydrogen or a nitrogen protecting group;

Z represents a methylene group optionally substituted by methyl;

X represents a bond or a C-^alkylene chain or an ethenylene chain;

A represents a C₂^alkylene chain or A is a chain of 2 to 4 members one of which is an oxygen or sulphur atom or the group NH or a substituted derivative thereof and the other members are methylene groups; and R₄ represents an optionally substituted phenyl group.

2. A compound as claimed in claim 1 wherein Ri R₂ and R₃ each represent hydrogen.

3. A compound as claimed in claim 1 or claim 2 wherein Z is methylene.

4. A compound as claimed in any of claims 1 to 3 wherein X is a bond.

5. A compound as claimed in any of claims 1 to 4 wherein A is a propylene chain.

6. A compound as claimed in any of claims 1 to 5 wherein R4 is phenyl or phenyl substituted by one or 2 groups selected from halogen, alkoxy, trifluoromethyl, carboxy, amino, nitro, methylenedioxy, alkylthio or SO₂NHR₇ wherein R₇ is methyl, allyl or benzyl.

7. A compound as claimed in any of claims 1 to 6 wherein R* is phenyl, or phenyl substituted by one or two groups selected from, chlorine, fluorine, nitro, methylenedioxy, alkylaminosulphonyl.

8. Compounds as defined in any of claims 2 to 7 for use in therapy.

9. The use of a compound as defined in any df claims 2 to 7 in the manufacture of a medicament for the treatment of bacterial infections.

10. A pharmaceutical composition comprising a compound as defined in any of claims 2 to 7 in admixture with one or more physiologically acceptable carriers as excipients.

11. A method of treatment of a human or non-human body to combact bacterial infections with method comprises administering to the body an effective amount of a compound as claimed in any of claims 2 to 7 or a physiologically acceptable salt thereof.

12. A process for the preparation of compounds as claimed in claim 1 which comprises cyclisation of a compound of formula (II)

(π)

in which Rι, R₂, R₃, X, Z and R, have the meanings defined in formula (I) or is a group convertible thereto, and X is oxygen or a phosphine group and if required or desired subjecting the compound, prior to or subsequently to any separation into its stereochemical isomers, to one or more of the following operations:

(a) removal of one or more protecting groups

(b) conversion of one compound of formula (I) into another compound of formula (I)