CA2252696A1 - Azetidinone derivatives for the treatment of atherosclerosis - Google Patents

Abstract

Selected compounds of formula (I) in which: R1 is hydrogen or a corresponding pharmaceutically acceptable ester or a pharmaceutically acceptable salt thereof; R2 and R3 which may be the same or different is each selected from hydrogen or optionally substituted C(1-6)alkyl; X is a group X'(CH2)m in which X' is CO, CONR4, COO, CONR4CO, CONHO or CH2O in which R4 is hydrogen or C(1-6)alkyl and m is 0 or an integer from 1 to 12; or a C(1-12)alkylene chain optionally interrupted by X'; and Y is an optionally substituted aryl group;
having the absolute configuration (4R,SS); are inhibitors of the enzyme Lp-PLA2 and thereof of use in treating atherosclerosis.

Description

CA 022~2696 1998-10-22 AZETIDINONE DERIVATIVES FOR THE TREATMENT OF ATHEROSCLEROSIS

The present invention relates to certain novel monocyclic ~-lactam compounds, processes for their preparation, intermediates useful in their preparation, pharmaceutical compositions ccnt~ining them and their use in therapy, in particular in the tre~tmen~ of atherosclerosis.
WO 95/00649 (Smith~Cline Reech~m plc) describe the phospholipase A2 enzyme Lipoprotein Associated Phospholipase A2 (Lp-PLA2), the sequence, isolation and purification thereof, icr)l~ted nucleic acids encoding the enzyme, and recombinant host cells transformed with DNA encoding the enzyme. Suggested therapeutic uses for inhibitors of the enzyme included atherosclerosis, diabetes, rhel~m~toid arthritis, stroke, myocardial infarction, reperfusion injury and acute and chronic infl~mm~rion. A subsequent publication from the same group further ~escr~ibes this enzyme (Tew D et al. Arterioscler Thromb Vas Biol 1996:16;591-9) wherein it is referred to as LDL-PLA2. A later patent application (WO
9S/0992 1, Icos Corporation) and a related publication in Nature (TjoeLker et al, vol 374, 6 April 1995, 549) describe the enzyme PAF-AH which has essenti~lly the same sequence as Lp-PLA2 and suggest that it may have potential as a therapuedc protein for reguladng pathological infl~m m~tory events.
It has been shown that Lp-PLA2 is responsible for the conversion of phosphaddylcholine to Iysophosphatidylcholine, duAng the conversion of low density lipoprotein (LDL) to its oxidised form. The enzyme is known to hydrolyse the sn-2 ester of the oxidised phosphatidylcholine to give Iysophosphatddylcholine and an oxidadvely modified fatty acid. Both products of Lp-PLA2 action are biologically active with lysophosphatidylcholine, a component of oxidised LDL, known to be a potent chemoattractant for circulating monocytes. As such, Iysophosphatidylcholine is thought play a signific~nt role in atherosclerosis by being responsible for the accumuladon of cells loaded with cholesterol ester in the arteAes. Inhibition of the Lp-PLA2 enzyme would therefore be expected to stop the build up of these macrophage enAched lesions (by inhibition of the formation of Iysophosphaddylcholine and oxidised free fatty acids) and so be useful in the treatrnent of atherosclerosis.
The increased Iysophosphatidylcholine content of oxidatively modified LDL is also thought to be responsible for the endothelial dysfunction observed in ~ . .

p~ti.o.nt.c with atherosclerosis. Inhibitors of Lp-PLA2 could therefore prove bçn~fi~ in the treatment of this phenomenon. An Lp-PLA2 inhibitor could also find utility in other disease states that exhibit endothelial dysfunction including bete-s, hypellension, angina pectoris and after ischaemia and reperfusion.
In addition, Lp-PLA2 inhibitnrs may also have a general application in any d~ordt;r that involves activated monocytes, macrophages or lymphocytes, as all of these cell types express Lp-PLA2. Examples of such disorders include psoriasis.
Furthermore, Lp-PLA2 inhibitors may also have a general application in any disorder that involves lipid peroxidation in conjunction with Lp-PLA2 activity to produce the two injurious products, lysophosphatidylcholine and oxidatively modif1ed fatty acids. Such conditions include the aforementioned conditions atherosclerosis, diabetes, rhPum~toid arthritis, stroke, myocardial infarction, l~lfusion injury and acute and chronic infl~mm~tion. Further such conditions include various neuropsychiatric disorders such as schizophrenia ~seePsychopharmacology Bulletin, 31, 159-165, 1995).
An earlier patent application ~VO 96/19451, SmithKline Beech~m plc) discloses compounds of the formula (A):

R,~S(O)nR3 o N~

(A) in which:
Rl and R2, which may be the same or different, is each selected from hydrogen, halogen or optionally substituted C(1 g)aLkyl;
R3 is aryl or arylC(1 4)alkyl which may be optionally substit~ltpr~:
X is a linker group;
Y is an optionally substituted aryl group; and nisO, 1 or2.
Such compounds of formula (A) are inhibitors of Lp-PLA2 and as such are expect~d to be of use in treating atherosclerosis and the other disease conditions noted above.

2 (SmithKline Beecham plc) describes further compounds having a substiuent such a methyl on the carbon attached to N- 1. In addtion, wO 97/41098 PC~/EP97/01898 PCT/EP96/05587 (SmithKline Beecham plc) discloses pro-drugs of compounds of formula (A) in which R3 is a 4-carboxybenzyl group.
Compounds of formula (A) exist in a number of stereoisomeric forms.
The C-4 carbon of the ,B-lactam ring is a chiral centre which will give rise to the pre~nce of stereoisomers. Furthermore, in compounds of formula (A) in which n ~ is 1, that is sulphoxide compounds, the presence of the SO moiety will introduce an additional chiral centre into the molecule and therefore give rise to the eYistence of extra stereoisomers. Preferred compounds of formula (A) are said tobe those in which the relative configurations at C-4 and the SO moiety are R,S
(4R,SS) and S,R (4S,SR) with the most preferred compounds having the absolute configuration (4R,SS).
Accordingly, the present invention provides for a compound of the formula (I):

~,~SOCH2~ CO2R' N~
~ CR2R3 XY
(I) in which:
R1 is hydrogen or a corresponding ph~rrn~ceu~ically acceptable ester or a pharm~e~ltic~lly acceptable salt thereof;
R2 and R3 which may be the same or different is each selected from hydrogen or optionally substituted C( 1 6)alkyl;
X is a group X'(CHz)m in which X' is CO, CoNR4, COO, CoNR4Co, CONHO
or CH20 in which R4 is hydrogen or C( 1 6)aLkyl and m is 0 or an integer from 1 to 12, or a C(1 12)alkylene chain optionally interupted by X'; and Y is an optionally substituted aryl group;
having the absolute configuration (4R,SS).
Representative examples of X include CO(CH2)m, CONH(CH2)m, COO(CH2)m, CON~CO(CH2)m. CoNHo(cH2)m CH2~(CH2)m and C(1 12)aLkylene. Preferably, X' is CO, CONR2 or CH20, more preferably CONH. Preferably, m is l, 2, 5, 6, 7 or 9, preferably 6. Suitably, X is CONH(CH2)6 or CH20(CH2)6. preferably CONH(CH2)6.
Suitably, R2 and R3 is each hydrogen or R2 is hydrogen and R3 is methyl.
Preferably, R2 and R3 is each hydrogen. It will be readily appreciated that when wo 97/41098 PCT/EP97/01898 R2 and R3 have different values, the carbon to which they are attached will be chiral. Preferably, the absolute configuration at this carbon is S. In such compounds of formula (1), the absolute configuration of the preferred enantiomeris (a-S, 4-R, S-S).
Suitably, Y is phenyl, optionally substituted by up to three further su~stituent~. Suitable substituen~s include halo, hydroxy, C(1 g)alkyl and C(l ~)alkoxy. Preferably, Y is phenyl optionally substituted by halo, more preferably 4-chloro or 4-fluoro-phenyl, most preferably, 4-auoro-phenyl.
Suitably X-Y is CONH(CH2)6Ph(4-F)/(4-Cl).
Suitable pharmaceutic~lly acceptable esters include C(1 6)alkyl or C(2 6)alkenyl esters, as well as ph~rm~eutically acceptable in vivo hydrolysableesters. The skilled person will appreciate that simple C(1 6)alkyl or C(2 6)alkenyl benzoate esters show little if any tendency to break down in the human body, to leave the parent acid or its salt, even though they may be susceptible to in vivo hydrolysis in ~nim~lc such as rabbits and dogs. The term "in vivo hydrolysable ester" is not therefore conventionally considered to include such esters.
Suitable C(1 6)alkyl or C(2 6)aL~cenyl esters include ethyl and allyl esters.
Suitable ph~rm~reutically acceptable in vivo hydrolysable ester groups for incorporation in R1 include those which break down readily in the human body to leave the parent acid or its salt.
Suitable values of R1 for use in vivo hydrolysable esters include:
-CH(Ra)O.CO.Rb;
-CH(Ra)O.CO.ORC;
-CH(Ra)CO.N~.eRf -RdNReRf;
-CH20Rg;
CH2 Rh 0~"0 o -CH(Ra)O.CO.C6H4Y 1 COCH(Ri)NH2; and 97/41098 PCTtEPg7/01898 -CH23J~o~Rb in which:
Ra is hydrogen, (C1-6)aLkyl, in particular methyl, (C3-7)cycloaLkyl, or phenyl, each of which may be optionally substituted;
Rb is (Cl-6)alkyl, (Cl-6)alkoxy(Cl-6)alkyl, phenyl, benzyl, (C3-7)cycloaL~cyl, (C 1 -6)alkyl(C3-7)cycloalkyl, 1 -amino(C 1 -6)aL~cyl, or 1-(Cl-6aL~cyl)amino(Cl-6)alkyl, each of which may be optionally subs~itut~d: or Ra and Rb together form a 1,2-phenylene group optionally substituted by one or two methoxy groups;
RC is (Cl-6)alkyl, (C3-7)cycloaLkyl, (Cl-6)aLkyl(C3-7)cycloaLkyl;
Rd is (C1-6)aLkylene optionally substituted with a methyl or ethyl group;
Re and Rf which may be the same or different is each (C1-6)aL~cyl; or aryl(Cl-4)aLkyl, optionally sl-bstituted with e.g. hydroxy;
Rg is (Cl-6)allcyl;
Rh is hydrogen, (Cl-6)alkyl or phenyl;
Ri is hydrogen or phenyl optionally substituted by up to three groups selected from halogen, (Cl-6)-aL~yl, or (Cl-6)alk~XY;
and yl is oxygen or NH.
Suitable values of R1 include:
(a) acyloxyallcyl groups such as acetoxymethyl, isobutyryloxymethyl, pivaloyloxymethyl, benzoyloxymethyl, a-acetoxyethyl, a-pivaloyloxyethyl, l-(cyclohexylcarbonyloxy)ethyl, (l-aminoethyl)carbonyloxymethyl, 2-methoxyprop-2-ylcarbonyloxymethyl, phenylcarbonyloxymethyl and 4-methoxyphenyl-carbonyloxymethyl;
~b) aL~oxy/cycloalkoxycarbonyloxyalkyl groups, such as ethoxycarbonyloxymethyl, t-butyloxycarbonyloxymethyl, cyclohexyloxycarbonyloxymethyl, l-methylcyclohexyloxycarbonyloxymethyl and a-ethoxycarbonyloxyethyl;
(c) dialkylaminoaL~cyl, especially di-loweraLkylamino aLkyl groups such as dimethylaminomethyl, dimethylaminoethyl, diethylaminomethyl or diethylaminoethyl;

. . .. . ...

CA 022~2696 1998-10-22 (d) S ~et ~mido groups such as N,N-dimethylaminocarbonylmethyl, N,N-(2-hydroxyethyl)aminocarbonylmethyl;
(e) lactone groups such as phthalidyl and dimethoxyphthalidyl;
(f) (5-methyl-2-oxo-1,3-dioxolen-4-yl)methyl; and (g) (2-methoxycarbonyl-E-but-2-en-yl)methyl.
Representative examples of Rl include:
(2-methoxycarbonyl-E-but-2-en-yl)methyl, isobutyryloxymethyl, 2-methoxyprop-2-ylcarbonyloxymethyl, phenylcarbonyloxymethyl, 4-methoxyphenyl-carbonyloxymethyl, t-butyloxycarbonyloxymethyl.
cyclohexyloxy-carbonyloxymethyl, 1-methylcyclohexyloxycarbonyloxymethyl, N,N-dimethylaminocarbonylmethyl, N-N-di-(2-hydroxyethyl)aminocarbonylmethyloxy and (S-methyl-2-oxo-1,3-dioxolen-4-yl)methyl.
Especially preferred compounds of Formula (I) include:
(4R, SS)-N-[6-(4-Fluorophenyl)hex-1-yl]-4-(4-carboxybenzylsulphinyl)-2-oxoazetidin-l-yl)flçet~mide and pharmaceuticaly acceptable salts thereof, in particular the sodium salt.
Since the compounds of Formula (I) are intended for use in pharmaceutical compositions, it will be understood that they are each provided in subst~nti~lly pure form. for example at least 50% pure, more suitably at least 75%
pure and preferably at least 95% pure (~o are on a wt/wt basis). ~mpure prepara~ions of the compounds of Formula (I) may be used for preparing the more pure forms used in the pharmaceutical compositions. Although the purity of interme~ S~ compounds of the present invention is less critical. it will be readily understood that the subst~n~i~lly pure form is preferred as for the compounds of Formula (I). Preferably, whenever possible. the compounds of the present invention are obtained in crystalline form.
When some of the compounds of this invention are allowed to crystallise or are recryst~lliced from organic solvents, solvent of cryst~llic~tion may be present in the crystalline product. This invention includes within its scope such solvates. Similarly, some of the compounds of this invention may be crystallisedor recrystallised from solvents containing water. In such cases water of hydration may be formed. This invention includes within its scope stoichiometric hydrates as well as compounds cont~ining variable arnounts of water that may be produced by processes such as Iyophilisation. In addition, different cryst~lIic~tion CA 022~2696 1998-10-22 W O97/41098 PCT~EP97/01898 conditiQns may lead to the formation of different polymorphic forms of crystalline products. This invention includes within its scope all polymorphic forms of the compounds of Formula (I).
Compounds of the present invention are inhibitors of the enzyme lipoprotein associated phospholipase A2 (Lp-PLA2) and as such are expected to be of use in therapy, in particular in the tre~tmPnt of atherosclerosis. In a further aspect therefore the present invention provides a compound of Formula (I) for use in therapy.
The compounds of Formula (I) are inhibitors of lysophosph~ti~ylcholine pro~luction by Lp-PLA2 and may therefore also have a general application in any disorder that involves endothelial dysfunction, for exarnple atherosclerosis, diabetes, hypertension, angina pectoris and after isch~emi~ and reperfusion. In addition, compounds of Formula (I) may have a general application in any disorder that involves lipid peroxidation in conjunction with enzyme activity, for el~mple in addition to conllitions such as atherosclerosis and diabetes, other conditions such as rheum~toid arthritis, stroke, infl~mm~tory conditions of the brain such as Alzheimer's Disease, myocardial infarction, reperfusion injury, sepsis, and acute and chronic infl~mm~tion. Further such conditions include various neuropsychiatric disorders such as schizophrenia (see Psychopharmacology Bulletin, 31, 159-165, 1995).
Further applications include any disorder that involves activated monocytes, macrophages or lymphocytes, as all of these cell types express Lp-PLA2. Examples of such disorders include psoriasis.
Accordingly, in a further aspect, the present invention provides for a method of treating a disease state associated with activity of the enzyme Lp-PLA2 which method involves treating a patient in need thereof with a therapeutically effective amount of an inhibitor of the enzyme. The disease state may be ~soci~d with the increased involvement of monocytes, macrophages or lymphocytes; with the formation of Iysophosphatidylcholine and oxidised free fatty acids; with lipid peroxidation in conjunction with Lp-PLA2 activity; or with endothelial dysfunction.
Compounds of the present invention may also be of use in treating the above mentioned disease states in combination with anti-hyperlipidaemic or anti-atherosclerotic or anti-diabetic or anti-anginal or anti-infl~mm~fory or anti-hypertension agents. Examples of the above include cholesterol synthesis CA 022~2696 1998-10-22 Wo 97/41098 PCT/EPg7/0}898 inhibitors such as statins, anti-oxidants such as probucol, insulin se~ tice c~lci~lm ch~nnel antagonists, and anti-infl:-mm~tQry drugs such as NSAIDs.
In therapeutic use, the compounds of the present invention are usually p"lmini.~tPred in a standard ph~rmaçeutical composition. The present invention therefore provides, in a further aspect, a pharmaceutical composition comprisinga compound of Formula {I) and a pharmaceutically acceptable carrier.
Suitable ph~rm~reuti~l compositions include those which are adapted for oral or parenteral ~mini~tration or as a suppository.
The compounds of Formula (~) which are active when given orally can be formulated as liquids, for example syrups, suspensions or emulsions, tablets, c~ps~ s and lozenges.
A liquid formulation will generally consist of a suspension or solution of the compound or pharmaceutically acceptable salt in a suitable liquid carrier(s)for example, ethanol, glycerine, non-aqueous solvent, for example polyethylene glycol, oils, or water with a suspending agent, preservative, flavouring or colouring agent.
A composition in the forrn of a tablet can be prepared using any suitable ph~rm~eutical carrier(s) routinely used for pfep~ing solid formulations.
Examples of such carriers include m~gn~ium stearate, starch, lactose, sucrose and cellulose.
A composition in the form of a capsule can be prepared using routine encars~ tion procedures. For example, pellets containing the active ingredient can be prepared using standard carriers and then f1lled into a hard gelatin capsule;
alternatively, a dispersion or suspension can be prepared using any suitable pharmaceutical carrier(s), for example aqueous gums, celluloses, silicates or oils and the dispersion or suspension then filled into a soft gelatin capsule.
Typical parenteral compositions consist of a solution or suspension of the compound of Formula (I) in a sterile aqueous carrier or parenterally acceptable oil, for example polyethylene glycol, polyvinyl pyrrolidone, lecithin, arachis oil or sesame oil. Alternatively, the solution can be lyophilised and then reconstituted with a suitable solvent jus~ prior to ~mini~tration.
A typical suppository formulall~n comprises a compound of Formula (I) which is active when ~mini.~tPred in this way, with a binding and/or lubricatingagent such as polymeric glycols, gelatins or cocoa butter or other low melting vegetable or synthetic waxes or fats.

wo 97/41098 pcTlEps7lol898 Preferably the composition is in unit dose form such as a tablel or c~rs~llP,.
Each dosage unit for oral ~(1mini~stration contains preferably from 1 to 500 mg (and for parenteral ~(1ministration contains preferably from 0.1 to 25 mg) of a compound of the Formula (I).
The daily dosage regimen for an adult patient may be, for example, an oral dose of between 1 rng and 1000 mg, preferably between 1 mg and 500 mg, or an intravenous, subcutaneous, or intramuscular dose of between 0.1 mg and 100 mg, preferably between 0.1 mg and 2~ mg, of the compound of the Forrnula (I), the compound being ~iminictered 1 to 4 times per day. Suitably the compounds will be ~dmini.ctered for a period of continuous therapy, for exarnple for a week or more.
Compounds of formula (I) may be prepared from convenient starting materials by adapting synthetic procedures well known in the art, with referenceto earlier applications WO 96/19451 and WO 97/02242 (SmithKline Beecham plc).
Preferred compounds of formula (I) in which X is an amide CONH may be prepared by a process which comprises treating a compound of forrnula (II):

~ SCH2~ CO2R1 o N~

(II) in which Rl is C(1 6)alkyl or C(2 6)a~kenyl and R2 and R3 are as hereinbefore ~Pfine~ and having the absolute configuration (4R,SS);
with an amine of the formula (III):

H2N(CH2)nY
(III) under suitable amide forming conditions, for instance in the presence of an activating agent such as N,N-dicyclohexylcarbodiimide and 1-hydroxybenzotriazole in a suitable solvent such as dry dimethylformamide;
and thereafter, and if neceSS~ry, . .

CA 022~2696 1998-10-22 W O 97/41098 PCT~EP97/01898 - (a) removing the ester group under suitable de-esterifying conditions to form the acid;
(b) converting the acid to a pharrnaceutically acceptable salt; and/or (c) converting the acid, a suitable salt, the ester or an activated derivative of the acid, to an in vivo hydrolysable ester by reaction with a compound of formula (IV):

RlR4 (IV) in which:
R4 is a reactive esterifying leaving group; and Rl is as hereinbefore defined;
under ester forming conditions.
In step (a) above, the free acid may be regenerated from a corresponding compound in which the carboxy group is protected as a C(1 6)aLkyl or C(2 6)alkenyl ester; using methods well known in the art for the particular protecting group, for in.ct~nce, when an allyl group, using palladium catalysed de-allylation (triphenylphosphine/ pyrrolidine/ tetrakis triphenyl-phosphinep~ m(o) in dichloromethane).
In step (b) above, salts by prepared by treating the corresponding acid with an appropriate base.
Suitable ester forming conditions for use in step (c) above are well known in the art and are described in, for instance, Comprehensive Organic Synthesi~c,Pergamon Press, 1991, 6, 323-380. Suitable ester forming conditions include:
(a) reacting a salt of the acid, for inct~nce, a sodium or a tertiary amine salt such as triethylamine, with a compound of formula (IV), in a polar aprotic solvent such as dimethyl formamide, dimethyl sulphoxide or acetonitrile, at moderate temperature, for instance in the range 0 to 100~C;
(b) reacting the acid with a compound of formula (IV) in the presence of a base such as an alkali metal carbonate or a tertiary amine, in a polar aprotic solvent and temperature as for (a);
(c) reacdng the acid with a compound of formula (IV) in which R7 is a hydroxyl group, under dehydrating conditions, for instance the Mitsunobu reaction employing an azodicarboxylate and a trivalent phosphorus reagent (Mitsunobu, Synthesis, 1981, 1); or ~ (d) reacting an activated derivative of the acid, for in.~t~n~e a mixed anhydride, for in.ct~nre an iso-butylcarbonic or a meth~ne sulphonic anhydride or a carbodiimide (DCC) adduct, with a compound of formula (IV) in which R7 is a hydroxyl group, in the plesence of a suitable base such as a tertiaryarnine, forin.ct~nce, triethylamine, in an aprotic solvent such as tetrahydrofuran, at a moderate temperature, preferably in the range -20 to +20~C, or alternatively, inthe ~bsellce of a base but using a preformed salt of the alcohol, for in.ct~nce the rn~JIesium or lithium ~lko~ifle, Preferred conditions include the use of the sodium salt of the acid in combination with a halide or sulphonate derivative of the compound of formula (IV).
Compounds of formula (II) are useful interme~ tes in the preparation of a compound of formula (I). Accordingly, in a further aspect, the present inventionprovides for a compound of formula (II) as hereinbefore defined.
Compounds of formula (II) are sulfoxides and may be readily prepared by oxidising a corresponding thio compound of formula (V):

SCH2~ CO2R' r~
o N~ CR2R3--C02H
(V) in which Rl, R2 and R3 are as hereinbefore defined, and having the absolute configuration (4R);
with a conventional oxidising agent such as m-chloroperbenzoic acid (mcpba) or ozone and thereafter and, if necess~ry~ isolating the required diastereoisomer having the desired absolute configuration (4R, SS), for in.ct~n(~e by fractionalcryst~lli.c~tion and/or chromatography.
Compounds of formula (V) are of use in preparing compounds of formula (I). Accordingly, in a further aspect, the present invention provides for compounds of formula (V) as hereinbefore defined.
Compounds of formula (I) may also be obtained by an alternative process in which the two steps hereinbefore described are reversed, that is a compound of formula (V) is first treated with a compound of formula (III), to form the amide -W O 97/41098 PCT~EP9~tO1898 ~ bond, and the reslllt~nt thio intprme~i~te then oxi~i.ced to the correspondingsulfoxide of formula (II), preferably using a chiral o~ icing system which yields the required isomer as the predominant product.
Compounds of formula (V) having the absolute configuration (4R) may be obtained from the cc~ ponding racemic compound of formula (VI):

O ~ ~ 2 3 (VI) in which R* is a carboxy protecting group, for in.ct~n-~e C(1 6)aLlcyl or C(2 6)aL~enyl and R2 and R3 are as hereinbefore defined;
via the formation of a diastereoisomeric salt with a chiral base such as (-)-cinchonidine; and thereafter:
(a) isolating the preferred diastereoisomeric salt may be obtained by fractionalcryst~ tiQn; and then (b) generating the en~ntiomeric free acid thelt:Çr~ l by aci~lific~tion Diastereoisomeric salts formed from a compound of formula (VI) and a chiral base are of use in preparing compounds of formula (I). Therefore, in a further aspect, the present invention provides for such salts.
Compounds of formula (VI) may be obtained according to the procedures described in earlier applications WO 96/19451 and WO 97/02242 (SmithKline Reech~m plc).
The process hereinbefore described for compounds of formula (I), as well as an ~ltern~tive process for preparing compounds of formula (II), in which R2 and R3 is each hydrogen is s~lmm~rised in the following scheme, in which R
co-lesponds to CH2C6H4CO2R1:

PCT/Er97/01898 a~c RS F'~ LC-CH,CO~ SP eh ~ SP~
NH NH Ib-~ CO~U- O CO~I O _ CO~I I

lo3 O 0.
F~SR H2N(Cll~)rPh-X r~SR

O --CONHICIl,)~ X o CO,R
(~) (Il) S~ S~e 1) 03 ~ SAc 1) AcN03, el3N 13~ SR / 1) FIN 2~ (~0)3~ ~ 2) LG-R
O _~CO~ O CO~ O CO~

Compounds of formula (I) in which R2 is hydrogen and R3 is aLkyl, for inst~nce methyl, may be usefully prepared by a corresponding process in which the aLkyl group is introduced at an early stage, by aLkylating an azetidinone acetate of forrnula (Vll):

SCH2C6H~--C~2R

o~N
CO2R~-(VII) in which R** is C(1 6)alkyl, for instance, methyl, and R* is as hereinbefore defined;
with an aLkylating agent under standard aLIcylating conditions; and thereafter, isolating the diastereoisomers thus obtained by fractional cryst~ ion and/or chromatography. Suitably, the compound of formula (VII) may be a single enantiomer, having the required absolute configuration (4R).
Suitable aLlcylating agents include an aLkyl iodide, in the presence of a suitable base such as sodium hydride or potassium hydroxide, optionally with a quaternary ammonium salt such tetrabutyl ammonium bromide, in a suitable alkylating solvent such as tetrahydrofuran ( I~IF), and at a temperature in the range -10 to 0~C. Other suitable conditions include lithium bis(trimethylsilyl)amide in THF, optionally with 1,3-dimethylimidazolidin-2-one, and at a temperature of about -70~C.

.
.

PCT/EPs7/018s8 wo 97/41098 The newly formed propionate ester may then be converted to the coQt;sponding acid, using basic conditions such as aqueous sodium hydroxide in THF, followed by amide bond formation and then oxidation of the thio group, as previously described. Enantiomers may be usefully isolated by chiral chromatography, for inst~nce hplc using a chiral stationary phase, on compounds of formula (I), at the alkyl/alkenyl ester stage.
The sequences can be readily adapted for other values of X, by reference to earlier applications WO 96/19451 and WO 97/02242 (SmithNine Beecham plc).
For inct~nce~ compounds of formula (I) in which the linker group X
cont~in.c an ether function may be plepa,ed by a suitable ether coupling reaction, for in.ct~nce, when X' is CH20, treating a compound of formula (VIII):

~C~S --CH2~co2R1 N

~ ~ CR2R3--CH L2 (vm in which Rl, R2 and R3 are as hereinbefore defined;
with a compound of formula (IX):

L3(CH2)mY
(IX) in which one of L2 and L3 is a halogen or another suitable leaving group such astriflate or tosylate and the other is OH or a suitable salt therof and m and Y are as hereinbefore defined; under standard ether forming conditions. The thus formed ether compound may then be treated with an oxidising agent to convert the thio group into a suIphinyl group, to give a compound of formula (I).
If the compound of formula (Vm) is a racemic compound, this will lead to a mixture of diastereoisomers. Oxidation of the thio group will create a furtherchiral centre and the resultant distereoisomers may be separated by fractional cryst~lli~inn and/or chromatography. Individual enatiomers may then be obtained by chiral chromatography.
-.. . . ~ .

Suitable compounds of formula (VIII) may be prepared by analogy with the pl~cesses described in WO 97/02242.
Compounds of formula (I~ which are in vivo hydrolysable ~ pharmaçeu~ y acceptable esters may be conveniently prepared from the corresponding parent acid by a process which compromises treating the corresponding parent acid or a salt, aL~cyl ester or activated derivative thereof;
The present invendon will now be illustrated by the following ex~mpl~.s.
Chiral compounds are described as 4R or 45, SR or SS where the 4 describes the centre at the C4 position in the azetidinone and the S describes the sulfoxide centre. Diastereoi.~omer 1 derived from a 4R sulfide has the configuration 4R, SR. The corresponding diastereoisomer 2 is 4R, SS. Such configurations are by extropolation, based on their lH NMR spectra, from configurations obtained initially by x-ray analysis of a limited number of compounds. The absolute configuration at the chiral o~-carbon, when one of R2 and R3 is hydrogen and theother is methyl, is described as oc-R or oc-S.

Example 1 - (R)-N-[6-(4-Fluorophenyl)hex-l-yl]-4-(4 ~ A~rbonylb.,..,i,~lll,io)-2-oYQq7eff~in l-yl~ m'~'~
a Allyl 4(br~ l)b~ te 4-(Bromomethyl)benzoic acid (103 g, 0.48 moles)was suspended in thionyl chloride (200 ml) and dimethylformamide (1 ml) was added. The mixture was heated under reflux until clear, evaporated and azeotroped with toluene (2 x 150ml). The res~llting oil was dissolved in dichloromethane and added dropwise to acooled solution of pyridine (42 ml) and allyl alcohol (40 ml) in dichlorom~th~ne.
The mixture was stirred at room temperature for 1 hour, then washed with water, 2M hydrochloric acid, sodium hydrogen carbonate solution and brine. The organic solution was dried and evaporated to give allyl 4-(bromomethyl)ben70~e as a clear oil (98g, 84% yield). 'H NMR d (CDC13) 4.61 (2H, s, CH2), 4.82 (2H, m, CH20), 5.34 (2H, m, CH2CH-), 6.05 (lH, m, CHCH2), 7.45 (2H, d, Ph-H), 8.03 (2H, d, Ph-H).
b. Allyl 4(aceblthiomethyl)b~n70~te Allyl 4-(bromo~ne~hyl)benzoate (98 g, 0.4 moles) in dry dimethylforrnamide (100 ml) was added dropwise to a cooled suspension of pot~c.cium thioacetate (46 g, 0.4 moles) in dry dimethylformamide (200 ml). The cooling bath was removed and the mixture was stirred overnight. The rection mixture was poured into waterand extracted with ethyl acetate (x3). The combined extracts were washed with water and brine. l'he mixture was dried and evaporated to give allyl 4-(acetylthiomethyl)benzoate as an orange oil (lOOg, 100% yield). 'H NMR d (CDCl3) 2.36 (3H, s, COCH3), 4.13 (2H, s, CH2), 4.82 (2H, m, C~20), 5.32 (2H, m, C~12CH-), 6.05 (lH, m, CHCH2), 7.35 (2H, d, Ph-H), 7.98 (2H, d, Ph-H).
c. 4(4(Allyloxycarbonyl)benzylthio)~7eti~1in-2-one Allyl alcohol (27 ml) in dry tetrahydrofuran (50 ml) was added dropwise to a solution of potassium tert-butoxide (4.93 g, 0.044 moles) in dry tetrahydrofuran(100 ml). After stirring for 5 minutes a solution of allyl 4-(acetylthiomethyl)benzoate (IO.1 g, 0.04 moles) in dry tetrahydrofuran (100 ml) was added dropwise. After stirring for 15 minutes a solution of 4-acetoxy~P~i~in-2-one (5.16 g, 0.04 moles) was added dropwise. The mixture was stirred for 1 hour and evaporated. The residue was partidoned between ethyl acetate and water and the aqueouse was extracted with ethyl acetate. The combined extracts were washed with brine, dried and evaporated. Flash chromatography (silica gel, ethyl acetate-petrol) gave 4-(4-allyloxycarbonylbenzylthio)azetidin-2-one as an oil (9.lg, 82% yield). 'H NMR
d (CDCl3) 2.84 (lH, dd, H3a~, 4.31 (lH, dd, H3b), 3.88 (2H, s, S-CH2), 4.68 (lH, dd, H4), 4.78 (2H, m, C_ 2~~' 5-35 (2H, m, C_2CH-), 6.05 (lH, m, CHCH2), 6.07 (lH, br. singlet, N-H), 7.40 (2H, d, Ph-H), 8.03 (2H, d, Ph-H).
d. Methyl 4-(4-(Allyloxycarbonyl)benzylthio)-2-oxo~7e~Air~ 1-y~ e~e -., , . . . . ,, . .. . ~ , .

W O 97141098 PCT~EP97/01898 To a stirring solution of 4-(4-(allyloxycarbonyl)benzylthio)~7Ptidin-2-one (2.55g, 9.2 mmol), tetrabutylammonium bromide (0.33 g, 1.02 mmol) and methyl bromo~Get~te (1.06 ml, 11.2 mmol) in dry tetrahydrofuran (40 ml) was added powdered potassium hydroxide (0.63 g, 11.2 mmol) keeping the reaction temperature below 30 by means of a cold water bath. After 2 h, the mixture was diluted with water and e~t~cted three times with ethyl acet~t~.. The combined extracts were dried (MgSO4) and evaporated and the residue chromatographed (fine silica, ethyl acetate-petrol) to give the title compound as a clear oil, yield 2.66 g (83%).
'H NMR ~ (CDC13) 2.97 (lH, dd, H3a),3.26, 4.07 (each lH, C~2CO, d), 3.42 (lH, dd, H3b), 3.70 (3H, s, C~3O), 3.81 (2H, s, SCH2), 4.83 (2H, m, C~2O), 4.93 (lH, dd, H4), 5.35 (2H, m, CE~2CH),6.03 (lH, m, CHCH2), 7.39 (2H, d, Ph-H), 8.02 (2H, d, Ph-H) e. (+/)-4(4(Allyloxycarbonyl)benzylthio)-2~oxo~7eff~lin-l-ylaceff~cacid To a solution of methyl 4-(4-(allyloxycarbonyl)benzylthio)-2-oxoa7PtiAin- 1-ylacetate (2.17 g, 6.21mmol) in tetrahydrofuran (20 ml) was added dropwise with cooling (ice bath) over 10 min a 1 molar aqueous solution of potassium hydroxide. After a further 30 min, the solution was diluted with water and extracted twice with ether. The aqueous layer was then acidified (dil.
hydrochloric acid) with cooling and the oil which precipited was extracted into ether. The combined extracts were dried (MgSO4) and evaporated to a clear oil which eventually crystallised under petrol and was filtered, washed and dried togive the title compound as white crystals, 1.87 g, 90% yield 'H NMR ~ (CDC13) 2.g8 (lH, dd, H3a), 3.34, 4.06 (each lH, CH2CO, d), 3.42 (lH, dd, H3b), 3.82 (2H, s, SCH2),4.82 (2H, m, C~2O), 4.92 (lH, dd, H4), 5.34 (2H, m, C.~2CH), 6.03 (1H, m, CHCH2),7.39 (2H, d, Ph-H), 8.02 (2H, d, Ph-H) f. (-)-(R)-4-(~(Allyloxycarbonyl)benzylthio)-2-o~o~7effAin-1-ylacetic acid 4-(4-(Allyloxycarbonyl)benzylthio)-2-oxoazetidin-1-ylacetic acid ( 3.41 g, 10.2 mmol) and cinchonidine (2.99 g,10.2 mmol) in ethanol (40 ml) were heated to boiling when a clear solution was obtained. On st~nd~ng for 90 min, the crystalline salt which had precipitated was filtered off, and recryst~ ed from ethanol (20 ml). The solid obtained was stirred vigorously with ether and water whilst acidifying with dil. hydrochloric acid, and when complete solution was obtained the layers were separated and the aqueous layer further extracted with ether. The combined extracts were dried (MgS04) and evaporated to an oil which crystallised on trituration with light petrol to give the title compound as white crystals, m.p. 74-6~C, 6.7 g, 50% yield aD2S = -24.2 (c. 0.7 w/v CHCl3,25~C) 'H NMR ~ (CDC13) 2.97 (lH, dd, H3a), 3.26, 4.07 (each lH, C~2CO, d), 3.42 (lH, dd, H3b),3.70 (3H, s, CH30), 3.81 (2H, s, SCH2), 4.B3 (2H, m, C~20), wo 97t41098 PCT/EPg7/01898 4.93 (lH, dd, H4), 5.35 (2H, m, C~2CH), 6.03 (lH, m, C~CH2), 7.39 (2H, d, Ph-H), 8.02 (2H, d, Ph-H) g. (R)-N~[6-(4~ oro~ henyl)hex-l-yl]-4-(4-alloxycarbonyll)c..L,rlll.io)-2-ox~ l-yl)acetamide To a cooled (ice bath) soludon of (R)-4-(4-(allyloxycarbonyl)benzylthio)-2-oxo~7~ti~in-l-ylacetic acid (12.51 g, 0.0373 mol), 1-hydroxybenzotriazole hydrate (5.04 g, 0.0373 mol) and 6-(4-fluorophenyl)hexylamine (0.0373 mol) in dry dimethylform~mi~e (150 ml) was added with stirring dicyclohexylcarbodiimide (7.29 g, 0.0373 mol). After 20 min the cooling bath was removed, and after a further 16 h, the solvent was evaporated under reduced plCiS~ and the residue treated with ethyl acetate and the insoluble precipitate filtered oK and discarded. The filtrate was further diluted with ethyl acetate, washed with 0.2 M hydrochloric acid, then saturated sodium hydrogen carbonate, dried and evaporated under reduced pressure. The residue was triturated with ethyl acetate/light petrol to give the title compound as white crystals, m.p. 54-7~C, 17 g, 89% yield 'H NMR ~ (CDC13) 1.30- 1.60 (8H, m, 4xC~2), 2.55 (2H, t, J=7.6 Hz, C~2Ph), 2.90, 2.97 (lH, dd, J=2.4, 15.4 Hz, H3), 3.23 (2H, m, NHCH2), 3.35, 3.41 (lH, dd, J=5.1, 15.4 Hz, H3), 3.53, 3.78 (each lH, d, J=16.6 Hz, NC_ 2)~ 3.86 (2H, s,SC~2), 4.83 (3H, m, CO2C~2, H,), 5.37 (2H, m, CH2=CH), 6.0 (2H, m, N~, CH2-C~D, 6.g4 (2H, m, 4-FPh-O, 7.10 (2H, m, 4-FPh-O, 7.39 (2H, d, J=8.3 Hz, 4-CO2allylPh-~I), 8.02 (2H, d, J=8.3 Hz, 4-CO2allylPh-O
Example 2 - (4R, SS)-N-[6-(4-Fluorophenyl)hex-l-yl]-4-(4-allo~,~ l.onylbenzylsulphinyl)-2-oxo~7eti~1;n-l-yl)~re~mi~e A soludon of (R)-N-[6-(4-fluorophenyl)hex-1-yl]-4-(4-alloxycarbonylbenzylthio)-2-oxoazetidin-1-yl)acetamide (16.36 g, 0.0319 mol) in dichloromethane (150 ml) was cooled to -65 to -70 and a solution of m-chlolopell,enzoic acid (6.61 g, 0.0383 mol) in dichloromethane (120 ml) added dropwise with stirring over 20 min. After 1 h, the mixture was washed with saturated sodium metabisulphite solution, then saturated sodium hydrogen ca~ nate, then dried (MgSO4) and evaporated to a solid which was recryst~lliced from ethyl acetate to give a mixture of diastereoisomers 2 and 1 in the ratio 3:2.
Chromatographic separation (HPLC) gave diastereomer 2 (4R, SS) as a white crystalline solid, m.p. 133-5~C, 3.3 g, 20% yield aD25~C = +74.0 (c. 0.6% w/v CHCI3, 25~C) 'H NMR ~ (CDC13) 1.30-1.60 (8H, m, 4xCH2), 2.56 (2H, t, J=7.6 Hz, C~2Ph), 2.91, 2.95 (lH, dd, J=2.4, 15.2 Hz, H3), 3.27 (3H, m, NHCH2, ~3), 3.94, 4.22 (each lH, d, J=17.2 Hz, NC~2), 4.04, 4.18 (each lH, d, J=12.8 Hz, SOC.~2), 4.65 (lH, m, ~,), 4.84 (2H, m, CO2CH2), 5.37 (2H, m, CH2=CH), 6.0 (lH, m, ... ...

WO 97/41098 rCTAEP97/01898 CH2=C~D, 6.95 (3H, m, 4-FPh~ ),), 7.10 (2H, m, 4-FPh-~), 7.36 (2H, m, 4~O2alIylPh-O, 8.09 (2H, m, 4-CO2allylPh-O.
Example 3 - (4R, SR)-N-t6-(4-Fluorophenyl)hex-l-yl]-4-(4-allG~C~ bOnyll)C.~ phinyl)-2.Q~Qc~>~ l;n.l.yl)- ~t~m~
From the HPLC chromatography described in Example 2 above the other ~ diastereoi~orner (Dia 1: 4R, S~) was obtained as a white crystalline solid (1.8 g, 11 % yield) m.p. 175-7~C
aD2~~C = -119.7 (c. 0.5% wJv CHCl3, 25~C) 'H NMR ~ (CDCl3) 1.30-1.60 (8H, m, 4xCH2), 2.55 (2H, t, J=7.6 Hz, CH2Ph), 2.95, 2.98 (lH, dd, J=4.8, 14.8 Hz, H3), 3.24 (2H, m, NHCH2), 3.42, 3.46 (lH, dd, J=2.4, 14.8 Hz, H3), 3.76, 4.09 (each lH, d, J=17.2 Hz, NCH2), 3.95, 4.01 (each lH, d, J=13.2 Hz, SOCH2), 4.59 (lH, m, H4), 4.84 (2H, m, CO2CH2), 5.37 (2H, m, CH2-CH), 6.0 (lH, m, CH2=CH), 6.53 (lH, m, NH), 6.95 (2H, m, 4-FPh-H), 7.10 (2H, m, 4-FPh-H), 7.36 (2H, d, J=8 Hz, 4-CO2allylPh-H), 8.09 (2H, d, J=8 Hz, 4-CO2allylPh-H).
EYample 4 ~ (4R, SS)~N-[6-(4-Fluorophenyl)hex-1-yl]-4-(4 ~bo~ lphinyl)-2-oYQ~7etlllin-l-yl)~c~t~ ~ide A solution of (4R, SS)-N-[6-(4-Fluorophenyl)hex-l-yl]-4-(4-alloxycarbonylbenzyl.clllphinyl)-2-oxoazetidin-1-yl)~cet~mide (Example 2) (0.185 g, 0.35 mmol), triphenylphosphine (0.092 g, 0.35 mmol), pyrrolidine (0.033 ml, 0.4 mmol) and tetralcis triphenylphosphinep~ dium(0) (0.012 g, 0.01 mmol) in dichlorometh~ne (10 ml) was stirred under nitrogen for 16 h. A further 0.012 g (0.01 mmol) of tetrakis triphenylphosphinep~ m(O) was added and after a further 4 h the reaction was complete. The solution was diluted with water, acidified (2N HCI), the layers separated and the aqueous layer further e~l,a~,led with dichloromethane. The combined extracts were dried (MgS04)and evaporated to a yellow oil, which was triturated with ether. A yellow solid was obtained which was filtered off and dissolved in sodium hydrogen carbonate solution. Sh~king with ether gave an emulsion which was separated by tre~tmp~nt with ethyl acetate and centrifugation. The aqueous layer was then acidified (2N
HCl) and extr~ctPd with dichloromethane, and the extracts dried (MgSO4)and evaporated. The residue was triturated with ether to give a white solid which was filtered, washed and dried to give the title compound as a white solid, m.p. 105-7~C, 0.1 g, 58% yield aD23~C = -31.7 (c. 0.5% w/v DMSO, 25~C) 'H NMR ~ (DMSO) 1.26 (4H, m, 2xC~2), 1.38 (2H, m, CH2), 1.50 (2H, m, CH2), 2.96, 2.99 (lH, dd, J=2, 15.2 Hz, H3), 3.06 (2H, m, NHC~2), 3.84, 4.09 (each lH, d, J=17.2 Hz, NC~l), 4.13, 4.31 (each lH, d, J=12.8 Hz, SOC.~2), 4.84 ... ~ .. .... ... .. . ... .... . .. ...

wo 97l41098 PCT/EPg7/01898 (lH, m, ~), 7.05 (2H, m, 4-FPh-~), 7.19 (2H, m, 4-FPh-0, 7.47 (2H, d, J=8 Hz, 4-CO2allylPh-~), 7.93 (2H, d, J=8 Hz, 4-CO2allylPh-H), 8.13 (lH, m, N~), 13 (lH, bs, CO~). --Example 5 - (-)-(4R, SS)-N-[6-(4-Fluorophenyl)hex-l-yl]-4-(4 CarbOXy~ phinyl)-2~oyQp7et~ n~l~yl)acetalTlide~5~~ -- salt A mixture of N-[6-(4-fluorophenyl)hex-1-yl]-4-(4-carboxybenzylsulphinyl)-2-o~t~q7Ptil1in-l-yl)~cet~mide (0.51 g) and sodium bicarbonate (0.088 g) in water (15 ml) was sonic~tP~l for S min, methanol added (20 ml) and the mixture sonic~te~l for a further 20 min. After filtration the solution was evapotated to a low volume, diluted with water and lyophili~ed to give the ~itle compound as a white powder (0.52 g), m.p. 238-40~C. aD = -31.7~ (c O.S, DMSO) Example 6 - 4R, SS-N-(6-(4-Fluorophenyl)hexyl)-4-(4 eth~ A~ 1benzylsulphinyl)-2-oxoazetidin-1-yl ~- e~miAe a Ethyl 4-(brorn~lnethyl)b~n7Q~te 4-(Bromomethyl)benzoic acid (25.75g, 0.1197moles) was suspended in thionyl chloride (SOml) and dimethylformamide (0.25ml) was added. The mixture was heated under reflux for 25 minutes until clear, evaporated and azeotroped with toluene (x2). The res~ ing oil was dissolved in dichloromethane (75ml) and added dropwise over 10 mimltps to a solution of absolute alcohol (8.6ml, 0.1465moles), pyridine (10.5ml, 0.1298moles) in dry dichloromPth~ne (50ml), cooled to 10~C. The ice ba~h was removed and the reaction was stirred for 45 minlltPs ~ then washed with water, 2NHCl, water, sodium hydrogen carbonate solution and brine. The organic solution was dried (MgS04) and evaporated to give a mixture of 60:40 ethyl 4-(bromomethyl)ben7o~te ethyl 4-(chloromethyl)bçn70~te as an oil (25.6g, 94%) lH nmr ~ (CDC13) 1.40 (3H, m, CH3), 4.40 (2H, m, CH20), 4.50, 4.61 (2H, 2xs, C~2CI/Br), 7.45 (2H, m, Ar-~), 8.01 (2H, m, Ar-H) b. Ethyl 4-(- ~ :trlll,iomethyl)ben7~te 60:40 Ethyl 4-(bromomethyl)ben70~te ethyl 4-(chloromethyl)ben7.o~te (25.0g, 0.11 lmoles) in dry dimethylformamide (lSOml), cooled to 5~C, was treated with potassium thioacetate (13.3g, 0.117moles) and the temperature rose to 20~C. The reaction was stirred at room temperature for 2 hours, poured into water (250ml) and extracted with diethyl ether (3xlOOml). The organic extracts were combined, washed with water, dried (MgS04), charcoaled and evaporated to give ethyl 4-(acetylthiomethyl)benzoate as a brown soild (26.0g, 99%), m.p. 36-37~C.
1H nmr ~ (CDC13) 1.38 (3H, t, J=7.1Hz, C~3), 2.36 (3H, s, COCH3), 4.14 (2H, s, C~2S), 4.36 (2H, q, CH20), 7.3~ (2H, d, J = 8.2Hz, Ar-H), 7.97 (2H, d, J=
8.2Hz, Ar-H) W O 97/41098 PCTAEP97tO1898 c. 4(4(Ethox,~c~bonyl)benzylthiop7et;~in-2.one A solution of sodium (1.87g, 0.0813moles) in absolute alcohol (300ml) was treated with a solution of ethyl 4-(acetylthiomethyl)benzoate (19.4g, 0.0814moles) in absolute alcohol (75ml) over 3 minut~~. The reaction was stirredat room temperature for 30 min~ltes, cooled to -5~C and treated with a solution of 4-~etoxyazetidin-2-one (lO.Og, 0.07745moles) over 5 minutes The cooling bath - was removed and reaction was stirred for 2 hours, evaporated to dryness and treated with brine (200ml) and extracted with ethyl acetate (200ml, lOOml). The organic extracts were combined washed with brine, dried (MgS04) and evaporated to a red oil. Purified by flash column chromatography on silica gel eluted with 3:1 to 1:2 petroleum ether 40-60~C:ethyl acetate to give 4-(4-(ethoxycarbonyl)benzylthio)~7eti-1in-2-one as an orange oil (18.64g, 91%).
lH nmr o (CDC13) 1.38 (3H, t, J=7.1Hz, C~3), 2.82, 2.89 (lH, 2xm, ~13). 3.29, 3.35 (lH, 2xm, ~3), 3.88 (2H, s, CH2S), 4.37 (2H, q, CH20), 4.70 (lH, m, H1), 5.70 (lH,bs, NO, 7.40 (2H, d, J = 8.3Hz, Ar-~O, 8.00 (2H, m, Ar-H) d. Methyl (4(4ethoxycarbonyl)be"Lylll.io)-2-oxoazetidin-1-yl~ret~te A stirred solution of 4-(4-(ethoxycarbonyl)benzylthio)azetidin-2-one (217.3g, 0.819mol), methyl bromo~cet~te (128.5g,0.84mol) and tetrabutylammonium bromide (25.8g,0.08mol) in dry THF (9OOml) was cooled in an ice bath to 20~C
and finely ground po~sium hydroxide (48.3g, 0.86mol) was added in one portion. The reaction exothermed to 45~C and was allowed to cool back to 30~C
when the ice bath was removed and stirring continued for lhr. More potassium hydroxide (2.4g, 0.043mol) was added and stirred 30mins when this addition was repeated. After a further 30mins, the reaction mixture was f~ltered through hyflo, washing with more THF. The combined organics were evaporated to a red oil.
Ether (11) was added an shaken well. The ether was decanted and the process repeated. The combined ether extracts were evaporated to give the title compound as a dark red oil (199.8g, 72% yield) H NMR ~ (CDC13), 1.40 (3H, t, J=7Hz, CH2C~3), 2.98 (lH, dd, J=2, lS Hz, ~3), 3.26, 4.03 (each lH, d, J=18 Hz, NCH2), 3,42 (lH, dd, J=5, lS Hz, ~3), 3.70(3H, s, OCH3), 3.81, (2H, m, SCH2), 4.38 (2H, q, J=7 Hz, OC~2), 4.93 (lH, m, ~,), 7.39 (2H, m, Ph-~ ), 7.99 (2H, m, Ph-H ).
e. (4-(4E~oxycarbonyl)benzylthio)-2-oxo~7et~ n-l-ylacetic acid Methyl (4-(4-ethoxycarbonyl)benzylthio)-2-oxoazetidin-1-ylacetate (169.8g,0.503mol) was disolved in THF (750ml), cooled to 0~C and a solution of potassium hydroxide (29.7g, 0.529mol) in water (SOOml) was added over lSmin at O - 5~C then the mixture was stirred at 0~C for 45mins. Ether (11) and water (21) were added, the layers separated and the aqueous washed with ether (11), then acidified with conc hydrochloric acid (55ml) and extracted with dichloromethane W O 97141098 PCT~EP97/01898 (2 x 11). The combined extracts were washed with brine, dried (MgS04) and evaporated to give the title compound as a green solid (128.4g, 79% yield).
f. (-)-R-(4(1 Ethox~c~ l onyl)benzylthio)-2-oxo~7eRAin-1-ylacetic acfd 4-(4-Ethoxycarbonyl)benzylthio)-2-oxoazeyidin-1-ylacetic acid (46.0g, 0.1422moles) and (-)-cinchonidine (41.88g, 0.1423moles) were dissolved in abolute alcohol (450ml). The solution was cooled for 1.5 hours, filtered, and dried to give the salt as a cream solid (33.15g). This solid was recryst~lli~d from absolute alcohol (300ml) to give 23.6g of salt which was mixed with water (SOOml) and diethyl ether (500ml) and acidi~led with dilute HCl (SOml). When all the solid had dissolved the layers were separated and the aquous layer was extracted witn ether (250ml). The organic extracts were combined, ethyl acetate (lOOml) was added and washed with water, dried (MgS04), filtered and evaporated to give R-(4-(4-ethoxycarbonyl)benzylthio)-2-oxoazetidin- 1 -ylaceticacid as a colourless solid. (10.93g, 23.8%) m.p. 135-137~C.
aD = -23.5 (c = 0.46 w/v in chloroform at 25~C) IH NMR ~ (CDC13) 1.39 (3H, t, J=7. lHz, C~3), 2.96, 3.02 (IH, dd, J =2.2, 15.3 Hz, ~3), 3.33, 4.05 (each lH, d, J=18.4 Hz, NCH2C02H), 3.40, 3.46 (lH, dd, J= 5.1, 15.3 Hz, H3), 3.82 (2H, s, SCH2), 4.37 (2H, q, C02CH2), 4.68 (lH, b, H), 4.92 (lH, m, ~4), 7.38 (2H, d, J=8.2Hz, Ph-H), 7.99 (2H, d, J=8.2Hz, Ph-g. 4R,SR-(4-(4-Ethoxycarbonyl)benzylsulphinyl)-2-oxQ~7eti~lin-l-,y'v~e'i~
acid (~liq~t~r~~cmer 1) h. 4R,SS-(4(4 Ethoxycarbonyl)benzylcl-lphinyl)-2-Ql~o~7et~ n-1-y~- -acid (.li&~ oisomer 2) A solution of (-)-R-(4-(4-ethoxycarbonyl)benzylthio)-2-oxoazetidin- 1 -ylacetic acid (10.81g, 0.03343moles) in d;y dichloromethane (400ml) at -70~C was treated with ozone until a blue colouration appeared. The reaction was allowed to warm to room temperature and dichlorometh~ne (SOml) was added to aid stirring.
The solution was evaporated to dryness and the resulting soild was mixed with chloroform (200ml). The colourless solid was collected by filtration to give 4R,SR-(4~(4-ethoxycarbonyl)benzylsulphinyl-2-oxoazetidin-1-ylacetic acid (4.11g,36%)m.p.162-164~C. (contains 15.8:1 dial:dia2) 'H NMR ~ (DMSO) 1.33 (3H, t, J=7Hz, C_3), 2.97, 3.04 (lH, dd, J=4.8,14.8Hz, ), 3.12, 3.16 (lH, dd, J= 1.6, 14.8Hz, H3), 3.83, 4.17 (each lH, d, J= 18Hz, NC~2C02H), 4.92, 4.24 (lH, d, J = 12.8Hz, S~~2), 4.32 (2H, q, C02C~2), 4.99 (lH, m, ~I4). 7.48 (2H, d, J=8.0Hz, Ph-O, 7.96 (2H, d, J=8.0Hz, Ph-~) The filtlate from the above was evaporated, mixed with diethyl ether and filtered to give 4R,SS-(4-(4-ethoxycarbonyl)benzylsulphinyl-2-oxoazetidin-1-ylacetic acid (6.42, 56%) m.p. 152-155~C. (contains 92:8 dia2:dial) ~ .. . , . . .. , ., . ~ _, .. . .

W O 97/41098 PCTAEP97/018g8 'H NMR ~ (DMSO) 1.33 (3H, t, J=7 Hz, C~3), 2.97, 3.01 (lH, dd, J=2.0,15.5Hz, ~I3), 3.35 (lH, m, ~3), 3.95, 4.17 (each lH, d, J= 18.2Hz, NC~I2C02H), 4.15 (lH, d, 1 of SOC~2), 4.32 (3H, m, 1 of SOC~2, C02C~12), 4.82 (lH, m, ,~4), 7.51 (2H, d, J=8.2Hz, Ph-O, 7.97 (2H, d, J=8.2Hz, Ph~
i. (+)-4R, SS-N-(6-(4nuorophenyl)hexyl)-4-(4 ethoA~c& bonylb~ 1)-2-oY~7e~Air~-l-ylacet~ idé
6-(4 Fluorophenyl)hexylamine (1.82g, 0.00932moles) in dry dimethylform~mide (75ml) was added to a mixture of 4R,SS -(4(4-ethoxycarbonyl)benzylclllrhin 2-oxoazetidin-1-ylacetic acid (3.15g, 0.00928moles), N,N,-dicyclohexylcarbo~iimide (1.92g, 0.00931moles) and l-hydroxybenzotriazole (1.25g, 0.00925moles). The reaction was stirred at room temperature for 3.5 hours, diluted with ethyl acetate (lOOml) and cooled. The mixture was filtered to remove urea and the filtrate was evaporated to dryness. The residue was dissolved in ethyl acetate (400ml), washed with sodium hydrogen carbonate solution, brine, dried and evaporated to a colourless solid (5.8g) which was recrys~l1ised from ethyl acetate (125 ml) to give the product (3.0g). Purif1c~tioll by column chromatography gave 4R,SS -(4-(4-ethoxycarbonyl)benzylsulphinyl-2-oxoazetidin-1-ylacetic acid as a colourless solid, 155-156~C, 1.8 g, 38% yield 'H NMR ~ (CDC13) 1.3-1.6 (1 lH, m, 4xCH2, C~3), 2.56 (2H, t, J = 7.6Hz, C~2Ph), 2.89, 2.96 (lH, dd, J=2.4,15.3Hz, ~3), 3.25 (3H, m, NHC~I2, EI3).
3.94, 4.22 (each lH, d, J=17Hz, NC~12CO), 4.03, 4.18 (each lH, d, J=12Hz, SOC~12), 4.39 (2H, q, C02CH2), 4.65 (lH, m, ~4), 6.9-7.12 (SH, m, N_,p-ClPh-~D, 7.35 (2H, d, J=8.3Hz, Ph-O, 8.07 (2H, d, J=8.3Hz, Ph-H) aD = +85.2 (c = 0.5% w/v in chloroform at 25~C) Found: C, 62.6; H, 6.3; N, 5.4%; Cz7H33FN205S requires: C, 62.8; H, 6.4; N, 5.4%

FYPr~PIe 7 (+)-4R~SS-N-(6-(4-ChIOrOPhenYI)heXYI)~4-(4-eth~x~ lb~ ulphinyl)-2-oyQ~7et~ n l yl~cet~rn;rle Tre~tment of 4R~ss-(4-(4-ethoxycarbonyi)benzylsulphinyl-2-oxo~7~tidin-l-ylacetic acid with 6-(4-chlorophenyl)hexylamine, N,N-dicyclohexylcarbodiimide, and l-hydroxybenzotriazole in dry dimethylformamide as described for Example 6 above, followed by the same work-up procedure gave the title compound as a colourless solid, 159-161~C, 45% yield 'H NMR ~ (CDC13) 1.3-1.6 (1 lH, m, 4xC~2, CH3), 2.56 (2H, t, J = 7.6Hz, C~2Ph), 2.89, 2.9S (lH, dd, J=2.4,15.3Hz, ~3), 3.25 (3H, m, NHC~12, H3), 3.94, 4.24 (each lH, d, J=17Hz, NCH2CO), 4.03, 4.18 (each lH, d, J=12Hz, SOC~i2), 4.36 (2H, q, C02CH2), 4.65 (lH, m, .~I4), 7.04 (lH, m, N~D, 7.06-7.26 (4H, m, p-ClPh-~), 7.35 (2H, d, J=8.3Hz, Ph-H), 8.07 (2H, d, J=8.3Hz, Ph-H) -W O 97141098 PCT~EP97/01898 aD = +83.9 (c = 1.0% w/v in chloroform at 25~C) Pound: C, 60.9; H, 6.1; N, 5.2%; C27H33ClN205S requires: C, 60.8; H, 6.2; N, 5.3%
Example 8 - (+)-4R, SS-N-(6-(4-Fluorophenyl)hexy1)-4-(4-ethoxycarbonylbc~ ulphinyl)-2-oYo~7eR~lin-l-yl~ret~mide a p-Metho~ l [(3S, 4R) 1 ~ce'ylll-io-3-bromo-2~ o~7eti~
yl]acetate Ozoni~ed oxygen was bubbled through a solution of p-methoxybenzyl 2-~(35, 4R)-4acetylthio-3-bromo-2-oxo~7~tir~in-1-yl]-3-methylbut-2-enoate (Osborne N.
F. et al., J. Chem. Soc., Perkin Trans. 1,1 994, 179) (20. 16 g, 0.0456 mol) in ethyl acetate (400 ml) at -65 to -70~C until a permanent blue solution was obtained. Excess ozone was removed by the passage of oxygen, then trimethyl phosphite (53.8 ml, 0.456 mol) was added dropwise. After 15 min. the solution was allowed to warm to room temperature, then stood for 16 hr. The solvents were evaporated and the residue reevaporated twice from toluene,thçn dissolved in ethyl acetate (300 ml) and stirred vigorously for l.S hr. with a solution of p-tol~Pn~Ps~lphonic acid (2 g) in water (100 ml). After dilution with water the organic layer was separated and the aqueous layer further extracted with ethyl acetate. The combined extracts were washed succescively with saturated aq.
sodium hydrogen carbonate and brine, then dried (MgS04) and evaporated.
Pllrtft~ation by flash chromatography (silica, ethyl acetate-pet. ether) gave the title compound as a light brown oil, yield 10.6 g (58%).
b. Silver (3S,4R)~3-bromo~ o-me&oxybenzyloxycarbonylmethyl)-2-oY-~7~ nP_4.~hi~ t~
A solution of p-methoxybenzyl [(35, 4R)-4-acetylthio-3-bromo-2-oxoazetidin-1-yl~acetate (4.13 g, 0.01 mol) in methanol (90 ml) was added with stirring in subdued light to a solution of silver nitrate (2.27 g, 0.0133 mol) in methanol (90 ml). Triethylamine (1.87 ml, 0.0133 mol) was then added with ice cooling, and stirring continl~ed for 1 hr. at S-10~C followed by 30 min. at room temperature.The .,.i,~lu~ was re-cooled (ice bath) and the precipitated solid filtered and washed twice with ice cold methanol then hexane to give the title compound, yield 4.6 g (96%).
c 4Carbe~ cA~benzyliodide Tre~-mçnt of 4-carboxybenzylbromide with thionyl chloride followed by ethanol in pyridine have a mixture of 4-carbethoxybenzyl chloride and bromide which was treated (14.6 g) with sodium iodide (39.8g) in acetone (lSOml) at reflux temperature for 20hrs. The mixture was cooled, filtered and the solvent evaporated off. The residue was taken up in ether (lSOml) and washed with ., .. . , .. , . ,, . ~, .

wo 97/41098 pcTlEps7lol898 water, aqueous sodium thios~lrh~te, water, brine, dried and evaporated to give the title compound as a pale yellow solid, m.p. 61-3~C, 17.5 g (91% yield).
'H NMR ~ (CDC13) 1.39(3H, t, J 7.1 Hz, CH3), 3.37(2H, q, J=7.1 Hz, C~I2CH3), 4.46(2H, s, C~I2I), 7.43 (2H, m, 3,5-Ph-H), 7.96(2H, m, 2,6-Ph-H) d. p-MethoA~ L~I [(3S, 4R)-4-(4-carbethoxy)be,.z,y111,io-3-bromo-2-o~7~ff~ -yl]PC~t~t~
A solution of silver (35,4R)-3-bromo- 1-(p-methoxybenzyloxycarbonylmethyl)-2-oxo~7~ti~in~-4-thiolate (2.34g, 0.005mol) in aoetonitrile (20 ml) was treated with a solution of 4-carbethoxybenzyl iodide (1.74g, 0.006mol) in ~cetonitnl~ (lOml) and the mixture stirred in subdued light for 2.5hrs. The ~ e was filtered through hyflo, the filtrate evaporated and the residue purified by flash chromatography (silica, ethyl acetate-pet. ether) to give the dtle compound as awhite solid, m.p. 97-9~C, 1.60g (61% yield).
lH NMR ~ (CDC13), 1.39(3H, t, J=7.1Hz, CH2CH3), 3.48, 4.07 (each lH, d, J=18 Hz, NC~2), 3.80 (SH, s, SCH2 + OC~3), 4.37 (2H, q, J=7.1Hz, CH2CH3), 4.~1, 4.91 (each lH, d, J=1.6 Hz, H3 + ~4), 5.03, 5.12 (2H, 2xd, J=11.8Hz, OC~2), 6.89 (2H, m, 3,5-(4-CH30Ph)-0, 7.26-7.41 (4H, m, 2,6-(4-C02Et)Ph-~I
+ 2,6-(4-CH30Ph)-H), 8.00 (2H, m, 3,5-(4-C02Et)Ph-El ).
e. p-Metiho~be.-L~I [(3S, 4R)-4-(4-carbethoxy)benzylsulphinyl-3-bromo-2-QYnq7~ l-yl]ar~et~te Diast 2: 1 75:25 p-Methoxybenzyl [(3S, 4R)-4-(4-carbethoxy)benzylthio-3-bromo-2-oxoazetidin-l-yl]acetate (15.2g, 0.029mol) was dissolved in dichloromethane (400ml), cooled to -65~C and a solution of m-ch}oroperbenzoic acid (8.9g,0.029mol) in dichloromethane (450 ml) added dropwise with stirring over 20 min. The cooling bath was removed and the mixture stirred at RT for 2hrs. The solution was shaken with a mixture of saturated aqueous sodium sulphite and saturated sodium hydrogen carbonate and the organic layer separated, washed with water, dried (MgS04) and evaporated to an oil.
Hot ethyl acetate (60ml) was added and after cooling in freezer overnight, p-methoxybenzyl [(3S, 4R)-4-(4-carbethoxy)benzylsulphinyl-3-bromo-2-oxo~7~ti~in-l-yl]acetate cryst~llised out as a white solid as a 75:25 mixture ofdiastereomers (2:1), 9.94g, (63% yield) lH NMR of major component: ~ (CDC13), 1.39(3H, t, J=7. lHz, CH2C~3), 3.80 (3H, OC~3), 4.0 - 4.2 (3H, m, SOCH2 + NCH2), 4.3 - 4.4 (3H, m, CH2CH3 +
NC~2), 4.50, 4.81 (each lH, d, J=1.7 Hz, H3 +~4), S.1 (2H, m, OCH2), 6.88 (2H, m, 3,5-(4-CH30Ph)-H), 7.26 (2H, m, 2,6-(4-CH30Ph)-H), 7.41 (2H, m, 2,6-(4-C02Et)Ph-~ ), 8.00 (2H, m, 3,5-(4-C02Et)Ph-H ).
f. [(4R)-4-(4-carbethoxy)benzylsulphinyl-2-oxoazetidin-1-yllacetic acid Diast 2 W O 97/41098 PCT~EP97/01898 p-Metho~ylJen~yl [(35, 4R)-4-(4-carbethoxy)benzylsulphinyl-3-bromo-2-oxo~7Pti-1in-l-yllacetate (8.75g, 16.25mmol) was dissolved by warming in ethanol (400ml) and a solution of sodium bicarbonate (4.0g, 48mmol) in water (40ml) was added. To the cloudy mixture was added 10% Pd on carbon (O.Sg) and the warm mixture (initial temp 40~C) was hydrogenated at 50psi, room temp for 2 hrs. More catalyst (l.Og) was added, the mixture warmed to 35~C and hydrogçn~tP~ as above for 2 hrs. The mixture was filtered through hyflo and evaporated to give a mixture of p-methoxybenzyl and ethyl esters of the des-bromo derivative as brown oil. This was dissolved in ethanol (SOml), cooled to 10~C and lN sodium hydroxide (17ml) was added dropwise over lOmin. The mixture was stirred at 10~C for 45mins then most of the ethanol was evaporated off and the residue dissolved in dichloromethane and water. The layers were separated and the aqueous washed with dichloromethane then acidified with conc hydrochloric acid (2ml) and extracted with dichloromethane. The extracts were dried (MgS04) with the addition of charcoal, filtered through hyflo and evaporated to a black foam. This was dissolved in chloroform and refrigerated overnight. A small amount of solid was filtered off and the filtrate evaporated to a black oil which solidified under ether to give the title compound as an off-white solid, 3.77g, (68% yield) (ratio of dia 2:dia 1 94:6). The chromatographic and spec~r~l characteristics of this material identified it as the same compound prepared in Example 6h.
'H NMR ~ (DMSO) 1.32 (3H, t, J=7Hz, CH3), 2.99 (lH, m, E~3), 3.33 (lH, m, ), 3.95 (lH, d, J= 18.2Hz, NC~2), 4.15 (2H, m, NCH2 + SOCH2), 4.34 (3H, m, SOC~2 + C02C~2), 4.82 (lH, m, ~4), 7.51 (2H, d, J=8.3Hz~ Ph-O, 7.97 (2H, d, J=8.3Hz, Ph-H) g. [(4R~4-(4-carbethoxy)benzylsulphinyl-2,oxoazetidin.1-yl]acetic acid D;ast 2 [(4R, SS)-4-(4-carbethoxy)benzylsulphinyl-2-oxoazetidin-1-yl]acetic acid (Diast 2) was also prepared by the following procedure: p-Methoxybenzyl [(35, 4R)-4-(4-carbethoxy)benzylsulphinyl-3-bromo-2-oxoazetidin- 1 -yl]acetate (Diast 2: 1 75:25) (4.59g, 8.52mmol) was dissolved in dichloromethane (SOml), cooled in an ice bath and activated zinc powder (1.1 lg, 17.05mmol) was added followed by glacial acetic acid (8ml). After 1 hr, the mixture was diluted with dichlorom~h~nl~-water, and the organic layer washed with saturated aq.
NaHC03, brine, dried (MgSO4) and evaporated to an oil which was purified by flash chromatography (silica, ethyl acetate-pet. ether) to give the title compound as an oil which solidified on standing.
h. (Jr)-4R~ SS-N-(6-(4-Fluorophenyl)hexyl)-4-(4-ethoxycarbonylbenzylsulphinyl)-2-oxoazel:idin~l-yl~re W O 97/41098 PCT~EP97/01898 Tre~tmP-nt of [(4R)-4-(4-carbethoxy)benzylslllrhinyl-2-oxoq-7etidin- l-yl]aceticacid with 6-(~-fluoropheny)lhexylamine under the conditions described for FY,qmple 6i gave, after chromatography, the title compound with clentical spectrql and chromatographic characteristics to that prepared in Example 6i.
aD~~C = +71.9 (c. 1.0% w/v CHCl3, 25~C) Example 9 - (4R, SS)-N-(6-[4-Fluorophenyl]hex-1-yl)-4-(4 (~yclohexyloxycarbonyloA,~",et~ loxycarbonyl)bc.~ h;~yl)-2 ;n-l-yl)r ~ ePnlide A solution of (4R, SS)-N-(6-[4-fluorophenyl]hex-1-yl)-4-(4-carboxybenzylsulphinyl)-2-oxoq7~ti-1in-l-yl)~et,qmide (see above, 4 g, 0.00819 mol) and iodomethyl cyclohexyl carbonate (3.49 g. 0.0123 mol) in N-methylpyrrolidinone (40 ml, dried over 4A molecular sieves) was treated with anhydrous potqssium carbonate (1.7 g, 0.0123 mol) and stirred for 16 h at ambient ~emrer~ture. The mixture was diluted with water and extracted thoroughly with ethyl acetate, and the combined extracts washed with 5% aq.
sodium thiosulphate, dried (MgSO4) and evaporated. The residue was purified by chromatography (fine silica, ethyl acetate) and the title compound obtained as white crystals m.p.105-7~C after trituration with ethertlight petrol, 4.16 g, 79%
yield lH NMR ~ (DMSO) 1.23-1.~1 (14H, m), 1.64 (2H, m), 1.84 (2H, m), 2.51 (2H, m), 3.04 (3H, m), 3.35 (lH, m), 3.83 and 4.09 (each lH, d), 4.15 and 4.35 (each lH, d), 4.58 (lH, m), 4.85 (l~I, m), 5.95 (2H, s), 7.05 (2H, m), 7.18 (2H, m), 7.53 (2H, d), 7.98 (2H, d), 8.11 (lH, bt).
Found: C, 61.6; H, 6.4; N,4.6%; C33H41FN2O8S requires: C, 61.5; H, 6.4; N, 4.3%

Example 10 - ~4R, SS)-N-(6-[4-Fluorophenyl]hex-l-yl)-4-(4-(tert-bUtyloA~ bon,~lo~ loxycarbonyl)l)&.lz,~lsulphinyl)-2-o,~ l;A:n-l-yl)acetam;de A sollltion of (4R, SS)-N-(6-[4-fluorophenyl]hex-1-yl)-4-(4-carboxybenzylsulphinyl)-2-oxoazetidin-1-yl)acet~mide (0.2g, 0.00041 mol) and iodome~hyl tert-butyl carbonate (0.49 g. 0.001 mol) in N-methylpyrrolidinone (2 ml, dried over 4A molecular sieves) was treated with anhydrous potassium carbonate (0.14 g, 0.001 mol) and stirred for 16 h at ambient temperature. The mixture was diluted with water and extracted thoroughly with ethyl acetate, and the combined extracts washed with aq. sodium sulphite, dried (MgSO4) and evaporated. The residue was crystallised by trituration with light petrol to give the title compound as white crystals m.p.102-4~C, 0.23 g, 92%yield ... ~ . .. . . .

W O 97/41098 PCT~EP97/01898 'H NMR ~ (DMSO) 1.25-1.52 (17H, m), 2.50 (2H, m), 3.06(3H, m), 3.35 (lH, m), 3.84 and 4.09 (each lH, d), 4.15 and 4.35 (each lH, d), 4.86 (lH, m), 5.90 (2H, s), 7.06 (2H, m), 7.18 (2H, m), 7.53 (2H, d), 7.97 (2H, d), 7.99 (lH, bt) Found: C, 60.1; H, 6.3; N, 4.7%; C31H39F~J208S requires: C, 60.2; H, 6.4; N, 4.5%
Example 11 - (4R, SS)-N-(6-[4-Fluorophenyl]hex,l-yl)~(4(1-methylcydohex~ ~ I,onylo~~ U.~ loxycarbonyl)b~ rhinyl)-2-oxoaze~din-l-yl)acetamide A sol~ltion of (4R, SS)-N-(6-[4-fluorophenyl]hex-1-yl)-4-(4-carboxybenzylsulphinyl)-2-oxoazetidin-l-yl)~cet~mide (0.2g, 0.00041 mol) and chloromethyl l-methylcyclohexyl carbonate (0.21 g. 0.001 mol) in N-methylpyrrolidinone (2 ml, dried over 4A molecular sieves) was treated with anhydrous potassium carbonate (0.14 g, 0.001 mol) and potassium iodide (0.166 g, 0.001 mol) and stirred for 16 h at ambient temperature. The mixture was diluted with water and extracted thoroughly with ethyl acetate, and the combinedextracts washed with aq. sodium thiosulphate, dried (MgSO4) and evaporated.
The residue was purified by flash chromatography (fine silica, ethyl acetate) and the material thus obtained cryst~lli.ced by trituration with ether, to give the title compound as yield white crystals, m.p. 92-3~C, 0.11 g, 41% yield lH NMR ~ (DMSO)1.25-1.48 (16H, m), 2.00 (2H, m), 2.5 (SH, m), 3.07 (3H, m), 3.36 (lH, m), 3.83 and 4.09 (each lH, d), 4.15 and 4.35 (each lH, d), 4.86 (lH, m), 5.91 (2H, s), 7.06 (2H, m), 7.18 (2H, m), 7.54 (2H, d), 7,98 (2H, d), 8.11 (lH, bt) FY~P'~ 12 - (4R, SS)-N-(6-[4-Fluorophenyl]hex-l-yl)-4-(4-(phenyl~ l.<..-~lox~ ll.yloxycarbonyl)be~L~ lphinyl)-~oxoq7e~ n-l-yl)ac~ ide A solution of (4R, SS)-N-(6-[4-fluorophenyl]hex-1-yl)-4-(4-carboxybenzylsulphinyl)-2-oxoazetidin-1-yl)~ce~mide (5 g, 0.0102 mol) and benzoyloxychlorometh~ne (2.62 g. 0.0154 mol) in N-methylpyrrolidinone (50 ml, dried over 4A molecular sieves) was treated with anhydrous potassium carbonate (2.12 g, 0.0154 mol) and potassium iodide (2.55 g, 0.0154 mol) and stirred for 16 h at ambient temperature. The mixture was diluted with water and extracted thoroughly with ethyl acetate, and the combined extracts washed with aq. sodium thiosulph~, dried (MgSO4) and evaporated. The residue was purifiled by flash chromatography (fine silica, ethyl acetate) and the material thus obtained cryst~lliced by trituration with ether to give the title compound as white crystals, m.p.l 17-9~C, 4 g, 52% yield .. .... .....

O 97/41098 PCr/Eps7lol898 lH NMR ~ (CDCl3) 1.33 (4H, m), 1.5-1,62 (4H, m), 2.56 (2H, t), 2.gS, (lH, dd), 3.22 (3 H, m), 3.94 and 4.20 (each lH, d), 4.04 and 4.16 (each lH, d), 4.65 (lH,m), 6.25 (2H, s), 6.94 (3H, m), 7.-11 (2H, m), 7.37 (2H, d), 7.46 (2H, m), 7.59 (lH, m), 8.1 (4H, m) Found: C, 63.7; H,5.5; N, 4.5%; C33H35FN2O7S requires:C, 63.7; H, 5.7; N, 4.5%

Example 13- (4R, SS)-N-(6-~4-~luorophenyllhex-1-yl)-4-(4(4 methoxyphenylcarbonylo~l.,el~.rloxycarbonyl)b~ lphinyl)-2-oY~!~7eti'~ yl)acetamide A solution of (4R, SS)-N-(6-[4-fluorophenyl]hex-1-yl)-4-(4-carboxybenzylclllrhinyl)-2-oxo~7Pti-~in-l-yl)~cet~mide (2 g, 0.0041 mol) and 4-methoxybenzoyloxymethyl chloride (2 g. 0.01 mol) in N-methylpyrrolidinone (20 ml, dried over 4A molecular sieves) was treated with anhydrous potassium carbonate (1.38 g, 0.01 mol) and potassium iodide (1.66 g, 0.01 mol) and stirredfor 16 h at ambient temperature. The mixture was diluted with water and extracted thoroughly with ethyl acetate, and the combined extracts washed with aq. sodium thiosulphate, dried (MgSO4) and evaporated. The residue was purified by flash chromatography (fine silica, ethyl acetate) and the m~teri~l thus obtained crystallised by trituration with ether, then recryst~lli.~d from dichlorome~ ne/light petrol, to give the title compound as white crystals, m.p.
115-8~C, 1.34 g, 50% yield lH NMR o (DMSO) 1.23-1.6 (8H, m), 2.50 (2H, m), 3.02-3.1(3H, m), 3.34 ( lH, m ), 3.8 and 4.09 (each lH, d), 3.84 (3H, s), 4.14 and 4.33 (each lH, d), 4.85 (lH, m), 6.17 (2H, s), 7.05 (4H, m), 7.16 (2H, m), 7.52 (2H, d), 7.97 (4H, m), 8.1 (lH, bt) Example 14- (4R, SS)-N-(6-[4-Fluorophenyl]hex-l-yl)-4-(4 (isobutyryloA,~ lhyloxycarbonyl~Je.,.~ ulphinyl)-2-oY~7eff~l:n_l-yl)acetamide A solution of (4R, SS)-N-(6-[4-fluorophenyl]hex-1-yl)-4-(4-carboxybenzylsulphinyl)-2-oxoazetidin-1-yl)~ce~mide (0.2g, 0.00041 mol) and isobutyryloxymethyl iodide (0.23 g. 0.001 mol) in N-methylpyrrolidinone (2 ml, dried over 4A molecular sieves) was treated with anhydrous potassium carbonate (0.14 g, 0.001 mol) and stirred for 16 h at ambient temperature. The mixture wasdiluted with water and extracted thoroughly with ethyl acetate, and the combinedextracts washed with aq. sodium thiosulphate, dried (MgS04) and evaporated.
The residue was purified by chromatography (fine silica, ethyl acetate) and the material thus obtained crystallised by trituration with ether, to give the titlecompound as white crystals, m.p. 104-6~C, 0.14 g, 58% yield W O 97/41098 PCT~EP97/01898 lH NMR ~ (DMSO)l.09 (6H, d), 1.25-1.52 (8H, m), 2.50 (2H, m), 2.61 (lH, m), 2.97-3.06 (3H, m), 3.34 (lH, m), 3.83 and 4.09 (each lH, d), 4.1S and 4.34 (eachlH, d), 4.84 (lH, m), 5.95 (2H, s~, 7.06 (2H, m), 7.16 (2H, m), 7.53 (2H, d), 7.97 (2H, d), 7.99 (lH, bt) Example 15- (4R, SS)-N-(6-[4~1uorophenyl]hex-1-yl)-4(4-(2-meth~x~rop-2-ylcarbo..,~lvx,~ loxyc~rbo..~l)~,~z~lsulphinyl)-2~xQ~7Pff~l;~-l-yl)acetamide A solution of (4R, SS)-N-(6-[4-Fluorophenyl]hex-l-yl)-4-(4-carbo~ybenzylsulphinyl)-2-0~0~7~ti~1in-1-yl)~cet~mi~ (3 g, 0.00614 mol) and 2-methoxyprop-2-ylcarbonyloxymethyl chloride (1.53 g. 0.00921 mol) in N-methylpyrrolidinone (30 ml, dried over 4A molecular sieves) was treated with anhydrous potassium carbonate 1.27 g, 0.00921 mol) and potassium iodide (1.53 g, 0.00921 mol) and stirred for 16 h at ambient temperature. The mixture was diluted with water and extracted thoroughly with ethyl acetate, and the combinedextracts washed with aq. sodium thiosulphate, dried (MgSO4) and evaporated.
The residue was purified by chromatography (fine silica, ethyl acetate) and the material thus obtained as a solution in ethyl acetate further washed with aq.
sodium thiosulph~te then stirrred for 10 min with MgS04 and decolourising charcoal. The solids were filtered off and the filtrate evaporated, and the residue cryst~lliced by trituration with ether/light petrol to give the title compound as white crystals m.p. 92-4~C, 2.6 g, 68~ yield lH NMR ~ (CDC13) 1.34 (4H, m), 1.45 (6H, s), 1.58 (4H, m), 2.56 (2H, t), 2.95 (lH, dd), 3.17-3.37 (3H, m + 3H, s), 3.96 and 4.10 (each lH, d), 4.05 and 4.22 (each lH, d), 4.69 (lH, m), 6.06 (2H, s), 6.93 (3H, m), 7.09 (2H, m), 7.39 (2H, d), 8.09 (2H, d) FY~n~P'C 16 - (4R, SS)-N (6-[4 FIUOrOPhenYI]heX-1-YI)-4 (4 ((5 methYI-2-OXO-1,3 dioY~le~ 1 yl)methyloxycarbonyl)be~ ul~hinyl)-2-oxo~7et~Ain-l-yl)r~t~ e Asolutionof (4R,SS)-N-(6-[4-fluorophenyl~hex-1-yl)-4-(4-carboxybenzylsulphinyl)-2-oxoazetidin-1-yl)acetamide (5 g, 0.0102 mol) and 4-bromomethyl-S-methyl-1,3-dioxol-2-one (2.96 g. 0.0154 mol) in N-methylpyrrolidinone (50 ml, dried over 4A molecular sieves) was treated with anhydrous potassium carbonate (2.12 g, 0.0154 mol) and stirred for 16 h at ambient temperature. The mixture was diluted with water and extracted thoroughly with ethyl acetate, and the combined extracts washed with water, brine, dried (MgS04) and evaporated. The residue was purified by chromatography (fine silica, ethyl acetate) and the material thus obtained W O 97/41098 PCT~EP97/01898 cryst~lli.~d by trituradon with ether/light petrol to give the dtle compound as white crystals, m.p. 84-7~C, 3.81 g, 62% yield.
'H NMR 8 (CDC13) 1.24-1.54 (8H, m), 2.22 (3H, s), 2.5 (2H, m), 2.95 (lH, dd), 3.05 (2H, m), 3.33 (lH, m), 3.84 and 4.07 (each lH, d), 4.15 and 4.32 (each lH, d), 4.85 (lH, m), 5.22 (2H, s), 7.06 (2H, m), 7.18 (2H, m), 7.51 (2H, d), 7.98 (2H, d), 8.1 (lH, bt) Found: C, 59.7; H, 5.5; N, 4.7%; C30H33FN208S requires:C, 60.0; H, 5.5; N, 4.7%

Example 17 - (4R, SS)-N-(6-[4-~luorophenyl]hex-1-yl)-4-(4-((2-h~Yycarbonyl.E-but-2-en-yl)~ l,yloxycarbonyl)benzyl.cl-lrhi~yl)-2-o~o~7e~din-l-yl)acetamide A solution of (4R, SS)-N-(6-~4-fluorophenyl]hex-1-yl~-4-(4-carboxybenzyl~ rhinyl)-2-oxoq7P-tidin-l-yl)acetqmide (0.195 g, 0.0004 mol) and methyl E-2-bromomethylbut-2-enoate (0.16 g. 0.0008 mol) in N-methylpyrrolidinone (2 ml, dried over 4A molecular sieves) was treated with anhydrous potassium carbonate (0.11 g, 0.0008 mol) and stirred for 16 h at ambient temperature. The mixture was diluted with water and extracted thoroughly with ethyl acetate, and the combined extracts dried (MgSO4) and evaporated. The residue was purified by chromatography (fine silica, ethyl acetate) and the mqtPri~l thus obtained crystallised by trituration with ether to give the title compound as white microprisms, 0.024 g, 10% yield 1H NMR ~ (CDCl3) 1.25-1.63 (8H, m), 1.99(3H, d), 2.56 (2H, t), 2.93 (lH, dd), 3.17-3.36 (3H, m), 3.79 (3H, s), 3.93 and 4.17 (each lH, d), 3.43 and 4.23 (eachlH, d), 4.64 (lH, m), 5.12 (2H, s), 6.91-7.26 (6H, m), 7.34 (2H, d), 8.04 (2H, d) FY~nP'e 18 - (4R, SS)-N-(6-[4-Fluorophenyl]hex-l-yl)-4-(4-( N-N-di~..ell.,~ rbonylmethyloxycarbonyl)benzylsulphinyl)-2-oxoazetidin-1-yl)acetamide A solution of (4R, SS)-N-(6-[4-fluorophenyl]hex-1-yl)-4-(4-carboxybenzyl.culphinyl)-2-oxoq7~ti~in-1-yl)acet~mide (0.2 g, 0.0004 mol) and a-chloro-N,N-dimethyl acetamide (0.17 g. O.OOl mol) in N-methylpyrrolidinone (2 ml, dried over 4A molecular sieves) was treated with anhydrous potassium carbonate (0.14 g, 0.001 mol) and stirred for 16 h at ambient temperature. The mixture was diluted with water and extracted thoroughly with ethyl acetate, and the combined extracts dried (MgSO4) and evaporated. The residue was crystqlliced by trituration with ether to give the title compound as white cryatals, m.p. 141-4~C, 0.01 g W O97/41098 PCT~EP97/01898 lH NMR ~ (CDC13) 1.25-1.75 (8H, m), 2.56 (2H, t), 2.93 (lH, dd), 2.99 (3H, s), 3.04 (3H, s), 3.17-3.36 (3H, m), 3.85-4.25 (4H, 4 x d), 4.65 (lH, m), 4.97 (2H, s), 6.93 (2H, m), 7.10 (3H, m), 7.36-(2H, d), 8.13 (2H, d).

Example 19 - (4R, SS)-N-(6-[4Fluorophenyl]hex-l-yl)-4-(4-( N-N-di-(2-h~lr~iAJ~ h bo,.~ .eUIrloxycarbonyl)bellL,yl~ hinyl)-2-~ o ~ in-1-yl)acetamide To a sr~lutiQn of N,N-(l,1-dihydroxyethyl)bromo~cet~mi~e (1.20g, 5.3mmol) in DMF (lOml) was added (4R, SS)-N-(6-[4-fluorophenyl]hex-1-yl)-4-(4-carboxybenzylsulphinyl)-2-oxoazetidin-1-yl)acet~mide (l.OOg, 2.05mmol) followed by c~Psium carbonate (1.3g, 4.0mmol). The mixture was stirred for 18 hrs then separated between ethyl acetate and 2M hydrochloric acid. Separation ofthe organics followed by washing with bdne, drying (MgS04) and concentration provided the crude product as a white solid. The solids were washed with chloroform, filtered and dried to yield the product as a white solid (0.97g, 75%) mp 119-121~C.
'H nmr ~ (DMSO~") 1.28-1.55(m,8H), 2.5(t, J=12Hz, 2H), 2.8(br d, J=16Hz, lH), 3.1(q, J=lOHz, 2H), 3.3-3.7(m, 9H), 3.9(d, J=27Hz, lH), 4.1(d, J=27Hz, lH), 4.15(d, J=20Hz, lH), 4.35(d, J=20Hz, lH), 4.7(br, lH), 4.85(m, lH), 4.95(br, lH), 5.2(s, 2H), 7.0(apparent t, J=14Hz, 2H), 7.15(dd, J=14, 9Hz, 2H), 7.5(d, J=13Hz, 2H), 8.05(d, J=13Hz, 2H), 8.2(t, J=8Hz, lH) Example 20 F~g~ mnrs of (R,S/S,R)-1-(2-(6-Fluorophenyl)hexyloxyhthyl-4(4allylo~c~rl,onylbel~yl.~ulrllinyl)-2-oxo~7pt~(line a 6-(4Fluorophenyl)hexyloxye~h~nol Ethylene glycol (41.26g) and 6-(4-Quorophenyl)hexyl bromide (17.22g) were added to a solution of sodium hydroxide (2.79g) in water (2.5ml) and the mixturewas heated at 110~for 24 hours. The mixture was cooled and partitioned between water (150ml) and diethyl ether (150ml). The layers were separated and the organic layer was washed with water, brine, dried (MgS04) and evaporated to an orange oil. Purification by column chromatography on silica gel eluted with [5:1]
to [1:1] P.E 40-60~C:ethyl acetate gave the product as an orange oil (9.9lg, 62%).
b. 6-(4-Fluorophenyl)hexyloxy triflate 6-(4-Fluorophenyl)hexyloxy ethanol (4.0g), pyridine (1.43g) and DMAP (40mg) were dissolved in dry dichloromethane (30ml), cooled to -10~C and triflic anhydride (5.6g) in dry dichloromethane (lOml) was added over 5 minutes keeping temperature below 0~C. The mixture was stirred at -10~C to 0~C over 60 rninll~es and then washed with water (SOml), brine (50ml), dried (MgS04) and evaporated to a brown oil ~6.17g, ~%).

wo 97/41098 pcTlEps7lol8s8 c. 1~ 1uorophenyl)hexyloxy)ethyl-4-(4-allyloxyc~ l)o~ lb~ io)-2 oxoazetidine A solution of 6-(4-fluolophenyl3hexyloxy triflate (7.0g), 4-(4-allyloxycarbonylben7tlthiQ)~7eti~in-2-one (5.08g) and tetrabutylammonium bromide (0.59g) in dry THF (lSOml) was cooled to 15~C under nitrogen and was treated with powdered potassium hydroxide (1.08g). The cooling bath was removed and the reaction was stirred for 30 minuteS, Powdered KOH (SOmg) was added and the reaction was stirred at room temperature for 30 minutes. The mixture was filtered through celite and washed through with ethyl acetate (lOOml). The filtrate was evaporated to a dark oil and purification by flash column chromatography on silica gel eluted with [3:2] to [1:1] P.E. 40-60~C:
ethyl acetate gave the product as an orange oil (4.65g, 51%).
lH NMR ~ (CDC13) 1.3-1.6 (8H, m, 4xCE~2,), 2.55 (2H, t, J = 7.6Hz, CH2Ph), 2.85, 2.90 (lH, dd, ~3), 3.05 (lH, m, lH), 3.26, 3.32(1H, dd, J=4.9, lS.lHz, ), 3.37-3.68 (SH, m, NCH2CH2, OC~2), 3.86 (2H, s, SCH2), 4.76 (lH, m, ), 4.81 (2H, m C02CH2), 5.36 (2H, m, CH=CH2), 6.03 (lH, m, CH=CH2), 6.94 (2H, m p-FPh-O, 7.07 (2H,m, p-FPh-O, 7.39 (2H, d, J=8.3Hz, Ph-H), 8.01 (2H, d, J=8.3Hz, Ph-O
d. (R,S/S,R)-1-(2-(6-Fluorophenyl)hexyloxy~ethyl-4-(4-allyloxycarbonylbe.,L,~ l~ulphinyl)-2-oxoazetidine (diast 2) A solution of 1-(2-(6-fluorophenyl)hexyloxy)ethyl-4-(4-allyloxycarbonylbenzylthio)-2-oxoazetidine (4.58g) in dichloromPth~n~ (lOOml), cooled to -70~C, was treated with a solution of mcpba (2.0g) in CH2C12 (125ml) dropwise over 1 hour m~int~ining the temperature at below -70~C. The cooling bath was removed and the reaction was stirred for 1.5 hours, washed with a solution of 10% aq.sodium hydrogen carbonate(lSOml) + 10% aq. sodium s-llphi~e (ISOml). The layers were separated and the organic layer was washed with brine, dried (MgS04) and evaporated to an oil which solidified on st:lnding. Repeat recyr~t~llis~tions from diethyl ether (3 times) gave diastereoisomer 2 as a colourless solid (0.7g,12.2%).m.p. 73-74~C
lH NMR o (CDCl3) 1.3-1.6 (8H, m, 4xCE~2,), 2.56 (2H, t, J = 7.8Hz, C~2Ph), 2.66, 2.69 (lH, dd, J=2,14.8Hz, H3), 3.38-3.68 (7H, m, H3, NCH2CH2, ocEI2)~
4.07 (2H, s, SOCEI2), 4.54 (lH, m, ~4), 4.83 (2H, m C02C_2)~ 5.36 (2H, m, CH=C~I2) 6.03 (lH, m, C~=CH2), 6.94 (2H, m, p-FPh-H), 7.10 (2H, m, p-FPh-~I), 7-38 (2H, d, J=8.4Hz, Ph-H~, 8.08 (2H, m, Ph-H) e. (~,RtS,S)-(2-(6-Fluorophenyl)hexyloxy)ethyl-4-(4-~llyloxycarbonyl~.,LyLs~llphinyl)-2-oxo~7eh~line (diast 1) A sample of diastereoisomer 1 (contains 21 % dia2) was obtained as a colourless solid, m.p.46-50~C.

. . .

wo 97/41098 PCT/EPg7/01898 lH NMR ~ (CDC13) 1.3-1.6 (8H, m, 4xCH2,), 2.55 (2H, t, J = 7.8Hz, C~2Ph), 2.66, 2.69 (lH, dd, J=4.8,14.8Hz, H3), 3.38-3.68 (7H, m, H3, NC~2C~12, OC~I2), 3.94 (2H, m, SOC~2), 4.55 (lH, m, ~4), 4.83 (2H, m C02C~2), 5.36 (2H, m, CH=C~2) 6.03 (lH, m, C.~=CH2), 6.94 (2H, m, p-FPh-~), 7.10 (2H, m, p-FPh-~), 7.38 (2H, d, J=8.4Hz, Ph-H), 8.08 (2H, m, Ph~
F.n~rltiom~.r~ of (R,S/S,R)-1-(2-(6-fluorophenyl)hexyloxy)ethyl-4-(~
allyloxycarbonylbenzylsulphinyl)-2-oxo~7Pti~in~ were separated by hplc on a Chiracel OD -20MM eluted with 60:40 ethanol hexane:
f. (+)-1-(2-(~Fluorophenyl)hexyloxy)ethyl-4-(4-allyloA~c~bo.-~lb~ ulphinyl)-2-oxo~7e~ ;ne (diast. 2) 164mg, colourless solid. M.p. 52-53~C
Fn~ntiomerically pure [O~]D = +57-5~ (c=0.5%w/v in ethanol at 25~C) lH NMR o (CDC13) 1.3-1.6 (8H, m, 4xCH2,), 2.56 (2H, t, J = 7.8Hz, C~2Ph), 2.66, 2.69 (lH, dd, J=2,14.8Hz, H3), 3.38-3.68 (7H, m, ~3, NCH2CH2, OC~2), 4.07 (2H, s, SOC~2), 4.54 (lH, m, H4), 4.83 (2H, m CO2CH2), 5.36 (2H, m, CH=C~12) 6.03 (lH, m, C~=CH2), 6.94 (2H, m, p-FPh-~), 7.10 (2H, m, p-FPh-~), 7.38 (2H, d, J=8.4Hz, Ph-H), 8.08 (2H, m, Ph-O
g. (-) 1-(2-(6-Fluorophenyl)hexyloxy)ethyl-4(4-allyloA,~c~l,onyll,&.u~ hinyl)-2-oxo~eh-line (diast. 2) 170mg, colourless solid M.p. Sl-S3~C
9g.69% desired enantiomer with 0.31 % of the other en~ntiorner present [a]D = -57.9~C (c=0.4%w/v in ethanol at 25~C) lH NMR ~ (CDC13) 1.3-1.6 (8H, m, 4xCH2,), 2.56 (2H, t, J = 7.8Hz, CH2Ph), 2.66, 2.69 (lH, dd, J=2,14.8Hz, H3), 3.38-3.68 (7H, m, ~3, NCH2CH2, OCH2), 4.07 (2H, s, SOCH2), 4.54 (lH, m, _4), 4.83 (2H, m CO2CH2), 5.36 (2H, m, CH=CH2) 6.03 (lH, m, C~=CH2), 6.94 (2H, m, p-FPh-H), 7.10 (2H, m, p-FPh-O, 7.38 (2H, d, J=8.4Hz, Ph-H), 8.08 (2H, m, Ph-O
FY~rle 21 (R,S/S,R)-1-(2-(6-Fluorophenyl)hexyloxy)ethyl-4(4-c~rl,oA,~ bulphinyl) 2-oYo~ti~l;ne (diast. 2) (R,S/S,R)- 1-(2-(6-Fluorophenyl)hexyloxy)ethyl-4-(4-allyloxycarbonylbenzylsulphinyl)-2-oxoazetidine (220mg), triphenylphosphine (1 lmg), tetrakis(triphenylphosphine)p~ lium (0) (lSmg) in dry CH2C12 (5ml) under nitrogen was treated with pyrrolidine (37~11) and the mixture was stirred at room temperature for 19 hours.. Purificadon by flash column chromalography on silica gel eluted with a CH2C12:acetone:acetic acid gradient elution gave the product as an oil. The oil was dissolved in CH2C12 (4ml) and diluted with diethyl W O 97/41098 PCT~EP97/01898 ether (75ml), washed with water, brine, dried (MgS04), filtered and evaporated ~ to an oil which soli~ifipd on cooling. Trituration with diethyl ether gave the product as a cream solid (lSSmg, 78%) m.p.9S-96~C
lH NMR ~ (CDC13) 1.25-1.6 (8H, m, 4xCH2,), 2.56 (2H, t, J=7.6Hz, CH2Ph), 2.69, 2.76,(1H, dd, J=2.1,15.1Hz,~3), 3.10, 3.17 (lH, dd, J=S.l, lS.l~lz), 3.37-3.74 (6H,m, NC~12C~2, 0C~12), 4.11 (2H, s, SOC~2), 4.64 (lH, m, ~4), 6.94 (21H, m, p-FPh-ED, 7.10 (2H, m, p-FPh-~O, 7.41 (2H, d, J=8Hz, Ph-H), 8.07 (2H, d, J=8Hz, Ph-~D
Ex~mple22 (-)-1-(2-(6-Fluorophenyl)hexyloxy)ethyl-4-(4 ho~b~.-L,~ ph;nyl) 2-oxo~7et~line (diast. 2) (-)- 1-(2-(6-Fluorophenyl)hexyloxy)ethyl-4-(4-allyloxycarbonylbenzyl.culphinyl)-2-oxo~7Pridine (145mg), triphenylphosphine (7.3mg), tetrakis(triphenylphosphine)p~ rn (O) (lO.Smg) in dry CH2C12 (4ml) under nitrogen was treated with pyrrolidine (23.5~1) and the mixture was stirred at room temperature for 6 hours. Pyrrolidine (5~1) was added and the reaction was stirred at room temperature for 19 hours. Pllnfica~ion by flash column chromatography on silica gel eluted with a CH2C12:acetone:acetic acid gr~di~nt elution gave thetitle compound as an oil. Trituration with diethyl ether gave the product as a cream solid (113mg, 85%) m.p.123-124~C, 1O~]D = -53-5~ ( c=O.S %w/v in ethanol al 25~C) lH NMR ~ (DMSO) 1.25-1.6 (8H, m, 4xCE[2,), 2.50 (2H, t, CH2Ph), 2.91, 2.95 (lH, dd, J=2,15.2Hz, ~3), 3.22-3.53 (7H,m, ~3, NCH2CH2, OC.~2), 4.16, 4.31 (each lH, 2xd, J=13Hz, SOC~2), 4.73 (lH, m, H4), 7.0 (2H, m, p-FPh-H), 7.19 (2H, m, p-FPh-O, 7.48 (2H, d, J=8~Iz, Ph-H), 7.94 (2H, d, J=8Hz, Ph-O

~YP~p'e 23 (+)-1-(2-(6-Fluorophenyl)hexyloxy)ethyl-4-(4-carbo~~ ulphinyl)-2-oyo~7~ ne (diast. 2) (+)- 1-(2-(6-Fluorophenyl)hexyloxy~ethyl-4-(4-allyloxycarbonylbenzylsulphinyl)-2-oxoazetidine (140mg), triphenylphosphine (7mg), tetrakis(triphenylphosphine)p~ m (O) (lOmg) in dry CH2C12 (4ml) under nitrogen was treated with pyrrolidine (23.5~u1) and the mixture was stirred at room temperature for 19 hours. I.Smg more catalyst and pyrrolidine (S~l) were added and the reaction was stirred for 2 hours, diluted with water (25ml) and CH2C12 (25ml) and acidified to pH 2 with HCl (2N). The layers were separated and the aquoues layer was washed with CH2C12 (25ml) The organic extracts were combined, dried (MgS04) and evaporated to a yellow oil. Purification by flash column chromatography on silica gel eluted with a CH2C12:acetone:acetic acid gradient elution gave the product as an oil. Trituration with diethyl ether gave the product as a colourless solid (109mg, 84%) m.p. l 18-120~C, [a]D = +so.oo ( c=0.5%w/v in ethanol at 25~C) lH NMR ~ (DMSO) 1.25-1.6 (8H, m, 4xC~2,), 2.50 (2H, t, CH2Ph), 2.91, 2.95 (lH, dd, J-2,15.2Hz,~3), 3-22-3.53 (7H,m, ~3, NCH2CE12~ 0C~12), 4-16, 4-31 (each lH, 2xd, J=13Hz, SOCH2), 4.73 (lH, m, H4), 7.0 (2H, m, p-FPh-EI), 7.19 (2H, m, p-FPh-~D, 7.48 (2H, d, J=8Hz, Ph-H), 7.94 (2H, d, J=8Hz, Ph-~) Example 24 Enanffomers of (R,S/S,R)-1.(2-(6.Chlorophenyl)hexyloxy)ethyl-4(4allyloxy~arbonylb~ lrhinyl)-2-oy~l~7eh~in~
a 6~ Chlorophenyl)hexyloxyeth"n"~
Ethylene glycol (51.lg) and 6-(4-chlorophenyl)hexyl bromide (22.7g) were added to a solution of sodium hydroxide (3.46g) in water (3.1ml) and the mixture was heated at 110~for 24 hours. The mixture was cooled and partitioned between water (300ml) and diethyl ether (300ml). The layers were separated and the aqueous layer was washed with ether (lSOml). The organic layers were combined and washed with water, brine, dried (MgS04) and evaporated to yellow oil.
Pllrific;~tion by column chromatography on silica gel eluted with [3:1~ to [2:1]P.E 40-60~C:ethyl acetate gave the product as a yellow oil (14.13g, 67%).
b. 6-(4Chlorophenyl)hexyloxy trinate 6-(4-Chlorophenyl)hexyloxy ethanol (7.6g), pyridine (2.53g) and DMAP (79mg) were dissolved in dry dichloromethane (60ml), cooled to -10~C and triflic anhydride (lO.Og) in dry dichlorometh~n~ (20ml) was added over 7 minutes keeping T below 0~C. The mixture was stirred at 0~C for 45 minutes, washed with water (60ml), brine (60ml), dried (MgS04) and evaporated to a dark oil (11.13g, 97%).
~ (2-(~Chlorophenyl)hexyloxy)ethyl~4.(4-allyloxycarbonylbenzylthio)-2-0~ ;r~A solution of 6-(4-chlorophenyl)hexyloxy triflate (11. lg), 4-(4-allyloxycarbonylbenztlthio)azetidin-2-one (7.69g) and tetrabutylarnmonium bromide (0.89g) in dry THF (200ml) was cooled to 10~C under nitrogen and was treated with powdered potassium hydroxide (1.63g). The cooling bath was removed and the reaction was stirred for 40 minllt~s Powdered KOH (163mg) was added and the reaction was stirred at room temperature for l.5h, partitionedbetween brine (600ml) and ethyl acetate (400ml). The mixture was filtered through hy-flo and the layers were separated. The organic layer was dried (MgS04) and evaporated to a dark oil. Purification by flash column chromatography on silica gel eluted with [2:1] to [l:l]-P.E. 40-60~C: ethyl acetate gave the product as an orange oil (7.28g, 51 ~0).
lH NMR ~ (CDC13) 1.3-1.6 (8H, m, 4xC~2,), 2.55 (2H, t, J = 7.6Hz, CH2Ph), 2.85, 2.90 (lH, dd, ~3), 3.05 (lH, m, lH), 3.26, 3.32(1H, dd, J=4.9, l5.1Hz, .. ...

W O 97/41098 PCT~EP97/01898 ), 3.37-3.68 (SH, m, NC~2C~2, OC~2), 3.86 (2H, s, SC~2), 4.76 (lH, m, ~I4), 4.81 (2H, m CO2C~12), 5.36 (2H, m, CH-C~E~2) 6.03 (lH, m, C~=CH2) 7.07 (2H, d, J=8.4Hz, p-ClPh-~l~, 7.23 (2H,m, p-ClPh-~), 7.39 (2H, d, J=8.3Hz, Ph-H), 8.01 (2H, d, J=8.3Hz, Ph-O
d. (R,S/S,R)-1-(2-(6~Chlorophenyl)hexyloxy)ethyl-4-(4-Jlr)hinyl)-2-o~o~7eR~line A solu~on of 1-(2-(6-Chlorophenyl)hexyloxy)ethyl-4-(4-allyloxycarbonylbenzylthio)-2-oxo~7~ti~linP (7.2g) in dichlorometh~nP (175ml), cooled to -70~C, was treated with a solution of mcpba (3.0g) in CH2C12 (175ml) dropwise over 1 hour m~int~inin~ the temperature at below -70~C. The cooling bath was removed and the reaction was stirred for 1.5 hours, washed with a solution of 10% aq.sodium hydrogen carbonate(200ml) + 10% aq. sodium sulphite (200ml). The layers were separated and the organic layer was washed with brine, dried (MgSO4~ and evaporated to an oil which soli~lifiPd on standing. Repeat ;y-~ li.~tions from diethyl ether (4 times) gave diastereoisomer 2 as a colourless solid (0.gg,12.2%).
lH NMR ~ (CDC13) 1.3-1.6 (8H, m, 4xC~2,), 2.56 (2H, t, J = 7.8Hz, C.E12Ph), 2.66, 2.69 (lH, dd, J=2.4,15.2Hz, ~3), 3.07, 3.11 (lH, dd, J=4.8, 15.2Hz,~3), 3.37-3.68 (6H, m, NC~2C~I2, OCH2), 4.07 (2H, s, SOCH2), 4-54 (lH, m, ~14), 4.83 (2H, m CO2CH2), 5.36 (2H, m, CH=CH2) 6.03 (lH, m, CH=CH2) 7.09 (2H, d, J=8.8Hz, p-ClPh-O, 7.23 (2H, d, J=8.8Hz, p-ClPh-~I), 7.39 (2H, d, J=8.4Hz, Ph-H), 8.07 (2H, m, Ph-O

The above racemic compound was separated by hplc on a Chiracel OD -20mmeluted with 80:20 ethanol:hexane to give the enantiomers:
e. (~ (2-(6-Chl~ olJhenyl)hexyloxy)ethyl-4-(4-allyloxycarbonylb~ p~l;nyl)-2-o~os7e~i~;ne 83mg, colourless solid. M.p. 57-59~C
99.95% desired enantiomer with 0.05% of the other enantiomer present [a]~ = +52.2~ (c=0.28%w/v in ethanol at 25~C) lH NMR ~ (CDC13) 1.3-1.6 (8H, m, 4xC~,), 2.56 (2H, t, J = 7.8Hz, CH2Ph), 2.66, 2.69 (lH, dd, J-2.4,15.2Hz, H3), 3.07, 3.11 (lH, dd, J=4.8, 15.2Hz, H3), 3.37-3.68 (6H, m, NC~I2CH2, OC~2), 4.07 (2H, s, SOC~2), 4.54 (lH, m, ~4), 4.83 (2H, m C02CH2), 5.36 (2H, m, CH=CH2) 6.03 (lH, m, C_=CH2) 7.09 (2H, d, J=8.8Hz, p-ClPh-~), 7.23 (2H, d, J=8.8Hz, p-ClPh-H), 7.39 (2H, d, J=8.4Hz, Ph-H), 8.07 (2H, m, Ph-H) f. (-)-1-(2-(6-Chlorophenyl)hexyloxy)ethyl-4-(4-allyloxycarbonylbenzylsulpl~inyl)-2-oxoazetidine 103mg colourless solid M.p. 58-59~C

W O97141098 PCT~EP97/01898 99.62% desired enantiomer with 0.38% of the other enantiomer present - [a}D = -61.9~C (c=0.06%w/v in ethanol at 25~C) lH NMR ~ (CDC13) 1.2-1.6 (8H, m, 4xC~2,), 2.56 (2H, t, J = 7.8Hz, C~2Ph), 2.66, 2.69 (lH, dd, J=2.4,15.2Hz, ~3), 3.07, 3.11 (lH, dd, J=4.8, 15.2Hz, E13), 3.37-3.68 (6H, m, NC~2C~I2, OC~2, 4.07 (2H, s, SOC~2), 4.54 (IH, m, ~14).
4.83 (2H, m C02C~2), 5.36 (2H, m, CH-C.~2) 6.03 (lH, m, C~=CH2) 7.09 (2H, d, J=8.8Hz, p-ClPh-~l), 7.23 (2H, d, J=8.8Hz, p-ClPh-O, 7.39 (2H, d, J=8.4Hz, Ph-H), 8.07 (2H, m, Ph-~) Example2S (-)-1-(2~ Chlorophenyl)hexyloxy)ethyl.4(1 c~bu~rl,enzylsulphinyl)-2-0~7eb~ (diast. 2) (-)- 1-(2-(6-Chlorophenyl)hexyloxy)ethyl-4-(4-allyloxycarbonylbenzyls-llrhinyl)-2-oxoa~tidine (86.2mg), triphenylphosphine (4.2mg), tetrakis(triphenylphosphine)palladium (0) (6mg) in dry CH2C12 (2ml) under nitrogen was treated with pyrrolidine (13.S~l) and the mixture was stirred at room temperature for 19 hours. Purification by flash column chromatography on silica gel eluted with a CH2C12:acetone:acetic acid gradient elution gave the product as an oil. Trituration with diethyl ether gave the title compound as a cream solid (59.8mg, 75%) m.p.102-102~C, [O~]D = -37.32~ ( c=0.209%w/v in ethanol at 2S~C) lH NMR ~ (CDC13) 1.25-1.6 (BH, m, 4xCH2,), 2.56 (2H, t, J = 7.6Hz, C~2Ph), 2.71, 2.74 (lH, dd, J-2,15Hz, H3), 3.12, 3.15 (lH, dd, J=5.2, lSHz, H3), 3.38-3.72 (6H, m, NCH2C~2, OCH2), 4.09 (2H, m, SOCH2), 4.60 (lH, m, H4), 7.09 (2H, d, J=8.8Hz, p-ClPh-H), 7.22 (2H, d, J=8.8Hz, p-ClPh-H), 7.40 (2H, d, J=8Hz, Ph-H), 8.08 (2H, d, J=8Hz, Ph-O
F.Y~rlple 26 (+)-1-(2-(6-Chlorophenyl)hexyloxy)ethyl-4-(~1 ca. b(,x~l,e-,L,~ -,yl)-2-oxoazeffdine (diast. 2) (+)- 1-(2-(6-Chlorophenyl)hexyloxy)ethyl-4-(4-allyloxycarbonylbenzylsulphinyl)-2-oxoaz~ddine (65.6mg), triphenylphosphine (3.2mg), tetrakis(triphenylphosphine)p~tl~dillm (O) (4.6mg) in dry CH2C12 (2ml) under nitrogen was treated with pyrrolidine (10.3111) and the mixture was stirred at room temperature for 19 hours. Purification by flash column chromatography on silica gel eluted with a CH2C12:acetone:acetic acid gradient elution gave the product as an oil. Azeotroping with water and acetone followed by trituration with diethyl ether gave the product as a cream solid (44.7mg, 73%) m.p.104-105~C, [Ot]D =
+51.92~ ( c=0.208%w/v in ethanol at 25~C) lH NMR ~ (CDC13) 1.25-1.6 (8H, m, 4xCH2,), 2.56 (2H, t, J = 7.6Hz, C_2Ph), 2.71, 2.74 (lH, dd, J=2,15Hz, H3), 3.12, 3.15 (lH, dd, J=S.2, lSHz, H3), 3.38-3.72 (6H, m, NC~2CH2, OCH2), 4.09 (2H, m, SOCH2), 4.60 (lH, m, ~4), 7.09 - 3~ -wo 97/41098 P~ 97101898 (2H, d, J=8.8Hz, p-ClPh-~l), 7.22 (2H, d, J=8.8Hz, p-ClPh-E:I), 7.40 (2H, d, J=8Hz, Ph-H), 8.08 (2H, d, J=8Hz, Ph-~) Example 27 (a-S, 4R, S-S)-N-[6-(4-lluorophenyl)hexyl]-2-[(4-allylo~ bo~ lphinyl)]-2-oYo~7,~t-din-l-ylpropionamide a R-Methyl-4~(4allyloA~ rbonyl)benzylthio]-2-o~7~ ;n-1-~ e~s~
A ~ pen.~ion of R~[(4-allylo~yc&ll,onyl)benzylthio~-2-oxo~tiAin-1-ylacetic acid (~rom F-~rnrlP lf, 21.55g) and anhydrous pot~c~ium carbonate (8.88g)in 1-methyl-2-pyrrnli~linonP (lOOml) was treated with methyl iodide (10.94g) and the ulG was stirred for 2h. Methyl iodide (l.Og) was added and after 30 nlinutPs the reaction was partitioned between brine (SOOml) and diethyl ether (500ml). The layers were separated and the aqueous layer was washed with diethyl ether (SOOml). The organic extracts were combined washed with water (x2), brine, dried (MgS04) and evaporated to an orange oil. Purification by flash column chromatography on silica gel eluted with [1:1] P.E 40-60~C:ethyl acetate gave the title compound as a yellow oil (contains 2% dimethyl ester) (20.0g, 89%).
lH NMR o (CDC13) 2.94, 3.01 (lH, dd,J= 2.1, 15.2Hz, ~3), 3.25 (lH, d, J=18Hz, 1 of NC~2), 3.39, 3.45 (lH, dd, J=5.1, 15.2Hz, EI3), 3.70 (3H, s, C~3),3.81 (2H, s, SCH2), 4.04 (lH, d, J=18Hz, 1 of NC~2), 4.83 (2H, m, C02C~2), 4.93 tlH, m, 4H), 5.35 (2H, m, CH=CH2), 6.04 (lH, m, CH=CH2), 7.39 (2H, d, J=8Hz, Ph-H), 8.02 (2H, d, J=8Hz, Ph-O
b. a-R,4-R-Methyl 2-{4-[(4-allyloxycarbonyl)benzylthio]-2.oYo~7~ in-l-yl}propionate and a-S,4-R-Methyl 2-{4-[(4-allyloxycarbonyl)ben_ylthiol-2-~,Acaz~tidin-l-yl}propionate A solution of R-methyl-4-[(4-allyloxycarbonyl)benzylthio]-2-oxo~7P~idin-l-yl~cet~te (13.2g) in dry tetrahydrofuran (250ml) at -75~C under nitrogen was treated with a lM solution of lithium bis(trimethylsilyl)amide in THF ~46.3ml~
over 10 minutes keeping the temperature below -70~C. 1,3-Dimethylimi~7olidin-2-one (30.5ml) was added keeping the temperature below -70~C. The resulting suspension was stirred at -75~C for 30 minutes and then treated with methyl iodide (4.3ml) over 1 minute and the temperature rose to -68~C. The reaction was stirred for 1.5 hours at -75~C and then allowed to warm to -20~C over 30 minutes. The reaction was cooled to -75~C and quenchPd with glacial acetic acid (3.5ml), partititioned between water (300ml) and diethyl ether (250ml). The layers were separated and the aqueous layer was washed with diethyl ether (250ml). The organic extracts were combined washed with brine (x3), dried (MgS04), and evaporated to a coloured oil. Ratio of 50% R,R
(diaA): 15% starting material :35% S,R (dia B). Repeat purification by flash column chromatography on silica gel eluted with P.E. 40-60~C:ethyl acetate gave the products as coloured oils.

W O 97141098 PCT~EP97/01898 R,R Diastereoisomer (A), 3.91g (29%) (contains 9% dia B) lH NMR ~ (CDC13) 2.9 (IH, dd, ~I3), 3.30 (lH, ~3), 3.75 (3H, s, C~3),3.88 (2H, s, SC~2), 4.4 (lH, m, C~ .83 (2H, m, C02C~12). 4-90 (lH, m~ 4H)~ 5-35 (2H, m, CH=C~2), 6.04 tlH, m, C~=CH2), 7.39 (2H, d, J=8Hz, Ph-H), 8.02 (2H, d, J=8Hz, Ph-~) S,R Diastereoisomer (B), 5.36g (39%) (contains some sm and 43% dia A) lH NMR o (CDCl3) 2.86, 2.92 (lH, dd, J=2.4, 15.2Hz, ~3), 3.28, 3.33 (lH,dd, J=5.1, 15.2Hz, ~3), 3.73 (3H, s, CH3),3.85 (2H, s, SCH2), 3.95 (lH, m, CH), 4.71 (lH, m, 4H), 4.83 (2H, m, C02CH2), 4.90 (lH, m, 4H), 5.35 (2H, m, CH=C~2), 6.04 (lH, m, CH=CH2), 7.40 (2H, d, J=8Hz, Ph-H), 8.02 (2H, d, J=8Hz, Ph-~) c a-s~4R-2-4{4-[(4-Allyloxycarbonyl)b~ ylll~io]-2-oxo~7eH lin-l-yl}propionic acid A solution of methyl 2-{4-[(4-allyloxycarbonyl)benzylthio]-2-oxoazetidin-1-yl~propionate (2.65g) (mixture of 5% dia A (R,R): 27% dia B (S,R): 65 %
des<xMe) in THF (SOml) at 3~C was treated with a lN sodium hydroxide solution (7.5ml) over 60 minut~c- The cooling bath was removed and the reaction was stirred for 30 minutes. lN NaOH (l.Oml) was added over 30 minutes and the reaction was then stirred for 30 minutes~ diluted with brine (75ml) and extracted with diethyl ether (75ml). The aqueous layer was acidified with lNHCl and extracted with diethyl ether (2x75ml). The organic extracts were combined washed with brine, dried (MgS04)~ and evaporated to give the title compound as an orange oil (2.5g, 98%).and as a mixture in 5%R,R: 27%R,S: 65% des aMe.
d. a-S,4R-N-[6-(~Fluorophenyl)hexyi]-2-[(4-allyloxycarbonyl)benzylthio]-2-o~ q7e~ in-l-ylpropion~m:-le 6-(4-Fluorophenyl)hexylamine (l.SSg) in dry DMF (50ml) was added to a mixture of 2-[(4-allyloxycarbonyl)benzylthio]-2-oxoazetidin-1-ylpropionic acid (2.70g) (above), l-hydroxybenzotriazole (0.95g), N~Nl-dicyclohexylcarbo~ mide (1.46g) and the mixture was sti~ed at room temperature for 4h. The suspension was diiuted with diethyl ether (lOOml) and filtered to remove urea. The filtratewas washed salurated aq.NaHC03, brine, dried (MgS04), and evaporated to an oil. Purification by flash column chromatography on silica gel eluted with [2: 1]
P.E.40-60~C:ethyl acetate gave the product a-S, 4-R diastereoisomer (B)(contains10% dia A) as a yellow oil (0.497g,13.4%).
lH NMR ~ (CDCl3) 1.25-1.7 (1 lH, m, 4xCH2, CH3),2.55 (2H, t, J=7.6Hz, C~12Ph), 2.83, 2.89 (lH, dd, H3), 3.25 (3H, m, NCH2, H3), 3.86 (2H, s, SCH2), 4.10 (lH, m, CH), 4.69 (lH, m, 4H), 4.83 (2H, m, C02C~I2), 5-4 (2H, m, CH=C~2), 6.04 (lH, m, C~=CH2), 6.45 (lH, m, NH), 6.94 (2H, m, p-FPh-O, 7.09 (2H, m, p-FPh-H), 7.39 (2H, d, J=8Hz, Ph-H), 8.028 (2H, m, Ph-H) wo 97/41098 PCT/EPg7/01898 Dia~.eoi~omer A was also isolated lH NMR ~ (cncl3) 1.25- 1.7 (1 lH, m, 4xCH2, CH3),2.55 (2H, t, J=7.6Hz, C~I2Ph), 2.78, 2.85 (lH, dd, J=2.3, 15.4Hz, ~3), 3.25 (3H, m, NC~I2, ~I3), 3.89 (2H,m, SC~2), 4.05 (lH, m, CH), 4.81 (3H, 4, C02CH2), 5.4 (2H, m, CH~ 2), 6.04 (lH, m, C~I=CH2), 6.48 (lH, m, NH), 6.94 (2H, m, p-FPh-a), 7.09 (2H, m, p-FPh-~D, 7.39 (2H, d, J=8Hz, Ph-H), 8.02 (2H, d, Ph-~) e. a-S,4-R,S-S-N-[6-(4Fluorophenyl)hexyl]-2-[(4-~llyl~ Lonyl)b~ l)]-2-oxoazetidin-l-ylpr A solution of S,R-N-[6-(4-fluorophenyl)hexyl]-2-[(4-allyloxycarbonyl)benzylthio~-2-oxo~7~ti-lin-l-ylpropionarnide (1.20g)(80:20 DiaB:A) in dichlorome~h~n~- (25ml), cooled to -75~C, was treated with a solutionof mcpba (0.71g) in CH2C12 (25ml) over 1 hour keeping the temperature at -75~C. The cooling bath was removed and the reaction was stirred for 2 hours, diluted with CH~C12 (25ml), washed with 10%aq. sodium sulphite (SOml), sat.NaHC03 (SOml),dried (MgS04) and evaporated to a coloured oil.
Purification by fla~sh column chromatography on silica gel eluted with ethyl acetate gave the title compound as a 60:40 mixture of dia B2 (a-S, 4-R, S-S):diaBl(a-S,4-R,S-R) Pllrific~tion on Kromasil S~lm silica (250mmx4.6mm) eluted with 50% hexane:
40% ethanol: 10% CHC13 gave the title compound Diastereoisomer B2: (oc-S, 4-R, S-S)-N-[6-(4-fluorophenyl)hexyl~-2-[(4-allyloxycarbonyl)benzylsulphinyl)]-2-oxo~7~tidin-1-ylpropionarnide as a colourless oil.
lH NMR S (CDC13) 1.25-1.7 (llH, m, 4xC~2, CH3).2.55 (2H, t, J=7.6Hz, C~2Ph), 2.83, 2.89 (lH, dd,J= 2, 15Hz, H3), 3.24 (3H, m, NCH2, H3), 4.02 (2H, m, SOC~2), 4~ 44 (lH, m, CH), 4.60 (lH, m, 4H), 4.83 (2H, m, C02CH2), 5.4 (2H, m, CH=C~2), 6.04 (lH, m, C~=CH2), 6.85 (lH, m, NH), 6.94 (2H, m, p-FPh-~), 7.09 (2H, m, p-FPh-O, 7.37 (2H, d, J=8H~, Ph-H), 8.08 (2H, d, J=8Hz, Ph-ED

Example 28 (~-S, 4R, S-S)-N-[6-(4-Fluorophenyl)hexyl]-2-r(4-l~o~ ~ylsulphinyl)-2-oxo~7eti~lin- l-ylpropior~mi~e A solution of (a-S, 4-R, S-S)-N-[6-(4-fluorophenyl)hexyl]-2-[(4-allyloxycarbonyl)benzylsulphinyl]-2-oxo~7eti~in- 1-ylpropionamide (dia B2) (240mg),triphenylphosphine (6mg), tetrakis(triphenylphosphine)palladium (O) (lSmg) in dry CH2C12 (5ml) under nitrogen was treated with pyrrolidine (39~
and the mixture was stirred at room temperature for 19 hours. The mixture was diluted with CH2C12 (50ml) and water (25ml) and acidified with 2NHCl. The layers were separated and the aqueous was washed with CH2C12 (2x50ml). ~he organic extracts were combined, dried (MgS04) and evaporated to a yellow gum.
pllrifir~tion by flash column chromatography on silica gel eluted with a W O 97/41098 PCT~EP97/01898 CH2C12:acetone:acedc acid gradient eludon gave the product as an oil. The oil was dissolved in CH2C12 (4ml) and diluted with diethyl ether (75ml), washed with water, brine, dried (MgS04), filtered and evaporated to a brown foam (123mg, 56%) [a]= No signifi~nt opdcal rotation (c=1.1%w/v in CHCl3) lH NMR ~ (CDC13) 1.32-1.6 (13H, m, CH3, 4xCH2), 2.55 (2H, t, J=7.6Hz, C~I2Ph), 2.84, 2.88 (lH, dd, J=2.4, 15.2Hz, ~3), 3.1-3.3 (3H, m, NHC~2, ~3), 4.(~4, 4.10 (2H, 2xd, J=13Hz, SOCH2), 4.4 (lH, q, CHCH3), 4.68 (lH, m, ~4), 6.94 (3H, m, N~, p-F-Ph-~D, 7.10 (2H, m, p-F-Ph-~), 7.39 (2H, m, Ar-O, 8.06 (2H, m, Ar-~l) Example 29 (a-R, 4R, S-R)- and (o~-R, 4-R, S-S)-N-r6-(4-Fluc ophenyl)hexyl]-2-[(4-allyloxycarbonyl)benzylsulp}linyl)~-2oY-7.7Pt.l;r.-1 -ylpropionamide A solutdon of (o~-R, 4-R)-N-[6-(4-fluorophenyl)hexyl]-2-[(4-allyloxycarbonyl)benzylthio]-2-oxoazeddin- l-ylpropionamide (1.25g)(diastereoisomer A) in dichloromethane (25ml), cooled to -75~C, was treated with a soludon of mcpba (0.75g) in CH2C12 (25ml) over 1 hour keeping the temperature at -75~C. The cooling bath was removed and the reacdon was stined for 2 hours, diluted with CH2C12 (SOml), washed with 10%aq. sodium sulphite (SOml), sat.NaHC03 (50ml), water, dried (MgS04) and evaporated to a coloured oil. The oil was dissolved in ethyl acetate (7.5ml) and cooled. The res-~lting solid was collected, washed with diethyl ether and dried to give (o~-R, 4-R, S-R)-N-[6-(4-fluorophenyl)hexyl]-2-[(4-allyloxycarbonyl)benzylsulphinyl)]-2-oxoazetidin-l-ylpropionamide (dia A1) as a colourless solid 0.125g,9.7%. The filtrate was purified by flash column chromatography on silica gel eluted with ethyl acetate: P.E. 40-60~C. Pure R,R,R fractions were combined and recrystallised from ethyl acetate/diethyl ether to give R,R,R-N-[6-(4-fluorophenyl)hexylJ-2-[(4-allyloxycarbonyl)benzylsulphinyl)]-2-oxoazetidin- 1-ylpropion~mide as a colourless solid (0.19Sg, 15%), m.p.139-140~C:
lH NMR ~ (CDCl3) 1.25-1.7 (1 lH, m, 4xCH2, CH3),2.56 (2H, t, J=7.6Hz, C~2Ph), 2.79, 2.83 (lH, dd,J= 5, l5Hz, H3), 3.20 (2H, m, NcEl2)~ 3-33, 3-39 (lH, dd,J= 2, l5Hz, H3), 3.94 (2H, m, SOC~2), 4.16 (lH, m, CH), 4.77 (lH, m, 4H), 4.83 (2H, m, C02C~2), 5.4 (2H, m, CH=C~2), 6.04 (lH, m, C_=CH2), 6.72 (lH, m, NH), 6.94 (2H, m, p-FPh-O, 7.09 (2H, m, p-FPh-H), 7.37 (2H, d, J=8Hz, Ph-H), 8.08 (2H, d, J=8Hz, Ph-~) Impure R,R,S fractions were combined and recrystallised from ethyl acetate/diethyl ether to give (oc-R, 4-R, S-S)-N-[6-(4-fluorophenyl)hexyl]-2-[(4-W O 97/4109B PCT~EP97/01898 allylo~cycarbonyl)benzylculphinyl)]-2-oxo~7~ti~in-l-ylpropio~mide as a colourless solid (0.204g,15.8%), m.p. 102~C:
lH NMR ~ (CDC13) 1.25-1.7 (1 lH, m, 4XCE~2~ CH3),2.56 (2H, t, J=7.6Hz, C~2Ph), 2.83 (lH, dd, .~13), 3.20 (3H, m, NC~2, ~3), 4.10 (3H, m, SOC~2, CH), 4.65 (lH, m, 4H), 4.83 (2H, m, CO2C~2), 5.4 (2H, m, CH-C~2), 6.04 (lH, m, C~l=CH2), 6.94 (2H, m, p-FPh-~), 7.09 (2H, m,, NH, p-FPh-EI), 7-37 (2H, d, J=8Hz, Ph-H), 8.08 (2H, d, J=8Hz, Ph-O

In an analagous manner the following chloro compounds were prepared from r~cemic 4-[(4-allyloxycarbonyl)benzylthio]-2-oxoazetidin-1-ylacetic acid.

Example 30 (-)-(a-S, 4-R, S-S)- and (+)-(a-R, 4S, S-R)-N-[6-(4 Chlorophenyl)hexyl]-2-[(4-allylo~ ,1,onyl)b~ ul~hinyl]-2-oxoazetidin-l-ylpropionamide a N-[6-(4-Chlorophenyl)hexyl]-Z-[(4-allyloxycarbonyl)benzylthio]-2-~!YO~ --l-ylpropionamide A mixture of methyl-2-( (4-allyloxycarbonyl)benzylthio]-2-oxo~7e~i~in- 1-yl3propionate(4.87g), hydroxybenzotriazole (1.88g), DCC (2.89g) in dry DMF
(60ml) was treated with 6-(4-chlorophenyl)hexylamine and stirred at room temperature for 6 days. The orange suspension was diluted with diethyl ether (200ml) and filtered to remove plecipiLate. Filtrate was washed with dil NaHC03 (200ml), brine, dried (MgSO4), and evaporated to an orange oil. Repeat flash column chromatography on silica gel using P.E 40-60~C:ethyl acetate gave:
Diastereoisomer A (R,RJS,S) (0.8 lg,l 1%) lH NMR ~ (CDC13) 1.2~-1.7 (1 lH, m, 4xC~I2, CH3),2.55 (2H, t, J=7.6Hz, C~2Ph), 2.85 (lH, dd, ~3), 3.25 (3H, m, NCH2, H3), 3.89 (2H,m, SCH2), 4.05 (lH, m, CH), 4.81 (3H, 4, CO2CH2), 5.4 (2H, m, CH=CH2), 6.04 (lH, m, C~I=CH2), 6.48 (lH, m, NH), 7.07 (2H, d, J=8.4Hz, p-ClPh-H), 7.22 (2H, d, J=8.8Hz, p-ClPh-~O, 7.39 (2H, d, J=8Hz, Ph-H), 8.08 (2H, d, Ph-H) Diasteroisomer B (R,S/S,R) (1.63g, 22%) lH NMR o (C~C13) 1.25-1.7 (1 lH, m, 4xC~2, CH3),2.55 (2H, t, J=7.6Hz, C~12Ph), 2.83, 2.89 (lH, dd, J= 2.3, 15.3Hz, ~3), 3.25 (3H, m, NC~12, ~3), 3.86 (2H, s, SCH2), 4.10 (lH, m, CH), 4.69 (lH, m, 4H), 4.83 (2H, m, CO2CH2), 5.4 (2H, m, CH=CEI2), 6.04 (lH, m, C~=CH2), 6.43 (lH, m, NH), 7.07 (2H, d, J=8.4Hz, p-ClPh-~[), 7.22 (2H, d, J=8.8Hz, p-ClPh-H), 7.39 (2H, d, J=8Hz, Ph-H), 8.08 (2H, m, Ph-~D
b. N-[6-(4-Chlorophenyl)hexyl]-2-[(4-allyloxycarbonyl)be..~ lphinyl3-2-oYn ~7~h~1;r l-ylpropion~m;~le A solution of (R,S/S,R)-N-[6-(4-chlorophenyl)hexyl]-2-((4-allyloxycarbonyl)-4-benzylthio)-2-oxoazetidin- 1 -ylpropionamide (1.60g)(diastereoisomer B) in W O 97/41098 rCT~EP97/01898 dichloromethane (30ml), cooled to -75~C, was treated with a solution of mcpba (0.92g) in CH2C12 (30ml) over 45 minutes keeping the temperature below -70~C.
The cooling bath was removed ar~d the reaction was stirred for 45 minut~
diluted with CH2C12 (20ml), washed with 10%aq. sodium sulphite (50ml), sat.NaHCO3 (50ml), dried (MgSO4) and evaporated to give the title compound as a mixture of stereoisomers as a coloured oil.(1.6g, 100%). (65%:35% sulfoxidediastereoisomer B2:Bl) This m~t~ri~l was separated by hplc: Sulfoxide dia B2 was separated from dia Bl using Beckman silica l5cm x 4.6mm eluted with 10:90 eth~nol heY~n~ and subsequent enantiomer separation of diasteroisomer B2 used Chiracel OD-4.6mm eluted with 25:75 ethanol:heY~ne ~ (-)-(a-S, 4R, S-S)-N-[6-(4-Chlorophen~l)hexyl]-2-l(4 allylo~ca~l,onyl)~l-L~lsulphinyl]-2-oxoazetidin-1-ylpropion~mi~le ((-)B2) Fn~ntiomerically pure, colourless oil, 148mg, [a]D= -4.2~ (c=0.4%w/v in CHC13) lH NMR o (CDC13) 1.25- 1.7 (11 H, m, 4xC~2, CH3),2.55 (2H, t, J=7.6Hz, C~Ph), 2.83, 2.89 (lH, dd,J= 2.4, 15.3Hz, ~3), 3.24 (3H, m, NCH2, ~3), 4.08 (2H, m), 4.60 (lH, m, CH), 4.69 (lH, m, 4H), 4.83 (2H, m, C02C.~2), 5.4 (2H, m, CH=C~2), 6.04 (lH, m, C_=CH2), 6.95 (lH, m, NH), 7.08 (2H, d, J=8.4Hz, p-ClPh-O, 7.22 (2H, d, J=8.8Hz, p-ClPh-~), 7.39 (2H, d, J=8Hz, Ph-H), 8.08 (2H, d, J=8Hz, Ph-H) d. (+)-(a-R, 4S, S~R)--N-[6-(4-Chlorophenyl)hexyl]-2-[(4-all,~Jc~c~bonyl)benzylsulphinyl~-2-oxo~7eti~1in-l.ylpropion~mi~e ((+~B2) Sarnple contains 0.83% of the other enantiomer, colourless oil,145mg, [a]D=
~4.3~ (c=0.4%w/v in CHC13) lH NMR ~ (CDCl3) 1.25-1.7 (1 lH, m, 4xCH2, CH3),2.55 (2H, t, J=7.6Hz, CH2Ph), 2.83, 2.89 (lH, dd,J= 2.4, 15.3Hz, H3), 3.24 (3H, m, NCH2, H3), 4.08 (2H, m), 4.60 (lH, m, CH), 4.69 (lH, m, 4H), 4.83 (2H, m, CO2CH2), 5.4 (2H, m, CH~C~2), 6.04 (1~, m, CH=CH2), 6.95 (lH, m, NH), 7.08 (2H, d, J=8.4Hz, p-ClPh-O, 7.22 (2H, d, J=8.8H~, p-ClPh-O, 7.39 (2H, d, J=8Hz, Ph-H), 8.08 (2H, d, J=8Hz, Ph-H) Example 31 (-)-(a-S, 4R~ S-S)-~6-(4-Chlorophenyl)hexyl]-2-(4-carboxy~ ulphinyl)-2-o~o~eff~ -ylpropion~mi~le A solution of (-)-(a-S, 4-R, S-S)-N-[6-(4-chlorophenyl)hexyl]-2-~(4-allyloxycarbonyl)benzylsulphinyl]-2-oxoazetidin-1-ylpropionamide (125mg), triphenylphosphine (6mg), tetrakis(triphenylphosphine)palladium (0) (8mg) in dry CH2C12 (2ml) under nitrogen was treated with pyrrolidine (19.5111) and the mixture was stirred at room temperature for 19 hours. Purification by flash column chromatography eluted with a CH2Cl2:acetone:acetic acid gradient elution gave the product as an oil. The oil was dissolved in CH2Cl2 (2ml) and diluted with diethyl ether (SOml), washed with, brine (x2), dried (MgSO4), ... .

W O 97/41098 PCT~EP97/01898 filtered and evaporated to givee the title compound as a yellow foam (104mg, 87%) [a]D= -3.7~ (c=0.5%w/v in CHCl3) lH NMR ~ (CDCl3) 1.25-1.6 (8H, m, 4xC~2,), 1.62 (3H, d, J=7.2Hz, CH3),2.55 (2H, t. J=7.6Hz, C~12Ph), 2.86, 2.89 (lH, dd, J=2.4, 15.2Hz,~3), 3.20 (3H, m, NC~2, ~3), 4.06 (2H, m, SOCH2), 4.45 (lH, m, CH), 4.66 (lH, m, 4H), 6.95 (lH, m, NH), 7.08 (2H, d, J=8.4Hz, p-ClPh-~), 7.22 (2H, d, J=8.8Hz, p-ClPh-~), 7.39 (2H, d, J=8Hz, Ph-H), 8.08 (2H, d, J=8Hz, Ph-O
Example 32 (+)-(a-R, 4-S, S-R)-N-[6.(4.Chlorophenyl)hexyl]-2-(4.
carboxybL~ Iph;nyl)-2.0xo~7eti~1ir l-ylpropi~n~m;~p A solution of (+)-(a-R, 4-S, S-R)-N-[6-(4-chlorophenyl)hexyl]-2-[(4-allyloxycarbonyl)benzylsulphinyl]-2-oxoazetidin-1-ylpropionamide (123mg), triphenylphosphine (6mg), tetrakis(triphenylphosphine)palladium (O) (8mg) in dry CH2C12 (2ml) under nitrogen was treated with pyrrolidine (19.5~11) and the mixture was stirred at room temperature for 19 hours. Purification by flash column chromatography eluted with a CH2Cl2:acetone:acetic acid gradient elution gave the product as an oiL The oil was dissolved in CH2C12 (2ml) and diluted with diethyl ether (SOml), washed with, brine (x2), dried ~MgS04), filtered and evaporated to a yellow foam (lOlmg, B9%) [OC]D= +3.3~ (c=0.3%w/v in CHC13) lH NMR ~ (CDCl3) 1.25-1.6 (8H, m, 4xC~2,), 1.62 (3H, d, J=7.2Hz, CH3),2.55 (2H, t, J=7.6Hz, CH2Ph), 2.88 (lH, dd, ~3), 3.20 (3H, m, NC~2, a3), 4.06 (2H, m,SOC~2),4.45(1H,m,CH),4.69(1H,m,4H),6.95(1H,m,NH),7.08(2H,d, J=8.4Hz, p-ClPh-O, 7.22 (2H, d, J=8.8Hz, p-ClPh-H), 7.39 (2H, d, J=8Hz, Ph-H), 8.08 (2H, d, J=8Hz, Ph-H) The following racemic compounds were also prepared. These may then be separated into enantiomers by hplc, using a chiral stationary phase, in a similar manner to separations hereinbefore described.
Descrip~on 1 (R,S/S,R)-1-(2-(6-Chlorophenyl)hexyloxy)ethyl-4-(4-carboA~ rhi~yl)-2-~xo~7Pt~Aine (diast 2) (R,S/S,R)- 1 -(2-(6-Chlorophenyl)hexyloxy)ethyl-4-(4-allyloxycarbonylbenzylsulphinyl)-2-oxoazetidine (220mg), triphenylphosphine (1 lmg), tetrakis(triphenylphosphine)palladium (O) (15mg) in dry CH2Cl2 (Sml) under nitrogen was treated with pyrrolidine (37~11) and the mixture was stirred at room temperature for 19 hours. Purification by flash column chromatography on silica gel eluted with a CH2Cl2:acetone:acetic acid gradient elution gave the product as an oil. The oil was dissolved in CH2C12 (4ml) and diluted with diethyl wo 97/41098 PCT/EP97/01898 ether (75ml), washed with water, brine, dried (MgS04), filtered and evaporated to an oil which solidif1ed on cooling. Trituration with diethyl ether gave the title compound as a cream solid (149mg, 75%) m.p.107-108~C
lH NMR o (CDC13) 1.25-1.6 (8H, m, 4xC~,), 2.56 (2H, t, J = 7.8Hz, C~2Ph), 2.71, 2.74 (lH, dd, J=2,15.2Hz, ~3), 3.12, 3.15 (lH, dd, J=5.2, 15.2Hz, ~3), 3.38-3.72 (6H, mS NC~I2CH2, 0C~2), 4.10 (2H, m, SOC~2), 4.63 (lH, m, ~4), 7.09 (2H, d, J=8.8Hz, p-ClPh-O, 7.22 (2H, d, J=8.8Hz, p-ClPh-.O, 7.40 (2H, d, J=8Hz, Ph-H), 8.08 (2H, d, J=8Hz, Ph-H) Description 2 (R,S/S,R)-1-(2-(6-Chlorophenyl)hexyloxy)ethyl-4-(4 ethox~,carl,onylbenzy~ rhinyl)-2-oy~7eh lin~ (dfast 2) a 1-(2-(6-Chlorophenyl)hexyloxy)ethyl-4(4-ethox~c~, l,onyl~ io)-2-t;~line A soludon of 6-(~chlorophenyl)hexyloxy triflate (3.62g), 4-(4-ethoxycarbonylben7tlthio)azet din-2-one (2.47g) and tetrabutylammonium bromide (O.~Og) in dry THF (70ml) was cooled to 10~C under nitrogen and was treated with powdered potassium hydroxide (0.62g). The cooling bath was removed and the reaction was stirred for 2 hours . The mixture was partidoned between brine (200ml) and ethyl acetate (200ml) and filtered through celite. Thelayers were separated and the aqueous was washed with ethyl acetate (lOOml).
The organic extracts were combined, dried (MgS04) and evaporated to a dark oil. Pllrific~tion by flash column chromatography on silica gel eluted with [2:1]
to [1:1] P.E. 40-60~C: ethyl acetate gave ~he title compound as a yellow oil (2.41g, 51%).
lH NMR ~ (CDCl3) 1.25-1.6 (1 lH, m, CH3, 4xCH2,), 2.55 (2H, t, J =7.6Hz, C~I2Ph), 2.85 (1~, dd, H3), 3.05 (lH, m), 3.25, 3.30 (lH, dd, ~3), 3.36-3.72 (5H, m, NC~2C~I2, oC~I2), 3.85 (2H, s, SCH2), 4.37 (2H, q, J=7.1Hz, C02C~3) 4.76 (lH, m, ~4), 7.07 (2H, d, J=8.4Hz, p-ClPh-.O, 7.22 (2H, m, p-ClPh-EDt 7.39 (2H, d, J=8Hz, Ph-H), 8.08 (2H, d, J=8Hz, Ph-O
b. (R,S/S,R)-1-(2-~6-Chlorophenyl)hexyloxy)ethyl-4-(4 etho~l,onylb~ nyl) 2-olro~7eH~linP (diast 2) A solution of 1-(2-(6-chlorophenyl)hexyloxy)ethyl-4-(4-ethoxycarbonylbenzylthio)-2-oxo~7eti~ine (2.~7g) in dichloromethane (75ml), cooled to -70~C, was treated with a solution of mcpba (l.Og) in CH2C12 (75ml) dropwise over 1 hour m~inl~ining the temperature at below -70~C. The cooling bath was removed and the reaction was stirred for 2 hours, washed with a solution of 10% aq.sodium hydrogen carbonate(50ml) + 10% aq. sodium sulphite (~Oml).
The layers were separated and the organic layer was washed with brine, dried (MgS04) and evaporated to an oil. Recryst~llic~tinn from diethyl ether (40ml) gave a colourless solid (0.9lg). ~lrifi~tion by flach column chromatography -- 4~ -W O971410g8 PCT~EP97/01898 eluted on silica gel with [2: 1] ethyl acetate:hexane and recyst~ ~tion of the less polar product from diethyl ether (lSml) gave diastereoisomer 2 as a colourless solid (0.40g,16%).m.p. 81-82~C -lH NMR ~ (CDC13) 1.25-1.6 (1 lH, m, CH3, 4xCH2,), 2.55 (2H, t, J =7.8Hz, C~2Ph), 2.63, 2.69 (lH, dd, J=2.2, 15.1Hz, H3), 3.05, 3.11 (lH, dd, J=5.1, 15.1Hz,.~3), 3.36-3.74 (6H, m, NC~2C~2, OC~2), 3.75 (2H, s, SOC~2)- 4-39 (2H, q, J=7.1Hz, C02C~3) 4.54 (lH, m, ~4), 7.09 (2H, d, J=8.4Hz, p-ClPh-H), 7.22 (2H, m, p-ClPh-~l), 7.37 (2H, d, J=8.3Hz, Ph-H), 8.06 (2H, m, Ph-O

W O 97/41098 PCT~EP97101898 Biological Data 1. &reen for Lp-PLA2 inhibition.

Enzyme acdvity was det~rmin~d by measuring the rate of turnover of theartificial substrate (A) at 37 ~C in 50mM HEPES (N-2-hydroxyethylpiperazine-N'-2-eth~nesulrhorlic acid) buffer cont~ining 150mM NaCl, pH 7.4.

~ ~ O(CH2)9CH3 N~2~ fl 01 O(CH2)2NMe3 (A) Assays were performed in 96 well dtre plates.

Lp-PLA2 was pardally purified by density gradient centrifugadon of human plasma. Acdve fractions were pooled and used as the source of Lp-PLA2. The enzyme was pre-incubated at ~7 ~C with vehicle or test compound for 10 min in a total volume of 180 ~Ll. The reacdon was then initi~te~ by the addidon of 20 ~ullOx substrate (A) to give a final substrate concentradon of 20 ~lM. The reacdon was followed at 405 nm for 20 minutes usir,g a plate reader with ~uto~ti~
mixing The rate of reaction was measured as the rate of change of absorbance.

Results:

The compounds of Examples 1 and 2, the corresponding carboxylic acid (+/-)-N-t6-(4-chlorophenyl)hex-l-yl]-4-(4-carboxybenzylsulphinyl)-2-oxo~7Pti~in- 1-yl)~cet~mide and the carboxylic acid (4R, SS)-N-[6-(4-fluorophenyl)hex-1-yl]-4-(~carboxybenzylsulphinyl)-2-oxoazetidin-1-yl)acetamide had ICso values in the range 4 to 20 nM. The compounds of Example 12,13 and 14 had ICso values in therange2to4nM.
2. Evaluation of Bioavailability The pro-drug esters were evaluated in dog and human liver microsomes according to standard procedures for their ability to be hydrolysed to the parent acid. The results are given in the table below.

. .

o~ ~0 O
~ ~ (CH,J,~3F

Example Acid production Buffer SGF
pH 75 pH 1.2 R Dog mic~llm~n mic tl/2h tV2h O 'O 9 100% 80% 3 3.5 0~_0 0 100% 75%

iO% 65% 3.5 2.5 o~o~ J 12 100% 60% >4 1.5 ,~OM~
o_,o~J 13 100% 50% 3 7 o~o~ 14 100% 70% 5~7 58 I o~
~'-~~< 15 100% 100% 8-9 25 ~~ 16 100% 45% 1.5 OEt 6 10% <2%

Acid production - % conversion of test ester to parent acid by dog or liver microsomes after incubation of lllm test compound for 15 min, determined by me~cl-ring the concentration of parent acid produced by HPLC detection of acid (100% = l~M acid produced). Figures are rounded to nearest 5%.

3. Evaluationofstability - 4g -W O 97/41098 PCT~EPg7/01898 Stability was estim ltP-d by determining half lives for decomposition test compound in pH 7.5 buffer (50 mM phosphate) and pH 1.2 buffer (USP simulated gastric fluid = NaCVHCl pH 1.2 + pepsin-). Initial concentration = 50 uM and compound monitored by HPLC. Figures rounded to nearest 0.5 h.

Preferred compounds are those exhibiting good conversion of ester to acid in biological systems, while showing good stability in buffers (e.g. examples 9, 11, 12, 15, 16).

Claims (28)

Claims

1. A compound of the formula (I):

in which:
R1 is hydrogen or a corresponding pharmaceutically acceptable ester or a pharmaceutically acceptable salt thereof;
R2 and R3 which may be the same or different is each selected from hydrogen or optionally substituted C(1-6)alkyl;
X is a group X'(CH2)m in which X' is CO, CONR4, COO, CONR4CO, CONHO
or CH2O in which R4 is hydrogen or C(1-6)alkyl and m is 0 or an integer from 1 to 12; or a C(1-12)alkylene chain optionally interupted by X'; and Y is an optionally substituted aryl group;
having the absolute configuration (4R,SS).

2. A compound as claimed in claim 1 in which R2 and R3 is each hydrogen or R2 is hydrogen and R3 is methyl.

3. A compound of formula (I) as claimed in claim 1 or 2 in which X is CO(CH2)m,CONH(CH2)m, COO(CH2)m, CONHCO(CH2)m, CONHO(CH2)m, CH2O(CH2)m. or C(1-12)alkylene.

4. A compound of formula (I) as claimed in any one of claims 1 to 3 in which X
is CONH(CH2)m or CH2O(CH2)m.

5. A compound of formula (I) as claimed in any one of claims 1 to 4 in which X
is CONH(CH2)6.

6. A compound of formula (I) as claimed in any one of claims 1 to 5 in which Y
is phenyl, optionally substituted by up to three further substituents

7. A compound of formula (I) as claimed in any one of claims 1 to 6 in which Y
is phenyl optionally substituted by halo.

8. A compound of formula (I) as claimed in any one of claims 1 to 7 in which X-Y is CONH(CH2)6Ph(4-F)/(4-Cl).

9. A compound of formula (I) as claimed in any one of claims 1 to 8 in which thepharmaceutically acceptable ester is a C(1-6)alkyl or C(2-6)alkenyl ester or a pharmaceutically acceptable in vivo hydrolysable ester.

10. A compound of formula (I) as claimed in 9 in which R1 for use in an in vivo hydrolysable ester is selected from:
-CH(Ra)O.CO.Rb;
-CH(Ra)O.CO.ORc;
-CH(Ra)CO.NReRf -RdNReRf;
-CH2ORg;

-CH(Ra)O.CO.C6H4Y1COCH(Ri)NH2; and in which:
Ra is hydrogen, (C1-6)alkyl, in particular methyl, (C3-7)cycloalkyl, or phenyl, each of which may be optionally substituted;
Rb is (C1-6)alkyl, (C1-6)alkoxy(C1-6)alkyl, phenyl, benzyl, (C3-7)cycloalkyl, (C1-6)alkyl(C3-7)cycloalkyl, 1-amino(C1-6)alkyl, or 1-(C1-6alkyl)amino(C1-6)alkyl, each of which may be optionally substituted; or Ra and Rb together form a 1,2-phenylene group optionally substituted by one or two methoxy groups;
R c is (C1-6)alkyl, (C3-7)cycloalkyl, (C1-6)alkyl(C3-7)cycloalkyl;

R d is (C1-6)alkylene optionally substituted with a methyl or ethyl group;
R e and R f which may be the same or different is each (C1-6)alkyl; or aryl(C1-4) alkyl, optionally substituted with e.g. hydroxy;
R g is (C1-6)alkyl;
R h is hydrogen, (C1-6)alkyl or phenyl;
R i is hydrogen or phenyl optionally substituted by up to three groups selected from halogen, (C1-6)-alkyl, or (C1-6)alkoxy;
and Y1 is oxygen or NH.

11. A compound of formula (I) as claimed in claim 10 in which R1 is:
(a) acyloxyalkyl groups such as acetoxymethyl, isobutyryloxymethyl, pivaloyloxymethyl, benzoyloxymethyl, .alpha.-acetoxyethyl, .alpha.-pivaloyloxyethyl, 1-(cyclohexylcarbonyloxy)ethyl, (1-aminoethyl)carbonyloxymethyl, 2-methoxyprop-2-ylcarbonyloxymethyl, phenylcarbonyloxymethyl and 4-methoxyphenyl-carbonyloxymethyl;
(b) alkoxy/cycloalkoxycarbonyloxyalkyl groups, such as ethoxycarbonyloxymethyl, t-butyloxycarbonyloxymethyl, cyclohexyloxycarbonyloxymethyl, 1-methylcyclohexyloxycarbonyloxymethyl and .alpha.-ethoxycarbonyloxyethyl;
(c) dialkylaminoalkyl, especially di-loweralkylamino alkyl groups such as dimethylaminomethyl, dimethylaminoethyl, diethylaminomethyl or diethylaminoethyl;
(d) acetamido groups such as N,N-dimethylaminocarbonylmethyl, N,N-(2-hydroxyethyl)aminocarbonylmethyl;
(e) lactone groups such as phthalidyl and dimethoxyphthalidyl;
(f) (5-methyl-2-oxo-1,3-dioxolen-4-yl)methyl; and (g) (2-methoxycarbonyl-E-but-2-en-yl)methyl.

12. Acompound of formula (I) as claimed in claim 11 in which R1 is:
(2-methoxycarbonyl-E-but-2-en-yl)methyl, isobutyryloxymethyl, 2-methoxyprop-2-ylcarbonyloxymethyl, phenylcarbonyloxymethyl, 4-methoxyphenyl-carbonyloxymethyl, t-butyloxycarbonyloxymethyl, cyclohexyloxy-carbonyloxymethyl, 1-methylcyclohexyloxycarbonyloxymethyl, N,N-dimethylaminocarbonylmethyl, or (5-methyl-2-oxo-1,3-dioxolen-4-yl)methyl.

13. A compound of formula (I) selected from:
(4R, SS)-N-[6-(4-Fluorophenyl)hex-1-yl]-4-(4-carboxybenzylsulphinyl)-2-oxoazetidin-1-yl)acetamide (and pharmaceuticaly acceptable salts thereof, in particular the sodium salt);
(4R, SS)-N-[6-(4-Fluorophenyl)hex-1-yl]-4-(4-alloxycarbonylbenzylsulphinyl)-2-oxoazetidin-1-yl)acetamide;
(4R, SS)-N-(6-(4-Fluorophenyl)hexyl)-4-(4-ethoxycarbonylbenzylsulphinyl)-2-oxoazetidin-1-ylacetamide;

(4R, SS)-N-(6-(4-Chlorophenyl)hexyl)-4-(4-ethoxycarbonylbenzylsulphinyl)-2-oxoazetidin-1-ylacetamide;

(-)-1-(2-(6-Fluorophenyl)hexyloxy)ethyl-4-(4-carboxybenzylsulphinyl)-2-oxoazetidine (diast. 2);
(+)-1-(2-(6-Fluorophenyl)hexyloxy)ethyl-4-(4-carboxybenzylsulphinyl)-2-oxoazetidine (diast. 2);
(+)-1-(2-(6-Fluorophenyl)hexyloxy)ethyl-4-(4-allyloxycarbonylbenzylsulphinyl)-2-oxoazetidine (diast. 2);
(-)-1-(2-(6-Fluorophenyl)hexyloxy)ethyl-4-(4-allyloxycarbonylbenzylsulphinyl)-2-oxoazetidine (diast. 2);

(-)-1-(2-(6-Chlorophenyl)hexyloxy)ethyl-4-(4-carboxybenzylsulphinyl)-2-oxoazetidine (diast. 2);
(+)-1-(2-(6-Chlorophenyl)hexyloxy)ethyl-4-(4-carboxybenzylsulphinyl)-2-oxoazetidine (diast. 2);
(+)-1-(2-(6-Chlorophenyl)hexyloxy)ethyl-4-(4-allyloxycarbonylbenzylsulphinyl)-2-oxoazetidine;
(-)-1-(2-(6-Chlorophenyl)hexyloxy)ethyl-4-(4-allyloxycarbonylbenzylsulphinyl)-2-oxoazetidine;

(.alpha.-S, 4-R, S-S)-N-[6-(4-fluorophenyl)hexyl]-2-[(4-allyloxycarbonyl)benzylsulphinyl)]-2-oxoazetidin-1-ylpropionamide;
(.alpha.-S, 4-R, S-S)-N-[6-(4-Fluorophenyl)hexyl]-2-[(4-carboxybenzylsulphinyl)-2-oxoazetidin-1-ylpropionamide;

(.alpha.-R, 4-R, S-R)--N-[6-(4-Fluorophenyl)hexyl]-2-[(4-allyloxycarbonyl)benzylsulphinyl)]-2-oxoazetidin-1-ylpropionamide;
(.alpha.-R, 4-R, S-S)-N-[6-(4-Fluorophenyl)hexyl]-2-[(4-allyloxycarbonyl)benzylsulphinyl)]-2-oxoazetidin-1-ylpropionamide;
(-)-(.alpha.-S, 4-R, S-S)-N-[6-(4-Chlorophenyl)hexyl]-2-[(4-allyloxycarbonyl)benzylsulphinyl]-2-oxoazetidin-1-ylpropionamide ((-)B2);
(-)-(.alpha.-S, 4-R, S-S)-[6-(4-Chlorophenyl)hexyl]-2-(4-carboxybenzylsulphinyl)-2-oxoazetidin-1-ylpropionamide;

(+)-(.alpha.-R, 4-S, S-R)-N-[6-(4-Chlorophenyl)hexyl]-2-[(4-allyloxycarbonyl)benzylsulphinyl]-2-oxoazetidin-1-ylpropionamide ((+)B2); and (+)-(.alpha.-R, 4-S, S-R)-N-[6-(4-Chlorophenyl)hexyl]-2-(4-carboxybenzylsulphinyl)-2-oxoazetidin-1-ylpropionamide.

14. A compound of formula (I) selected from:
(4R, SS)-N-(6-[4-Fluorophenyl]hex-1-yl)-4-(4-(cyclohexyloxycarbonyloxymethyloxycarbonyl)benzylsulphinyl)-2-oxoazetidin-1-yl)acetamide;
(4R, SS)-N-(6-[4-Fluorophenyl]hex-1-yl)-4-(4-(tert-butyloxycarbonyloxymethyloxycarbonyl)benzylsulphinyl)-2-oxoazetidin-1-yl)acetamide;
(4R, SS)-N-(6-[4-Fluorophenyl]hex-1-yl)-4-(4-(1-methylcyclohexyloxycarbonyloxymethyloxycarbonyl)benzylsulphinyl)-2-oxoazetidin-1-yl)acetamide;
(4R, SS)-N-(6-[4-Fluorophenyl]hex-1-yl)-4-(4-(phenylcarbonyloxymethyloxycarbonyl)benzylsulphinyl)-2-oxoazetidin-1-yl)acetamide;
(4R, SS)-N-(6-[4-Fluorophenyl]hex-1-yl)-4-(4-(4-methoxyphenylcarbonyloxymethyloxycarbonyl)benzylsulphinyl)-2-oxoazetidin-1-yl)acetamide;
(4R, SS)-N-(6-[4-Fluorophenyl]hex-1-yl)-4-(4-(iso-butyryloxymethyloxycarbonyl)benzylsulphinyl)-2-oxoazetidin-1-yl)acetamide;
(4R, SS)-N-(6-[4-Fluorophenyl]hex-1-yl)-4-(4-(2-methoxyprop-2-ylcarbonyloxymethyloxycarbonyl)benzylsulphinyl)-2-oxoazetidin-1-yl)acetamide;
(4R, SS)-N-(6-[4-Fluorophenyl]hex-1-yl)-4-(4-((5-methyl-2-oxo-1,3-dioxolen-4-yl)methyloxycarbonyl)benzylsulphinyl)-2-oxoazetidin-1-yl)acetamide;

(4R, SS)-N-(6-[4-Fluorophenyl]hex-1-yl)-4-(4-((2-methoxycarbonyl-E-but-2-en-yl)methyloxycarbonyl)benzylsulphinyl)-2-oxoazetidin-1-yl)acetamide;
(4R, SS)-N-(6-[4-Fluorophenyl]hex-1-yl)-4-(4-(N-N-dimethylaminocarbonylmethyloxycarbonyl)benzylsulphinyl)-2-oxoazetidin-1-yl)acetamide: and (4R, SS)-N-(6-[4-Fluorophenyl]hex-1-yl)-4-(4-(N-N-di-(2-hydroxyethyl)aminocarbonylmethyloxycarbonyl)benzylsulphinyl)-2-oxoazetidin-1-yl)acetamide;

15. A pharmaceutical composition comprising a compound of formula (I) as defined in any of the preceding claims and a pharmaceutically acceptable carrier.

16. A compound of formula (I) as defined in claim 1 for use in therapy.

17. The use of a compound of formula (I) as defined in claim 1 in the manufacture of a medicament for treating atheroscelrosis.

18. The use of a compound of formula (I) as defined in claim 1 in the manufacture of a medicament for treating diabetes, hypertension, angina pectoris, after ischaemia, reperfusion, rheumatoid arthritis, stroke, myocardial infarction, reperfusion injury, sepsis, and acute and chronic inflammation, inflammatory conditions of the brain such as Alzheimer's Disease, neuropsychiatric disorders such as schizophrenia, and psoriasis.

19. A method of treating a disease state associated with activity of the enzyme Lp-PLA2 which method involves treating a patient in need thereof with a therapeutically effective amount of an inhibitor of the enzyme.

20. A method as claimed in claim 19 in which the disease state is associated with the increased involvement of monocytes, macrophages or lymphocytes.

21. A method as claimed in claim 19 in which the disease state is associated with the formation of lysophosphatidylcholine and oxidised free fatty acids.

22. A method as claimed in claim 19 in which the disease state is associated with lipid peroxidation in conjunction with Lp PLA2 activity.

23. A method as claimed in claim 19 in which the disease state is associated with endothelial dysfunction.

24. A process for preparing a compound of formula (I) as defined in claim 1 and in which X is an amide CONH which comprises treating a compound of formula (II):

in which R1 is C(1-6)alkyl or C(2-6)alkenyl and R2 and R3 are as defined in claim 1, and having the absolute configuradon (4R,SS);
with an amine of the formula (III):

H2N(CH2)nY
under suitable amide forming conditions, for instance in the presence of an activating agent such as N,N-dicyclohexylcarbodiimide and 1-hydroxybenzotriazole in a suitable solvent such as dry dimethylformamide;
and thereafter, and if necessary, (a) removing the ester group under suitable de-esterifying conditions to form the acid;
(b) converting the acid to a pharmaceutically acceptable salt; and/or (c) converting the acid, a suitable salt, the ester or an activated derivative of the acid, to an in vivo hydrolysable ester by reaction with a compound of formula (IV):

in which:
R4 is a reactive esterifying leaving group; and R1 is as hereinbefore defined;
under ester forming conditions.

25. A compound of formula (V):

in which R1, R2 and R3 are as defined in claim 1, and having the absolute configuration (4R);
having the absolute configuration (4R).

26. A diastereoisomeric salt formed from a compound of formula (VI):

in which R* is a carboxy protecting group, for instance C(1-6)alkyl or C(2-6)alkenyl and R2 and R3 are as defined in claim 1; and a chiral base.

27. A process for resolving an intermediate compound of formula (VI):

in which R* is carboxy protecting group, for instance a C(1-6)alkyl or C(2-6)alkenyl and R2 and R3 are as defined in claim 1;

- which process comprises the formation of a diastereoisomeric salt with a chiral base such as (-)-cinchonidine.

28. A process for preparing a compound of formula (I) as defined in claim 1 and in which X is an amide CONH which comprises the process defined in claim 27.