Title of Invention	FURANONE DERIVATIVES AND METHODS OF MAKING SAME
Abstract	Novel synthesis methods, to the products of such novel methods, and to uses of these products. In particular, the present invention provides methods for the reactions of furanones, in particular fimbrolides, with amines. The invention has particular application in the synthesis of halogenated l,5-dthydro-pyrrol-2-one, 5-halomethylene substituted l,5-dihydropyrrol-2-ones (laetam analogues of fimbrolides), 5-amino substituted furanones and 5-ammomethylene-2(5H)-furanones and their synthetic analogues. The invention also relates to novel compounds and uses thereof.

Title of Invention

FURANONE DERIVATIVES AND METHODS OF MAKING SAME

Abstract

Novel synthesis methods, to the products of such novel methods, and to uses of these products. In particular, the present invention provides methods for the reactions of furanones, in particular fimbrolides, with amines. The invention has particular application in the synthesis of halogenated l,5-dthydro-pyrrol-2-one, 5-halomethylene substituted l,5-dihydropyrrol-2-ones (laetam analogues of fimbrolides), 5-amino substituted furanones and 5-ammomethylene-2(5H)-furanones and their synthetic analogues. The invention also relates to novel compounds and uses thereof.

Full Text	"Furanone derivatives and methods of making same" Technical Field The present invention relates to novel synthesis methods, to the products of such novel methods, and to uses of these products. In particular, the present invention provides methods for the reactions of furanones, tn particular fimbralides, with amines. The invention has particular application in the synthesis of halogenated 1,5-dihydro-pynrol-2-one, 5-halomethylene substituted 1,5-dihydropyrrol-2-ones (lactam analogues of fimbrolides) 5-amino substituted furanones and 5-aminomethylene-2(5H)-furanones and their synthetic analogues. The invention also relates to novel compounds and uses thereof. Background Fimbrolides (halogenated 5-methylene-2(5H)-furanones) possess a wide range of important biological properties including antifungal and antimicrobial properties (see WO 96/29392 and WO 99/53916, the disclosures of which are incorporated herein by cross-reference) These metabolites can be isolated from red marine algae Delisea fimbnata, Delisea elegans and Del/sea puichra. Despite their biological activity very few hetero atom containing analogues of these molecules have been reported in the literature. The majority of the published syntheses of fimbrolides focus on the preparation of naturally occurring fimbrolides themselves. Recently we have developed methods that yield both the natural and non-natural fimbrolides in good yields (see WO 99/54323 and WO 0200639 the disclosure of which is incorporated herein by cross-reference). We have now found that, surprisingly, fimbrolides undergo reactions with amines under mild conditions. We have found this discovery to be particularly useful in the synthesis of 5-hydroxy-5-aIkyl substituted 1,5-dihydro-pyrro!-2-one, 5-amino-5-alkyI substituted 2(5H)-furanones and 5-aminomethyIene substituted 2(5H)-furanones. Furthermore 5-hydroxy-5-halomethyl substituted 1,5-dihydro-pyiTol-2-one generated under these conditions can be dehydrated to yield 5-halomethylene substituted 1,5-dihydropyrrol-2-ones (lactam analogues of fimbrolides), and the 5-amino-5-bromomethyl substituted 2(5H)-furanones can be dehydrobrommated to yield a range of 5-aminomethylene W0 2004/016588 PCT/AU2003/001053 2 Substituted 2(5H)-furanones. These furanones can be further functionahsed to yield a range of novel analogues. Summary of the Invention In a first aspect, the present invention provides a method for the preparation of compound of formula 11 wherein R1 and R2 are independently selected from the group H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophilic; R3 and R4 are independently selected from the group H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl; Rs is selected from the group consisting of H, hydroxy, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophilic, or forms part of an amino acid, or is a nudeoside, an oligomer, a polymer, a dendnmer, a substrate or a surface; the method comprising reacting a compound of formula I WO 2004/016588 PCT/AU2003/001053 wherein R1 and R2 are independently H, halogen, alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyt, substituted or unsubstituted aikenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophilic; R3 and R4 are independently H, halogen, alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl or arylalkyl; and R is hydroxy, halogen; and "--- " represents a single bond in which case R is absent, or a double bond, provided that at least one of R1, R2, R3 and R4 is halogen, with a compound of formula R5NH2 wherein R5 is selected from the group consisting of H, substituted or unsubstituted alkyl, hydroxy, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophllic, or forms part of an amino acid, or is a nucleoside, an oiigomer, a polymer, a dendnmer, a substrate or a surface. The reaction may optionally be carried out in the presence of solvent. Preferably, in the compound of formula II, at feast one of R1, R2, R3 and R4 is halogen. In the structural formulae descnbed herein, a particular geometry is not to be taken as specified. For example, the formulae covers both 2- and E-isomers. The reaction may be performed In the presence or absence of a solvent. The solvent may be any suitable solvent. Preferable solvents in the present invention include alkyl acetates, aromatic hydrocarbons, chionnated alkanes, cyclic or open chain ethers such as tetrahydrofuran, diethyl ether, dioxane, and WO 2004/016588 PCT/AU2003/001053 4 C1-C3 acids. More preferably, the solvents are aromatic hydrocarbons and chlorinated alkanes. Most preferably, the solvent is dichloromethane, as well as dtchloroethane and trichloroethane. The reaction Is preferably earned out at mild temperatures. Preferably the cyclisation reaction is performed at a temperature in the range of 20-150°C Where a solvent is present, the cyclisation may be performed at reflux temperature, for example, at the reflux temperature of dichloromethane. Optionally the reaction may be carried out below reflux temperature under pressure. The reaction time may vary from about 2 hours to 12 hours or more and is typically about 2 hours or more, It will be appreciated that reaction conditions may be varied depending upon the individual nature of the substrate and the desired rate of the reaction. Non-limiting examples of compounds of formula II, which may be described as 5-aikyl-5-hydroxy substituted 1,5-dihydro-pyrrol-2-one5, that can be synthesised by the method of the invention Include: WO 2004/016588 PCT/AU2003/001053 5 In a second aspect, the present invention provides a compound of formula II. wherein R1 and R2 are independently H, halogen, alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substrtuted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophilic; R3 and R4 are independently H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl or arylalkyl; R5 is selected from the group consisting of H, hydroxy, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substrtuted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fiuorophilic, or forms part of an amino acid, or is a nucleoside, an oligomer, a polymer, a dendrimer, a substrate or a surface.. Particularly preferred are compounds of formula II in which at least one of R1, R2, R3 and R4 is halogen. The Inventors have found the 5-alkyM5-riydroxy substituted 1,5-dihydro-pyrrol-2-one of formula II can be dehydrated to yield a range of 5-(halomethylene)- 1,5-dihydro-pynrol-2-one, 5-(dihalomomethylene)-1,5-dihydro-pyrrol-2-one. Accordingly in a third aspect, the present invention provides a method for the dehydration of a compound of formula II above, to prepare a compound of formula III; WO 2004/016588 PCT7AU2003/001053 wherein R1 and R2 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted ior unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophilic; R3 and R4 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl; and R5 is a defined above, the .method comprising contacting a compound of formula II with a dehydrating agent Preferably at (east one of R1, R2. R3 and R4 in formula HI is halogen; Examples of suitable dehydrating agents inciude phosphorus pentoxide, silica gel, molecular sieves, alumina, acidic resins and polymers, phosphorus oxychlonde, acetic anhydride, N.N'-dicyclohexylcarbodiimide (DCC), fnfluoroacetic- acid, sulfuric acid, tnfluoroacetic anhydride, trifluorosulfomc acid anhydride (trffiic anhydride). Preferably dehydration is carried out using phosphorus pentoxide in the presence of a solvent. The solvent may be any suitable solvent. Preferable solvents in the present invention include alkyl acetates, aromatic hydrocarbons, chlorinated alkanes, tetrahydrofuran, diethyl ether, dioxane and C1-C3 acids. More preferably, the solvents are aromatic hydrocarbons and chlorinated alkanes. Most preferably, the solvent is dichloromethane, as well as dichloroethane and tnchloroethane. The reaction is preferably carried out at mild temperatures. Preferably the dehydration reaction is performed at a temperature in the range of from about 20-150°C WO 2004/036588 PCT/AU2003/001053 7 Where a solvent is present, the cyclisation may be performed at reflux temperature of the solvent, for example, at the reflux temperature of dichloromethana The reaction time may range from about 2 hours to 12 hours or more and is typically about 2 hours or more. It will be appreciated that reaction conditions may be varied depending on the individual nature of the substrate and the desired rate of the reaction. Non-limiting examples of furanones (III) that can be synthesised by this procedure are listed below. We believe that the 1,5-dihydro-pyrrol-2-ones prepared of formula HI are novel compounds- Thus, in a fourth aspect, the present invention provides a compound of formula III: WO 2004/016588 PCT/AU2003/001053 wherein R1 and R2 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straightcham or branched chain, hydrophilic or fluoraphilic; R3 and R4 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl or arylalkyl; and R5 is as defined above. Preferably at least one of R1, R2, R3 and R4 is halogen. Furthermore the present inventors have also found that furanones of formula (I) when treated with certain amines can yield 5-amino substituted or 5-ammomsthylene substituted furanones. Alternatively, the compounds of formula I can be treated with an alcohol to yield 5' alkoxy substituted furanones. For example when 4-bromo-5-bromomethylene-2(5H)uranone was treated with aniline it gave 4-bromo-5-phenylaminomethyIene-2(5H)-fuiranone in good yields. In contrast, the reaction of 4-bromo-5-bromomethylene-2(5H)-furanone with benzyl amine, gave the corresponding 5-benzylamino-4-bromo-5-bromomethyl-2(5H)-furanone. Accordingly, in a fifth aspect, the present invention provides a method for the preparation of a compound of formula IV WO 2004/016588 PCT/AU2003/001053 9 wherein R1 and R2 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalky!, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophilic; R3 and R4 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, R5 is as defined above, X is O or NR, where R5 may be R1, the method comprising reacting a compound of formula I wherein R3 is a hydrogen and "--- " represents a double bond. Preferably at least one of R1 R2, R3 and R4 is halogen. Preferably R6 is H. Representative examples of furanones (IV) that can be synthesised by this procedure are listed below. In yet a sixth aspect, the present invention provides a compound of formula IV WO 2004/016588 PCT/AU2003/001053 10 wherein R1 and R2 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstrtuted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophilic; R3 and R4 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl; and R5 and X are as defined above. Preferably, at least one of R1, R2. R3 and R4 is halogen. Accordingly a seventh aspect the present invention provides for a method for preparation of a compound of formula V. wherein R1 and Rz are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstrtuted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstrtuted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl( optionally Interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophilic; R3 is selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl or WO 2004/016588 PCT/AU 2003/001053 11 arylalkyl, wherein R5 is H, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted aikenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophilic; X is O or NR6, where R6 is as defined above; and R6 is as defined above. Non-limfting examples of furanones of formula (V) that can be synthesised by this procedure are listed below In an eighth aspect, the present invention provides a compound of formula V: wherein R1 and R2 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted aikenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophilic; R3 is selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl or arylalkyl; WO 2004/016588 PCT/AU20O3/001053 12 X is O or NR6, where Re is as defined above, and R5 is as defined above. In yet a ninth aspect the present invention provides a compound of formula (VI) wherein R1 and R2 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substitutGd or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic orfluorophilic, R3 and R4 are independently selected from H, halogen, alkyl, substituted or unsubstituted aryl or arylalkyl; wherein R5 is H, substituted or unsubstituted alky), substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic orfluorophilic; R5 is defined as above; and Z is selected from the group R2, halogen, OC(0)R2 =O, amine azide, thiol, R2 mercaptoaryl, arylalkoxy, mercaptoarylalkyl, SC(O)R2, OS(O)2R2. NHC(O)R2. =NR2 or NHR2. The compounds of formula V! may be prepared by functionahzing a fimbrolide of formula (III) wherein, R1, R2, R3 and FUare as defined above, with a reagent described in WO 99/54323, (the disclosure of which is incorporated herein by cross-reference). Reagents for introduction and manipulation of the Z group include halogenating and oxidising agents (N-halosuccinimide, iead tetraacetate, selenium dioxide, Jones reagent), nucleophlles (Including organic metal carboxylates, organic alcohols, dimethyl sulfoxide and organonrtnles) and electrophiles including (organrc acids, isocyanates, carboxylic or sulfonic acid hahdes and diethylammosulfurtnfluoride) WO 2004/016588 PCT/AU 2003/001053 13 Non-limiting examples of furanones of formula (VI) that can be synthesised by this procedure are listed below. In a tenth aspect, the present invention provides an oligomer or polymer formed by oligomerising or polymerising a compound of formula It - VI, described herein directly or with one or more other monomers The one or more other monomer may be any suitable polymerisable copolymer e.g. acrylate ester such as alkyl, hydroxyaikyl, aminoalkyl, or substituted substituted or unsubstituted aryl acrylates or methacrylates, WO 2004/016588 PCT7AU2003/U01053 14 crotonates, substituted or unsubstituted acrylonitriles, vinyl alcohols or acetates, styrene and siloxanes. R5 may be a residue of a natural or synthetic compound. R6 may be a biological or non-biological compound. For example, R5 may be a coenzyme or cofactor. R5 may be an oligomer or a polymer, which may be biological or synthetic. For example, the oligomer or polymer may be a peptide or polyamide. The polymer may be a protein, for example, an enzyme or a receptor, R5 may be an oligomer or polymer comprising nucleic acid residues. The polymer may be a polynucleotide, for example, DNA or RNA. R5 may form part of or be bonded to a nucleoside. The nucleoside may be a D- or L-nudeosfde. R5 may be linked to a sugar moiety of the nucleoside. R5 may be a surface or substrate with which the nftrogen atom of is associated. The association may be chemical bonding, for example covalent bonding. The surface or substrate may be biological or synthetic. Altematfvely, the association may be by means of adsorption. Methods for forming such associations are described in more detail below. R5 may also be a dendrimer. A review of dendrimers is provided in Klajnert, B. and Bryszewska, M. (2001) Dendrimers.-properties and applications, Acta Biochemica Polonica Vol. 48 No.1/2001, the disclosure of which is incorporated herein by reference. A plurality of compounds in accordance with the invention may be carried by the dendrimer. The compound may be immobilised directly onto at least part of the surface of the material of the substrate or via one or more intermediate layers interposed between the substrate material and the immobilised layer. The intermediate layer (s) may be bonding layer (s). The substrate may be shaped or non-shaped. The substrate may be solid, semi-solid or flexible. The substrate may be a woven or non-woven film or sheet. The substrate may be a natural or synthetic filament or fibre The substrate may be a natural material, for example, a plant seed. The material from which the substrate is formed may be selected to suit the particular application For example, In the case of a shaped blomedical device the material may meet other specifications of the application, such as mechanical and optical properties. The substrate may be -3 shaped article including, but are not limited, medical devices, for example, implantable biomedical devices such as unnary WO 2004/016588 PCT7AU2003/001053 15 catheters, percutaneous access catheters, stents, as well as non-implantable devices such as contact lenses, contact lens storage cases, and the like. The matenal from which the article is formed can be a metal, a ceramic, a solid synthetic polymer, or a solid natural polymer, for example a solid biopolymer. Examples of useful materials for this invention are titanium, hydroxyapatite, polyethylene (which are useful materials for orthopaedic implants), polyurethanes, organosiloxane polymers, perfluorinated polymers (which are useful matenals for instance for catheters, soft tissue augmentation, and blood contacting devices such as heart valves), acrylic hydrogel polymers and siloxane hydrogel polymers (for instance for contact tens and intraocular lens applications), and the like, and any combination thereof. The surfaces of these materials can be chemically inert or contain reactive functional groups. Further examples of substrates include archival documents, antiques and art, rare and valuable seeds intended for storage (e. g seed banks of conservation groups), etc In which case the substrate may be paper, material or other natural or synthetic material. The substrate may be a shell fish or aquaculture apparatus, for example, that described in PCT/AU98/00508, the disclosure of which is incorporated herein by reference. As mentioned above, R5 may be associated with a surface of substrate. If necessary, the surface of the substrate may be optionally treated at least in part to activate the surface, to which the compounds of the present invention may be reacted to immobilise the compound. Reference to at least part of the surface of the substrate includes a surface of one or more intermediate layers applied to the substrate. The compounds may be Immobilised on the substrate surface by any suitable technique. Immobilization may be by covalent or non covalent means Preferably, the compounds are immobilised on the substrate surface by means of covalent bonds. The immobilization of furanone compounds on to the substrate prevents their loss from the surface, thus ensuring long-lasting antimicrobial action. The association between the compounds of the invention and the substrate may be charactensed by the formula : X-Y-Z where X is a substrate. Y is an optional chemical linking noiety and Z is a compound In accordance with the present invention. The linking moiety, if present, may be a homobifunctional or heterobifunctional linking moiety. Y may be a simple WO 2004/016588 PCT/AU2003/001053 16 component (eg a short molecule) or it may comprise a plurality of units or components that may be the same of different. Y may comprise a number of components or units that may be "built up"in a stepwise fashion. The formation of a covalent interfacial linkage is much preferable to an ionic bond since in biological media where the salt content is such that ionic bonds are interfered with and ironically attached molecules can be displaced from a surface. In the context of substrates that are medical devices, covalent anchoring of the compound(s) also serves to eliminate concerns regarding possible deleterious effects that compounds might cause at sites distant from the device, such as in the liver, brain, or kidney tissues of a living human organism. In medical applications it is important to anchorthe furanone compound (s) via an interfacial covalent bond that is not subject to cleavage in the host environment that the biomedical device is to be placed in., Methods for the covalent immobilization of organic molecules onto solid surfaces are well known to those skilled in the art Interfacial reactions leading to the formation of covalent interfacial bonds are denved from well known organic-synthetic reactions. The choice of immobilization reaction depends on both the nature of the substrate material and the chemical composition of the furanone derivative (s) that are desired for a particular application For example, a compound that contains a hydroxyl group in a side chain distal to the ring system, can be linked covalently onto surfaces using epoxide chemistry analogous to the reaction pathway described forthe immobilization of polysaccharides onto epoxidated surfaces in Li et a/., Surface Modification of Polymeric Biomateriais (BD Ratner and DG Castner, Eds), Plenum Press, NY, 1996 pages 165-173 (the disclosure of which is incorporated herein in its entirety), through isocyanate groups attached to the surface to produce stable urethane linkages through thermal processes, or through carboxylic acid groups or their equivalents, such as acid chlorides, on the surface to produce ester linkages. A compound that contains an aldehyde group can be linked onto surface amine groups using a reductive animation reaction. A compound that contains a carboxylic acid group can be linked onto surface amine groups using carbodumide chemistry interfacial coupling reactions must of course be selected not only for their ability to achieve the desired covalent linkage but also for avoidance of adverse effects on the furanone compound (s) to be attached. Particularly, the furanone WO 2004/016588 PCT/AU20O3/001053 17 nng system tends to be labile to alkaline conditions. Such limitations are well known to those skilled in the art. Among the many possible interfacial coupling reactions known in the art, there is sufficient scope for selection of reactions that proceed in a suitable pH range and with furanones substituted with various functional groups in various positions Some solid substrate matenals possess reactive surface chemical groups that can undergo chemical reactions with a partner group on a compound and thereby form a covalent interfacial linkage directly. Alternatively, in situ covalent linkage can be made directly through the addition of a doubly functionaiised linker molecule to the active surface in the presence of an appropriate compound, or stepwise by sequential addition of doubly functionaiised linker molecules and then an appropriate compound It is not always possible to immobilize furanone compounds directly onto solid substrate materials; in these cases, surface activation or one or more interfaciaf bonding layer (s) is used to effect covalent immobilization of the compounds. Such surface activation is essential when immobilizing compounds onto polymeric materials such as fluoropolymers and polyolefins. Surface activation of solid substrate matenals can be achieved in a number of ways. Examples are corona discharge treatment or low pressure plasma treatment of polymers. These methods are well known to introduce a vanety of functional groups onto polymeric surfaces. An alternative approach is to provide an interfacial bonding layer interspersed between the solid substrate material or medical device and the compound layer. The application of a thin fnterfacial bonding layer can be done using methods such as dip coating, spin coating, or plasma polymerization. The chemistry of the bonding layer is selected such that appropriate reactive chemical groups are provided on the surface of this layer, groups that then are accessible for reaction with compound of the invention. Particularly versatile is the subsequent application of multiple thin interfacial bonding layers; this method can provide a very wide range of desired chemical groups on the surface for the immobilization of a wide range of functionalized furanones and enables usage of compounds optimized for their biological efficacy. By providing a thin, surface-coated layer of compounds, the optical quality of antibacterial devices of this invention is not reduced, which makes the WO 2004/016588 PCT/AU200J/001053 18 invention applicable to transparent ophthalmic devices such as contact lenses and intraocular lenses. The present invention provides thin surface coatings that provide antimicrobial properties and/or antifungal properties to solid materials onto which the coatings have been applied More particularly, the coatings may be designed to reduce or prevent colonization of biomedical devices by bactena that cause adverse effects on the health of human users of biomedical devices when such devices are colonized by bacteria. The active antibacterial layer comprises one or a plurality of furanorie compounds selected for both their antibacterial activity and absence of cytotoxicity as well as any other adverse biomedical effect on the host environment that the coated device contacts. In an eleventh aspect, the present invention provides incorporation of compounds produced by the methods according to the first, third, fifth, seventh, ninth, or tenth aspects either in surface coatings or polymers through any part of the molecule, for example, newly Introduced functionality on the alkyl chain or the alkyl chain or the halomethylene functionality itself via direct polymerisation or copolymerisation with suitable monomers. In an twelfth aspect, the present invention provides a compound produced by the method according to the first, third, fifth, seventh, ninth, or eleventh aspects of the present invention. In a thirteenth aspect, the present invention provides the use of a compound produced according to the present invention. The present inventors have found that many of the 1,5-dihydro-pyrrol-2-one derivatives and furanones having the formula (II), (III), (IV), (V) and (VI) have antimicrobial and/or antifouling properties. Accordingly, the flmbrolide derivatives are suitable for use as antimicrobial and/or antifouling agents. Thus in a fourteenth aspect the present invention provides methods of use of compounds of formula (II), (III), (IV), (V) and (VI) in medical, scientific and/or biological applications. For these and other applications, the compounds of the present invention may be formulated as a composition. In a fifteenth aspect, the present invention provides a composition comprising at least one compound of formula (II), (III), (IV), (V) or (VI) The compositions of the third aspect of the invention may be in any suitable form. The composition may include a carrier or diluent The carrier may WO 2004/016588 PCT/AU2003/001053 19 be liquid or solid. For example, the compositions may be in the form of a solution or suspension of at least one of the compounds in a liquid. The liquid may be an aqueous solvent or a non-aqueous solvent The liquid may consist of or comprise a one or more organic solvents. The liquid may be an ionic liquid. Particular examples of carrier or diluents include, but are not limited to, water, polyethylene glycol, propylene glycol, cyclodextnn and derivatives thereof The composition may be formulated for delivery in an aerosol or powder form. The composition may include organic or inorganic polymeric substances. For example, the compound of the invention may be admixed with a polymer or bound to, or adsorbed on to, a polymer When the composition is to be formulated as a disinfectant or cleaning formulation, the composition may include conventional additives used in such formulations. Non-limiting examples of the physical form of the formulations include powders, solutions, suspensions, dispersions, emulsions and gels Formulations for pharmaceutical uses may incorporate pharmaceutically acceptable earners, diluents and excipients known to those skilled in the art. The compositions make be formulated for parenteral or non-parenteral administration. The composition of the invention may be formulated for methods of introduction including, but not limited to, topical, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, ophthalmic, and oral routes. It may be formulated for administration by any convenient route, for example by infusion or bolus injection, by absorption through epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa, etc.) and may be administered together with other biologically active agents. Administration may be localized or systemic The composition may be formulated forintraventncular and intrathecal injection. Pulmonary administration can also be employed, e.g., by use of an inhaler or nebulizer, and formulation with an aerosolizing agent. In certain preferred embodiments the composition further comprises other active agents such as antibiotics and cleaning agents. In a sixteenth aspect, the present invention provides a method of treating an infection in a human or animal subject the method comprising administration to the subject of an effective amount of the compound of the invention. The treatment may therapeutic arid/or prophylactic. WO 2004/016588 PCT/AU2003/001053 20 The compounds of the present invention can act as quorum sensing inhibitors and therefore find use in any application where such as effect is desired. For example, the compounds of the present invention may have use in preventing the establishment and expression of virulence by microorganisms through the inhibition of quorum sensing systems and/or other extracellular systems (eg see, International patent application No. PCT/AU01/01621, the disclosure of which is incorporated herein in its entirety). The present invention is suitable for biofilms onginating from a single type of organism and for mixed biofilms By "mixed biofilms" is meant biofilms created by more than one type of microorganism Most preferably, it is envisioned that biofilms will be created by at least two organisms from the group consisting of bacteria, algae, fungi, and protozoa. The effects of treating biofilms with homoserine lactones have been demonstrated with Pseudomonas aeruginosa. The HSLs have generally been isolated from a wide range of bacteria known to produce biofiims. Among these are the enterobacteria. The presence of the HSLs In a wide range of bactena indicates that the compounds of the present invention can be used to effectively treat not only Pseudomonas sp. biofiims but also mixed biofilms containing Pseudomonas sp. and biofilms composed of bacteria other than Pseudomonas aeruginosa. The following is a list of groups of Gram-Negative bacteria that have members which use homoserine lactones for cell-cefl communication: anaerobic Gram Negative Straight, Curved and Helical Rods; Bacteroidaceae; The Rickettsias and Chlamydias, Dissirrrilatory Sulfate-or Sulfur-Reducing Bacteria; the Mycoplasmas; The mycobacteria; Budding and/or Appendaged Bactena; Sheathed Bactena; Nocardioforms; and Actinomycetes, for example. See Bergey's Manual of Systematic Bacteriology, First Ed. John G. Holt, Editor in Chief (1984), incorporated herein by reference. The method of the sixteenth aspect may be used to treat an infection or condition in a subject that is characterised by biofilm formation. Non-limiting examples of human infections involving biofilms include dental canes, penodontitis, otitis media, muscular skeletal infections, necrotismg fascitis, biliary tract Infection, ostsomyelitis, bacterial prostatitis , native valve endocarditis, cystic fibrosis pneumonia, meloidosis, and nosocomial infections such as ICU pneumonia, sutures, exit sites, arteriovenous sites, scleral buckles, contact lenses, urinary catheter cystitis, peritonea! dialysis (CAPD) WO 2004/016588 PCT/AU2003/00J053 21 pentonitis, UDs, endotracheal tubes, Hickman catheters, central venous catheters, mechanical heart valves, vascular grafts, biliary stent blockage, and orthopedic devices, penile prostheses. Further applications are described in Costerton J et al, (1999) Vol. 284, Science pp1318-i322 and Costerton J and Steward, (2001) Battling Biofilms, Scientific American pp 75-81, the disclosures of which are incorporated herein by reference. Other locations in which biofilms may form included dnnking water pipes, which may lead to corrosion or disease, household drains, dental plaque which may lead to gum disease and cavities, which may lead to gun disease or cavities, contact lenses which may lead to eye infections, ears which may lead to chronic infection and lungs which may lead to pneumonia. The condition may be cystic fibrosis. The infection may be that resulting from a skin .infection, burn infection and/or wound infection. The method and composition of the invention may be particularly suitable for the treatment of infection in immuno compromised individuals. In yet a seventeenth aspect, the present invention provides a method for treating biofilm formation on a surface by contacting the surface with a compound in accordance with the present invention. The term "surface" as used herein relates to any surface which may be covered by a biofilm layer. The surface may be a biologtcal (eg tissue, membrane, skin etc) or non-biological surface. The surface may be that of a natural surface, for example, plant seed, wood, fibre etc The surface or substrate may be any hard surface such as metal, organic and inorganic polymer surface, natural and synthetic elastomers, board, glass, wood, paper, concrete, rock, marble, gypsum and ceramic materials which optionally are coated, eg with paint, enamel etc; or any soft surface such as fibres of any kind (yams, textiles, vegetable fibres, rock wool, hair etc.); or porous surfaces; skin (human or animal); keratinous materials (nails etc.). The hard surface can be present in process equipment or components of cooling equipment, for example, a cooling tower, a water treatment plant, a dairy, a food processing plant, a chemical or pharmaceutical process plant. The porous surface can be present in a filter, eg. a membrane filter. Particular examples of surfaces, that may be treated in accordance with the invention include, but are not limited to, toilet bowls, bathtubs, drains, WO 2004/016588 PCT/AU2003/001053 22 highchairs, counter tops, vegetables, meat processing rooms, butcher shops, food preparation areas, air ducts, air-conditioners, carpets, paper or woven product treatment, nappies (diapers), personal hygiene products (eg sanitary napkins) and washing machines. The cleaning composition may be in the form of a toilet drop-in or spray-on devices for prevention and removal of soil and under rim cleaner for toilets. The compositions and methods of the present invention also have applications in cleaning of Industrial surfaces such as floors, benches, walls and the like and these and other surfaces in medical establishments such as hospitals (eg surfaces in operating theatres), veterinary hospitals, and in mortuanes and funeral parlours. A compound of the invention may be incorporated into epidermal bandages and lotions. Alternatively, the compounds of the invention may be incorporated into cosmetic formulations^ for example, aftershave lotions. Compositions of the present invention may be m the form of an aqueous solution or suspension containing a cleaning-effective amount of the active compound described above. The cleaning composition may be in the form of a spray, a dispensable liquid, or a toilet tank drop-in, under-rim product for prevention, removal and cleaning of toilets and other wet or intermittently wet surfaces in domestic or industrial environments. The compositions of the present invention may additionally comprise a surfactant selected from the group consisting of anionic, non-ionic, amphotenc, biological surfactants and mixtures thereof. Most preferably, the surfactant is sodium dodecyl sulfate. One or more adjuvant compounds may be added to the cleaning solution of the present invention. They may be selected from one or more of biocides, fungicides, antibiotics, and mixtures thereof to affect planktonics. pH regulators, perfumes, dyes or colorants may also be added. By "cleaning-effective" amount of active compound, it is meant an amount of the compound which is necessary to remove at least 10% of bacteria from a biofilm as determined by a reduction in numbers of bacteria within the biofilm when compared with a biofilm not exposed to the active compound. The cleaning methods of the present invention are suitable for cleaning surfaces. They may be used to treat hard, rigid surfaces such as dra»n pipes, glazed ceramic, porcelain, glass, metal, wood, chrome, plastic, vinyl and formica or soft flexible surfaces such as shower curtains, upholstery, laundry WO 2004/016588 PCT/AU2003/001053 23 and carpeting. It is also envisioned that both woven and non woven and porous and non-porous surfaces would be suitable. In other embodiments of the present invention, the composition of the invention may be formulated as a dentifrice, a mouthwash or a composition for the treatment of dental caries. The composition may be formulated for acne treatment or cleaning and disinfecting contact lenses (eg as a saline solution). The method of the invention may be used to treat medical devices, In yet a further aspect, the present invention extend to a medical device having a least one surface associated with a compound(s) in accordance with the present invention. The method of the invention may be used to treat implanted devices that are permanent such as an artificial heart valve or hip joint, and those that are not permanent such as indwelling catheters, pacemakers, surgical pins etc. The method may further be used in situations involving bacterial infection of a host, either human or animal, for example fn a topical dressing for burn patients. An example of such a situation would be the infection by P. aeruginosa of superficial wounds such as are found in burn patients or in the lung of a cystic fibrosis patient. In other forms, the present invention can be used to treat integrated circuits, circuit boards or other electronic or microelectronic devices. In yet another aspect, the present invention provides a method for the inhibition of a biological pathway is a dell, the method compnsing administering to the ceil a compound in accordance with the present invention. Terminology The term "alkyl" is taken to mean both straight chain alkyl groups such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tertiary butyl, and the like. Preferably the alkyl group is a lower alkyl of 1 to 6 carbon atoms The alkyl group may optionally be substituted by one or more groups selected tram alkyl, cycloalkyl, alkenyl, alkynyi, halo, carboxyl, haloalkyl, haloalkynyl, hydroxy, substituted or unsubstituted alkoxy, alkenyloxy, haloaikoxy, haloalkenyloxy, nitro, ammo, nitroalkyi, nitroaikenyl, nitroalkynyl, nftroheterocyclyl, alkylammo, dialkylamino, aikenylamtne, alkynylamino, acyl, alkenoyi, alkynoyl, acylamino, diacylamino, acyloxy, alkylsulfonytoxy, heterocyclyl, heterocycioxy, heterocydamino, haloheterocyclyl, alkylsulfenyl, alkylcarbonyioxy, alkylthio, acylthio, phosphorus-containing groups' such as phosphono and phosphinyl. WO 2004/016588 PCT/AU2003/001053 24 The term "alkoxy" denotes straight chain or branched alkyloxy, preferably C1-10 alkoxy. Examples include methoxy, ethoxy, n-propoxy, isopropoxy and the different butoxy isomers The term "alkenyl" includes groups formed from straight chain, branched or mono- or polycyclic alkenes and polyene. Substituents include mono- or poly-unsaturated alkyl or cycloalkyl groups as previously defined, preferably C2-10 alkenyl. Examples of alkenyl include vinyl, allyl, 1-methylvinyl, butenyl, iso-butenyl, 3-rnethyl-2-butenyl, 1-pentenyl, cyclopentenyl, 1-methyi-cyclopentenyl, 1-hexenyl, 3-hexenyl, cyclohexenyl, 1-heptenyl, 3-heptenyl, 1-octenyl, cyclooctenyl, 1-nonenyI, 2-nonenyI, 3-nonenyl, 1-decenyl, 3-deceny], 1,3-butadfenyl, 1,4-pentadienyl, 1,3-cyclopentadienyl, 1,3-hexadienyl, 1,4-hexadienyl, 1,3-cyclohexadienyl, 1,4-cyclohexadienyl, 1,3-cycloheptadienyl, 1,3,5-cycloheptatrienyl, or 1,3,5,7-cyclooctatetraenyl. The term "halogen" includes fluonne, chlorine; bromine or iodine, preferably bromine or fluorine. The term "heteroatoms" denotes O, N, S or Si. The term "acyl" used either alone or in compound words such as "acyloxy", "acylthio", "acylamino" or diacylamrno" denotes an alkanoyl, aroyl, heteroyl, carbamoyl, alkoxycarbonyl, alkanesulfonyl, arysulfonyl, and is preferably a C1-10 alkanoyl. Examples of acyl include carbamoyl; straight chain or branched alkanoyl, such as forrnyl, acetyl, propanoyl, butanoyl, 2- methylpropanoyl, pentanoyl, 2,2-dimethylpropanoyI, hexanoyl, heptanayl, octanoyl, nonanoyl, decanoyl; alkoxycarbonyl, such as methoxycarbonyl, ethoxycarbonyl, t-butoxycarbonyl, t-pentyioxycarbonyl or heptyloxycarbonyl, cycloalkanecarbonyl such as cyclopropanecarbonyl cyclobutanecarbonyl, cyclopentanecarbonyl or cyclohexanecarbonyl; alkanesulfonyl, such as rnethanesulfonyl or ethanesulfonyl; alkoxysulfonyl, such as methoxysulfonyl or ethoxysulfonyl; heterocycloalkanecarbonyl; heterocyclyoalkanoyi, such as pyrrolidinylacetyl, pyrrolidinylpropanoyl, pyrrolinylacletyl, pyrrolylacetyl, pyrrolidinylbutanoyl, pyrrolidinylpentanoyl, pyrrolidinylhexanoyl or thiazohdinylacetyl; heterocyclylalkenoyl, such as heterocyclylpropenoyl, heterocyclylbutenoyl, heterocyclylpentenoyl or heterocyclylhexenoyl; or hetsrocyclylglyoxyloyl, such as, thiazolidinyJglyoxytoyl or pyrrolidinylglyoxyloyl. The term "aryl" refers to aryl groups havjng 6 through 10 carbon atoms and includes, for example, phenyl, naphthyl, indenyi. Typically the aryl group WO 2004/016588 PCT/AU2003/001053 25 will be phenyl or naphthyl as compounds having such groups are more readily available commercially than other aryl compounds, The term "substituted aryl" refers to aryl groups having 1 through 3 substituents independently selected ifrom the group of lower alkyl, lower substituted or unsubstituted alkoxy, halonitro, or haloalkyl having 1 through 3 carbon atoms and 1 through 3 halo atoms. Typical substituted aryl groups include, for example, 2-fluorophenyI, 2-chlorophenyl, 2,e-dimethylphenyl, 4-fiuoroprienyl, 2-methylphenyl, 2-chloro, 3-chloromethylphenyl, 2-nitro, 5-methylphenyl, 2,6-dichlorophenyl, 3-trifluoromethylphenyl, 2-methoxyphenyl, 2-bromonaphth-1-yl, 3-methoxyinden-1-yl, and the like. Carboxyaryl eg carboxy phenyl, aminoaryl eg ammophenyl The term "fluorophilic" is used to jindicate the highly attractive interactions between certain groups, such as highly fluorinated alkyl groups of C4-C10 chain length, towards perfluoroalkanes and perfiuoroalkane polymers. The term "arnino acid" as used herein includes any compound having at least one ammo group and at least one! carboxyl group. The amino acid may be a naturally occurring ammo acid or it may be a non-naturally occurring amlno acid. The amines used in this invention may be soluble in the reaction medium or insoluble in the reaction medium. Examples of soluble amines include ammonia, alkyl(, aryl-, arylalkyl-, and heterocyclic amines. Examples of insoluble amines include basic amine resins and amine containing biological and synthetic polymers. The term "optionally substituted" includes, but is not limited to such groups as halogen; hydroxy; hydroxy substituted alkyl; substituted or unsubstituted S(O)m alkyl or S(O)m aryl wherein m is 0,1 or 2, such as methyl thio, methylsulfinyl or methyl suffonyl; amino, mono and di-substituted ammo; alkyl, cycloalkyl, or cycloalkyl alkyl group; halosubstituted alkyl, such as CF3; an optionally substituted aryl, optionally substituted arylalkyl, such as benzyl or phenethyl, wherein these aryl moieties may also be substituted one to two times by halogen; hydroxy; hydroxy substituted alkyl; alkoxy; S(0)m alkyl, ammo, mono and di- alkyl substituted amino, substituted or unsubstituted alkylSIO- for example (CH3)3SiO- The term "Medical devices" as used herein includes disposable or permanent catheters, (e.g., central venous catheters, dialysis catheters, long-term tunneled central venous catheters, short-term central venous catheters, WO 2004/016588 PCT/AU2003/001053 26 peripherally inserted central catheters, peripheral venous catheters, pulmonary artery Swan-Ganz catheters, urinary catheters, and peritoneal catheters), long-term urinary devices, tissue bonding urinary devices, vascular grafts, vascular catheter ports, wound drain tubes, ventricular catheters, hydrocephalus shunts heart valves, heart assist devices (e.g., left ventricular assist devices), pacemaker capsules, incontinence devices, penile implants, small or temporary joint replacements, urinary dilator, cannulas, elastomers, hydrogels, surgical instruments, dental instruments, tubings, such as intravenous tubes, breathing tubes, dental water lines, dental drain tubes, and feeding tubes, fabrics, paper, indicator stnps (e.g., paper indicator stnps or plastic Indicator stnps), adhesives (e.g., hydrogel adhesives, hot-melt adhesives, or solvent-based adhesives), bandages, orthopedic implants, and any other device used in the medical field. "Medical devices" also include any device which may be inserted or implanted into a human being or other animal, or placed at the insertion or implantation site such as the skin near the insertion or implantation site, and which include at least one surface which is susceptible to colonization by biofilm embedded microorganisms, Medical devices also include any other surface which may be desired or necessary to prevent biofilm embedded microorganisms from growing or proliferating on at least one surface of the medical device, or to remove or clean biofilm embedded microorganisms from the at least one surface of the medical device, such as the surfaces of equipment in operating rooms, emergency rooms, hospital rooms, clinics, and bathrooms In one specific embodiment, the biofilm penetrating composition is integrated into an adhesive, such as tape, thereby providing an adhesive which may prevent growth or proliferation of biofilm embedded microorganisms on at least one surface of the adhesive. Implantable medical devices include orthopedic implants. Insertable medical devices include catheters and shunts which. The medical devices may be formed of any suitable metallic matenats or non-metallic materials known to persons skilled in the art. Examples of metallic matenals include, but are not limited to, tivanium, titanium, and stainless steel, and derivatives or combinations thereof. Examples of non-metallic materials include, but are not limited to, thermoplastic or polymeric materials such as rubber, plastic, polyesters, polyethylene, polyurethane, silicone, Gortex (tm) (Polytetrafluoroethylene), Dacron (tm) (polyethylene tetraphthalate), Teflon WO 2004/016588 PCT/AU2O03/001053 27 (Polytetrafluoroethylene), latex, elastomers and Dacron(tm) sealed with gelatin, collagen or albumin, and derivatives or combinations thereof. The present invention also extends to a method of regulating a cells characterised by AHL-mediated quorum sensing or an AI-2 pathway comprising contacting the cells with a compound in accordance with the present invention. Throughout this specification the word "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps. Reference is made to patent applications PCT/AU01/01621, PCT/AU02/00797, PCT/AU99/00284. PCT/AU99/00285, PCT/AU00/01553, PCT/AU01/00296, PCT/AU01/00295, PCT/AU01/00407. PCT/AU89/00508 and PCT/AU01/00751 which relate to furanones and analogues and to uses of these compounds and the entire disclosures of which are incorporated herein by reference. WO 2004/016588 PCT/AU2003/001053 28 Modes for Carrying Out the Invention The invention is further described in and illustrated by the following examples. The examples are not to be construed as limiting the invention in any way. EXPERIMENTAL DETAILS Genera/. Melting points are uncorrected. Microanalyses were performed by Dr H.P, Pham of The University of New South Wales Microanalytical Laboratory. 1H NMR spectra were obtained in CDCI3 on a Bruker AC300F (300 MHz) or a Bruker DMX50Q (500 MHz) spectrometer. 13C NMR were obtained in the same solvent on a Bruker AC300F (75.5 MHz) or a Bruker DMX500 (125.8 MHz) spectrometer Chemical shifts were measured on the 8 scale internally referenced to the solvent peaks: CDCI3 (5 7.26, 5 77.04). Ultraviolet spectra were measured on an Hitachi U-3200 spectrophotometer and refer to solutions in absolute MeOH. Infrared spectra were recorded on a Perkin-Etmer 298 or a Perkin-Eimer 580B spectrophotometer and refer to paraffin mulls. The electron impact mass spectra were recorded on an VG Quattro mass spectrometer at 70eV lonisation voltage and 200°C ion source temperature. FAB spectra were recorded on an AutoSpecQ mass spectrometer. Column chromatography was carried out using Merck silica gel 60H (Art. 7736), whilst preparative thin layer chromatography was performed on 2 mm plates using Merck silica gel 60GF254 (Art. 7730). 3-Butyl-5-dibromomethyI-5'-hydroxy-1-phenyl-'1,5-dihydropyrrol-2-one A solution of 3-butyl-5-dibromomethylene-2(5H)fijranone (0.20 g; 0-65 mmol) in aniline (5 ml) was allowed to stand at room temperature for 24 h. The mixture was diluted with dichloromethane (25 ml) and washed with aqueous hydrochloric acid (2M, 20 ml). The organic phase was dried over sodium sulfate and evaporated to yield a yellow viscous oil (0.30 g). The crude product was chromatographed on silica using dichloromethane/ethylacetate (19:1; v:v) as the eluent. The major product, a pale yellow band, was collected and recrystallised from light petroleum to yield 3-butyl-5-dibromomethyl-5-hydroxy-i-phenyl-1,5-dihydropyrroi-2-one as colourless pnsms (0.24 g, 92%), m.p. 96-98°Cvmax 3211, 2957, 1679, 1597, 1500, 1417, 1117, 1058, 760, 698 cm-1. ?w: 263nm (Emax2,955), 202 (2,464). 1H n.m.r. ( (CDCI3): 7.54-7.37, m, Ph; WO 2004/016588 PCT/AU2003/001053 29 6.82, 1Hf s, C4-H; 5.56, 1H, s, -CHBr2; 3 42, S, C5-0H, 2.43-2.41, m. 2H, CH2, 1.64-0.97, m, C3-chain. 13C n.m.r. 6 (CDC/3): 13.7, 14.0, 22.3, 46.6, 92.2, 126.7,127.4,129.0, 134.0,135.5,136.-0,144.3, 169.0. 5-Dibromomethyl-3-hexyI-5-hydroxy-1-phenyl-1,5-dihydropyrrol-2-one A mixture of 3-hexyl-5-dtbromomethyiene-2(5H)furanone (0.40g, 1.16 mmol) and aniline (1 ml) in ethanol (6 ml) was refluxed tor 3h. The solvent was evaporated off and the residue extracted with dichlorornethane (25 ml). The organic phase was washed with aqueous hydrochloric acid (2M. 2 x 20 ml), dried over sodium sulfate and evaporated to yield a seml-soltd (0.39 g) The crude product was chromatographed on silica using dichloromethane/ethylacetate (19:1, v;v) as the eluent. The major product, a pale yellow band, was collected and recrystallised from light petroleum to yield 5-dibromom6thy1-3-hexyl-5-hydroxy-1 -phenyl-1 .S-dihydropyrrol^-one as a semi-crystalline solid (23%), rap. 43-45° (Found (HRESMS) 451.982217. C17H21Br2NO2Na+ (78Br) requires 451.983106). vmm: 3186, 2926, 1680, 1659, 1492, 1372, 1095, 1059, 897, 850, 766, 747, 699, 671 cm-1. Xmax: 261nm (Emax 4051), 206 (26.550). 1H n.m.r. ( (CDCI3). 7.5-7.25, m, 5H, Ph; 6.8, s, 1H, C4-H; 5.5, s, 1H, -CHBr?: 3.77, brs, 1H, C5-OH; 2.44-2.34, m. 2H, CH2; 2.03-0.91, 11H, C3-cham. 13C n.m.r. ( ( CDCI3): 14.2, 25.3, 29 0, 31 0, 46 6, 92.0, 104.8, 126.7, 127.0, 136.0, 144.0,168.0, 172.0. 1-Benzyl-3-butyl-5-dibromomethyl-5-hydroxy-1,5-dihydropyrroI-2-one A solution of 3-butyl-5-dibromomethylene-2(5H)furanone (1.03 g: 3.32 mmol) in benzyl amine (2ml) was allowed to stand at room temperature for 1 h during which time the reaction mixture solidified. The solid was dissolved in dichlorornethane (25 ml) and washed with aqueous hydrochloric acid (2M, 20 ml). The organic phase was dried over sodium sulfate and evaporated to yield a yellow viscous oil. The crude product was tnturated with light petroleum to yield a white solid (1.0 g; 74%) which was recrystallised from light petroleum to yield 1-benzyl-3-bUtyl-5-dlbromom9thyl-5-hydroxy-1,5-d!hydropyrrol-2-one as colourless needles, m.p. 92-93 (Found (HRESMS) m/z 479.974243 C18H21Br2NO3Na+ (70Br) requires 479.978123) vmax: 2987, 2953, 2920. 1677, 1650, 1449, 1424, 1069 cm-1. (max 207 (88,250). 1Hn.m.r. ( (CDCl3): 7.4-7.29, m, 5H, Ph, 6.72, s, 1H, C4-H; 5.56, s, 1H, -CHBr2; 4.54, bs, 2H, CH2Ph; 3.0, WO 2004/016588 PCT/AU2003/001053 30 1H, C-5 OH; 1.54-0.97, m, C-3 chain. 13C n m.r.5 (CDCI3): 13.7, 22 3, 29.3, 42.6, 46.8, 91:5, 127.6,128.3,128.7,136-9. 137 0,160.8/'170.6. 1-Ben2yI-5-dibromomethyI-3-hexyl-5-hydroxy-1,5-d'ihydropyrrol-2-one Method A A solution of 3-hexyl-5-dibromomethylene-2(5H)furanone (1.03 g: 3.32 mrnol) in benzyl amine (2 ml) was stirred at room temperature for 0.5h. Dichloromethane (15 ml) was added to the reaction mixture and the precipitated solid was filtered off. The filtrate was washed with aqueous hydrochloric acid (2M, 20 ml), dned over sodium sulfate and evaporated to yield a yellow viscous oil (0.36g) The crude product was chromatographed on silica using dichloromethane/ethyl acetate (1:19) as the eluent and recrystallised from light petroleum to yield 1-benzyI-5-dlbromorriethyl-3-hexyl-5-hydroxy-1,5-dihydropyrro!-2-one (0 11 g) as colourless needles m.p.105-10S°C. (Found (HRESMS) m/z 465 994011. C18H23Br2NO2NA+ (79Br) requires 465.998758. vms>: 3195, 2987, 2924, 2858, 1676, 1649, 1425, 1153, 1068, 968, 845, 730, 599 cm-1. (max 205 (emax 7740) nm. 1H n.m,r.8(CDCl3): 7.39-7.26, m, 5H, Ph; 6.7, s, 1H, C4-H; 5.6, s, 1H, -CHBr2; 4.54, d, J 15 Hz, 2H, CH2Ph; 2.89-2.35, m, 2H, CH2; 1.60^0.87, m, 13H, C3-chain. 13C n.m.r, S (CDCI3): 14, 22.45, 25, 27, 28.8, 31.4, 42,5, 46.7, 91.5, 127.6, 128.5, 128.6, 136.6, 136.7,144.0,170.0 Method B A mixture of 3-hexyl-5-dibromomethylene-2(5H)furanone (1.03 g- 3.32 mrnol) and benzyl amine (2ml) in ethanol (5 ml) was stirred at room temperature for 2.6h The cnjde product was isolated and purified as described above to yield 1-benzyl-5-dibromomethyl-3-hexyl-5-hydroxy-1,5-dihydropyrrol-2-one in (72%) yield. 1-Butyl-5-dibromomethyl-3-hexyl-5-hydrOxy-1,5-dihydropyrrol-2-one n-Butyiamme (0.272 g; 3.72 mmol) was added dropwise to a solution of 5-dibromomethylene-3-hexyI-2(5H)furanone (0,314 g; 0.93 mmol) m CH2CI2 (10 ml). The mixture was stirred at roomi temperature for 5 hrs. Column chromatography on silica with CH2CI2 followed by CH2Cl2/EtOAc (19;1) afforded the major product as a colourless oil (0.20 g) which upon WO 2004/016588 PCT/AU2003/001053 31 recrystallisation from petrol gave 1-butyl-5-dibromomethyI-3-hexyl-5-hydroxy-1,5-dihydropyrrol-2-one (52%) as colourless needles, rn.p. 85-86°. (Found(HRESMS) rn/z 432.013664. C15H25Br2NO2Na+ (79Br) requires 432 014407). vmM; 3230, 2957, 2859, 1672, 1650, 1458, 1422. 1375, 1270, 1233, 1139, 1079, 1023, 728. 666, 612 cm-1. W: 259 (emax 945), 206 (9658) nm. 1H n.m.r. 6 (CDCI3): 6.68, s, 1H, C4-H; 5.8, s, 1Hf CHBrz; 3.45, m, 1H, N-CH2; 3 10, m, 1H, N-CH2;-3.15. bs, OH; 3 20. m, 2H, -CH2; 2.33-2.31, m, -CH2-chain, 1.65-0.88, m, 14H, alkyl chain. 13C n.m.r. ( (CDCi3): 13.6, 20, 22, 25, 27, 29, 30.75, 31, 39, 46.6, 91.4,136,144.5,170. N-(2-Hydroxyethyl)-3-bulyl-5-dibrommethyl-5-hydroxy-2(5H)pyrolinone A solution of ethanolamlne (1 13g, 18.5 mmol) in CH2Cl2 (5 ml) was added dropwise to an ice-cooled solution of 3-butyl-5-dibromomethylen-2(5H)-furanone (1 -0g; 9.25 mmol) in dichlorornethane. The mixture was stirred at this temperature for 1 hr and then at room temperature for further 1h. The mixture was washed with waster (3 x 50 ml), dned over sodium sulfate and evaporated to yield a viscous oil (0.63g). The cuude product was chromatographed on silica using EtOAc as an eluent to yield N-(2-hydroxyethyl)-3-butyl-5-dibrommethyl-5-hydroxy-2(5H)pyrolinorie as an oil which solidified on keeping. Crystallisation from (CH2Cl2/petrol) afforded the title compound as colurless needles, m p. 68-70°. vmax: 3439, 3105, 3065, 2957, 2927, 1701, 1593, 1496. 1465, 1370, 1189, 1139, 1095, 1069, 10^7, 945, 835, 763 cm-1. ( max 203 nm 1H nmr ( (CDCI3) 0.93, t, 3H, CH3; 1.25-1.45, m, 4H, CH2; 2.35, m, 2H, CH2; 3.10, m, 1H, NCH2CH2OH; 3.84, m\| 2H, NCH2CHZOH; 4.10, m, 1H, NGH2CH2OH; 5.43, bs, 1H, OH; 5.84, s, lH, CHBr2; 6,78, s, 1H, H4.13C n.m.r ( (CDCI3): 13.73, 22.2, 24 9, 29.3 41.3, 45.7, 61.3, 90.3, 137.5, 142.8,171.1. 5-Dibromomethyl-3-hexyI-5-hydroxy-1,5-dihydropyrroI-2-one Liquid ammonia (5ml) was added to 5-dibromomethylene-3-hexyl-2(5H)furanone (0,50 g; 1,48 mmol) in a sealed tube held in a acetone/liquid nitrogen bath. The reaction mixture was allowed to warm up gradually and kept at room temperature overnight. After gradual evaporation of ammonia the product was extracted with EtOAc (20 ml), washed wfth water, dried over Na2SO4, and evaporated to yield a solid (0.30g). The crude product was purified on a silica column using first CH2CI2 as the eluent followed by EtOAc/MeOH (41). The yellow band upon solvent removal and crystallisation WO 2004/016588 PCT/AU2003/001O53 32 from petrol afforded a yellow crystalline solid (Q.07g ) of 5-dibromamethyI-3-hexyl-5-hydroxy-1,5 dihydropyrrol-2-one, m.p. 106-109. 1H n.m.r ( (CDCI3): 6.61; s, 1H, C4-H, 6.26, s, 1H, -NH; 5.68, s, 1H, CHBr2;,3.2, s, C5-OH; 2 28-2.23, m, -CH2,chain; 1.55-0.91, m, 11 H, chain. 13C n.m.r. ( (CDCI3): 13.9, 22, 25.5, 27, 29, 31.4,129, 140, 142, 170.5. 4-bromo-5-hydlroxy-5-hydroxyrnethy[-1,5-dihydropyrrol-2-one A suspension of 4-bromo-5-bromomethylene-2(5H)-furanone (1.30g, 5.16 mmol) in aqueous ammonia solution (20% w/w) was stirred at room temperature for 1/2 h. During this time a complete dissolution of furanone was observed. The solution was evaporated to dryness in vacuo at ca 35-40 °C, and finally under high vacuum at room temperature. The resulting solid (1.70 g) was recrystallised from ethanol to yield 4-bromo-5-hydroxy-5 -hydroxymethyl-1,5-dihydropyrrol-2-one as colourless granules (1.0 g). mp 14O-142 °C (decornp); vmax: 3259, 3100, 2949, 1667, 1592, 1419, 1370, 1152, 1076, 981, 872, 563 cm-1. Xmax 220 (( max 6077). 1H n.m.r. 5 (CDCI3): 8.09, s, -NH; 6.22, d, 2 Hz, H3; 4.97, t, 2 Hz, -CH2OH; 3.37, q.J2 Hz, OH; 2.48, d, 2Hz, -CH2OH. 13C n,m r. 5 (CDCl3): 69 6, 84.3, 132,8, 152,3, 174.1. 4-Bromo-3-hexyl-5-hydroxy-5-hydroxymethyM,5-dihydropyrrol-2-one A suspension of 4-bromo-3-hexyl-5-bromomethylene-2(5HHuranone (0.50 g; 1.48 mmol) in aqueous ammonia solution (30 mls; 28%) was stirred at room temperature for 2h, during which time the solid completely dissolved. The solution was evaporated to dryness,, and the residue extracted with dichloromethane (25 ml). The organic phase was dried over anhydrous sodium sulfate and evaporated to yield a red viscous oil. Chromatography on silica using ethyl acetate followed by ethyl acetate/methanol (4:1). gave a solid which upon recrystalhsation from light petroleum yielded 4-bromo-3-hexyl-5-hydroxy-5-hydroxymethyM,5-dihydropyrrol-2-one as colourless granules (O.16g; 36%), m.p. 134-135 °. vmax: 3304, 3256, 3185, 2961, 1670. 1589, 1441, 1350, 1136, 1069, 983 cm-1; (.max: 221 ((max 6,678), 196 (3,415) nm. 5-Ethyl-5-hydroxy-4-methy-1,5-dihydropytrol-2-one A mixture of 5-ethy!idene-4-methyl-2(5H)furanone (0,02 g; 0.162 mmol) in aqueous ammonia solution ( 5 m!; 28% w/w) was stirred at room temperature for 1.5 h during which time all of the furanone dissolved. The solution was WO 2004/016588 PCT/AU200J/001053 33 evaporated in vacuo to dryness eaving 5-ethyl-5-hydroxy-4-methyl-1,5-dihydropyrrol-2-one as a white solid (0.015g; 65%), m.p. 1B2-186°C (Found(HRESMS) m/z 184.067448 C6H11NO2Na+ requires 160 06815). vmax 3204, 29805 1698, 1664, 1633.5, 144si 1167, 1080, 1016, 983, 852, 769, 578. W 207 ((max23,180 ) nm. 1H n.m.r. ( (DMSO)-d6) 7 97, s, 1H. -NH; 5.55, s, 1H, C3-H; 3.18, s, 1H, C5-OH; 1.79, s, 3H, C4-Me; 1.69-1.52, m, 2H, C5-CH2-Me; 0.34, t, 3H, Me. 13C n mx ( (DMSO-d6): 7.9, 11.9, 29.2, 90.2, 121,7, 162, 171.6. 1-Benzyl-5-ethyl-5-hydroxy-4-methyl-1,5-dihydropyrrol-2-one A solution of 5-ethylidene-4-methyl-2(5H)furanone (0124 g; 10 mmol) in benzytemine (0.128g; 12 mmol) was the left to stand at room temperature for 72 hrs, during which time a solid precipitated from the reaction. The reaction mixture was triturated with CH2Cl2/petrol (1:3) and the precipitated solid was filtered and recrystallised from EtOAc/petrol to yield1-benzyI-5-ethyl-5-hydroxy-4-methyi-1,5-dihydropyrrol-2-one as colourless crystals m.p. 129-132° (70%). vmax 3247, 3082, 2964, 1669, 1638. 1496, 1353, 1101, 1053, 902, 708 cm-1. W 276 ((max 2,101), 237 (16,321), 243 (39,646) nm.1H n.m.r. ( (CDCIa): 7.4-7.24. m, 5H, Ph; S.79, ,s, 1H, C3-H, 4.46. 2 d, J 15 Hz, -CH2Ph; 3.81, bs, C5-OH; 1.92, s, C4-Me; 1.83-168, m. C5-CH2Me, 0.34, t, J 7.51 Hz, C5-CH2Me-13C n.rn.r. ( (CDCI3): 6.8, 11.85, 26, 41.9. 94, 122, 127, 128, 128 5, 138f 159, 170. 5-Aminomethyl-4-heptyI-5-hyc(roxy-1,5-dlhydropyrrol-2-one 5-Bromo-5-bromomethyl-4-heptyl-2(5H)furanone (0.50 g; 1.47 mmol) was dissolved in liquid ammonia in a sealed tube, and left to stand at room temperature for 72 h. Ammonia was allowed to gradually evaporated leaving behind a yellow crystalline solid. The solid was dissolved in not ethylacetate (ca 25) ml to remove ammonium bromide and the clear filtrate was concentrated to a small volume (ca 7 ml), to yield 5-aminomethyl-4-heptyl-5-hydroxy-1,5-dihydropyrrol-2-one as a crystalline solid, (0.1 g; 34%); m.p. 176°C. vmax: 3370, 3248, 2956, 2926, 2855, 1674, 1627, 1469, 1350., 1227,1095, 1082, 954, 555 cm'1. (max, 208 ((max 6845), 291 (2754) nm. 1H n.m.r 6 (CDCI3): 7.53, s, -NH; 5.49, d, C5-CH2NH2, 3.35, 3H, m, -C5-OH and -CH2NH2. 2.23-2.0, m, CH2; 1,52-0.85, m, 13H, alkyl chain. 13C ( (CDCl3): 14, 22.4,26, 26.5, 28.9,29, 31.6, 66,73, 78, 120.6,167.5,171 6. WO 2004/016588 PCT/AU2003/001053 34 5-Bromomethyl-4-heptyi-5-hydro3cy-1-phenyM35-dihydropyrrol-2-one 5-Bromo-5"bromomethyI-4-heptyl-2(6H)furanone (0.51g; 1.5 mmol) was dissolved in dry aniline (5 ml). The mixture soon solidified; it was allowed to stand at room temperature for 24 h Dichloromethane (25 mls) was added to the mixture and the organic phase was washed with aqueous hydrochloric acid (2M) and brine. The dried (Na2SO4) organic phase was evaporated to yield a yellow solid (0.50 g, 91%). Recrystallisation from light petroleum gave 5- bromonnethyl-4-heptyl-5-hydroxy-1-phenyl-1,5-dihydropyrrol-2-one as colourless needles, m.p. 152-154°C. vmax: 3194, 2956, 1930, 2854,1676,1626, 1589, 1502, 1494, 1393, 1246, 1141, 836, 758, 692 cm-1. Xmax: 257((max 3947), 202 (27,313) nm. 1H n.m r. ( (CDCI3): 7.55-7.26, m, 5H, Ph; 5.79, s, C3-H. 4.52, 1H, C5-OH; 3.39, d, 2H, C5-CH2Br; 2 27-2,12, m, 2H, chain; 1.6-0.91, tn, 13H, chain. 13C n.m.r. ( (CDCl3). 14, 22.5, 25.6, 25.8, 29, 29.2, 30 4, 31.6, 121.6, 126f 126.7, 130,134.6, 163, 170.5. 1-Benzyl-5-bromomethyl-4-heptyl-5-hydroxy-1,5-dihydropyrroI-2-one A mixture of 5-bromo-5-bromomethyl-4-r\|eptyi-2(5H)furanone (0.51g, 1.5 mmol) in benzylamme (0,30g; 2.82 mmol) in ethanol (6 ml) was stirred at room temperature for 1 h. Dichloromethane (25 ml) was added to the reaction mixture and the organic phase was washed with aqueous hydrochloric acid (2M) followed by brine. After drying over sodium sulfate, the solvent was evaporated in vacuo to yield 1-benzy!-5-bromomethyl-4-heptyl-5-hydroxy-1,5-dihydropyrrol' 2-one as a viscous oil (0 52 g; 97%) which solidrfied on standing in the fridge. Colourless needles from light petroleum; m.p. 94-96°.Vmax: 3270, 3062, 3033, 2957, 2854, 1667, 1637, 1607, 1496, 1416, 1335, 1297, 1257, 1190, 1161, 1140,1109, 1030. 950, 884, 865,769 cm-1. ( max 251 (( max2391), 206 (18,974) nm. 1H n.m.r. ( (CDCI3): 7.36-7.28, m, 5H, Ph; 5.85, s, C3-H; 4.54 and 3,42,2d, 2H each, C5-CH2Br and CH2Ph; 3-42, bs, 1H, C5-OH, 2.31-2.15, m, 2H, CH2; 1.62-0 88, m, 13H, alkyl chain.13C, n.m.r. 5(CDCI3). 14, 22,5,25.5, 26, 29, 29.2, 30.87, 41.9, 122, 127,128.3,137 5.163, 171. Synthesis of 3-alkyl-5-halom ethyl ene-135-di hydro pyrrol-2-one 3-ButyI-5-dibromomethylene-1-phenyl-1,5-dihydropyrrol-2-ohe WO 2004/016588 PCT/AU2003/001U53 35 Phosphorus pentoxide was added to a solution of 3-butyl-5-dibromomethy!-5-hydroxy-1-phenyM,5-dihydropyrrol-2-one in chloroform. The resulting mixture was stirred overnight at room temperature and passed through a pad of Celite. The crude product was chromatographed on silica and recrystallised from light petroleum to yield 3-butyl-5-dibromomethylene-1-phenyl-1-5-dihydropyrrol-2-one as orange needles (78%), orange crystals from petrol. (Found(HRESMS) m/z 419.954622. C16H17Br2NONa+ (7eBr) requires 419.955896). W 202 ((max 8137), 195 (3850) nm. 1H n.m.r. BCCDCls): 7.22-7.17, m. 5H, Ph; 7.17, s, C4-H; 2 38-2.36, m, 2H, CH2; 1.65-0.96, m, C3-chain. 13C n,m r. ( (Cody. 13.6, 22,3. 25.2, 29,5,128.3,128.8, 132.1,139, 140. 171.8. 3-Hexyl-5-dibromomethylene-1-phenyl-1,5-dihydropyrrol-2-one 3-Hexy!-S-dibromomethylene-1-pheny[-1 5-dihydropyrrol-2-one was prepared from 3-hexyl-5-dibrornomethyl-5-hydroxy-1-pheriyl-1,5-dihydropyrrol-2-one as descnbed above. Yellow granules from petrol. vmax: 3378, 2957, 2925, 2854, 1692, 1598, 1501, 1492, 1445, 1122, 1081, 743, S77 cm-1. Xmax: 309 (emax 19,681) nm. 1H n m.r. ( (CDCl3): 7 4-7.17, m, 6H, Ph and H4; 2.37-2.34, m, 2H, CH2, 1.57-0.89, m, 11H, C3-cham. 1-Benzyl-3-butyI-5-dibromomethylene-1,5-dihydropyrro(-2-one 1-Benzyl-3-butyl-5-dibromomethyl-5-hydroxy-1,5-dihydropyrroI-2-one was dehydrated with P2O5 in CHCI3 at room temperature for 72 hrs. The mixture was filtered through celite and the solyent evaporated in vacuo to yield a viscous oil, which solidified on keeping in a refrigerator. The solid was recrystallised from methanol/water to yield 1-benzyl-3-butyI-5~ dibromomethylene-1,5-dihydropyrrol-2-one as colourless plates, m.p. 56-58°C (91%). vmax: 2954, 1706, 1626, 1495, 1453, 1494, 1435, 1386, 1352, 1269, 1235, 1095, 765 cm-1. (max 324 ((max 5985), 283 (16,201), 206 (10,972) nm.1H n.m.r. 5 (CDCI3) 7.3-7.07, m, 6H, Ph and H4; 5.26, s, CH2Ph, 2.4-2.36, m, 2H, CH2; 1.6-0.95, m, C3-cham. 13C n.m.r. S (CDCI3): 13.7, 22,25, 29.6, 44.2, 74.7, 89.25,126, 127, 128, 132, 137.8,138 8, 140, 172.1 1-Benzyl-5-dibromomeihylene-3-hexyM,5-dihydropyrroI-2-one This compound was prepared according to the procedure described for 1-benzyl-3-butyl-5-dibromomethylene-1,5:dihydropyrrol-2-one. vmax 2960, 2848, 2923, 2854, 1696, 1592, 1496, 1453, 1354, 1316, 977, 830, 738, 630 cm-1. 1H WO 2004/016588 PCT/AU20O3/001053 36 n.mr. (CDCl3): 7.3-7.08, m, 5H, Ph, 7 26, s, 1H, H4; 5,26, 2H, -CHgPh; 2.4-2.36, m, 2H, CH2; 1.56-1.32, m, C3-chain. 1-Butyl-5-dibromomethylene-3-hexyI-1,5-dihydropyrrol-2-one This compound was prepared according to the procedure described for 1-benzyl-3-butyl-5-dibrornomethyIene-1,5HdihydropyrroI-2-one. Yield (30%). vmax: 2956, 2928, 2858, 1705, 1586, 1452,1360, 1335, 1194, 1135, 1058, 846, 829 741 cm-1 ; (max 290 ((max 18,927), 203 (9,409) nm. 1H n.m.r. 8(CDCI3)- 7.0, s, 1H, C4-H,; 3.99-3 93, t, 2H, -CH2N-; 2.3, t, -CH2- chain; 1.56-0 88, rn, 16H, chain 13C n.m.r. 5 (CDCl3), 13.7, 14, 19.7, 22.4, 25, 27/29, 31.4, 32.1, 40.6, 132,137,139,140.6,172.0. 5-Dibromomethylene-3-hexyl-1,5-dihydrapyrrol-2-one This product was prepared by the dehydration of 5-dibromomethyl-3-hexyI-5-hydroxy-1,5-dihydropyrroI-2-one as described above, m.p, 103-105°. 5-EthyIidene-4-methyl-1,5-dihydropyrrol-2-one 5-Ethyl-5-hydroxy-4-methyl-2(5H)pyrrolinpne was dehydrated to 5-ethylidene-4-methyl-1,5-dihydropyrrol-2-one with P2O5 in dichloromethane. vmax: 3158, 3093, 3036, 1670, 1495, 1434, 1397, 1381, 1356, 1279, 956, 867, 796, 639. (max: 173 ((max 33,010) nm. 1H n.m.r. 5(CDCI3): 8.94, s, 1H, -NH; 5.85, 1H, s. C3-H; 5 33, q, J 7 53 Hz, -=CHCH3, 2.1, s, 3H, C4-Me; 1,92, d, J 7.53, C5-Me-CH= 13C n.m.r. 5 (CDCI3) 11.7, 12.9,107,120 5, 140,148,172.0. 1 -BanzyI-5-ethylidene-4-methyM ,5-di hydro pyrroI-2-ons 1-Benzy!-5-etriyIIdene-4-methy!-1,5-dihydropyrrol-2-one was prepared by the dehydration of 1-benzyl-5-ethyl-5-hydroxy-4-methyl-1,5-dihydropyrrol-2-one as described before. (max: 206nm. 5-Bromomethylene-4-hepty-1-pheny\|-1,5-dihydropyrrol-2-one p-Toluenesulfonic acid (0.05g) was added to a solution of 5-bromomethyi-5-hydroxy-4-heptyl-1-phenyi-1,5-dihydropyrrol-2-one in toluene The mixture was refluxed for 1/2h and after cooling, washed with sat NaHCO3. The organic phase was dned over Na2SO4, and evaporated to yield an.E,Z mixture of 5-bromomethylene-4-heptyl-1-phenyl-1,5-dihydropyrroJ-2-one as a colourless oil which solidified on standing, m.p. 63-65°. vmax: 3414, 3080, 2952, 2853, 1695, WO 2004/016588 PCT/AU2003/001053 37 1627, 1597. 1499, 1446, 1382, 1269, 1074, 907, 831 cm"1; (max. 317 ((max 22,834), 278 (43,910), 204 (46,925) nm; 1H n.m S(CDCI3): 7.4-7.24, nv5H, Ph, 6.04 and 5.94, 2 s, 1H each, =CHBr and C3-H; 2.45, m, 2H, CHZ; 1.65-0.9, m, 13H, alkyl chain. 1 -Benzyl-5-bromomethylene-4-hepty 1-1,5-dihydropyrroI-2-one 1-Benzyl-5-bromomethyl-4-hepy!"5-hydrpxy-1,5-cIihyclropyrrol-2-one dehydrated smoothly to an E and Z mixture of 1-benzyl-5-bromomethylene-4-heptyl-1,5-dihydropyrrol-2-one upon heating a solution of 1-benzyl-5-bramomethyl-4-heptyl-5-hydroxy-1,5-dihydropyrrol-2-one with p-toluenesulfonic acid in toluene; m.p. 52-55°; vmax: 3096, 2927, 2857, 1704, 1630, 1387, 1357, 954, 855, 843 cnV\W: 319 ((max10,220), 276 (19,433), 206 (17,040) nm; 1H n.m,r. 5 (CDCI3): 7.29-7.15, m, 5H, Ph, 6.15 and 5.98, 2s, each 1H, =CHBr and C3-H; 2.39, m, 2H, CH2; 1.7-0.89, m, 13H, alkyl chain. Reaction of N-(2-HydroxyethyI)-3-butyl-2{5H)pyroIinone with acetic anhydride and Triethylamine N-(2-Acetoxyethy1)-3-butyl-5-(dibrombmethylene)-2(5H)pyrolinone A mixture of N-(2-hydroxyethyI)-3-butyl-5-clibroniomethyl-5-hydroxy-2(5H)pyrrolinone (0.2g, 0.54 mmol), acetic anhdride (0.44g; 4.4 mmol) and tnethylamme (0.44g; 4.4 mmol) in dry dichloromethane (10 ml) was refluxed for 2 hr. After cooling to room temperature, the mixture was washed with aqueous sodium bicarbonate and bnne The organic phase was dried over anhydrous sodium sulfate and evaporated to yield a viscous oil. 1H n.m.r showed it to be a mixture of the mono- (88%) and di-acetate (12%) derivatives. Chromatography on silica using EtOAc/CH2C!2 (5:1) as an eluent yielded 5-acetoxy-N-(2- acetoxyethyl)-3-butyl-5-dibrornomethyl-2(5H)pyrrolinone (12%) as an oil. vmax- 2957, 2931, 2875, 1766, 1720, 1433, 1369, 1236,.1044, 1013. 855, 707 cm-1 W: 217 ((max 1692), 268 (738) nm. 1H n m.r. 6(CDCl3) 0.91 (t, 3H, CH3); 1.38 (m, 2H, CH2); 1.55 (m, 2H, CH2); 2.05 and 2.10 (each s, 3H, CH3), 2.34 (m, 2H, CH2); 3.61 (m, 1H, NCH2CH2); 3.64 (m, 1H, NCH2CH2); 4.27 (m, 2H, NCH2CH3); 6.26 (s, 1H, CHBrs); 6.B3 (&, 1H, H4). 13C n.m.r. 5(CDCI3) 13.7, 20 S, 21.2, 22.1, 24.9r 38.7, 44.1, 61.5, 94.1,134.2, 144.3, 168.4, 170.6, 171 0. N-(2-acetoxyethy)-3-butyi-5-dibromomethyl-5-hydroxy-2(5H)pyrrolinone (88%) 1H n.m.r. 8(CDC!3) 0.93 (t, 3H, CH3); 1,38 (m, 2H, CH2); 1,55 (m, 2H, CH2); WO 2004/016588 PCT/AU2003/001053 38 2 21 (s, 3H, CH3); 2.34 (m. 2H, CHZ), 3.27 (m, 1H, NCH2CH2); 4.04 (m, 2H, NCH2CH2), 4,30 (s, 1H, OH); 4.62 (m, 1H, NCH2CH2); 5.85 (s, 1H, CHBr2); 6.73 (s, 1H, H4). 13C n.m.r. 5(CDCf3) 13.7, 20.9, 22.2, 24.9, 29.3, 38.1, 45.9, 62.5, 91.0, 137.4,143.3,170.4,171.9 Dehydration of N-(2-acetoxyethyl)-3-buty!-5-dibromomethyl'5-hydroxy-2(5H)pyrrolinone with p-toluenesulfonic a'cid in toluene gave quantitatively N-(2-acetoxyethy!)-3-butyl-5-(dibrornomethyleiie)-2(5H)pyroJinone. vmax 2957, 2929, 2870, 1744, 1705, 1441; 1368, 1229, 1177, 1161, 1130, 1035, Q30. 764 cm-1. 1H n.m.r. 5(COCi3) 0.93 (t, 3H, CH3); 1,36 (m, 2H, CH2), 1.55 (m, 2H, CH2); 2 02 (s, 3H, CH3); 2.32 (m, 2H, CH2); 4.25-4.31 (m, 4H, NCH2CH2); 7.05 (s, 1Ht H4). 13C n.m.r.'S(CDC13) 13-7, 20.7, 22.3, 25.1, 29.5, 39.&, 62.3, 73.8, 132.4, 138.6,140.3,170 6,172.0. Hydrolysis of N-(2-acetoxyethyl)~3-butyl-2(5H)pyrolinone N-(2-hydroxyethy!)-3-butyI-5-(dibromomethyIene)-2(5H)pynofinone A solution of potassium carbonate (1g) in water (3 ml) was added dropwise to a solution of N-(2-acetoxyethyl)-3-butyl-(dibromornethylene)-2(5H)pyrohnone (0.2g, 0.51 mmol) in methanol (7ml). After stirring the mixture at room temperature for 20 mins, methanol was removed in vacuo and the product extracted with ethylacetate (2 x 40 ml). The resulting extracts were combined, washed with brine, dried (Na2SO4), and evaporated to yield an oil (0 18 g; 94.5%), which solidified' upon standing in the fridge. Crystallisation from light petroleum gave N-(2-hydroxyethyI)-3-butyl-5-dibromom&thyten9- 2(5H)pyroimone as colurless granules, nrp. 48-50° max 3404, 2957, 2930, 2880, 1720, 1651, 1465, 1348, 1207, 1081, 1054, 1018, 936, 850, 716 cm"1. vmax 206 ((max 25.389), 239 (6,753), 288 (2,186) niTL 1H n.m.r. SCCDCb) 0.91 (t, 3H, CH3)l 136(171, 2H, CH2): 1.54 (rn, 2H, CH2); 2.29 (m; 2H, CH2); 3.B3 (m, 2H, NCH2CH2); 4.20 (m. 2H, NCH2CH2), 7.02 (s, 1H, H4), 13C n.m r S (CDCI3) 13.7, 22.3, 24.9, 29.5, 43.3, 46.8, 61.9, 74.5,132 3, 138.6,140.5, 173.2 Synthesis of 5-phenylaminomethylene-2(5H)furanone 4-Bromo-5~phenylaminomethylene-2(5H)furanone A solution of 4-brorno-5-bromomethylene-2(5H)-fu'anone (0.30 g; 0,79 mmol) was dissolved in aniline (5 ml), and left to stand at room temperature for 3 hrs, WO 2004/016588 PCT/AU2003/OOJ053 39 dunng which time the mixture solidified The solid was triturated with CH2Cl2/petroi (1.1; v/v, 20 m!) and filtered. The resulting solid was dned and recrystallised from ethanol to yield 4-bromo-5-pheny!aminomethylene-2(5H)furanone (0,24g, 49%) as yellow needles, m.p. 200-202 (decomp). (Found (HRESMS) m/z 287.963053. CnH8BrNO2Na+ (79Br) requires m/z 287 963840). v max 3233, 3127, 1730, 1697, 1595, 1498, 1276, 1195, 932, 798, 756 cm-1. (max 397 nm ((max 50,686); 246 (12,769), 202 (15,961), 1H n m r. 5 (CDCI3): 9.99, d, J 10.44 Hz, 1H, -NHPh; 7.31-6.99, m, Ph; 7.07, dr J 10.44 Hz, 1H, -CHNHPh; 6.16, s, C3-H. 13C n.m.r. 5 (CDCl3): 109.0 116 2, 117.9, 129.9, 129.6,133.9; 167.5. 5-Phenylaminomethylene-4-bromo-3,-butyl-2(SH)-furanone A solution of 4-bromo-3-butyl-5-bromomethylene-2(5H)-furanone (0.25 g; 0.81 mmol) in aniline (0.082 g; 0.88 rnmal) was left to stand at room temperature for 72 h. The mixture was diluted with CH2CI2 (50 mi), washed with aqueous hydrochloric acid (2M) and dned over anhydrous sodium sulfate. The solvent was removed in vacuo leaving behind a brown viscous oil (0.29 g). The crude product was chromatographed on silica using dichloromethane to yield 5-phenylammomethylene-4-bromo-3-butyl-2(5H)-turanone as a yellow solid 1H n.m.r ( (CDCl3): 7.40-6.80, m, 5H, Ph; 6.70, d J 12.5 Hz, =CH(NH)Ph; 2.42-2.40, m, 2H, CH2-chain; 1.7-1.2, m, 4H, CH2-chain; 0.95, t J 7.3 Hz, CH3. (Found (HRESMS) m/z 344.021931. C15H16BrNO2Na+ (79Br) requires m/z 344.021891). 4-Bromo-5-phenylaminomGthylene-3-hexyl-2(5H)furanone A mixture of 4-bromo-3-hexyl-5-bromomethylene-2(5H)-furanone (0.50g; 1.48 mmol) and aniline (1ml) in ethanol (10 ml) was heated at reflux for 2 h. After cooling to room temperature, the mixture was evaporated to dryness and the residue extracted with dichloromethane (20 mi). The organic phase was washed with aqueous hydrochloric acid (2M) and dried over anhydrous sodium sulfate. Removal of the solvent and recrystallisation of the solid from light petroleum gave 3-bromo-5-phenylaminomethyIene-3-hcxyl-2(5H)furanone (0.50g. 100%) as yellow needles; m.p. 147-148°C. vmax 3242, 3161, 3109, 29212, 2842, 1728, 1683, 1600, 1581, 1'SOO, 1350, 1236, 1055, 960, 750, 673 cm-1. (max: 394 ((max 26,287) , 247 (8002) nm. 1H n.m.r 5(CDCI3): 7.32-6.97, m, 5H, Ph: 6.9B, s. -NMPh; 6.73, s, C5 =CH-NHPh; 2.4, t, -CH2cham; 1.61- WO 2004/016588 PCT/AU2003/001053 40 0 88, m, 11H, chain. 13C n.m.r.5(CDCI3): 14.0, 22.0, 24.8, 27.6, 29, 310, 103.0, 113,0,115.0,122.5, 124 0, 129.5 129.5, 131.0.139.9, 167.0. 5-Pheny[aminomethylene-4-heptyl-2{5H)furanone 5-Phenylaminomethyi-4-heptyi-S-hydroxy-2(5H)pyrra!inone 5-Bromomethylene-4-heptyI-2(5H)furanone (0.44g; 1.61 mmol) was dissolved in dry aniline (2 ml) and left to stand at room temperature for 24 h. Dichloromethane (10 ml) was added to the reaction mixture and the organic phase was washed with aqueous hydrochionc (2M) followed by water. After drying over sodium sulfate, the solvent was evaporated off to yield a pale yellow solid. The crude product was chromatographed on silica column using dichloromethane followed by CH2CI2/Et0Ac (2:1; v:v) as the eluents to yield 5-phenylaminomethyi-4-hepty!-5-hydroxy-2(5H)furanone (0.43g; 88%) as a pale yellow solid, m.p, 172-174°C. vmax: 3192, 3037, 2957, 2931, 2953. 1676, 1643, 1598, 1502, 1493, 1336, 1250. 1160, 923, 757 cm'1. (max 278 ((max 7188). 203 (8609) nm. 1H n.m.r, ( (CDCI3): 7.53-7.26, 6H, Ph and -NHPh; 5.73, s, 1H,C3-H, 5.11,s, 1HT\|C5-OH; 3 37, d, 2H, -CJigNHPh; 2.2-2.0, m, 2H, -CHz-chain; 1.25-0.91. mt 13H, Chain. 13C n.mr.5 (CDCIa): 14.0, 22 6, 25.5, 25.3, 25.8, 29 0, 29.2, 30.4, 31 6, 93.4, 121.8, 126.0, 126.7, 129.0,134.6, 163.0,170.4. 5-PhenytaminQmethytene-4-heptyl~2(5H)furanone A sample of 5-phenyIaminomethyl-4-heptyl-5-hydroxy-2(5H)furanone was dehydrated using p-toluenesulfonic acid in toluene to yield an E and Z mixture of 5-phenylaminomethylene-4-heptyI-2(5H)furanone as a colourless oil which solidified on standing in the fridge. vmax 3088(-NH), 3052, 2927, 2856, 1712, 1626, 1598, 1499, 1454, 1264, 1195, 759, 699 cm-1 (max 292 ((max 7623), 204 (4728) nm. 1H n.m.r. ( (CDCIa) 7.4-7.25, 6H, Ph and -NHPh; 6.19-6.1, d, 1H, C3-H; 5.93-6.0, 1H, d, C5- =CMNHPh; 1.68-0.90, 15H, chain. 13C n.m r.6(CDCI3). 14, 22.5, 26.4, 28 1, 28.9, 29.0, 29.2, 30.0, 31.6, 31 7, 88.7, 93.0, 118.5, 1226, 127.8, 128.2, 128.4. 128.6, 128.7, 129.3, 129 5, 134.0, 135.0,142.0,143.0, 152.0, 153.2,168.0. 4-Methyl-5-(1-phenylamIno-ethylidene)~5W-furan-2-one A solution of 5-ethylidene-4-methyl-2(5H)furanone (0,31 g; 2.5 mmol) in aniline (0 26 g; 2.75 mmol) was left to stand at r.t for 3 hrs, during which time a solid precipitated from the reaction. The reaction mixture was triturated with WO 2004/016588 PCT/AU2003/001053 41 CH2CI2/petro] (1:1) and the solid filtered and recrystallised from EtOAc/petroi to yield 5-ethyl-5-hydroxy-4-inethyJ-1 -phenyl-1,5-dihydropyrrol-2-one as colourless crystals (70% ); m.p. 97-100°. vmax: 3287, 1884, 1704, 1530, 1496, 1353. 1101, 1053, 971, 897, 790, 756, 688. 638 cm"1. (max 273 ((max 15,256), 226 (16,382), 243 (39,646) nm. 1H n.m r. S (DMSO-de) 10.11, S, 1H, -NH; 7.57, d, 2H, ArH, 7.30, t, 3H, ArH; 6.08, S, 1H, C3-H; 3 27, s, 3H, CH3; 1-96, s, 3H, CH3. 13C ri.m.r. ( (CDCl3): 20.9, 119.6, 123.7, 123.8, .129.1, 139.3. 142.9, 163.1. 170.4. Synthesis of 5-arylamino and arylalkylamino-2(5H)furanones 4-bromo-5-benzylam!ao-5-bromomethyl-2(5H)furanone Benzyl amine (0.10 g, 0.95 mmol) was added with stirring to an, ice-cooled solution of the 4-bromo-5-(bromornethyIene)-2(5H)furanone (0.16 g, 0.64 mmol) in dichloromethane (10 ml). The mixture was stirred at room temperature for 2.5 h, washed with aqueous hydrochloric acid solution (1M, 10 ml), dried (Na2SO4), and evaporated to yield a brown oil. The crude product was chromatographed on silica using dichloramethane/ethyl acetate (1:4; vv) as the eiuent and recrystallfsed from dichloromethane/light petroleum to yield 4~ bromo-5-benzyIamino-5-brornomethy!-2(5H)fiiranorte as orange flakes m.p. 137-139 ° (Found (HRESMS) m/z 381.901032. C21H11 Br2NO2Na+ (79Br) requires 381.904812). vmax 3256, 1674, 1655, 1431, 1413, 1352, 1072, 1054, 699 cm-1. (max 257 ((max 2879) nm 1H n.m.r. ( (CDCl3): 7.38,d, J 11 Hz, 1H,-NHCH2-; 7.37-7.29, m, Ph; 6.38, s, C3-H, 4.65, d, J 15 Hz; 1H, -CH2Bn 4.44. d, J 15 Hz, 1H, -CH2Br and 3.58-3.44, dd, J 15 Hzt CH2Ph. 13C n rn.r.5 (CDCI3): 30.6, 42.8, 53.0, 92.2, 128.0,128.2,12819, 137.0, 142.0, 168.0 4-Bromo-5-benzybminO-5-bromomethyl-3-hexyl-2(5H)furanone Benzylamine (0.32g; 2.96 mmol) was added with stirring to a solution of 4-bromo-3-hexyl-5-bromomethylene-2(5H)-furanone (0.50 g- 1 48 mmol) in ethanol (6 ml). The mixture was stirred at room temperature for 1 h and evaporated to dryness.The residue was extracted with dichloromethane (20 ml) and the dichloromethane extract washed with aqueous hydrochloric acid (2M). After drying over anhydrous sodium sulfate, removal, of the solvent gave a thick viscous oil, Column chromatography on silica gel using dichloromethane followed by dichloromethane/ethy! acetate (19:1) as the eluents afforded 4-bromo-5-benzylamino-5-bromomethyl-3-hexyl-2(5H)ftiranone (0.36g; 56%) as a WO 2004/016588 PCT/AU2003/001053 42 Viscous oil, m.p. 72-75°.vmax3277, 3065, 3032, 2954, 2928, 2657,1681, 1496, 1411, 1355, 1151, 1064, 1104, 1030, 988, 907, 726, 698. (maX: 277 (( max 39,542), 205 (38,034) nm. 1H n.m.r. 5(CDCl3): 7.4-7.26, m,' Ph; 4.8-4.74. d, and 4.4, d, C5-CH2Bn 3.6 and 3.53, d, CB-NHCH2Ph; 2.42-2.33. m. -CH2, chain; 1.56-0,85, m, 11H, chain. 13C n.m.r. ( (CDCI3): 22.0, 25:0. 27.0, 28.8, 31.4, 42.9, 46.7, 49.5, 90.6, 91.6, 127.0, 128.0, 129.0, 136.0, 136.7, 136.9, 138 0, 140.0,144.0,168.0,170.6 5-Phenylamino-3,5-dimethyI-2(5H)-furanone Method A A solution of 3,5-dimethyl-5-hydroxy-2(5H)-furanone (0.13g; 1.02 mmol) in dry aniline (2 mls) was stirred at room temperature for 1 hr. A thin layer chromatography analysis of the mixture (developing solvent; CH2CI2) indicated completion of the reaction as indicated toy the disappearance of the starting material. Dichloromethane (25 mls) were added to the mixture and the solution washed with aqueous hydrochloric acid solution (1M, 3 x 20 mls). The organic layer was dried over anhydrous sodium sulfate and evaporated to yield 5-phenylarnino-3,5-dimethyl-2(5H)-furanone as a viscous oil which solidified on keeping (0 013 g), A sample was recrystallised from dichloromethane/light petroleum to yield the furanone as colouriiees needles v max 3360, 3088, 2965, 1770, 1601, 1570,1536,1294> 1246, 1132,1040, 999, 867, 756, 697 cm-1. W 236 nm. 1H n.m.r. ( (CDCl3): 7.18, t, 2H Ph; 6.90, t, 1H, ArH, 6.89, s, 1H, H4. 6.83, d, 2H, ArH; 4.24, bs, 1h, OH; 1,91, s, 3H, C3-Me;1.75. s, 3H, -Me. 13C n.m.r. ( (CDCI3): 10.4, 26.2, 95.5, 121.3, 122.7, 128 9, 132.4, 133.5, 141.9, 148.8,156.5, 171.9. Method B A mixture of 3,5-dimethyl-5-hydroxy-2(5H)-furanone (0.13g; 1,02 mmol) and aniline (2 ml) in dry toluene (10 ml) was refluxed for 5h. The mixture was cooled and evaporated. The residue was dissolved in dichloromethane (25 ml) and the solution washed with aqueous hydrochlonc acid solution (1M; 3 x 20 ml). The organic layer was dried over anhydrous sodium sulfate and evaporated to yield 5-phenylarnino-3,5-dimethyl-2(5H)-furanone as a viscous oil. The crude product was chromatographed on silica using dichloromethane/ethyl acetate (19'1) as the eluent (Yield 58-0%). 5-Phenylamino-5-methy!-4-phenyl-2(5H)-furanone WO 2004/016588 PCT/AU2003/001053 43 A mixture of 5-hydroxy-5-methyl-4-phenyl-2(5H)-furanone (0.13g; 1.02 mmol) and aniline (2 ml) in dry toluene (10 ml) was refluxed for 5h. The mixture was cooled and washed with aqueous hydrochloric acid solution (2M; 3 x 20(mls). The organic layer was dried over anhydrous sodium sulfate and evaporated to yield a viscous oil. The crude product was chromatographed on silica using dichloromethane/ethyl acetate (19:1) as the eluent and recrystallised from dichloromethane/Iight petroteurn to yield 5-phenyiamino-5-rnethyI-4-phenyI-2(5H)-furanone (0.10 g; 72%) as colourless flakes m.p. 15S-160°Cvmax: 3355, 1724, 1608, 1534, 1501, 1320, 1291, 1376, 1030. 943, 846. 770, 756! 691, 639 (max 276((max 7056), 238 (5615)nm. H n.m.r. S(CDCl3): 7.94-7 44, m' 5H, -Ph; 7.14-6.82, m, 5H,Ph; 6.4, s, 1H, C3-H; 4.53, bs, 1H, -N.HPh; 1.9, s, C5-Me. 13Cn.m.r.5(CDCl3) 117.3,120,122,5,128, 129,5,131, 142,159,166,170. 5-BenzylaminomethyI-3-methyl-2{5H)furanone Phosphorus pentoxide (2g) was added to a solution of 3.5-dimethyl-5-hydlraxy-2(5H)-furanone (0.50g; 2.15 mmol) in dichtoromethane (25 ml). The mixture was refluxed for 2h and the cooled solution was filtered through celite and evaporated in vacuo to yield 3-methyl-5-methylene-2(5H)-furanone as a colourless oil (0.37 g; 82%) The methylene product was dissolved in dichloromethane (5 ml) and benzyiamme (1.15 g; 10.8 mmol) was added at room temperature. The mixture was stirred at room temperature for 1 h. After evaporation of the solvent the crude product was chromatographed on silica using dichloromethane/light petroleum as the eluent to yield 5-benzylamtnomeihyI-3-methyI2(5H)furandne as a colourless oil (0.12 g; 26%). vmax: 2929, 2854, 1788, 1747, 1715, 1618, 1456, 1388, 1373, 845, 712 cm"1. W: 308 nm ((max 1462), 260 (5243).1H n.m.r.S (CDCl3): 7.29-7.21, m, 6H, Ph and-NHCH2Ph; 6.65, s, 1H, C4-H; 4.82,s, 2H, -CH2Ph; 4.70, d, -CHgNHPh; 2.02, s, C3-Me. 13C n.m.r. 5 (CDCI3): 10,8, 25.9, 42.9, 95.0.105.3, 126.9,127.1, 128.5,131.2,134.2,137.2, 148.4. Side-chain functionalization 3-(1-Bromohexyl)-1-butyl-5-dibromomethylene-1,5-dihydropyrroI-2-one N-Bromosuccinimide (0.32g; 1.79 mmol) was added to a solution of 1-butyl-5-dibromomethyl-3-'hexyl-1(5-dihydropyrrol-2-one (0.64 g; 1.63 mmol) containing few crystals of benzoyl peroxide in CCl4 (25 ml). The mixture was heated at reflux under a 100 watt fluorescent lamp for 24 h. The reaction mixture was WO 2004/036588 PCT/AU2O03/001053 44 cooled and passed through a pad of Celite. The filtrate was evaporated to dryness to yield a brown oil which was chromatographed on a silica column using CH2Cl2/petrol (1;1) as the eluent to yield 3-(r-Bromohexy!)-1-Butyl-5-dibromomethylene-1,5-dihydropyrml-2-one (0,46 g; 59.8%) as a pale yetlow oil. (Found): HRESMS): m/z 483.849575. C15H14NB4O3) requires 483,651758 vmax 3017,2950,1709ri598,1593, 1480, 12,15, 1194, 845, 695, 668 cm-1. (max: 326 ((max 4070) nm 1H n.m.r. 8(CDCI3)- 7.28, s, 1H, H4; 4.7$, t, -CHBr-chain; 2.19-2.11,m, -CH2-Chain; 1.53-0.98, m, alky, chain. 13C n.rn.r. ( (CDCI3): 13.12, 20.95, 26.8, 39, 43.9, 79.5, 95,128.6, 128.9.129.4, 133.8, ,134.5, 138.2, 139.6, 168.7 3-(1 '-Bromobutyl)-! -butyl-5»dibromomethylene-1,5-dihydropyrrol-2-one N-Bromosuccinimide (0 32g; i .79 mmol) was added to a spiution of N-butyl-5-dlbromomethyl-3-hexyl-2(5H)pyrrolinone (0.64 g; 1.63 mmol) containing few crystals of benzbyl peroxide (0.01 g) in CCI4 (25 ml). The mixture was heated at reflux under a 100 watt fluorescent lamp for 24 h. The reaction mixture was cooled and passed through a pad of Cel'rte. The filtrate was evaporated to dryness to yield a semi-solid (0.69g) which was chromatographed on a silica column using CH2Cl2/petrol (1:1) as the eluent to yield 3-(1'-bromobutyl)-1-butyl-5-dibromomethylene-i.s-dihydropyrrbl-2-one (0.4Bg; 60%) as a pale yellow oil. (max 2930, 2957, 2871, 1705, 1584, 1357, 1192, 1055, 902, 769, 651 cm-1; (max: 325 ((max 11,669), 202 (9,879) nm. 1H n.m.r, 5(CDCl3): 7.29, d, 1H, C4-H; 4.76, t. 1Ht C3-CiHBr- chain; 3.98,t, 2H, >NCH2-; 2.10, m, -CH2-chain; 1.58-0 93. m, 12H, chain; 13C n.m.r. 5(CDCI3): 13-781 13-96, 19,8, 22.4, 27.4, 31, 32.2, 37. 41, 43.9, 98.7. 132.5, 138 2, 140, 169.0, 3-(1 '-Bromobuty\|)-5-dibromomethylene- N-phenyl-1,5-dihydropyrrol-2-one N-Bromosuccinimide (0.056g; 0.316 mmol) was added to a solution of 5-dibromomethyl-3-buty!-1-phenyl-1,5-dihydropyrroI-2-one (0-64 g; 1.63 mmot) containing few crystals of benzoyl peroxide (0.01 g) in CCI4 (10 ml). The mixture was heated at reflux under a 100 watt fluorescent lamp for 24 h. The reaction mixture was cooled and passed through a pad of Celite. The filtrate was evaporated to dryness to yield a brown oil (0.17g) which was chromatographed on a silica column using CH2CI2/petrol (1:1) as the eluent to yield 3-(1'-bromobutyl)-5-dibromomethylene-1-phenyM,5-dihydropyrroI-2-one as a pale viscous oil (0.10g). (Found.'HRESMS) m/z: 483.849575, O 2004/016588 PCT/AU2003/00I053 45 C15H14Br3NONa+ (Br79) requires 483.851758. vmax: 3017, 2950, 1709, 1598, 1593, 1480, 1215, 1194, 1122. 845, 756, 695, 668 Cm-1. (.maj(; 326 ((max 3,896), 202 (5,566) nrn. 1H n.m.r. 8CDCl3): 7.45, m, 6H. Ph and C3-H; 4.86, t, 1H, C3-CHBr- chain; 2.16, m, -CH2 chain: 1.53-0.98, m, 5H, alkyl chain. 13C n.m.r.5(CDCi3): 13, 21, 26.8, 39, 43, 79.5, 95, 107. 128.6, 129.4, 134, 134.5, 138, 139.6,169. N-Phenyl-3-(1-hydroxybutyl)-5-dlbromomethylene-2(5H}pyrrolinone A solution of N-Phenyl-3-(1-bromobutyl)-6-dibromomethyIene-2(5H)pyroIinone (0.0194mol) in, DMSO (60 ml) containing few drops of water was left to stand aside at room temperature for 6 days. The mixture was diluted with dichioromethane (100 m!) and the resulting solution washed with brine (3 x 120 ml). The organic phase was dried over anhydrous sodium sulfate and evaporated to yield a pale yellow oil (9.08g). The Crude product was purified on a silica column using initially dichioromethane followed by dichloromethane/ethyl acetate to afford N-phenyI-5-dibrommeihylene-3(1 -hydroxybutyI)-2(5H)pyroltnone (6.83g; 88%) as pale yellow needles (dichloromethane/light petroleum), m p. 93-95°. vmax: 3439, 3065, 2957, 2927, 2871, 1701, 1496, 1455, 1370, 1189, 1139, 1095, 1069, 1038, 945, 835, 763, 697 cm-1. (max203 (w 11,968), 313 (10,707) nm. 1H n.m.r. 5 (CDCI3) 0.97 (t, 3H, CH3), 1.39 (m, 2H, CH2); 1.79 (m, 2H, CH2), 4.61, m, 1H, H1'; 2.71, bs, 1H, OH, 7.23 (s, 1H, H4); 7.21-7.44 (m. 5H. ArH). 13C n.m.r. 5(CDCI3) 13.7, 14, 18.5. 37 8, 67.4,128.6,128.9, 128.3, 129.5, 131.5,134.5,139.8, 170.8. N-Phenyl-3-(1'-acetoxybutyl)-5-dibromomethylene-2(5H)pyrrolinone A solution of acetyl chloride (0.25 ml, 3.2 mmol) in dichioromethane (3 ml) was added dropwise to an ice-cooled solution of N-Phenyl-3-(1-hydroxybutyl)-5-dibromomethylene-2(5H)pyrolinone (0.1g, 0.25mmo!) in dichioromethane (10 ml) containing triethylamine (0.25 ml, 2.47 mmol). The mixture was stirred in ice for 1h and then at room temperature overnight. The mixture was poured into saturated sodium bicarbonate solution (20 ml) and extracted with dichioromethane (3 x 30 ml). The organic phase washed with water (3 x 20 ml), dried over anhydrous sodium sulfate and evaporated to yield a pale yellow oil (0.11g). The crude product was purified or) a silica column using dichloromethane/ethyl acetate (15:1) to afford N-phenyl-5-dibrommethylene-3(1'-acetoxybutyt)-2(5H)pyro!inone (0.1g) as a viscous oil Vmax: 2960, 2931, WO 2004/016588 PCT/AU20U3/001053 46 2873, 1753,1712, 1592, 1494, 1454, 1362, 1262, 1232. 1193, 1148, 1096, 1060, 836, 753, 698 cm-1. (max 281, 321 nm. 1H n.m.r. 5,(CDCI3) 0.96 (t, 3H, CH5): 1.38 (m,2H, CH2); 1 91 (mt 2H, CH^; 5.72, m, 1H, HV; 7.21 (s, 1H, H4); 7 2i-7.40(m,5H,ArH). Malonic acid mono-[1-(5-dibromomethyl6ne-2-oxo-1-phcnyl-235-dihydro-1H-pyrrol-3-yl)-butyl] ester A solution of N-Phenyl-3-(1-hydroxybutyi)-5-dibromomethy!ene-2(5H)pyrolinone (0-5g, 1.25 mmol) in dichloromethane (15 mi) containing triethylamine (0.25 ml, 2.47 mmol) was added drop wise over a period of 9h to an ice-cooled solution of malonyl dichlonde (0.36g, 25 mmol) in .dichloromethane (10 ml), The mixture was allowed to stand at room temperature overnight, washed with brine (3 x 20 ml), dned over anhydrous sodium sulfate and evaporated to yield a brown viscous oil. The crude product was purified on a silica column using ethyl acetate/methanol (1:4) to afford malonic acid mono-[1-(5-dibromornethyIerie-2-oxo-1-phenyl-2,5-dihydror1H-pyrrol-3-yl)-butyl] ester. (0.48g) as a viscous oil vmax 3470, 2959, 2873,1709, 1594, 1494, 1454, 1362, 1245, 1194,1148,1096, 1060. 835, 753, 698 cm-1, (max 271, 303 nm. 1H n.m.r 8 (CDCI3) 0.96 (t, 3H, CH3); 1.46 (m, 2H, CH2); 1.88 (m. 2H, CHa); 3.01, m, 2H, COCH2CO; 5 67, m, 1H. H1; 7.23, m, 2H, ArH; 7.43 (m, 3H, ArH); 7,59, s, 1H, H4. Preparation of 2(5H)pyrrolinone-polystyrene copolymer A mixture of styrene (7.13g), 3-(1'-Bromobuty!)-5-dibrarnornethylene- N-phenyl-1,5-dihydropyrrol-2-one (0.37g) and AlBN (0.026g) was degassed for 1/2h bby purging with argon gas and then heated at 65°C for 3h. After completion of polymerisation, the mixture was poured into methanol and precipitated polymer was filtered, washed extensively with methanol and dried to yield the copolymer (2.389, 32%) Surface attachment of 2(5H)pyrrolinone A layer of malonic acid rnono-[1-(5-dibromomethylene-2-oxo-i-phenyl-2.5-dihydro-1H-pyrrol-3-yl)-butyl] ester was covalently attached to a surface containing ammo groups by immersing the surface in a solution of 2(5H)pyrrolinone (2mg/ml) in acetonitrile/water containing, NHS, N-hydroxy succinimide. The mixture was shaken for 10 minutes and EDC, N-(3- WO 2004/016588 PCT/AU2003/001053 47 dimethylaminopropyi)-N'-ethylacrbodlimide hydrochloride, was added to the solution to give a final concentration of (2mg/ml), After shaking the solution for 24h, the surface was taken out of the solution and washed thoroughly with water and dried. The surface analysis was performed using XPS and %bromme was used a s a marker for determining the extent of covalent attachment. Biological activity of furanones Effect of furanones as inhibitor of AHL-mediated quorum sensing, Ai-2 pathway and growth of S. aureus Methods Gfp assay Briefly, the Gfp assay determines the relative effectiveness of a compound as an inhibitor of AHL mediated quorum sensing. The assay is dependent on a bacterial strain that carries a reporter plasmid. This plasmid expresses the green fluorescent protein (Gfp) in the presence of AHLs (2). The presence of a competitor will prevent AHL mediated Gfp expression of the reporter. The assay can be used to generate an index of inhibition for each compound. The results here, presented as good, moderate, or poor, are based on the index of each of the compounds as an inhibitor of AHL mediated quorum sensing using this bioassay. Attachrnent/Biofiim formation The ability of furanones to inhibit biofilm formation or attachment has been determined using a modification of the 96 well microtitre method described by Christensen et al. ((1)). The furanones are added to the wells of the microplate and the solvent is allowed to evaporate, leaving the furanones adsorbed onto the plate. Then a suspension of the monitor bacterium, Pseudomonas aerugmosa, is added to each well and incubated for 24h. Following incubation, the wells are nnsed to remove unattached or loosely adhered cells. The attached wells are fixed with formaldehyde and subsequently stained with cyrstal violet. Following extensive washing to remove the crystal violet, the wells are read at 600 nm. The attachment/biofilrn formation in the presence of the furanones is calculated as the percentage of the controls, which are not exposed to the furanones. WO 2004/016588 PCT/AU2003/001053 48 Two-Component signal transaction Assays Taz-1 Assay The Taz-assay carried out according to the method of Jin and Inouye (1993) with the following alterations. E coli RU1012 (pYT0301) were grownovernight In M9 medium at 37°C supplemented with 100 ug/ml ampicillin and 50 ug/ml kanamycin This overnight culture was then used to inoculate 50 ml M9 medium in side-arm flasks which were then incubated at 37°C and shaken at 180 rpm The OD610 of the growing cultures was monitored regularly and when the OD610 = 0.2 the cultures were placed on ice. Aspartate was added to side-arm flasks to give a final concentration of 3 mM (aspartate stock solution made up in M9 salts). The test compound or mixtures of, compounds were dissolved in ethanol and added to cultures to give the required final concentrations. Negative controls were prepared with equal volumes of ethanol. Cultures were then placed in a 37°C incubator and shaken for 4 hours (OD6io approximately 0.7) before being removed and put on ice. Samples were then removed for eta-galactosidase assays carried out according to the method of Miller (1972). V. harveyl bioassay for the detection of AI-2 activity The V. harveyi bioassay was performed as described previously (Surette and Bassler, 1998). The V. harveyi reporter strain BB170 was grown for 16 hours at 30°C with shaking in AB medium. Ceils were diluted 1 ;5,000 into 30°C prewarmed AB medium and 90 ul of the diluted suspension was added to wells containing supernatant. Furanones were added to the wells to achieve the desired final concentrations and the final volume in each well was adjusted with stenle medium to 100 ul. Ten ul of V. BB152 (AM-, AI-2+) supernatant was used as a positive control and 10 ul of E. coli DH5a supernatant or sterile media was used as a negative control. This strain of E. coll has previously been shown to harbor a mutation in the AI-2 synthase gene, ygaG, which results in a truncated protein wfth no Al-2 activtty (Surette et al. 1998). The microtiter plates were incubated at 30°C with shaking at 175 rpm. Hourly determinations of the total luminescence were quantified using the cherniluminescent setting on a Wallac (Gaithersburg, MD) model 1450 Microbeta Plus liquid scintillation counter. The V, harveyi cell density was WO 2004/016588 PCT/AU2003/001053 49 monitored by the use of a microplate reader (Bio-Rad, Hercules, CA). Activity is reported as the percentage of activity obtained from V. harveyi BB152 cell-free supernatant. While the absolute values of luminescence varied considerably between experiments, the pattern of results obtained was reproducible. Growth of Staphylococcus aureus Material and methods The growth of Staphylococcus aureus against furanones was tested in sidearm flasks. One percent of an overnight culture was added to the growth media, Nutnent Broth, containing furanones at the concentrations 1-50 ug/ml. The bacteria were incubated at 37C and growth was measured at 610 nm. The results of these experiments are summarised in the table 1. Table 1. Summary of activity for lactam and other N containing analogues as inhibitor of AHL-mediated quorum sensing, Al-2 pathway and growth of S. aureus. Compound AHL Al-2 (% of control) S. aureus (% of control) 26%, 50 ug/ml 57%, 10ug/ml 80%, 5ug/ml NE at 50 ug/ml 21 %, 50 ug/ml NE at 50 ug/ml +++ NE at 50 ug/ml 0% growth at ug/ml for 10hrs WO 2004/016588 PCT/AU2003/001053 50 39% (100ug/ml) 102% (25 Mg/ml) +++ 2% (50 Mg/ml) 104% (50 Mg/ml) 61 % (20 Mg/ml) No effect 50M9/mt ++ 50% (100 Mg/ml) No effect 50M9/ml +++ No effect 50MS/ml +++ +++ ++ Noeffect (50M9/ml) No effect (50pg/ml) WO 2004/016588 PCT/AU2003/001053 51 ++ No effect (50ug/mi) Other signal regulated Phenotypes 40-50% reduction 30% 40% reduction in in swimming reduction in cholera toxin motility in V. attachment production by V cholerae and V. by V. cholemo vulntficus vulnificus 40-50% reduction 50-80% reduction 30% 40% reduction fn in swimming in protease reduction in cholera toxin motility in V. production by V. attachment production by V. cholerae and V. Vulnificus by V. cholerae vulntficus vulnificus 40-50% reduction 30% 40% reduction in in swimming reduction in cholera toxin motility in V. attachment production by V- cholerae and V. by V. cholerae vulntficus vulnificus Christensen, G. D., W. A. Simpson, J. J. Younger, L. M. Baddour, F. F. Barrett, D. M. Meiton, and E H. Beachey. 1985. Adherence of coagulase-negative staphylococci to plastic tissue culture plates: a quantitative model for the adherence of staphylococci io medical devices. J. Ciin. Microbiol. 22(6):996-1006. Andersen, J. B., C. Sternberg, L. K. Pouisen, S. P. Bjom, M. Givskov, and S. Molm. 1998. New unstable vanants of green fluorescent protein for studies of WO 2004/016588 PCT/AU2003/001053 52 transient gene expression in bacteria. Appl. Environ. Microbiol. 64(6):2240-2246. Jin, T and M. inouye. 1993. Ligand binding to the receptor domain regulates the ratio of kinase to phosphatase activities of the signalling domain of the hybrid Eschenchia coll transmembrane receptor, Taz1. J. Mol. Biol. 232: 484-49 Miller, J. H. 1972 Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor,. N.Y. Surette, M. G., and B. L. Bassler. 1998. Quorum sensing in Eschenchia coh and Salmonella typhimurium. Proc. Nat!. Acad. Sci, USA 95:7046-7050 Surette, M. G., M. B. Miller, and B. L. Bassler. 1999. Quorum sensing in Eschenchta coh, Salmonella typhimurium, and Vibrio han/eyi: a new family of genes responsible for autoinducer production. Proc, Natl. Acad. Sci., USA 96:1639-1644. Any description of prior art documents herein is not to be taken as an admission that the documents form part of the common general knowledge of the relevant art. It will be appreciated by persons skilled in the art that numerous variations and/or modifications may be made to the invention as shown in the specific embodiments without departing from the spirit or scope of the invention as broadly described. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive. Any description of prior art documents herein is not to be taken as an admission that the documents form part of the common general knowledge of the relevant art. 53 WE CLAIM: 1 A method for the preparation of a compound of formula II wherein R1 and R2 are independently selected from the group H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted substituted or unsubstituted oxoalkyl, substituted or unsubstituted substituted or unsubstituted alkenyl, substituted or unsubstituted substituted or unsubstituted aryl or arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophihc, R3 and R4 are independently selected from the group H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, R5 is selected from the group consisting of H, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted alkylsilyl, substituted or unsubstituted substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophihc, or forms part of an ammo acid, or is a nucleoside, an ohgomer, a polymer, a dendnmer, a substrate or a surface, the method comprising reacting a compound of formula I 54 wherein R1 and R2 are independently H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophihc, R3 and R4 are independently H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl or arylalkyl, and R is hydroxy, halogen, and "===" represents a double bond, in which case R is absent, or a single bond, provided that at least one of R1, R2, R3 and R4 is halogen, with a compound of formula R5NH2 wherein R5 is selected from the group consisting of H, substituted or unsubstituted alkyl, hydroxy, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophilic, or forms part of an ammo acid, or is a nucleoside, an oligomer, a polymer, a dendnmer, a substrate or a surface 2 A method as claimed in claim 1 wherein R5 is a residue of a naturally occurring compound 3 A method as claimed in claim 1 wherein R5 is a biomolecule 4 A method as claimed in claim 3, where R5 is a coenzyme or cofactor 5. A method as claimed in any one of claims 1 to 5, wherein R5 is an oligomer or a polymer and the oligomer or polymer is a biomolecule 6 A method as claimed in claim 5, wherein R5 is a peptide or polyamide 7 A method as claimed in any one of claims 1 to 5, wherein R5 is a protein residue 6 55 8 A method as claimed in claim 7, where R5 is an enzyme or a receptor 9. A method as claimed in any one of claims 1 to 5, wherein R5 is an ohgomer or polymer comprising nucleic acid residues. 10 A method as claimed in claim 9, wherein R5 is a polynucleotide 11 A method as claimed in claim 10, wherein the polynucleotide is DNA or RNA 12. A compound of formula II wherein R1 and R2 are independently H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or iluorophilic; R3 and R4 are independently H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, R5 is selected from the group consisting of H, hydroxy, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophilic, or forms part of an amino acid, or is a nucleoside, an ohgomer, a polymer, a dendnmer, a substrate or a surface 56 13 A compound as claimed in claim 12, wherein at least one of R1, R2, R3 and R4 is halogen 14 A method for preparing a compound of formula III, the method comprising dehydration a compound of formula II as claimed in claim 12 or 13: wherein R1 and R2 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophihc, R3 and R4 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl or arylalkyl, and R5 is selected from the group consisting of H, substituted or unsubstituted alkyl, hydroxy, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophihc, or forms part of an amino acid, or is a nucleoside, an oligomer, a polymer, a dendrimer, a substrate or a surface 15 A method as claimed in claim 14, wherein at least one of R1, R2, R3 and R4 in formula III is halogen 16 A method as claimed in claim 14 or 15, wherein the dehydration is carried out in the presence of a dehydrating agent 15 57 17 A compound of formula III wherein R1 and R2 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophihc, R3 and R4 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl or arylalkyl, and R5 is selected from the group consisting of H, substituted or unsubstituted alkyl, hydroxy, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophihc, or forms part of an amino acid, or is a nucleoside, an ohgomer, a polymer, a dendnmer, a substrate or a surface 18 A compound as claimed in claim 17, wherein at least one of R1, R2, R3 and R4 is halogen 19 A compound as claimed in claim 17 or 18, selected from the group consisting of 58 20 A compound of formula (VI) wherein R1 and R2 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophihc; R3 and R4 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted aryl or arylalkyl, R5 is selected from the group consisting of H, substituted or unsubstituted alkyl, hydroxy, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted 59 or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophihc, or forms part of an amino acid, or is a nucleoside, an ohgomer, a polymer, a dendnmer, a substrate or a surface; and Z is selected from the group R2, halogen, OC(O)R2, =0, amine azide, thiol, R2, mercaptoaryl, arylalkoxy, mercaptoarylalkyl, SC(O)R2, OS(O)2R2, NHC(O)R2, =NR2 orNHR2 21 An ohgomer or a polymer formed by oligomensing or polymerising a compound of formula II, III or VI directly or with one or more other monomers 22 An ohgomer or a polymer as claimed in claim 21, wherein the one or more other monomer is selected from the group acrylate ester such as substituted or unsubstituted alkyl, hydroxyalkyl, armnoalkyl, or substituted substituted or unsubstituted aryl acrylates or methacrylates, crotonates, substituted or unsubstituted acrylonitnles, vinyl alcohols or acetates, styrene and siloxanes. 23. A surface coating or polymer having incorporated therein a compound as claimed in claim 12 or claim 17 24. A composition comprising an effective amount of a compound as claimed in claim 12 or claim 17 for treating an infection in a human or animal subject 25 A composition comprising an effective amount of a compound as claimed in claim 12 or claim 17 for treating an infection or condition in a subject that is characterised by biofilm formation. 26. A composition as claimed in claim 25, wherein the condition is cystic fibrosis 27. A composition as claimed in claim 25, wherein the condition is dental caries, periodontitis, otitis media, muscular skeletal infections, necrotising fascitis, biliary tract infection, osteomyelitis, bacterial prostatitis , native valve endocarditis, cystic fibrosis pneumonia, 26. 6o meloidosis 28 A composition as claimed in claim 25, wherein the condition is nosocomial infection 29. A composition as claimed in claim 25, wherein the infection is ICU pneumonia or an infection associated with sutures, exit sites, artenovenous sites, scleral buckles, contact lenses, urinary catheter cystitis, peritoneal dialysis (CAPD) peritonitis, lUDs, endotracheal tubes, Hickman catheters, central venous catheters, mechanical heart valves, vascular grafts, biliary stent blockage, and orthopaedic devices, penile prostheses 30. A composition as claimed in claim 25, wherein the infection is selected from the group of a skin infection, burn infection and wound infection 31 A composition as claimed in any one to claims 25 to 30, wherein the subject is an immunocompromised individual 32. A composition composing an effective amount of a compound as claimed in claim 12 or claim 17 for treating or preventing biofilm formation on a surface 33. A composition as claimed in claim 32, wherein the surface is a biological surface. 34 A composition as claimed in claim 33, wherein the surface is a surface of a tissue, membrane or skin 35 An implant device having at least one surface associated with a compound as claimed in claim 12 or claim 17 36 An implant device as claimed in claim 35, wherein the device is an artificial heart valve or hip joint, an indwelling catheter, pacemaker, surgical pin and the like 34 61 37 A biofilm removing or inhibiting composition comprising a compound as claimed in claim 12 or claim 17 and a vehicle or carrier, wherein the amount of the mixture is effective to remove or disrupt a bacterial biofilm or inhibit normal biofilm formation 38 A composition composing a cleaning-effective amount of a compound as claimed in claim 12 or claim 17 for removing a biofilm from a biofilm-containing surface 39 A composition comprising an effective amount of a compound as claimed in claim 12 or claim 17 wherein the amount is effective to prevent biofilm formation on a surface 40. A method of removing a biofilm from a non-biological surface comprising the step of administering a cleaning-effective amount of a compound as claimed in claim 12 or claim 17, to a biofilm-containing non-bio logical surface 41 A method of preventing biofilm formation on a non-biological surface comprising the step of administering an effective amount of a compound as claimed in claim 12 or claim 17 to a non-biological surface, wherein the amount is effective to prevent biofilm formation. Novel synthesis methods to the products of such novel methods and to uses of these products. In particular, the present invention provides methods for the reactions of furanones, in particular fimbrolides, with amines. The invention has particular application in the synthesis of halogenated l,5-dthydro-pyrrol-2-one, 5-halomethylene substituted l,5-dihydropyrrol-2-ones (laetam analogues of fimbrolides), 5-amino substituted furanones and 5-ammomethylene-2(5H)-furanones and their synthetic analogues. The invention also relates to novel compounds and uses thereof.

Full Text

"Furanone derivatives and methods of making same"
Technical Field
The present invention relates to novel synthesis methods, to the products of such novel methods, and to uses of these products. In particular, the present invention provides methods for the reactions of furanones, tn particular fimbralides, with amines. The invention has particular application in the synthesis of halogenated 1,5-dihydro-pynrol-2-one, 5-halomethylene substituted 1,5-dihydropyrrol-2-ones (lactam analogues of fimbrolides) 5-amino substituted furanones and 5-aminomethylene-2(5H)-furanones and their synthetic analogues. The invention also relates to novel compounds and uses thereof.
Background
Fimbrolides (halogenated 5-methylene-2(5H)-furanones) possess a wide range of important biological properties including antifungal and antimicrobial properties (see WO 96/29392 and WO 99/53916, the disclosures of which are incorporated herein by cross-reference) These metabolites can be isolated from red marine algae Delisea fimbnata, Delisea elegans and Del/sea puichra.
Despite their biological activity very few hetero atom containing analogues of these molecules have been reported in the literature. The majority of the published syntheses of fimbrolides focus on the preparation of naturally occurring fimbrolides themselves. Recently we have developed methods that yield both the natural and non-natural fimbrolides in good yields (see WO 99/54323 and WO 0200639 the disclosure of which is incorporated herein by cross-reference).
We have now found that, surprisingly, fimbrolides undergo reactions with amines under mild conditions. We have found this discovery to be particularly useful in the synthesis of 5-hydroxy-5-aIkyl substituted 1,5-dihydro-pyrro!-2-one, 5-amino-5-alkyI substituted 2(5H)-furanones and 5-aminomethyIene substituted 2(5H)-furanones. Furthermore 5-hydroxy-5-halomethyl substituted 1,5-dihydro-pyiTol-2-one generated under these conditions can be dehydrated to yield 5-halomethylene substituted 1,5-dihydropyrrol-2-ones (lactam analogues of fimbrolides), and the 5-amino-5-bromomethyl substituted 2(5H)-furanones can be dehydrobrommated to yield a range of 5-aminomethylene

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Substituted 2(5H)-furanones. These furanones can be further functionahsed to yield a range of novel analogues. Summary of the Invention
In a first aspect, the present invention provides a method for the preparation of compound of formula 11

wherein R1 and R2 are independently selected from the group H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophilic;
R3 and R4 are independently selected from the group H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl;
Rs is selected from the group consisting of H, hydroxy, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophilic, or
forms part of an amino acid, or
is a nudeoside, an oligomer, a polymer, a dendnmer, a substrate or a surface;
the method comprising reacting a compound of formula I

WO 2004/016588 PCT/AU2003/001053

wherein R1 and R2 are independently H, halogen, alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyt, substituted or unsubstituted aikenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophilic;
R3 and R4 are independently H, halogen, alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl or arylalkyl; and R is hydroxy, halogen; and
"--- " represents a single bond in which case R is absent, or a double bond,
provided that at least one of R1, R2, R3 and R4 is halogen,
with a compound of formula R5NH2
wherein R5 is selected from the group consisting of H, substituted or unsubstituted alkyl, hydroxy, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophllic, or
forms part of an amino acid, or
is a nucleoside, an oiigomer, a polymer, a dendnmer, a substrate or a surface.
The reaction may optionally be carried out in the presence of solvent.
Preferably, in the compound of formula II, at feast one of R1, R2, R3 and R4 is halogen.
In the structural formulae descnbed herein, a particular geometry is not to be taken as specified. For example, the formulae covers both 2- and E-isomers.
The reaction may be performed In the presence or absence of a solvent. The solvent may be any suitable solvent. Preferable solvents in the present invention include alkyl acetates, aromatic hydrocarbons, chionnated alkanes, cyclic or open chain ethers such as tetrahydrofuran, diethyl ether, dioxane, and

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4
C1-C3 acids. More preferably, the solvents are aromatic hydrocarbons and chlorinated alkanes. Most preferably, the solvent is dichloromethane, as well as dtchloroethane and trichloroethane.
The reaction Is preferably earned out at mild temperatures. Preferably the cyclisation reaction is performed at a temperature in the range of 20-150°C
Where a solvent is present, the cyclisation may be performed at reflux temperature, for example, at the reflux temperature of dichloromethane. Optionally the reaction may be carried out below reflux temperature under pressure.
The reaction time may vary from about 2 hours to 12 hours or more and is typically about 2 hours or more, It will be appreciated that reaction conditions may be varied depending upon the individual nature of the substrate and the desired rate of the reaction.
Non-limiting examples of compounds of formula II, which may be described as 5-aikyl-5-hydroxy substituted 1,5-dihydro-pyrrol-2-one5, that can be synthesised by the method of the invention Include:

WO 2004/016588 PCT/AU2003/001053
5
In a second aspect, the present invention provides a compound of formula II.

wherein R1 and R2 are independently H, halogen, alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substrtuted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophilic;
R3 and R4 are independently H, halogen, substituted or unsubstituted
alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl or arylalkyl;
R5 is selected from the group consisting of H, hydroxy, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substrtuted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fiuorophilic, or
forms part of an amino acid, or
is a nucleoside, an oligomer, a polymer, a dendrimer, a substrate or a surface..
Particularly preferred are compounds of formula II in which at least one of R1, R2, R3 and R4 is halogen.
The Inventors have found the 5-alkyM5-riydroxy substituted 1,5-dihydro-pyrrol-2-one of formula II can be dehydrated to yield a range of 5-(halomethylene)- 1,5-dihydro-pynrol-2-one, 5-(dihalomomethylene)-1,5-dihydro-pyrrol-2-one.
Accordingly in a third aspect, the present invention provides a method for the dehydration of a compound of formula II above, to prepare a compound of formula III;

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wherein R1 and R2 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted ior unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophilic;
R3 and R4 are independently selected from H, halogen, substituted or
unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or
unsubstituted aryl or substituted or unsubstituted arylalkyl; and
R5 is a defined above,
the .method comprising contacting a compound of formula II with a dehydrating agent
Preferably at (east one of R1, R2. R3 and R4 in formula HI is halogen;
Examples of suitable dehydrating agents inciude phosphorus pentoxide, silica gel, molecular sieves, alumina, acidic resins and polymers, phosphorus oxychlonde, acetic anhydride, N.N'-dicyclohexylcarbodiimide (DCC), fnfluoroacetic- acid, sulfuric acid, tnfluoroacetic anhydride, trifluorosulfomc acid anhydride (trffiic anhydride).
Preferably dehydration is carried out using phosphorus pentoxide in the presence of a solvent. The solvent may be any suitable solvent. Preferable solvents in the present invention include alkyl acetates, aromatic hydrocarbons, chlorinated alkanes, tetrahydrofuran, diethyl ether, dioxane and C1-C3 acids. More preferably, the solvents are aromatic hydrocarbons and chlorinated alkanes. Most preferably, the solvent is dichloromethane, as well as dichloroethane and tnchloroethane.
The reaction is preferably carried out at mild temperatures. Preferably the dehydration reaction is performed at a temperature in the range of from about 20-150°C

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Where a solvent is present, the cyclisation may be performed at reflux temperature of the solvent, for example, at the reflux temperature of dichloromethana
The reaction time may range from about 2 hours to 12 hours or more and is typically about 2 hours or more. It will be appreciated that reaction conditions may be varied depending on the individual nature of the substrate and the desired rate of the reaction.
Non-limiting examples of furanones (III) that can be synthesised by this procedure are listed below.

We believe that the 1,5-dihydro-pyrrol-2-ones prepared of formula HI are novel compounds-
Thus, in a fourth aspect, the present invention provides a compound of formula III:

WO 2004/016588 PCT/AU2003/001053

wherein R1 and R2 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straightcham or branched chain, hydrophilic or fluoraphilic;
R3 and R4 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl or arylalkyl; and
R5 is as defined above.
Preferably at least one of R1, R2, R3 and R4 is halogen.
Furthermore the present inventors have also found that furanones of formula (I) when treated with certain amines can yield 5-amino substituted or 5-ammomsthylene substituted furanones. Alternatively, the compounds of formula I can be treated with an alcohol to yield 5' alkoxy substituted furanones. For example when 4-bromo-5-bromomethylene-2(5H)uranone was treated with aniline it gave 4-bromo-5-phenylaminomethyIene-2(5H)-fuiranone in good yields. In contrast, the reaction of 4-bromo-5-bromomethylene-2(5H)-furanone with benzyl amine, gave the corresponding 5-benzylamino-4-bromo-5-bromomethyl-2(5H)-furanone.
Accordingly, in a fifth aspect, the present invention provides a method for the preparation of a compound of formula IV

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wherein R1 and R2 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalky!, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophilic;
R3 and R4 are independently selected from H, halogen, substituted or
unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl,
R5 is as defined above,
X is O or NR, where R5 may be R1,
the method comprising reacting a compound of formula I wherein R3 is a hydrogen and "--- " represents a double bond.
Preferably at least one of R1 R2, R3 and R4 is halogen. Preferably R6 is H.
Representative examples of furanones (IV) that can be synthesised by this procedure are listed below.

In yet a sixth aspect, the present invention provides a compound of formula IV

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wherein R1 and R2 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstrtuted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophilic;
R3 and R4 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl; and
R5 and X are as defined above.
Preferably, at least one of R1, R2. R3 and R4 is halogen.
Accordingly a seventh aspect the present invention provides for a method for preparation of a compound of formula V.

wherein R1 and Rz are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstrtuted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstrtuted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl( optionally Interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophilic;
R3 is selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl or

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arylalkyl, wherein R5 is H, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted aikenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophilic;
X is O or NR6, where R6 is as defined above; and
R6 is as defined above.
Non-limfting examples of furanones of formula (V) that can be synthesised by this procedure are listed below

In an eighth aspect, the present invention provides a compound of formula V:

wherein R1 and R2 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted aikenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophilic;
R3 is selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl or
arylalkyl;

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X is O or NR6, where Re is as defined above, and R5 is as defined above.
In yet a ninth aspect the present invention provides a compound of formula (VI)

wherein R1 and R2 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substitutGd or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic orfluorophilic,
R3 and R4 are independently selected from H, halogen, alkyl, substituted or unsubstituted aryl or arylalkyl; wherein R5 is H, substituted or unsubstituted alky), substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic orfluorophilic;
R5 is defined as above; and
Z is selected from the group R2, halogen, OC(0)R2 =O, amine azide, thiol, R2 mercaptoaryl, arylalkoxy, mercaptoarylalkyl, SC(O)R2, OS(O)2R2. NHC(O)R2. =NR2 or NHR2.
The compounds of formula V! may be prepared by functionahzing a fimbrolide of formula (III) wherein, R1, R2, R3 and FUare as defined above, with a reagent described in WO 99/54323, (the disclosure of which is incorporated herein by cross-reference).
Reagents for introduction and manipulation of the Z group include halogenating and oxidising agents (N-halosuccinimide, iead tetraacetate, selenium dioxide, Jones reagent), nucleophlles (Including organic metal carboxylates, organic alcohols, dimethyl sulfoxide and organonrtnles) and electrophiles including (organrc acids, isocyanates, carboxylic or sulfonic acid hahdes and diethylammosulfurtnfluoride)

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Non-limiting examples of furanones of formula (VI) that can be synthesised by this procedure are listed below.

In a tenth aspect, the present invention provides an oligomer or polymer formed by oligomerising or polymerising a compound of formula It - VI, described herein directly or with one or more other monomers
The one or more other monomer may be any suitable polymerisable copolymer e.g. acrylate ester such as alkyl, hydroxyaikyl, aminoalkyl, or substituted substituted or unsubstituted aryl acrylates or methacrylates,

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crotonates, substituted or unsubstituted acrylonitriles, vinyl alcohols or acetates, styrene and siloxanes.
R5 may be a residue of a natural or synthetic compound. R6 may be a biological or non-biological compound. For example, R5 may be a coenzyme or cofactor. R5 may be an oligomer or a polymer, which may be biological or synthetic. For example, the oligomer or polymer may be a peptide or polyamide. The polymer may be a protein, for example, an enzyme or a receptor, R5 may be an oligomer or polymer comprising nucleic acid residues. The polymer may be a polynucleotide, for example, DNA or RNA. R5 may form part of or be bonded to a nucleoside. The nucleoside may be a D- or L-nudeosfde. R5 may be linked to a sugar moiety of the nucleoside.
R5 may be a surface or substrate with which the nftrogen atom of is associated. The association may be chemical bonding, for example covalent
bonding. The surface or substrate may be biological or synthetic. Altematfvely,

the association may be by means of adsorption. Methods for forming such
associations are described in more detail below.
R5 may also be a dendrimer. A review of dendrimers is provided in Klajnert, B. and Bryszewska, M. (2001) Dendrimers.-properties and applications, Acta Biochemica Polonica Vol. 48 No.1/2001, the disclosure of which is incorporated herein by reference. A plurality of compounds in accordance with the invention may be carried by the dendrimer.
The compound may be immobilised directly onto at least part of the surface of the material of the substrate or via one or more intermediate layers interposed between the substrate material and the immobilised layer. The intermediate layer (s) may be bonding layer (s).
The substrate may be shaped or non-shaped. The substrate may be solid, semi-solid or flexible. The substrate may be a woven or non-woven film or sheet. The substrate may be a natural or synthetic filament or fibre The substrate may be a natural material, for example, a plant seed. The material from which the substrate is formed may be selected to suit the particular application For example, In the case of a shaped blomedical device the material may meet other specifications of the application, such as mechanical and optical properties.
The substrate may be -3 shaped article including, but are not limited, medical devices, for example, implantable biomedical devices such as unnary

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catheters, percutaneous access catheters, stents, as well as non-implantable devices such as contact lenses, contact lens storage cases, and the like.
The matenal from which the article is formed can be a metal, a ceramic, a solid synthetic polymer, or a solid natural polymer, for example a solid biopolymer. Examples of useful materials for this invention are titanium, hydroxyapatite, polyethylene (which are useful materials for orthopaedic implants), polyurethanes, organosiloxane polymers, perfluorinated polymers (which are useful matenals for instance for catheters, soft tissue augmentation, and blood contacting devices such as heart valves), acrylic hydrogel polymers and siloxane hydrogel polymers (for instance for contact tens and intraocular lens applications), and the like, and any combination thereof. The surfaces of these materials can be chemically inert or contain reactive functional groups.
Further examples of substrates include archival documents, antiques and art, rare and valuable seeds intended for storage (e. g seed banks of conservation groups), etc In which case the substrate may be paper, material or other natural or synthetic material.
The substrate may be a shell fish or aquaculture apparatus, for example, that described in PCT/AU98/00508, the disclosure of which is incorporated herein by reference.
As mentioned above, R5 may be associated with a surface of substrate. If necessary, the surface of the substrate may be optionally treated at least in part to activate the surface, to which the compounds of the present invention may be reacted to immobilise the compound.
Reference to at least part of the surface of the substrate includes a surface of one or more intermediate layers applied to the substrate.
The compounds may be Immobilised on the substrate surface by any suitable technique. Immobilization may be by covalent or non covalent means Preferably, the compounds are immobilised on the substrate surface by means of covalent bonds.
The immobilization of furanone compounds on to the substrate prevents their loss from the surface, thus ensuring long-lasting antimicrobial action.
The association between the compounds of the invention and the substrate may be charactensed by the formula : X-Y-Z where X is a substrate. Y is an optional chemical linking noiety and Z is a compound In accordance with the present invention. The linking moiety, if present, may be a homobifunctional or heterobifunctional linking moiety. Y may be a simple

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component (eg a short molecule) or it may comprise a plurality of units or components that may be the same of different. Y may comprise a number of components or units that may be "built up"in a stepwise fashion.
The formation of a covalent interfacial linkage is much preferable to an ionic bond since in biological media where the salt content is such that ionic bonds are interfered with and ironically attached molecules can be displaced from a surface.
In the context of substrates that are medical devices, covalent anchoring of the compound(s) also serves to eliminate concerns regarding possible deleterious effects that compounds might cause at sites distant from the device, such as in the liver, brain, or kidney tissues of a living human organism. In medical applications it is important to anchorthe furanone compound (s) via an interfacial covalent bond that is not subject to cleavage in the host environment that the biomedical device is to be placed in.,
Methods for the covalent immobilization of organic molecules onto solid surfaces are well known to those skilled in the art Interfacial reactions leading to the formation of covalent interfacial bonds are denved from well known organic-synthetic reactions. The choice of immobilization reaction depends on both the nature of the substrate material and the chemical composition of the furanone derivative (s) that are desired for a particular application
For example, a compound that contains a hydroxyl group in a side chain distal to the ring system, can be linked covalently onto surfaces using epoxide chemistry analogous to the reaction pathway described forthe immobilization of polysaccharides onto epoxidated surfaces in Li et a/., Surface Modification of Polymeric Biomateriais (BD Ratner and DG Castner, Eds), Plenum Press, NY, 1996 pages 165-173 (the disclosure of which is incorporated herein in its entirety), through isocyanate groups attached to the surface to produce stable urethane linkages through thermal processes, or through carboxylic acid groups or their equivalents, such as acid chlorides, on the surface to produce ester linkages. A compound that contains an aldehyde group can be linked onto surface amine groups using a reductive animation reaction. A compound that contains a carboxylic acid group can be linked onto surface amine groups using carbodumide chemistry
interfacial coupling reactions must of course be selected not only for their ability to achieve the desired covalent linkage but also for avoidance of adverse effects on the furanone compound (s) to be attached. Particularly, the furanone

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nng system tends to be labile to alkaline conditions. Such limitations are well known to those skilled in the art. Among the many possible interfacial coupling reactions known in the art, there is sufficient scope for selection of reactions that proceed in a suitable pH range and with furanones substituted with various functional groups in various positions
Some solid substrate matenals possess reactive surface chemical groups that can undergo chemical reactions with a partner group on a compound and thereby form a covalent interfacial linkage directly.
Alternatively, in situ covalent linkage can be made directly through the addition of a doubly functionaiised linker molecule to the active surface in the presence of an appropriate compound, or stepwise by sequential addition of doubly functionaiised linker molecules and then an appropriate compound It is not always possible to immobilize furanone compounds directly onto solid substrate materials; in these cases, surface activation or one or more interfaciaf bonding layer (s) is used to effect covalent immobilization of the compounds. Such surface activation is essential when immobilizing compounds onto polymeric materials such as fluoropolymers and polyolefins.
Surface activation of solid substrate matenals can be achieved in a number of ways. Examples are corona discharge treatment or low pressure plasma treatment of polymers. These methods are well known to introduce a vanety of functional groups onto polymeric surfaces.
An alternative approach is to provide an interfacial bonding layer interspersed between the solid substrate material or medical device and the compound layer. The application of a thin fnterfacial bonding layer can be done using methods such as dip coating, spin coating, or plasma polymerization. The chemistry of the bonding layer is selected such that appropriate reactive chemical groups are provided on the surface of this layer, groups that then are accessible for reaction with compound of the invention.
Particularly versatile is the subsequent application of multiple thin interfacial bonding layers; this method can provide a very wide range of desired chemical groups on the surface for the immobilization of a wide range of functionalized furanones and enables usage of compounds optimized for their biological efficacy.
By providing a thin, surface-coated layer of compounds, the optical quality of antibacterial devices of this invention is not reduced, which makes the

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invention applicable to transparent ophthalmic devices such as contact lenses and intraocular lenses.
The present invention provides thin surface coatings that provide antimicrobial properties and/or antifungal properties to solid materials onto which the coatings have been applied More particularly, the coatings may be designed to reduce or prevent colonization of biomedical devices by bactena that cause adverse effects on the health of human users of biomedical devices when such devices are colonized by bacteria.
The active antibacterial layer comprises one or a plurality of furanorie compounds selected for both their antibacterial activity and absence of cytotoxicity as well as any other adverse biomedical effect on the host environment that the coated device contacts.
In an eleventh aspect, the present invention provides incorporation of compounds produced by the methods according to the first, third, fifth, seventh, ninth, or tenth aspects either in surface coatings or polymers through any part of the molecule, for example, newly Introduced functionality on the alkyl chain or the alkyl chain or the halomethylene functionality itself via direct polymerisation or copolymerisation with suitable monomers.
In an twelfth aspect, the present invention provides a compound produced by the method according to the first, third, fifth, seventh, ninth, or eleventh aspects of the present invention.
In a thirteenth aspect, the present invention provides the use of a compound produced according to the present invention. The present inventors have found that many of the 1,5-dihydro-pyrrol-2-one derivatives and furanones having the formula (II), (III), (IV), (V) and (VI) have antimicrobial and/or antifouling properties. Accordingly, the flmbrolide derivatives are suitable for use as antimicrobial and/or antifouling agents.
Thus in a fourteenth aspect the present invention provides methods of use of compounds of formula (II), (III), (IV), (V) and (VI) in medical, scientific and/or biological applications.
For these and other applications, the compounds of the present invention may be formulated as a composition.
In a fifteenth aspect, the present invention provides a composition comprising at least one compound of formula (II), (III), (IV), (V) or (VI)
The compositions of the third aspect of the invention may be in any suitable form. The composition may include a carrier or diluent The carrier may

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be liquid or solid. For example, the compositions may be in the form of a solution or suspension of at least one of the compounds in a liquid. The liquid may be an aqueous solvent or a non-aqueous solvent The liquid may consist of or comprise a one or more organic solvents. The liquid may be an ionic liquid. Particular examples of carrier or diluents include, but are not limited to, water, polyethylene glycol, propylene glycol, cyclodextnn and derivatives thereof
The composition may be formulated for delivery in an aerosol or powder form.
The composition may include organic or inorganic polymeric substances. For example, the compound of the invention may be admixed with a polymer or bound to, or adsorbed on to, a polymer
When the composition is to be formulated as a disinfectant or cleaning formulation, the composition may include conventional additives used in such formulations. Non-limiting examples of the physical form of the formulations include powders, solutions, suspensions, dispersions, emulsions and gels
Formulations for pharmaceutical uses may incorporate pharmaceutically acceptable earners, diluents and excipients known to those skilled in the art. The compositions make be formulated for parenteral or non-parenteral administration. The composition of the invention may be formulated for methods of introduction including, but not limited to, topical, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, ophthalmic, and oral routes. It may be formulated for administration by any convenient route, for example by infusion or bolus injection, by absorption through epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa, etc.) and may be administered together with other biologically active agents. Administration may be localized or systemic The composition may be formulated forintraventncular and intrathecal injection. Pulmonary administration can also be employed, e.g., by use of an inhaler or nebulizer, and formulation with an aerosolizing agent.
In certain preferred embodiments the composition further comprises other active agents such as antibiotics and cleaning agents.
In a sixteenth aspect, the present invention provides a method of treating an infection in a human or animal subject the method comprising administration to the subject of an effective amount of the compound of the invention.
The treatment may therapeutic arid/or prophylactic.

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The compounds of the present invention can act as quorum sensing inhibitors and therefore find use in any application where such as effect is desired. For example, the compounds of the present invention may have use in preventing the establishment and expression of virulence by microorganisms through the inhibition of quorum sensing systems and/or other extracellular systems (eg see, International patent application No. PCT/AU01/01621, the disclosure of which is incorporated herein in its entirety).
The present invention is suitable for biofilms onginating from a single type of organism and for mixed biofilms By "mixed biofilms" is meant biofilms created by more than one type of microorganism Most preferably, it is envisioned that biofilms will be created by at least two organisms from the group consisting of bacteria, algae, fungi, and protozoa.
The effects of treating biofilms with homoserine lactones have been demonstrated with Pseudomonas aeruginosa. The HSLs have generally been isolated from a wide range of bacteria known to produce biofiims. Among these are the enterobacteria. The presence of the HSLs In a wide range of bactena indicates that the compounds of the present invention can be used to effectively treat not only Pseudomonas sp. biofiims but also mixed biofilms containing Pseudomonas sp. and biofilms composed of bacteria other than Pseudomonas aeruginosa.
The following is a list of groups of Gram-Negative bacteria that have members which use homoserine lactones for cell-cefl communication: anaerobic Gram Negative Straight, Curved and Helical Rods; Bacteroidaceae; The Rickettsias and Chlamydias, Dissirrrilatory Sulfate-or Sulfur-Reducing Bacteria; the Mycoplasmas; The mycobacteria; Budding and/or Appendaged Bactena; Sheathed Bactena; Nocardioforms; and Actinomycetes, for example. See Bergey's Manual of Systematic Bacteriology, First Ed. John G. Holt, Editor in Chief (1984), incorporated herein by reference.
The method of the sixteenth aspect may be used to treat an infection or condition in a subject that is characterised by biofilm formation. Non-limiting examples of human infections involving biofilms include dental canes, penodontitis, otitis media, muscular skeletal infections, necrotismg fascitis, biliary tract Infection, ostsomyelitis, bacterial prostatitis , native valve endocarditis, cystic fibrosis pneumonia, meloidosis, and nosocomial infections such as ICU pneumonia, sutures, exit sites, arteriovenous sites, scleral buckles, contact lenses, urinary catheter cystitis, peritonea! dialysis (CAPD)

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pentonitis, UDs, endotracheal tubes, Hickman catheters, central venous catheters, mechanical heart valves, vascular grafts, biliary stent blockage, and orthopedic devices, penile prostheses. Further applications are described in Costerton J et al, (1999) Vol. 284, Science pp1318-i322 and Costerton J and Steward, (2001) Battling Biofilms, Scientific American pp 75-81, the disclosures of which are incorporated herein by reference.
Other locations in which biofilms may form included dnnking water pipes, which may lead to corrosion or disease, household drains, dental plaque which may lead to gum disease and cavities, which may lead to gun disease or cavities, contact lenses which may lead to eye infections, ears which may lead to chronic infection and lungs which may lead to pneumonia.
The condition may be cystic fibrosis. The infection may be that resulting from a skin .infection, burn infection and/or wound infection. The method and composition of the invention may be particularly suitable for the treatment of infection in immuno compromised individuals.
In yet a seventeenth aspect, the present invention provides a method for treating biofilm formation on a surface by contacting the surface with a compound in accordance with the present invention.
The term "surface" as used herein relates to any surface which may be covered by a biofilm layer. The surface may be a biologtcal (eg tissue, membrane, skin etc) or non-biological surface.
The surface may be that of a natural surface, for example, plant seed, wood, fibre etc
The surface or substrate may be any hard surface such as metal, organic and inorganic polymer surface, natural and synthetic elastomers, board, glass, wood, paper, concrete, rock, marble, gypsum and ceramic materials which optionally are coated, eg with paint, enamel etc; or any soft surface such as fibres of any kind (yams, textiles, vegetable fibres, rock wool, hair etc.); or porous surfaces; skin (human or animal); keratinous materials (nails etc.). The hard surface can be present in process equipment or components of cooling equipment, for example, a cooling tower, a water treatment plant, a dairy, a food processing plant, a chemical or pharmaceutical process plant. The porous surface can be present in a filter, eg. a membrane filter.
Particular examples of surfaces, that may be treated in accordance with the invention include, but are not limited to, toilet bowls, bathtubs, drains,

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highchairs, counter tops, vegetables, meat processing rooms, butcher shops, food preparation areas, air ducts, air-conditioners, carpets, paper or woven product treatment, nappies (diapers), personal hygiene products (eg sanitary napkins) and washing machines. The cleaning composition may be in the form of a toilet drop-in or spray-on devices for prevention and removal of soil and under rim cleaner for toilets. The compositions and methods of the present invention also have applications in cleaning of Industrial surfaces such as floors, benches, walls and the like and these and other surfaces in medical establishments such as hospitals (eg surfaces in operating theatres), veterinary hospitals, and in mortuanes and funeral parlours.
A compound of the invention may be incorporated into epidermal bandages and lotions. Alternatively, the compounds of the invention may be incorporated into cosmetic formulations^ for example, aftershave lotions.
Compositions of the present invention may be m the form of an aqueous solution or suspension containing a cleaning-effective amount of the active compound described above. The cleaning composition may be in the form of a spray, a dispensable liquid, or a toilet tank drop-in, under-rim product for prevention, removal and cleaning of toilets and other wet or intermittently wet surfaces in domestic or industrial environments.
The compositions of the present invention may additionally comprise a surfactant selected from the group consisting of anionic, non-ionic, amphotenc, biological surfactants and mixtures thereof. Most preferably, the surfactant is sodium dodecyl sulfate.
One or more adjuvant compounds may be added to the cleaning solution of the present invention. They may be selected from one or more of biocides, fungicides, antibiotics, and mixtures thereof to affect planktonics. pH regulators, perfumes, dyes or colorants may also be added.
By "cleaning-effective" amount of active compound, it is meant an amount of the compound which is necessary to remove at least 10% of bacteria from a biofilm as determined by a reduction in numbers of bacteria within the biofilm when compared with a biofilm not exposed to the active compound.
The cleaning methods of the present invention are suitable for cleaning surfaces. They may be used to treat hard, rigid surfaces such as dra»n pipes, glazed ceramic, porcelain, glass, metal, wood, chrome, plastic, vinyl and formica or soft flexible surfaces such as shower curtains, upholstery, laundry

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and carpeting. It is also envisioned that both woven and non woven and porous and non-porous surfaces would be suitable.
In other embodiments of the present invention, the composition of the invention may be formulated as a dentifrice, a mouthwash or a composition for the treatment of dental caries. The composition may be formulated for acne treatment or cleaning and disinfecting contact lenses (eg as a saline solution).
The method of the invention may be used to treat medical devices,
In yet a further aspect, the present invention extend to a medical device having a least one surface associated with a compound(s) in accordance with the present invention.
The method of the invention may be used to treat implanted devices that are permanent such as an artificial heart valve or hip joint, and those that are not permanent such as indwelling catheters, pacemakers, surgical pins etc. The method may further be used in situations involving bacterial infection of a host, either human or animal, for example fn a topical dressing for burn patients. An example of such a situation would be the infection by P. aeruginosa of superficial wounds such as are found in burn patients or in the lung of a cystic fibrosis patient.
In other forms, the present invention can be used to treat integrated circuits, circuit boards or other electronic or microelectronic devices.
In yet another aspect, the present invention provides a method for the inhibition of a biological pathway is a dell, the method compnsing administering to the ceil a compound in accordance with the present invention.
Terminology
The term "alkyl" is taken to mean both straight chain alkyl groups such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tertiary butyl, and the like. Preferably the alkyl group is a lower alkyl of 1 to 6 carbon atoms The alkyl group may optionally be substituted by one or more groups selected tram alkyl, cycloalkyl, alkenyl, alkynyi, halo, carboxyl, haloalkyl, haloalkynyl, hydroxy, substituted or unsubstituted alkoxy, alkenyloxy, haloaikoxy, haloalkenyloxy, nitro, ammo, nitroalkyi, nitroaikenyl, nitroalkynyl, nftroheterocyclyl, alkylammo, dialkylamino, aikenylamtne, alkynylamino, acyl, alkenoyi, alkynoyl, acylamino, diacylamino, acyloxy, alkylsulfonytoxy, heterocyclyl, heterocycioxy, heterocydamino, haloheterocyclyl, alkylsulfenyl, alkylcarbonyioxy, alkylthio, acylthio, phosphorus-containing groups' such as phosphono and phosphinyl.

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The term "alkoxy" denotes straight chain or branched alkyloxy, preferably C1-10 alkoxy. Examples include methoxy, ethoxy, n-propoxy, isopropoxy and the different butoxy isomers
The term "alkenyl" includes groups formed from straight chain, branched or mono- or polycyclic alkenes and polyene. Substituents include mono- or poly-unsaturated alkyl or cycloalkyl groups as previously defined, preferably C2-10 alkenyl. Examples of alkenyl include vinyl, allyl, 1-methylvinyl, butenyl, iso-butenyl, 3-rnethyl-2-butenyl, 1-pentenyl, cyclopentenyl, 1-methyi-cyclopentenyl, 1-hexenyl, 3-hexenyl, cyclohexenyl, 1-heptenyl, 3-heptenyl, 1-octenyl, cyclooctenyl, 1-nonenyI, 2-nonenyI, 3-nonenyl, 1-decenyl, 3-deceny], 1,3-butadfenyl, 1,4-pentadienyl, 1,3-cyclopentadienyl, 1,3-hexadienyl, 1,4-hexadienyl, 1,3-cyclohexadienyl, 1,4-cyclohexadienyl, 1,3-cycloheptadienyl, 1,3,5-cycloheptatrienyl, or 1,3,5,7-cyclooctatetraenyl.
The term "halogen" includes fluonne, chlorine; bromine or iodine, preferably bromine or fluorine.
The term "heteroatoms" denotes O, N, S or Si.
The term "acyl" used either alone or in compound words such as "acyloxy", "acylthio", "acylamino" or diacylamrno" denotes an alkanoyl, aroyl, heteroyl, carbamoyl, alkoxycarbonyl, alkanesulfonyl, arysulfonyl, and is preferably a C1-10 alkanoyl. Examples of acyl include carbamoyl; straight chain
or branched alkanoyl, such as forrnyl, acetyl, propanoyl, butanoyl, 2-
methylpropanoyl, pentanoyl, 2,2-dimethylpropanoyI, hexanoyl, heptanayl, octanoyl, nonanoyl, decanoyl; alkoxycarbonyl, such as methoxycarbonyl, ethoxycarbonyl, t-butoxycarbonyl, t-pentyioxycarbonyl or heptyloxycarbonyl,
cycloalkanecarbonyl such as cyclopropanecarbonyl cyclobutanecarbonyl, cyclopentanecarbonyl or cyclohexanecarbonyl; alkanesulfonyl, such as rnethanesulfonyl or ethanesulfonyl; alkoxysulfonyl, such as methoxysulfonyl or ethoxysulfonyl; heterocycloalkanecarbonyl; heterocyclyoalkanoyi, such as pyrrolidinylacetyl, pyrrolidinylpropanoyl, pyrrolinylacletyl, pyrrolylacetyl,
pyrrolidinylbutanoyl, pyrrolidinylpentanoyl, pyrrolidinylhexanoyl or
thiazohdinylacetyl; heterocyclylalkenoyl, such as heterocyclylpropenoyl,
heterocyclylbutenoyl, heterocyclylpentenoyl or heterocyclylhexenoyl; or
hetsrocyclylglyoxyloyl, such as, thiazolidinyJglyoxytoyl or pyrrolidinylglyoxyloyl.
The term "aryl" refers to aryl groups havjng 6 through 10 carbon atoms
and includes, for example, phenyl, naphthyl, indenyi. Typically the aryl group

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will be phenyl or naphthyl as compounds having such groups are more readily available commercially than other aryl compounds,
The term "substituted aryl" refers to aryl groups having 1 through 3 substituents independently selected ifrom the group of lower alkyl, lower substituted or unsubstituted alkoxy, halonitro, or haloalkyl having 1 through 3 carbon atoms and 1 through 3 halo atoms. Typical substituted aryl groups include, for example, 2-fluorophenyI, 2-chlorophenyl, 2,e-dimethylphenyl, 4-fiuoroprienyl, 2-methylphenyl, 2-chloro, 3-chloromethylphenyl, 2-nitro, 5-methylphenyl, 2,6-dichlorophenyl, 3-trifluoromethylphenyl, 2-methoxyphenyl, 2-bromonaphth-1-yl, 3-methoxyinden-1-yl, and the like. Carboxyaryl eg carboxy phenyl, aminoaryl eg ammophenyl
The term "fluorophilic" is used to jindicate the highly attractive interactions between certain groups, such as highly fluorinated alkyl groups of C4-C10 chain length, towards perfluoroalkanes and perfiuoroalkane polymers.
The term "arnino acid" as used herein includes any compound having at least one ammo group and at least one! carboxyl group. The amino acid may be a naturally occurring ammo acid or it may be a non-naturally occurring amlno acid.
The amines used in this invention may be soluble in the reaction medium or insoluble in the reaction medium. Examples of soluble amines include ammonia, alkyl(, aryl-, arylalkyl-, and heterocyclic amines.
Examples of insoluble amines include basic amine resins and amine containing biological and synthetic polymers.
The term "optionally substituted" includes, but is not limited to such groups as halogen; hydroxy; hydroxy substituted alkyl; substituted or unsubstituted S(O)m alkyl or S(O)m aryl wherein m is 0,1 or 2, such as methyl thio, methylsulfinyl or methyl suffonyl; amino, mono and di-substituted ammo; alkyl, cycloalkyl, or cycloalkyl alkyl group; halosubstituted alkyl, such as CF3; an optionally substituted aryl, optionally substituted arylalkyl, such as benzyl or phenethyl, wherein these aryl moieties may also be substituted one to two times by halogen; hydroxy; hydroxy substituted alkyl; alkoxy; S(0)m alkyl, ammo, mono and di- alkyl substituted amino, substituted or unsubstituted alkylSIO- for example (CH3)3SiO-
The term "Medical devices" as used herein includes disposable or permanent catheters, (e.g., central venous catheters, dialysis catheters, long-term tunneled central venous catheters, short-term central venous catheters,

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peripherally inserted central catheters, peripheral venous catheters, pulmonary artery Swan-Ganz catheters, urinary catheters, and peritoneal catheters), long-term urinary devices, tissue bonding urinary devices, vascular grafts, vascular catheter ports, wound drain tubes, ventricular catheters, hydrocephalus shunts heart valves, heart assist devices (e.g., left ventricular assist devices), pacemaker capsules, incontinence devices, penile implants, small or temporary joint replacements, urinary dilator, cannulas, elastomers, hydrogels, surgical instruments, dental instruments, tubings, such as intravenous tubes, breathing tubes, dental water lines, dental drain tubes, and feeding tubes, fabrics, paper, indicator stnps (e.g., paper indicator stnps or plastic Indicator stnps), adhesives (e.g., hydrogel adhesives, hot-melt adhesives, or solvent-based adhesives), bandages, orthopedic implants, and any other device used in the medical field. "Medical devices" also include any device which may be inserted or implanted into a human being or other animal, or placed at the insertion or implantation site such as the skin near the insertion or implantation site, and which include at least one surface which is susceptible to colonization by biofilm embedded microorganisms, Medical devices also include any other surface which may be desired or necessary to prevent biofilm embedded microorganisms from growing or proliferating on at least one surface of the medical device, or to remove or clean biofilm embedded microorganisms from the at least one surface of the medical device, such as the surfaces of equipment in operating rooms, emergency rooms, hospital rooms, clinics, and bathrooms In one specific embodiment, the biofilm penetrating composition is integrated into an adhesive, such as tape, thereby providing an adhesive which may prevent growth or proliferation of biofilm embedded microorganisms on at least one surface of the adhesive.
Implantable medical devices include orthopedic implants. Insertable medical devices include catheters and shunts which. The medical devices may be formed of any suitable metallic matenats or non-metallic materials known to persons skilled in the art. Examples of metallic matenals include, but are not limited to, tivanium, titanium, and stainless steel, and derivatives or combinations thereof. Examples of non-metallic materials include, but are not limited to, thermoplastic or polymeric materials such as rubber, plastic, polyesters, polyethylene, polyurethane, silicone, Gortex (tm) (Polytetrafluoroethylene), Dacron (tm) (polyethylene tetraphthalate), Teflon

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(Polytetrafluoroethylene), latex, elastomers and Dacron(tm) sealed with gelatin, collagen or albumin, and derivatives or combinations thereof.
The present invention also extends to a method of regulating a cells characterised by AHL-mediated quorum sensing or an AI-2 pathway comprising contacting the cells with a compound in accordance with the present invention.
Throughout this specification the word "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements,
integers or steps.
Reference is made to patent applications PCT/AU01/01621, PCT/AU02/00797, PCT/AU99/00284. PCT/AU99/00285, PCT/AU00/01553, PCT/AU01/00296, PCT/AU01/00295, PCT/AU01/00407. PCT/AU89/00508 and PCT/AU01/00751 which relate to furanones and analogues and to uses of
these compounds and the entire disclosures of which are incorporated herein
by reference.

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Modes for Carrying Out the Invention
The invention is further described in and illustrated by the following examples. The examples are not to be construed as limiting the invention in any way.
EXPERIMENTAL DETAILS
Genera/. Melting points are uncorrected. Microanalyses were performed by Dr H.P, Pham of The University of New South Wales Microanalytical Laboratory. 1H NMR spectra were obtained in CDCI3 on a Bruker AC300F (300 MHz) or a Bruker DMX50Q (500 MHz) spectrometer. 13C NMR were obtained in the same solvent on a Bruker AC300F (75.5 MHz) or a Bruker DMX500 (125.8 MHz) spectrometer Chemical shifts were measured on the 8 scale internally referenced to the solvent peaks: CDCI3 (5 7.26, 5 77.04).
Ultraviolet spectra were measured on an Hitachi U-3200 spectrophotometer and refer to solutions in absolute MeOH. Infrared spectra were recorded on a Perkin-Etmer 298 or a Perkin-Eimer 580B spectrophotometer and refer to paraffin mulls. The electron impact mass spectra were recorded on an VG Quattro mass spectrometer at 70eV lonisation voltage and 200°C ion source temperature. FAB spectra were recorded on an AutoSpecQ mass spectrometer. Column chromatography was carried out using Merck silica gel 60H (Art. 7736), whilst preparative thin layer chromatography was performed on 2 mm plates using Merck silica gel 60GF254 (Art. 7730).
3-Butyl-5-dibromomethyI-5'-hydroxy-1-phenyl-'1,5-dihydropyrrol-2-one
A solution of 3-butyl-5-dibromomethylene-2(5H)fijranone (0.20 g; 0-65 mmol) in aniline (5 ml) was allowed to stand at room temperature for 24 h. The mixture was diluted with dichloromethane (25 ml) and washed with aqueous hydrochloric acid (2M, 20 ml). The organic phase was dried over sodium sulfate and evaporated to yield a yellow viscous oil (0.30 g). The crude product was chromatographed on silica using dichloromethane/ethylacetate (19:1; v:v) as the eluent. The major product, a pale yellow band, was collected and recrystallised from light petroleum to yield 3-butyl-5-dibromomethyl-5-hydroxy-i-phenyl-1,5-dihydropyrroi-2-one as colourless pnsms (0.24 g, 92%), m.p. 96-98°Cvmax 3211, 2957, 1679, 1597, 1500, 1417, 1117, 1058, 760, 698 cm-1. ?w: 263nm (Emax2,955), 202 (2,464). 1H n.m.r. ( (CDCI3): 7.54-7.37, m, Ph;

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6.82, 1Hf s, C4-H; 5.56, 1H, s, -CHBr2; 3 42, S, C5-0H, 2.43-2.41, m. 2H, CH2, 1.64-0.97, m, C3-chain. 13C n.m.r. 6 (CDC/3): 13.7, 14.0, 22.3, 46.6, 92.2, 126.7,127.4,129.0, 134.0,135.5,136.-0,144.3, 169.0.
5-Dibromomethyl-3-hexyI-5-hydroxy-1-phenyl-1,5-dihydropyrrol-2-one
A mixture of 3-hexyl-5-dtbromomethyiene-2(5H)furanone (0.40g, 1.16 mmol) and aniline (1 ml) in ethanol (6 ml) was refluxed tor 3h. The solvent was evaporated off and the residue extracted with dichlorornethane (25 ml). The organic phase was washed with aqueous hydrochloric acid (2M. 2 x 20 ml), dried over sodium sulfate and evaporated to yield a seml-soltd (0.39 g) The crude product was chromatographed on silica using
dichloromethane/ethylacetate (19:1, v;v) as the eluent. The major product, a pale yellow band, was collected and recrystallised from light petroleum to yield 5-dibromom6thy1-3-hexyl-5-hydroxy-1 -phenyl-1 .S-dihydropyrrol^-one as a semi-crystalline solid (23%), rap. 43-45° (Found (HRESMS) 451.982217. C17H21Br2NO2Na+ (78Br) requires 451.983106). vmm: 3186, 2926, 1680, 1659, 1492, 1372, 1095, 1059, 897, 850, 766, 747, 699, 671 cm-1. Xmax: 261nm (Emax 4051), 206 (26.550). 1H n.m.r. ( (CDCI3). 7.5-7.25, m, 5H, Ph; 6.8, s, 1H, C4-H; 5.5, s, 1H, -CHBr?: 3.77, brs, 1H, C5-OH; 2.44-2.34, m. 2H, CH2; 2.03-0.91, 11H, C3-cham. 13C n.m.r. ( ( CDCI3): 14.2, 25.3, 29 0, 31 0, 46 6, 92.0, 104.8, 126.7, 127.0, 136.0, 144.0,168.0, 172.0.
1-Benzyl-3-butyl-5-dibromomethyl-5-hydroxy-1,5-dihydropyrroI-2-one
A solution of 3-butyl-5-dibromomethylene-2(5H)furanone (1.03 g: 3.32 mmol) in benzyl amine (2ml) was allowed to stand at room temperature for 1 h during which time the reaction mixture solidified. The solid was dissolved in dichlorornethane (25 ml) and washed with aqueous hydrochloric acid (2M, 20 ml). The organic phase was dried over sodium sulfate and evaporated to yield a yellow viscous oil. The crude product was tnturated with light petroleum to yield a white solid (1.0 g; 74%) which was recrystallised from light petroleum to yield 1-benzyl-3-bUtyl-5-dlbromom9thyl-5-hydroxy-1,5-d!hydropyrrol-2-one as colourless needles, m.p. 92-93 (Found (HRESMS) m/z 479.974243 C18H21Br2NO3Na+ (70Br) requires 479.978123) vmax: 2987, 2953, 2920. 1677, 1650, 1449, 1424, 1069 cm-1. (max 207 (88,250). 1Hn.m.r. ( (CDCl3): 7.4-7.29, m, 5H, Ph, 6.72, s, 1H, C4-H; 5.56, s, 1H, -CHBr2; 4.54, bs, 2H, CH2Ph; 3.0,

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1H, C-5 OH; 1.54-0.97, m, C-3 chain. 13C n m.r.5 (CDCI3): 13.7, 22 3, 29.3, 42.6, 46.8, 91:5, 127.6,128.3,128.7,136-9. 137 0,160.8/'170.6.
1-Ben2yI-5-dibromomethyI-3-hexyl-5-hydroxy-1,5-d'ihydropyrrol-2-one
Method A
A solution of 3-hexyl-5-dibromomethylene-2(5H)furanone (1.03 g: 3.32 mrnol) in benzyl amine (2 ml) was stirred at room temperature for 0.5h. Dichloromethane (15 ml) was added to the reaction mixture and the precipitated solid was filtered off. The filtrate was washed with aqueous hydrochloric acid (2M, 20 ml), dned over sodium sulfate and evaporated to yield a yellow viscous oil (0.36g) The crude product was chromatographed on silica using dichloromethane/ethyl acetate (1:19) as the eluent and recrystallised from light petroleum to yield 1-benzyI-5-dlbromorriethyl-3-hexyl-5-hydroxy-1,5-dihydropyrro!-2-one (0 11 g) as colourless needles m.p.105-10S°C. (Found (HRESMS) m/z 465 994011. C18H23Br2NO2NA+ (79Br) requires 465.998758. vms>: 3195, 2987, 2924, 2858, 1676, 1649, 1425, 1153, 1068, 968, 845, 730, 599 cm-1. (max 205 (emax 7740) nm. 1H n.m,r.8(CDCl3): 7.39-7.26, m, 5H, Ph; 6.7, s, 1H, C4-H; 5.6, s, 1H, -CHBr2; 4.54, d, J 15 Hz, 2H, CH2Ph; 2.89-2.35, m, 2H, CH2; 1.60^0.87, m, 13H, C3-chain. 13C n.m.r, S (CDCI3): 14, 22.45, 25, 27, 28.8, 31.4, 42,5, 46.7, 91.5, 127.6, 128.5, 128.6, 136.6, 136.7,144.0,170.0
Method B
A mixture of 3-hexyl-5-dibromomethylene-2(5H)furanone (1.03 g- 3.32 mrnol)
and benzyl amine (2ml) in ethanol (5 ml) was stirred at room temperature for
2.6h The cnjde product was isolated and purified as described above to yield
1-benzyl-5-dibromomethyl-3-hexyl-5-hydroxy-1,5-dihydropyrrol-2-one in (72%)
yield.
1-Butyl-5-dibromomethyl-3-hexyl-5-hydrOxy-1,5-dihydropyrrol-2-one
n-Butyiamme (0.272 g; 3.72 mmol) was added dropwise to a solution of 5-dibromomethylene-3-hexyI-2(5H)furanone (0,314 g; 0.93 mmol) m CH2CI2 (10 ml). The mixture was stirred at roomi temperature for 5 hrs. Column chromatography on silica with CH2CI2 followed by CH2Cl2/EtOAc (19;1) afforded the major product as a colourless oil (0.20 g) which upon

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recrystallisation from petrol gave 1-butyl-5-dibromomethyI-3-hexyl-5-hydroxy-1,5-dihydropyrrol-2-one (52%) as colourless needles, rn.p. 85-86°. (Found(HRESMS) rn/z 432.013664. C15H25Br2NO2Na+ (79Br) requires 432 014407). vmM; 3230, 2957, 2859, 1672, 1650, 1458, 1422. 1375, 1270, 1233, 1139, 1079, 1023, 728. 666, 612 cm-1. W: 259 (emax 945), 206 (9658) nm. 1H n.m.r. 6 (CDCI3): 6.68, s, 1H, C4-H; 5.8, s, 1Hf CHBrz; 3.45, m, 1H, N-CH2; 3 10, m, 1H, N-CH2;-3.15. bs, OH; 3 20. m, 2H, -CH2; 2.33-2.31, m, -CH2-chain, 1.65-0.88, m, 14H, alkyl chain. 13C n.m.r. ( (CDCi3): 13.6, 20, 22, 25, 27, 29, 30.75, 31, 39, 46.6, 91.4,136,144.5,170.
N-(2-Hydroxyethyl)-3-bulyl-5-dibrommethyl-5-hydroxy-2(5H)pyrolinone
A solution of ethanolamlne (1 13g, 18.5 mmol) in CH2Cl2 (5 ml) was added dropwise to an ice-cooled solution of 3-butyl-5-dibromomethylen-2(5H)-furanone (1 -0g; 9.25 mmol) in dichlorornethane. The mixture was stirred at this temperature for 1 hr and then at room temperature for further 1h. The mixture was washed with waster (3 x 50 ml), dned over sodium sulfate and evaporated to yield a viscous oil (0.63g). The cuude product was chromatographed on silica using EtOAc as an eluent to yield N-(2-hydroxyethyl)-3-butyl-5-dibrommethyl-5-hydroxy-2(5H)pyrolinorie as an oil which solidified on keeping. Crystallisation from (CH2Cl2/petrol) afforded the title compound as colurless needles, m p. 68-70°. vmax: 3439, 3105, 3065, 2957, 2927, 1701, 1593, 1496. 1465, 1370, 1189, 1139, 1095, 1069, 10^7, 945, 835, 763 cm-1. ( max 203 nm 1H nmr ( (CDCI3) 0.93, t, 3H, CH3; 1.25-1.45, m, 4H, CH2; 2.35, m, 2H, CH2; 3.10, m, 1H, NCH2CH2OH; 3.84, m| 2H, NCH2CHZOH; 4.10, m, 1H, NGH2CH2OH; 5.43, bs, 1H, OH; 5.84, s, lH, CHBr2; 6,78, s, 1H, H4.13C n.m.r ( (CDCI3): 13.73, 22.2, 24 9, 29.3 41.3, 45.7, 61.3, 90.3, 137.5, 142.8,171.1.
5-Dibromomethyl-3-hexyI-5-hydroxy-1,5-dihydropyrroI-2-one
Liquid ammonia (5ml) was added to 5-dibromomethylene-3-hexyl-2(5H)furanone (0,50 g; 1,48 mmol) in a sealed tube held in a acetone/liquid nitrogen bath. The reaction mixture was allowed to warm up gradually and kept at room temperature overnight. After gradual evaporation of ammonia the product was extracted with EtOAc (20 ml), washed wfth water, dried over Na2SO4, and evaporated to yield a solid (0.30g). The crude product was purified on a silica column using first CH2CI2 as the eluent followed by EtOAc/MeOH (41). The yellow band upon solvent removal and crystallisation

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from petrol afforded a yellow crystalline solid (Q.07g ) of 5-dibromamethyI-3-hexyl-5-hydroxy-1,5 dihydropyrrol-2-one, m.p. 106-109. 1H n.m.r ( (CDCI3): 6.61; s, 1H, C4-H, 6.26, s, 1H, -NH; 5.68, s, 1H, CHBr2;,3.2, s, C5-OH; 2 28-2.23, m, -CH2,chain; 1.55-0.91, m, 11 H, chain. 13C n.m.r. ( (CDCI3): 13.9, 22, 25.5, 27, 29, 31.4,129, 140, 142, 170.5.
4-bromo-5-hydlroxy-5-hydroxyrnethy[-1,5-dihydropyrrol-2-one
A suspension of 4-bromo-5-bromomethylene-2(5H)-furanone (1.30g, 5.16 mmol) in aqueous ammonia solution (20% w/w) was stirred at room temperature for 1/2 h. During this time a complete dissolution of furanone was observed. The solution was evaporated to dryness in vacuo at ca 35-40 °C, and finally under high vacuum at room temperature. The resulting solid (1.70 g) was recrystallised from ethanol to yield 4-bromo-5-hydroxy-5 -hydroxymethyl-1,5-dihydropyrrol-2-one as colourless granules (1.0 g). mp 14O-142 °C (decornp); vmax: 3259, 3100, 2949, 1667, 1592, 1419, 1370, 1152, 1076, 981, 872, 563 cm-1. Xmax 220 (( max 6077). 1H n.m.r. 5 (CDCI3): 8.09, s, -NH; 6.22, d, 2 Hz, H3; 4.97, t, 2 Hz, -CH2OH; 3.37, q.J2 Hz, OH; 2.48, d, 2Hz, -CH2OH. 13C n,m r. 5 (CDCl3): 69 6, 84.3, 132,8, 152,3, 174.1.

4-Bromo-3-hexyl-5-hydroxy-5-hydroxymethyM,5-dihydropyrrol-2-one
A suspension of 4-bromo-3-hexyl-5-bromomethylene-2(5HHuranone (0.50 g; 1.48 mmol) in aqueous ammonia solution (30 mls; 28%) was stirred at room temperature for 2h, during which time the solid completely dissolved. The solution was evaporated to dryness,, and the residue extracted with dichloromethane (25 ml). The organic phase was dried over anhydrous sodium sulfate and evaporated to yield a red viscous oil. Chromatography on silica using ethyl acetate followed by ethyl acetate/methanol (4:1). gave a solid which upon recrystalhsation from light petroleum yielded 4-bromo-3-hexyl-5-hydroxy-5-hydroxymethyM,5-dihydropyrrol-2-one as colourless granules (O.16g; 36%), m.p. 134-135 °. vmax: 3304, 3256, 3185, 2961, 1670. 1589, 1441, 1350, 1136, 1069, 983 cm-1; (.max: 221 ((max 6,678), 196 (3,415) nm.

5-Ethyl-5-hydroxy-4-methy-1,5-dihydropytrol-2-one
A mixture of 5-ethy!idene-4-methyl-2(5H)furanone (0,02 g; 0.162 mmol) in aqueous ammonia solution ( 5 m!; 28% w/w) was stirred at room temperature for 1.5 h during which time all of the furanone dissolved. The solution was

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evaporated in vacuo to dryness eaving 5-ethyl-5-hydroxy-4-methyl-1,5-dihydropyrrol-2-one as a white solid (0.015g; 65%), m.p. 1B2-186°C (Found(HRESMS) m/z 184.067448 C6H11NO2Na+ requires 160 06815). vmax 3204, 29805 1698, 1664, 1633.5, 144si 1167, 1080, 1016, 983, 852, 769, 578. W 207 ((max23,180 ) nm. 1H n.m.r. ( (DMSO)-d6) 7 97, s, 1H. -NH; 5.55, s, 1H, C3-H; 3.18, s, 1H, C5-OH; 1.79, s, 3H, C4-Me; 1.69-1.52, m, 2H, C5-CH2-Me; 0.34, t, 3H, Me. 13C n mx ( (DMSO-d6): 7.9, 11.9, 29.2, 90.2, 121,7, 162, 171.6.
1-Benzyl-5-ethyl-5-hydroxy-4-methyl-1,5-dihydropyrrol-2-one
A solution of 5-ethylidene-4-methyl-2(5H)furanone (0124 g; 10 mmol) in benzytemine (0.128g; 12 mmol) was the left to stand at room temperature for 72 hrs, during which time a solid precipitated from the reaction. The reaction mixture was triturated with CH2Cl2/petrol (1:3) and the precipitated solid was filtered and recrystallised from EtOAc/petrol to yield1-benzyI-5-ethyl-5-hydroxy-4-methyi-1,5-dihydropyrrol-2-one as colourless crystals m.p. 129-132° (70%). vmax 3247, 3082, 2964, 1669, 1638. 1496, 1353, 1101, 1053, 902, 708 cm-1. W 276 ((max 2,101), 237 (16,321), 243 (39,646) nm.1H n.m.r. ( (CDCIa): 7.4-7.24. m, 5H, Ph; S.79, ,s, 1H, C3-H, 4.46. 2 d, J 15 Hz, -CH2Ph; 3.81, bs, C5-OH; 1.92, s, C4-Me; 1.83-168, m. C5-CH2Me, 0.34, t, J 7.51 Hz, C5-CH2Me-13C n.rn.r. ( (CDCI3): 6.8, 11.85, 26, 41.9. 94, 122, 127, 128, 128 5, 138f 159, 170.
5-Aminomethyl-4-heptyI-5-hyc(roxy-1,5-dlhydropyrrol-2-one
5-Bromo-5-bromomethyl-4-heptyl-2(5H)furanone (0.50 g; 1.47 mmol) was dissolved in liquid ammonia in a sealed tube, and left to stand at room temperature for 72 h. Ammonia was allowed to gradually evaporated leaving behind a yellow crystalline solid. The solid was dissolved in not ethylacetate (ca 25) ml to remove ammonium bromide and the clear filtrate was concentrated to a small volume (ca 7 ml), to yield 5-aminomethyl-4-heptyl-5-hydroxy-1,5-dihydropyrrol-2-one as a crystalline solid, (0.1 g; 34%); m.p. 176°C. vmax: 3370, 3248, 2956, 2926, 2855, 1674, 1627, 1469, 1350., 1227,1095, 1082, 954, 555 cm'1. (max, 208 ((max 6845), 291 (2754) nm. 1H n.m.r 6 (CDCI3): 7.53, s, -NH; 5.49, d, C5-CH2NH2, 3.35, 3H, m, -C5-OH and -CH2NH2. 2.23-2.0, m, CH2; 1,52-0.85, m, 13H, alkyl chain. 13C ( (CDCl3): 14, 22.4,26, 26.5, 28.9,29, 31.6, 66,73, 78, 120.6,167.5,171 6.

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5-Bromomethyl-4-heptyi-5-hydro3cy-1-phenyM35-dihydropyrrol-2-one
5-Bromo-5"bromomethyI-4-heptyl-2(6H)furanone (0.51g; 1.5 mmol) was
dissolved in dry aniline (5 ml). The mixture soon solidified; it was allowed to
stand at room temperature for 24 h Dichloromethane (25 mls) was added to
the mixture and the organic phase was washed with aqueous hydrochloric acid
(2M) and brine. The dried (Na2SO4) organic phase was evaporated to yield a
yellow solid (0.50 g, 91%). Recrystallisation from light petroleum gave 5-
bromonnethyl-4-heptyl-5-hydroxy-1-phenyl-1,5-dihydropyrrol-2-one as
colourless needles, m.p. 152-154°C. vmax: 3194, 2956, 1930, 2854,1676,1626, 1589, 1502, 1494, 1393, 1246, 1141, 836, 758, 692 cm-1. Xmax: 257((max 3947), 202 (27,313) nm. 1H n.m r. ( (CDCI3): 7.55-7.26, m, 5H, Ph; 5.79, s, C3-H. 4.52, 1H, C5-OH; 3.39, d, 2H, C5-CH2Br; 2 27-2,12, m, 2H, chain; 1.6-0.91, tn, 13H, chain. 13C n.m.r. ( (CDCl3). 14, 22.5, 25.6, 25.8, 29, 29.2, 30 4, 31.6, 121.6, 126f 126.7, 130,134.6, 163, 170.5.
1-Benzyl-5-bromomethyl-4-heptyl-5-hydroxy-1,5-dihydropyrroI-2-one
A mixture of 5-bromo-5-bromomethyl-4-r|eptyi-2(5H)furanone (0.51g, 1.5 mmol) in benzylamme (0,30g; 2.82 mmol) in ethanol (6 ml) was stirred at room temperature for 1 h. Dichloromethane (25 ml) was added to the reaction mixture and the organic phase was washed with aqueous hydrochloric acid (2M) followed by brine. After drying over sodium sulfate, the solvent was evaporated in vacuo to yield 1-benzy!-5-bromomethyl-4-heptyl-5-hydroxy-1,5-dihydropyrrol' 2-one as a viscous oil (0 52 g; 97%) which solidrfied on standing in the fridge. Colourless needles from light petroleum; m.p. 94-96°.Vmax: 3270, 3062, 3033, 2957, 2854, 1667, 1637, 1607, 1496, 1416, 1335, 1297, 1257, 1190, 1161, 1140,1109, 1030. 950, 884, 865,769 cm-1. ( max 251 (( max2391), 206 (18,974) nm. 1H n.m.r. ( (CDCI3): 7.36-7.28, m, 5H, Ph; 5.85, s, C3-H; 4.54 and 3,42,2d, 2H each, C5-CH2Br and CH2Ph; 3-42, bs, 1H, C5-OH, 2.31-2.15, m, 2H, CH2; 1.62-0 88, m, 13H, alkyl chain.13C, n.m.r. 5(CDCI3). 14, 22,5,25.5, 26, 29, 29.2,
30.87, 41.9, 122, 127,128.3,137 5.163, 171.
Synthesis of 3-alkyl-5-halom ethyl ene-135-di hydro pyrrol-2-one 3-ButyI-5-dibromomethylene-1-phenyl-1,5-dihydropyrrol-2-ohe

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Phosphorus pentoxide was added to a solution of 3-butyl-5-dibromomethy!-5-hydroxy-1-phenyM,5-dihydropyrrol-2-one in chloroform. The resulting mixture was stirred overnight at room temperature and passed through a pad of Celite. The crude product was chromatographed on silica and recrystallised from light petroleum to yield 3-butyl-5-dibromomethylene-1-phenyl-1-5-dihydropyrrol-2-one as orange needles (78%), orange crystals from petrol. (Found(HRESMS) m/z 419.954622. C16H17Br2NONa+ (7eBr) requires 419.955896). W 202 ((max 8137), 195 (3850) nm. 1H n.m.r. BCCDCls): 7.22-7.17, m. 5H, Ph; 7.17, s, C4-H; 2 38-2.36, m, 2H, CH2; 1.65-0.96, m, C3-chain. 13C n,m r. ( (Cody. 13.6, 22,3. 25.2, 29,5,128.3,128.8, 132.1,139, 140. 171.8.
3-Hexyl-5-dibromomethylene-1-phenyl-1,5-dihydropyrrol-2-one
3-Hexy!-S-dibromomethylene-1-pheny[-1 5-dihydropyrrol-2-one was prepared from 3-hexyl-5-dibrornomethyl-5-hydroxy-1-pheriyl-1,5-dihydropyrrol-2-one as descnbed above. Yellow granules from petrol. vmax: 3378, 2957, 2925, 2854, 1692, 1598, 1501, 1492, 1445, 1122, 1081, 743, S77 cm-1. Xmax: 309 (emax 19,681) nm. 1H n m.r. ( (CDCl3): 7 4-7.17, m, 6H, Ph and H4; 2.37-2.34, m, 2H, CH2, 1.57-0.89, m, 11H, C3-cham.
1-Benzyl-3-butyI-5-dibromomethylene-1,5-dihydropyrro(-2-one
1-Benzyl-3-butyl-5-dibromomethyl-5-hydroxy-1,5-dihydropyrroI-2-one was dehydrated with P2O5 in CHCI3 at room temperature for 72 hrs. The mixture was filtered through celite and the solyent evaporated in vacuo to yield a viscous oil, which solidified on keeping in a refrigerator. The solid was recrystallised from methanol/water to yield 1-benzyl-3-butyI-5~ dibromomethylene-1,5-dihydropyrrol-2-one as colourless plates, m.p. 56-58°C (91%). vmax: 2954, 1706, 1626, 1495, 1453, 1494, 1435, 1386, 1352, 1269, 1235, 1095, 765 cm-1. (max 324 ((max 5985), 283 (16,201), 206 (10,972) nm.1H n.m.r. 5 (CDCI3) 7.3-7.07, m, 6H, Ph and H4; 5.26, s, CH2Ph, 2.4-2.36, m, 2H, CH2; 1.6-0.95, m, C3-cham. 13C n.m.r. S (CDCI3): 13.7, 22,25, 29.6, 44.2, 74.7, 89.25,126, 127, 128, 132, 137.8,138 8, 140, 172.1
1-Benzyl-5-dibromomeihylene-3-hexyM,5-dihydropyrroI-2-one
This compound was prepared according to the procedure described for 1-benzyl-3-butyl-5-dibromomethylene-1,5:dihydropyrrol-2-one. vmax 2960, 2848, 2923, 2854, 1696, 1592, 1496, 1453, 1354, 1316, 977, 830, 738, 630 cm-1. 1H

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n.mr. (CDCl3): 7.3-7.08, m, 5H, Ph, 7 26, s, 1H, H4; 5,26, 2H, -CHgPh; 2.4-2.36, m, 2H, CH2; 1.56-1.32, m, C3-chain.
1-Butyl-5-dibromomethylene-3-hexyI-1,5-dihydropyrrol-2-one
This compound was prepared according to the procedure described for 1-benzyl-3-butyl-5-dibrornomethyIene-1,5HdihydropyrroI-2-one. Yield (30%). vmax: 2956, 2928, 2858, 1705, 1586, 1452,1360, 1335, 1194, 1135, 1058, 846, 829 741 cm-1 ; (max 290 ((max 18,927), 203 (9,409) nm. 1H n.m.r. 8(CDCI3)- 7.0, s, 1H, C4-H,; 3.99-3 93, t, 2H, -CH2N-; 2.3, t, -CH2- chain; 1.56-0 88, rn, 16H, chain 13C n.m.r. 5 (CDCl3), 13.7, 14, 19.7, 22.4, 25, 27/29, 31.4, 32.1, 40.6, 132,137,139,140.6,172.0.
5-Dibromomethylene-3-hexyl-1,5-dihydrapyrrol-2-one
This product was prepared by the dehydration of 5-dibromomethyl-3-hexyI-5-hydroxy-1,5-dihydropyrroI-2-one as described above, m.p, 103-105°.
5-EthyIidene-4-methyl-1,5-dihydropyrrol-2-one
5-Ethyl-5-hydroxy-4-methyl-2(5H)pyrrolinpne was dehydrated to 5-ethylidene-4-methyl-1,5-dihydropyrrol-2-one with P2O5 in dichloromethane. vmax: 3158, 3093, 3036, 1670, 1495, 1434, 1397, 1381, 1356, 1279, 956, 867, 796, 639. (max: 173 ((max 33,010) nm. 1H n.m.r. 5(CDCI3): 8.94, s, 1H, -NH; 5.85, 1H, s. C3-H; 5 33, q, J 7 53 Hz, -=CHCH3, 2.1, s, 3H, C4-Me; 1,92, d, J 7.53, C5-Me-CH= 13C n.m.r. 5 (CDCI3) 11.7, 12.9,107,120 5, 140,148,172.0.
1 -BanzyI-5-ethylidene-4-methyM ,5-di hydro pyrroI-2-ons
1-Benzy!-5-etriyIIdene-4-methy!-1,5-dihydropyrrol-2-one was prepared by the dehydration of 1-benzyl-5-ethyl-5-hydroxy-4-methyl-1,5-dihydropyrrol-2-one as described before. (max: 206nm.
5-Bromomethylene-4-hepty-1-pheny|-1,5-dihydropyrrol-2-one
p-Toluenesulfonic acid (0.05g) was added to a solution of 5-bromomethyi-5-hydroxy-4-heptyl-1-phenyi-1,5-dihydropyrrol-2-one in toluene The mixture was refluxed for 1/2h and after cooling, washed with sat NaHCO3. The organic phase was dned over Na2SO4, and evaporated to yield an.E,Z mixture of 5-bromomethylene-4-heptyl-1-phenyl-1,5-dihydropyrroJ-2-one as a colourless oil which solidified on standing, m.p. 63-65°. vmax: 3414, 3080, 2952, 2853, 1695,

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1627, 1597. 1499, 1446, 1382, 1269, 1074, 907, 831 cm"1; (max. 317 ((max 22,834), 278 (43,910), 204 (46,925) nm; 1H n.m S(CDCI3): 7.4-7.24, nv5H, Ph, 6.04 and 5.94, 2 s, 1H each, =CHBr and C3-H; 2.45, m, 2H, CHZ; 1.65-0.9, m, 13H, alkyl chain.
1 -Benzyl-5-bromomethylene-4-hepty 1-1,5-dihydropyrroI-2-one
1-Benzyl-5-bromomethyl-4-hepy!"5-hydrpxy-1,5-cIihyclropyrrol-2-one dehydrated smoothly to an E and Z mixture of 1-benzyl-5-bromomethylene-4-heptyl-1,5-dihydropyrrol-2-one upon heating a solution of 1-benzyl-5-bramomethyl-4-heptyl-5-hydroxy-1,5-dihydropyrrol-2-one with p-toluenesulfonic acid in toluene; m.p. 52-55°; vmax: 3096, 2927, 2857, 1704, 1630, 1387, 1357, 954, 855, 843 cnV\W: 319 ((max10,220), 276 (19,433), 206 (17,040) nm; 1H n.m,r. 5 (CDCI3): 7.29-7.15, m, 5H, Ph, 6.15 and 5.98, 2s, each 1H, =CHBr and C3-H; 2.39, m, 2H, CH2; 1.7-0.89, m, 13H, alkyl chain.
Reaction of N-(2-HydroxyethyI)-3-butyl-2{5H)pyroIinone with acetic anhydride and Triethylamine
N-(2-Acetoxyethy1)-3-butyl-5-(dibrombmethylene)-2(5H)pyrolinone
A mixture of N-(2-hydroxyethyI)-3-butyl-5-clibroniomethyl-5-hydroxy-2(5H)pyrrolinone (0.2g, 0.54 mmol), acetic anhdride (0.44g; 4.4 mmol) and tnethylamme (0.44g; 4.4 mmol) in dry dichloromethane (10 ml) was refluxed for
2 hr. After cooling to room temperature, the mixture was washed with aqueous
sodium bicarbonate and bnne The organic phase was dried over anhydrous
sodium sulfate and evaporated to yield a viscous oil. 1H n.m.r showed it to be a
mixture of the mono- (88%) and di-acetate (12%) derivatives. Chromatography
on silica using EtOAc/CH2C!2 (5:1) as an eluent yielded 5-acetoxy-N-(2-
acetoxyethyl)-3-butyl-5-dibrornomethyl-2(5H)pyrrolinone (12%) as an oil. vmax-
2957, 2931, 2875, 1766, 1720, 1433, 1369, 1236,.1044, 1013. 855, 707 cm-1
W: 217 ((max 1692), 268 (738) nm. 1H n m.r. 6(CDCl3) 0.91 (t, 3H, CH3); 1.38
(m, 2H, CH2); 1.55 (m, 2H, CH2); 2.05 and 2.10 (each s, 3H, CH3), 2.34 (m, 2H,
CH2); 3.61 (m, 1H, NCH2CH2); 3.64 (m, 1H, NCH2CH2); 4.27 (m, 2H,
NCH2CH3); 6.26 (s, 1H, CHBrs); 6.B3 (&, 1H, H4). 13C n.m.r. 5(CDCI3) 13.7,
20 S, 21.2, 22.1, 24.9r 38.7, 44.1, 61.5, 94.1,134.2, 144.3, 168.4, 170.6, 171 0.
N-(2-acetoxyethy)-3-butyi-5-dibromomethyl-5-hydroxy-2(5H)pyrrolinone (88%)
1H n.m.r. 8(CDC!3) 0.93 (t, 3H, CH3); 1,38 (m, 2H, CH2); 1,55 (m, 2H, CH2);

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2 21 (s, 3H, CH3); 2.34 (m. 2H, CHZ), 3.27 (m, 1H, NCH2CH2); 4.04 (m, 2H, NCH2CH2), 4,30 (s, 1H, OH); 4.62 (m, 1H, NCH2CH2); 5.85 (s, 1H, CHBr2); 6.73 (s, 1H, H4). 13C n.m.r. 5(CDCf3) 13.7, 20.9, 22.2, 24.9, 29.3, 38.1, 45.9, 62.5, 91.0, 137.4,143.3,170.4,171.9
Dehydration of N-(2-acetoxyethyl)-3-buty!-5-dibromomethyl'5-hydroxy-2(5H)pyrrolinone with p-toluenesulfonic a'cid in toluene gave quantitatively N-(2-acetoxyethy!)-3-butyl-5-(dibrornomethyleiie)-2(5H)pyroJinone. vmax 2957, 2929, 2870, 1744, 1705, 1441; 1368, 1229, 1177, 1161, 1130, 1035, Q30. 764 cm-1. 1H n.m.r. 5(COCi3) 0.93 (t, 3H, CH3); 1,36 (m, 2H, CH2), 1.55 (m, 2H, CH2); 2 02 (s, 3H, CH3); 2.32 (m, 2H, CH2); 4.25-4.31 (m, 4H, NCH2CH2); 7.05 (s, 1Ht H4). 13C n.m.r.'S(CDC13) 13-7, 20.7, 22.3, 25.1, 29.5, 39.&, 62.3, 73.8, 132.4, 138.6,140.3,170 6,172.0.
Hydrolysis of N-(2-acetoxyethyl)~3-butyl-2(5H)pyrolinone
N-(2-hydroxyethy!)-3-butyI-5-(dibromomethyIene)-2(5H)pynofinone
A solution of potassium carbonate (1g) in water (3 ml) was added dropwise to a
solution of N-(2-acetoxyethyl)-3-butyl-(dibromornethylene)-2(5H)pyrohnone
(0.2g, 0.51 mmol) in methanol (7ml). After stirring the mixture at room
temperature for 20 mins, methanol was removed in vacuo and the product
extracted with ethylacetate (2 x 40 ml). The resulting extracts were combined,
washed with brine, dried (Na2SO4), and evaporated to yield an oil (0 18 g;
94.5%), which solidified' upon standing in the fridge. Crystallisation from light
petroleum gave N-(2-hydroxyethyI)-3-butyl-5-dibromom&thyten9-
2(5H)pyroimone as colurless granules, nrp. 48-50° max 3404, 2957, 2930, 2880, 1720, 1651, 1465, 1348, 1207, 1081, 1054, 1018, 936, 850, 716 cm"1. vmax 206 ((max 25.389), 239 (6,753), 288 (2,186) niTL 1H n.m.r. SCCDCb) 0.91 (t, 3H, CH3)l 136(171, 2H, CH2): 1.54 (rn, 2H, CH2); 2.29 (m; 2H, CH2); 3.B3 (m, 2H, NCH2CH2); 4.20 (m. 2H, NCH2CH2), 7.02 (s, 1H, H4), 13C n.m r S (CDCI3) 13.7, 22.3, 24.9, 29.5, 43.3, 46.8, 61.9, 74.5,132 3, 138.6,140.5, 173.2

Synthesis of 5-phenylaminomethylene-2(5H)furanone

4-Bromo-5~phenylaminomethylene-2(5H)furanone
A solution of 4-brorno-5-bromomethylene-2(5H)-fu'anone (0.30 g; 0,79 mmol) was dissolved in aniline (5 ml), and left to stand at room temperature for 3 hrs,

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dunng which time the mixture solidified The solid was triturated with CH2Cl2/petroi (1.1; v/v, 20 m!) and filtered. The resulting solid was dned and recrystallised from ethanol to yield 4-bromo-5-pheny!aminomethylene-2(5H)furanone (0,24g, 49%) as yellow needles, m.p. 200-202 (decomp). (Found (HRESMS) m/z 287.963053. CnH8BrNO2Na+ (79Br) requires m/z 287 963840). v max 3233, 3127, 1730, 1697, 1595, 1498, 1276, 1195, 932, 798, 756 cm-1. (max 397 nm ((max 50,686); 246 (12,769), 202 (15,961), 1H n m r. 5 (CDCI3): 9.99, d, J 10.44 Hz, 1H, -NHPh; 7.31-6.99, m, Ph; 7.07, dr J 10.44 Hz, 1H, -CHNHPh; 6.16, s, C3-H. 13C n.m.r. 5 (CDCl3): 109.0 116 2, 117.9, 129.9, 129.6,133.9; 167.5.
5-Phenylaminomethylene-4-bromo-3,-butyl-2(SH)-furanone
A solution of 4-bromo-3-butyl-5-bromomethylene-2(5H)-furanone (0.25 g; 0.81 mmol) in aniline (0.082 g; 0.88 rnmal) was left to stand at room temperature for 72 h. The mixture was diluted with CH2CI2 (50 mi), washed with aqueous hydrochloric acid (2M) and dned over anhydrous sodium sulfate. The solvent was removed in vacuo leaving behind a brown viscous oil (0.29 g). The crude product was chromatographed on silica using dichloromethane to yield 5-phenylammomethylene-4-bromo-3-butyl-2(5H)-turanone as a yellow solid 1H n.m.r ( (CDCl3): 7.40-6.80, m, 5H, Ph; 6.70, d J 12.5 Hz, =CH(NH)Ph; 2.42-2.40, m, 2H, CH2-chain; 1.7-1.2, m, 4H, CH2-chain; 0.95, t J 7.3 Hz, CH3. (Found (HRESMS) m/z 344.021931. C15H16BrNO2Na+ (79Br) requires m/z 344.021891).
4-Bromo-5-phenylaminomGthylene-3-hexyl-2(5H)furanone
A mixture of 4-bromo-3-hexyl-5-bromomethylene-2(5H)-furanone (0.50g; 1.48 mmol) and aniline (1ml) in ethanol (10 ml) was heated at reflux for 2 h. After cooling to room temperature, the mixture was evaporated to dryness and the residue extracted with dichloromethane (20 mi). The organic phase was washed with aqueous hydrochloric acid (2M) and dried over anhydrous sodium sulfate. Removal of the solvent and recrystallisation of the solid from light petroleum gave 3-bromo-5-phenylaminomethyIene-3-hcxyl-2(5H)furanone (0.50g. 100%) as yellow needles; m.p. 147-148°C. vmax 3242, 3161, 3109, 29212, 2842, 1728, 1683, 1600, 1581, 1'SOO, 1350, 1236, 1055, 960, 750, 673 cm-1. (max: 394 ((max 26,287) , 247 (8002) nm. 1H n.m.r 5(CDCI3): 7.32-6.97, m, 5H, Ph: 6.9B, s. -NMPh; 6.73, s, C5 =CH-NHPh; 2.4, t, -CH2cham; 1.61-

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0 88, m, 11H, chain. 13C n.m.r.5(CDCI3): 14.0, 22.0, 24.8, 27.6, 29, 310, 103.0, 113,0,115.0,122.5, 124 0, 129.5 129.5, 131.0.139.9, 167.0.
5-Pheny[aminomethylene-4-heptyl-2{5H)furanone
5-Phenylaminomethyi-4-heptyi-S-hydroxy-2(5H)pyrra!inone 5-Bromomethylene-4-heptyI-2(5H)furanone (0.44g; 1.61 mmol) was dissolved in dry aniline (2 ml) and left to stand at room temperature for 24 h. Dichloromethane (10 ml) was added to the reaction mixture and the organic phase was washed with aqueous hydrochionc (2M) followed by water. After drying over sodium sulfate, the solvent was evaporated off to yield a pale yellow solid. The crude product was chromatographed on silica column using dichloromethane followed by CH2CI2/Et0Ac (2:1; v:v) as the eluents to yield 5-phenylaminomethyi-4-hepty!-5-hydroxy-2(5H)furanone (0.43g; 88%) as a pale yellow solid, m.p, 172-174°C. vmax: 3192, 3037, 2957, 2931, 2953. 1676, 1643, 1598, 1502, 1493, 1336, 1250. 1160, 923, 757 cm'1. (max 278 ((max 7188). 203 (8609) nm. 1H n.m.r, ( (CDCI3): 7.53-7.26, 6H, Ph and -NHPh; 5.73, s, 1H,C3-H, 5.11,s, 1HT|C5-OH; 3 37, d, 2H, -CJigNHPh; 2.2-2.0, m, 2H, -CHz-chain; 1.25-0.91. mt 13H, Chain. 13C n.mr.5 (CDCIa): 14.0, 22 6, 25.5, 25.3, 25.8, 29 0, 29.2, 30.4, 31 6, 93.4, 121.8, 126.0, 126.7, 129.0,134.6, 163.0,170.4.
5-PhenytaminQmethytene-4-heptyl~2(5H)furanone
A sample of 5-phenyIaminomethyl-4-heptyl-5-hydroxy-2(5H)furanone was dehydrated using p-toluenesulfonic acid in toluene to yield an E and Z mixture of 5-phenylaminomethylene-4-heptyI-2(5H)furanone as a colourless oil which solidified on standing in the fridge. vmax 3088(-NH), 3052, 2927, 2856, 1712, 1626, 1598, 1499, 1454, 1264, 1195, 759, 699 cm-1 (max 292 ((max 7623), 204 (4728) nm. 1H n.m.r. ( (CDCIa) 7.4-7.25, 6H, Ph and -NHPh; 6.19-6.1, d, 1H, C3-H; 5.93-6.0, 1H, d, C5- =CMNHPh; 1.68-0.90, 15H, chain. 13C n.m r.6(CDCI3). 14, 22.5, 26.4, 28 1, 28.9, 29.0, 29.2, 30.0, 31.6, 31 7, 88.7, 93.0, 118.5, 1226, 127.8, 128.2, 128.4. 128.6, 128.7, 129.3, 129 5, 134.0, 135.0,142.0,143.0, 152.0, 153.2,168.0.
4-Methyl-5-(1-phenylamIno-ethylidene)~5W-furan-2-one
A solution of 5-ethylidene-4-methyl-2(5H)furanone (0,31 g; 2.5 mmol) in aniline
(0 26 g; 2.75 mmol) was left to stand at r.t for 3 hrs, during which time a solid
precipitated from the reaction. The reaction mixture was triturated with

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CH2CI2/petro] (1:1) and the solid filtered and recrystallised from EtOAc/petroi to yield 5-ethyl-5-hydroxy-4-inethyJ-1 -phenyl-1,5-dihydropyrrol-2-one as colourless crystals (70% ); m.p. 97-100°. vmax: 3287, 1884, 1704, 1530, 1496, 1353. 1101, 1053, 971, 897, 790, 756, 688. 638 cm"1. (max 273 ((max 15,256), 226 (16,382), 243 (39,646) nm. 1H n.m r. S (DMSO-de) 10.11, S, 1H, -NH; 7.57, d, 2H, ArH, 7.30, t, 3H, ArH; 6.08, S, 1H, C3-H; 3 27, s, 3H, CH3; 1-96, s, 3H, CH3. 13C ri.m.r. ( (CDCl3): 20.9, 119.6, 123.7, 123.8, .129.1, 139.3. 142.9, 163.1. 170.4.
Synthesis of 5-arylamino and arylalkylamino-2(5H)furanones 4-bromo-5-benzylam!ao-5-bromomethyl-2(5H)furanone
Benzyl amine (0.10 g, 0.95 mmol) was added with stirring to an, ice-cooled solution of the 4-bromo-5-(bromornethyIene)-2(5H)furanone (0.16 g, 0.64 mmol) in dichloromethane (10 ml). The mixture was stirred at room temperature for 2.5 h, washed with aqueous hydrochloric acid solution (1M, 10 ml), dried (Na2SO4), and evaporated to yield a brown oil. The crude product was chromatographed on silica using dichloramethane/ethyl acetate (1:4; vv) as the eiuent and recrystallfsed from dichloromethane/light petroleum to yield 4~ bromo-5-benzyIamino-5-brornomethy!-2(5H)fiiranorte as orange flakes m.p. 137-139 ° (Found (HRESMS) m/z 381.901032. C21H11 Br2NO2Na+ (79Br) requires 381.904812). vmax 3256, 1674, 1655, 1431, 1413, 1352, 1072, 1054, 699 cm-1. (max 257 ((max 2879) nm 1H n.m.r. ( (CDCl3): 7.38,d, J 11 Hz, 1H,-NHCH2-; 7.37-7.29, m, Ph; 6.38, s, C3-H, 4.65, d, J 15 Hz; 1H, -CH2Bn 4.44. d, J 15 Hz, 1H, -CH2Br and 3.58-3.44, dd, J 15 Hzt CH2Ph. 13C n rn.r.5 (CDCI3): 30.6, 42.8, 53.0, 92.2, 128.0,128.2,12819, 137.0, 142.0, 168.0
4-Bromo-5-benzybminO-5-bromomethyl-3-hexyl-2(5H)furanone
Benzylamine (0.32g; 2.96 mmol) was added with stirring to a solution of 4-bromo-3-hexyl-5-bromomethylene-2(5H)-furanone (0.50 g- 1 48 mmol) in ethanol (6 ml). The mixture was stirred at room temperature for 1 h and evaporated to dryness.The residue was extracted with dichloromethane (20 ml) and the dichloromethane extract washed with aqueous hydrochloric acid (2M). After drying over anhydrous sodium sulfate, removal, of the solvent gave a thick viscous oil, Column chromatography on silica gel using dichloromethane followed by dichloromethane/ethy! acetate (19:1) as the eluents afforded 4-bromo-5-benzylamino-5-bromomethyl-3-hexyl-2(5H)ftiranone (0.36g; 56%) as a

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Viscous oil, m.p. 72-75°.vmax3277, 3065, 3032, 2954, 2928, 2657,1681, 1496, 1411, 1355, 1151, 1064, 1104, 1030, 988, 907, 726, 698. (maX: 277 (( max 39,542), 205 (38,034) nm. 1H n.m.r. 5(CDCl3): 7.4-7.26, m,' Ph; 4.8-4.74. d, and 4.4, d, C5-CH2Bn 3.6 and 3.53, d, CB-NHCH2Ph; 2.42-2.33. m. -CH2, chain; 1.56-0,85, m, 11H, chain. 13C n.m.r. ( (CDCI3): 22.0, 25:0. 27.0, 28.8, 31.4, 42.9, 46.7, 49.5, 90.6, 91.6, 127.0, 128.0, 129.0, 136.0, 136.7, 136.9, 138 0, 140.0,144.0,168.0,170.6
5-Phenylamino-3,5-dimethyI-2(5H)-furanone
Method A
A solution of 3,5-dimethyl-5-hydroxy-2(5H)-furanone (0.13g; 1.02 mmol) in dry aniline (2 mls) was stirred at room temperature for 1 hr. A thin layer chromatography analysis of the mixture (developing solvent; CH2CI2) indicated completion of the reaction as indicated toy the disappearance of the starting material. Dichloromethane (25 mls) were added to the mixture and the solution washed with aqueous hydrochloric acid solution (1M, 3 x 20 mls). The organic layer was dried over anhydrous sodium sulfate and evaporated to yield 5-phenylarnino-3,5-dimethyl-2(5H)-furanone as a viscous oil which solidified on keeping (0 013 g), A sample was recrystallised from dichloromethane/light petroleum to yield the furanone as colouriiees needles v max 3360, 3088, 2965, 1770, 1601, 1570,1536,1294> 1246, 1132,1040, 999, 867, 756, 697 cm-1. W 236 nm. 1H n.m.r. ( (CDCl3): 7.18, t, 2H Ph; 6.90, t, 1H, ArH, 6.89, s, 1H, H4. 6.83, d, 2H, ArH; 4.24, bs, 1h, OH; 1,91, s, 3H, C3-Me;1.75. s, 3H, -Me. 13C n.m.r. ( (CDCI3): 10.4, 26.2, 95.5, 121.3, 122.7, 128 9, 132.4, 133.5, 141.9, 148.8,156.5, 171.9. Method B
A mixture of 3,5-dimethyl-5-hydroxy-2(5H)-furanone (0.13g; 1,02 mmol) and aniline (2 ml) in dry toluene (10 ml) was refluxed for 5h. The mixture was cooled and evaporated. The residue was dissolved in dichloromethane (25 ml) and the solution washed with aqueous hydrochlonc acid solution (1M; 3 x 20 ml). The organic layer was dried over anhydrous sodium sulfate and evaporated to yield 5-phenylarnino-3,5-dimethyl-2(5H)-furanone as a viscous oil. The crude product was chromatographed on silica using dichloromethane/ethyl acetate (19'1) as the eluent (Yield 58-0%).
5-Phenylamino-5-methy!-4-phenyl-2(5H)-furanone

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A mixture of 5-hydroxy-5-methyl-4-phenyl-2(5H)-furanone (0.13g; 1.02 mmol) and aniline (2 ml) in dry toluene (10 ml) was refluxed for 5h. The mixture was cooled and washed with aqueous hydrochloric acid solution (2M; 3 x 20(mls). The organic layer was dried over anhydrous sodium sulfate and evaporated to yield a viscous oil. The crude product was chromatographed on silica using dichloromethane/ethyl acetate (19:1) as the eluent and recrystallised from dichloromethane/Iight petroteurn to yield 5-phenyiamino-5-rnethyI-4-phenyI-2(5H)-furanone (0.10 g; 72%) as colourless flakes m.p. 15S-160°Cvmax: 3355, 1724, 1608, 1534, 1501, 1320, 1291, 1376, 1030. 943, 846. 770, 756! 691, 639 (max 276((max 7056), 238 (5615)nm. H n.m.r. S(CDCl3): 7.94-7 44, m' 5H, -Ph; 7.14-6.82, m, 5H,Ph; 6.4, s, 1H, C3-H; 4.53, bs, 1H, -N.HPh; 1.9, s, C5-Me. 13Cn.m.r.5(CDCl3) 117.3,120,122,5,128, 129,5,131, 142,159,166,170.
5-BenzylaminomethyI-3-methyl-2{5H)furanone
Phosphorus pentoxide (2g) was added to a solution of 3.5-dimethyl-5-hydlraxy-2(5H)-furanone (0.50g; 2.15 mmol) in dichtoromethane (25 ml). The mixture was refluxed for 2h and the cooled solution was filtered through celite and evaporated in vacuo to yield 3-methyl-5-methylene-2(5H)-furanone as a colourless oil (0.37 g; 82%) The methylene product was dissolved in dichloromethane (5 ml) and benzyiamme (1.15 g; 10.8 mmol) was added at room temperature. The mixture was stirred at room temperature for 1 h. After evaporation of the solvent the crude product was chromatographed on silica using dichloromethane/light petroleum as the eluent to yield 5-benzylamtnomeihyI-3-methyI2(5H)furandne as a colourless oil (0.12 g; 26%). vmax: 2929, 2854, 1788, 1747, 1715, 1618, 1456, 1388, 1373, 845, 712 cm"1. W: 308 nm ((max 1462), 260 (5243).1H n.m.r.S (CDCl3): 7.29-7.21, m, 6H, Ph and-NHCH2Ph; 6.65, s, 1H, C4-H; 4.82,s, 2H, -CH2Ph; 4.70, d, -CHgNHPh; 2.02, s, C3-Me. 13C n.m.r. 5 (CDCI3): 10,8, 25.9, 42.9, 95.0.105.3, 126.9,127.1, 128.5,131.2,134.2,137.2, 148.4.
Side-chain functionalization 3-(1-Bromohexyl)-1-butyl-5-dibromomethylene-1,5-dihydropyrroI-2-one
N-Bromosuccinimide (0.32g; 1.79 mmol) was added to a solution of 1-butyl-5-dibromomethyl-3-'hexyl-1(5-dihydropyrrol-2-one (0.64 g; 1.63 mmol) containing few crystals of benzoyl peroxide in CCl4 (25 ml). The mixture was heated at reflux under a 100 watt fluorescent lamp for 24 h. The reaction mixture was

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cooled and passed through a pad of Celite. The filtrate was evaporated to dryness to yield a brown oil which was chromatographed on a silica column using CH2Cl2/petrol (1;1) as the eluent to yield 3-(r-Bromohexy!)-1-Butyl-5-dibromomethylene-1,5-dihydropyrml-2-one (0,46 g; 59.8%) as a pale yetlow oil. (Found): HRESMS): m/z 483.849575. C15H14NB4O3) requires 483,651758 vmax 3017,2950,1709ri598,1593, 1480, 12,15, 1194, 845, 695, 668 cm-1. (max: 326 ((max 4070) nm 1H n.m.r. 8(CDCI3)- 7.28, s, 1H, H4; 4.7$, t, -CHBr-chain; 2.19-2.11,m, -CH2-Chain; 1.53-0.98, m, alky, chain. 13C n.rn.r. ( (CDCI3): 13.12, 20.95, 26.8, 39, 43.9, 79.5, 95,128.6, 128.9.129.4, 133.8, ,134.5, 138.2, 139.6, 168.7
3-(1 '-Bromobutyl)-! -butyl-5»dibromomethylene-1,5-dihydropyrrol-2-one
N-Bromosuccinimide (0 32g; i .79 mmol) was added to a spiution of N-butyl-5-dlbromomethyl-3-hexyl-2(5H)pyrrolinone (0.64 g; 1.63 mmol) containing few crystals of benzbyl peroxide (0.01 g) in CCI4 (25 ml). The mixture was heated at reflux under a 100 watt fluorescent lamp for 24 h. The reaction mixture was cooled and passed through a pad of Cel'rte. The filtrate was evaporated to dryness to yield a semi-solid (0.69g) which was chromatographed on a silica column using CH2Cl2/petrol (1:1) as the eluent to yield 3-(1'-bromobutyl)-1-butyl-5-dibromomethylene-i.s-dihydropyrrbl-2-one (0.4Bg; 60%) as a pale yellow oil. (max 2930, 2957, 2871, 1705, 1584, 1357, 1192, 1055, 902, 769, 651 cm-1; (max: 325 ((max 11,669), 202 (9,879) nm. 1H n.m.r, 5(CDCl3): 7.29, d, 1H, C4-H; 4.76, t. 1Ht C3-CiHBr- chain; 3.98,t, 2H, >NCH2-; 2.10, m, -CH2-chain; 1.58-0 93. m, 12H, chain; 13C n.m.r. 5(CDCI3): 13-781 13-96, 19,8, 22.4, 27.4, 31, 32.2, 37. 41, 43.9, 98.7. 132.5, 138 2, 140, 169.0,
3-(1 '-Bromobuty|)-5-dibromomethylene- N-phenyl-1,5-dihydropyrrol-2-one
N-Bromosuccinimide (0.056g; 0.316 mmol) was added to a solution of 5-dibromomethyl-3-buty!-1-phenyl-1,5-dihydropyrroI-2-one (0-64 g; 1.63 mmot) containing few crystals of benzoyl peroxide (0.01 g) in CCI4 (10 ml). The mixture was heated at reflux under a 100 watt fluorescent lamp for 24 h. The reaction mixture was cooled and passed through a pad of Celite. The filtrate was evaporated to dryness to yield a brown oil (0.17g) which was chromatographed on a silica column using CH2CI2/petrol (1:1) as the eluent to yield 3-(1'-bromobutyl)-5-dibromomethylene-1-phenyM,5-dihydropyrroI-2-one as a pale viscous oil (0.10g). (Found.'HRESMS) m/z: 483.849575,

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C15H14Br3NONa+ (Br79) requires 483.851758. vmax: 3017, 2950, 1709, 1598, 1593, 1480, 1215, 1194, 1122. 845, 756, 695, 668 Cm-1. (.maj(; 326 ((max 3,896), 202 (5,566) nrn. 1H n.m.r. 8CDCl3): 7.45, m, 6H. Ph and C3-H; 4.86, t, 1H, C3-CHBr- chain; 2.16, m, -CH2 chain: 1.53-0.98, m, 5H, alkyl chain. 13C n.m.r.5(CDCi3): 13, 21, 26.8, 39, 43, 79.5, 95, 107. 128.6, 129.4, 134, 134.5, 138, 139.6,169.
N-Phenyl-3-(1-hydroxybutyl)-5-dlbromomethylene-2(5H}pyrrolinone
A solution of N-Phenyl-3-(1-bromobutyl)-6-dibromomethyIene-2(5H)pyroIinone (0.0194mol) in, DMSO (60 ml) containing few drops of water was left to stand aside at room temperature for 6 days. The mixture was diluted with dichioromethane (100 m!) and the resulting solution washed with brine (3 x 120 ml). The organic phase was dried over anhydrous sodium sulfate and evaporated to yield a pale yellow oil (9.08g). The Crude product was purified on a silica column using initially dichioromethane followed by dichloromethane/ethyl acetate to afford N-phenyI-5-dibrommeihylene-3(1 -hydroxybutyI)-2(5H)pyroltnone (6.83g; 88%) as pale yellow needles (dichloromethane/light petroleum), m p. 93-95°. vmax: 3439, 3065, 2957, 2927, 2871, 1701, 1496, 1455, 1370, 1189, 1139, 1095, 1069, 1038, 945, 835, 763, 697 cm-1. (max203 (w 11,968), 313 (10,707) nm. 1H n.m.r. 5 (CDCI3) 0.97 (t, 3H, CH3), 1.39 (m, 2H, CH2); 1.79 (m, 2H, CH2), 4.61, m, 1H, H1'; 2.71, bs, 1H, OH, 7.23 (s, 1H, H4); 7.21-7.44 (m. 5H. ArH). 13C n.m.r. 5(CDCI3) 13.7, 14, 18.5. 37 8, 67.4,128.6,128.9, 128.3, 129.5, 131.5,134.5,139.8, 170.8.
N-Phenyl-3-(1'-acetoxybutyl)-5-dibromomethylene-2(5H)pyrrolinone
A solution of acetyl chloride (0.25 ml, 3.2 mmol) in dichioromethane (3 ml) was added dropwise to an ice-cooled solution of N-Phenyl-3-(1-hydroxybutyl)-5-dibromomethylene-2(5H)pyrolinone (0.1g, 0.25mmo!) in dichioromethane (10 ml) containing triethylamine (0.25 ml, 2.47 mmol). The mixture was stirred in ice for 1h and then at room temperature overnight. The mixture was poured into saturated sodium bicarbonate solution (20 ml) and extracted with dichioromethane (3 x 30 ml). The organic phase washed with water (3 x 20 ml), dried over anhydrous sodium sulfate and evaporated to yield a pale yellow oil (0.11g). The crude product was purified or) a silica column using dichloromethane/ethyl acetate (15:1) to afford N-phenyl-5-dibrommethylene-3(1'-acetoxybutyt)-2(5H)pyro!inone (0.1g) as a viscous oil Vmax: 2960, 2931,

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2873, 1753,1712, 1592, 1494, 1454, 1362, 1262, 1232. 1193, 1148, 1096, 1060, 836, 753, 698 cm-1. (max 281, 321 nm. 1H n.m.r. 5,(CDCI3) 0.96 (t, 3H, CH5): 1.38 (m,2H, CH2); 1 91 (mt 2H, CH^; 5.72, m, 1H, HV; 7.21 (s, 1H, H4);
7 2i-7.40(m,5H,ArH).
Malonic acid mono-[1-(5-dibromomethyl6ne-2-oxo-1-phcnyl-235-dihydro-1H-pyrrol-3-yl)-butyl] ester
A solution of N-Phenyl-3-(1-hydroxybutyi)-5-dibromomethy!ene-2(5H)pyrolinone (0-5g, 1.25 mmol) in dichloromethane (15 mi) containing triethylamine (0.25 ml, 2.47 mmol) was added drop wise over a period of 9h to an ice-cooled solution of malonyl dichlonde (0.36g, 25 mmol) in .dichloromethane (10 ml), The mixture was allowed to stand at room temperature overnight, washed with brine (3 x 20 ml), dned over anhydrous sodium sulfate and evaporated to yield a brown viscous oil. The crude product was purified on a silica column using ethyl acetate/methanol (1:4) to afford malonic acid mono-[1-(5-dibromornethyIerie-2-oxo-1-phenyl-2,5-dihydror1H-pyrrol-3-yl)-butyl] ester.
(0.48g) as a viscous oil vmax 3470, 2959, 2873,1709, 1594, 1494, 1454, 1362, 1245, 1194,1148,1096, 1060. 835, 753, 698 cm-1, (max 271, 303 nm. 1H n.m.r
8 (CDCI3) 0.96 (t, 3H, CH3); 1.46 (m, 2H, CH2); 1.88 (m. 2H, CHa); 3.01, m, 2H,
COCH2CO; 5 67, m, 1H. H1; 7.23, m, 2H, ArH; 7.43 (m, 3H, ArH); 7,59, s, 1H,
H4.
Preparation of 2(5H)pyrrolinone-polystyrene copolymer
A mixture of styrene (7.13g), 3-(1'-Bromobuty!)-5-dibrarnornethylene- N-phenyl-1,5-dihydropyrrol-2-one (0.37g) and AlBN (0.026g) was degassed for 1/2h bby purging with argon gas and then heated at 65°C for 3h. After completion of polymerisation, the mixture was poured into methanol and precipitated polymer was filtered, washed extensively with methanol and dried to yield the copolymer (2.389, 32%)
Surface attachment of 2(5H)pyrrolinone
A layer of malonic acid rnono-[1-(5-dibromomethylene-2-oxo-i-phenyl-2.5-dihydro-1H-pyrrol-3-yl)-butyl] ester was covalently attached to a surface containing ammo groups by immersing the surface in a solution of 2(5H)pyrrolinone (2mg/ml) in acetonitrile/water containing, NHS, N-hydroxy succinimide. The mixture was shaken for 10 minutes and EDC, N-(3-

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dimethylaminopropyi)-N'-ethylacrbodlimide hydrochloride, was added to the solution to give a final concentration of (2mg/ml), After shaking the solution for 24h, the surface was taken out of the solution and washed thoroughly with water and dried. The surface analysis was performed using XPS and %bromme was used a s a marker for determining the extent of covalent attachment.
Biological activity of furanones
Effect of furanones as inhibitor of AHL-mediated quorum sensing, Ai-2 pathway and growth of S. aureus Methods Gfp assay
Briefly, the Gfp assay determines the relative effectiveness of a compound as an inhibitor of AHL mediated quorum sensing. The assay is dependent on a bacterial strain that carries a reporter plasmid. This plasmid expresses the green fluorescent protein (Gfp) in the presence of AHLs (2). The presence of a competitor will prevent AHL mediated Gfp expression of the reporter. The assay can be used to generate an index of inhibition for each compound. The results here, presented as good, moderate, or poor, are based on the index of each of the compounds as an inhibitor of AHL mediated quorum sensing using this bioassay.
Attachrnent/Biofiim formation
The ability of furanones to inhibit biofilm formation or attachment has been
determined using a modification of the 96 well microtitre method described by
Christensen et al. ((1)). The furanones are added to the wells of the microplate and the solvent is allowed to evaporate, leaving the furanones adsorbed onto the plate. Then a suspension of the monitor bacterium, Pseudomonas aerugmosa, is added to each well and incubated for 24h. Following incubation, the wells are nnsed to remove unattached or loosely adhered cells. The attached wells are fixed with formaldehyde and subsequently stained with cyrstal violet. Following extensive washing to remove the crystal violet, the wells are read at 600 nm. The attachment/biofilrn formation in the presence of the furanones is calculated as the percentage of the controls, which are not exposed to the furanones.

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Two-Component signal transaction Assays Taz-1 Assay
The Taz-assay carried out according to the method of Jin and Inouye (1993) with the following alterations. E coli RU1012 (pYT0301) were grownovernight In M9 medium at 37°C supplemented with 100 ug/ml ampicillin and 50 ug/ml kanamycin This overnight culture was then used to inoculate 50 ml M9 medium in side-arm flasks which were then incubated at 37°C and shaken at 180 rpm The OD610 of the growing cultures was monitored regularly and when the OD610 = 0.2 the cultures were placed on ice. Aspartate was added to side-arm flasks to give a final concentration of 3 mM (aspartate stock solution made up in M9 salts).
The test compound or mixtures of, compounds were dissolved in ethanol and added to cultures to give the required final concentrations. Negative controls were prepared with equal volumes of ethanol. Cultures were then placed in a 37°C incubator and shaken for 4 hours (OD6io approximately 0.7) before being removed and put on ice. Samples were then removed for eta-galactosidase assays carried out according to the method of Miller (1972).
V. harveyl bioassay for the detection of AI-2 activity
The V. harveyi bioassay was performed as described previously (Surette and Bassler, 1998). The V. harveyi reporter strain BB170 was grown for 16 hours at 30°C with shaking in AB medium. Ceils were diluted 1 ;5,000 into 30°C prewarmed AB medium and 90 ul of the diluted suspension was added to wells containing supernatant. Furanones were added to the wells to achieve the desired final concentrations and the final volume in each well was adjusted with stenle medium to 100 ul. Ten ul of V. BB152 (AM-, AI-2+) supernatant was used as a positive control and 10 ul of E. coli DH5a supernatant or sterile media was used as a negative control. This strain of E. coll has previously been shown to harbor a mutation in the AI-2 synthase gene, ygaG, which results in a truncated protein wfth no Al-2 activtty (Surette et al. 1998). The microtiter plates were incubated at 30°C with shaking at 175 rpm. Hourly determinations of the total luminescence were quantified using the cherniluminescent setting on a Wallac (Gaithersburg, MD) model 1450 Microbeta Plus liquid scintillation counter. The V, harveyi cell density was

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monitored by the use of a microplate reader (Bio-Rad, Hercules, CA). Activity is reported as the percentage of activity obtained from V. harveyi BB152 cell-free supernatant. While the absolute values of luminescence varied considerably between experiments, the pattern of results obtained was reproducible.
Growth of Staphylococcus aureus
Material and methods
The growth of Staphylococcus aureus against furanones was tested in sidearm
flasks. One percent of an overnight culture was added to the growth media,
Nutnent Broth, containing furanones at the concentrations 1-50 ug/ml. The
bacteria were incubated at 37C and growth was measured at 610 nm.
The results of these experiments are summarised in the table 1.
Table 1. Summary of activity for lactam and other N containing analogues as inhibitor of AHL-mediated quorum sensing, Al-2 pathway and growth of S.
aureus.

Compound
AHL
Al-2 (% of control)
S. aureus (% of control)

26%, 50 ug/ml 57%, 10ug/ml 80%, 5ug/ml
NE at 50 ug/ml

21 %, 50 ug/ml
NE at 50 ug/ml

+++

NE at 50 ug/ml

0% growth at ug/ml for 10hrs

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39%
(100ug/ml)
102% (25 Mg/ml)

+++
2% (50 Mg/ml)
104% (50 Mg/ml)

61 % (20 Mg/ml)
No effect 50M9/mt

++
50% (100 Mg/ml)
No effect 50M9/ml

+++

No effect 50MS/ml

+++

+++

++

Noeffect (50M9/ml)

No effect
(50pg/ml)

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++

No effect (50ug/mi)

Other signal
regulated
Phenotypes

40-50%
reduction

30%
40% reduction in

in
swimming

reduction in
cholera
toxin

motility
in V.

attachment
production
by V

cholerae
and V.

by V.
cholemo

vulntficus

vulnificus

40-50%
reduction
50-80% reduction
30%
40% reduction fn

in
swimming
in protease
reduction in
cholera
toxin

motility
in V.
production by V.
attachment
production
by V.

cholerae
and V.
Vulnificus
by V.
cholerae

vulntficus

vulnificus

40-50%
reduction

30%
40% reduction in

in
swimming

reduction in
cholera
toxin

motility
in V.

attachment
production
by V-

cholerae
and V.

by V.
cholerae

vulntficus

vulnificus

Christensen, G. D., W. A. Simpson, J. J. Younger, L. M. Baddour, F. F. Barrett, D. M. Meiton, and E H. Beachey. 1985. Adherence of coagulase-negative staphylococci to plastic tissue culture plates: a quantitative model for the adherence of staphylococci io medical devices. J. Ciin. Microbiol. 22(6):996-1006.
Andersen, J. B., C. Sternberg, L. K. Pouisen, S. P. Bjom, M. Givskov, and S. Molm. 1998. New unstable vanants of green fluorescent protein for studies of

WO 2004/016588 PCT/AU2003/001053
52
transient gene expression in bacteria. Appl. Environ. Microbiol. 64(6):2240-2246.
Jin, T and M. inouye. 1993. Ligand binding to the receptor domain regulates the ratio of kinase to phosphatase activities of the signalling domain of the hybrid Eschenchia coll transmembrane receptor, Taz1. J. Mol. Biol. 232: 484-49
Miller, J. H. 1972 Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor,. N.Y.
Surette, M. G., and B. L. Bassler. 1998. Quorum sensing in Eschenchia coh and Salmonella typhimurium. Proc. Nat!. Acad. Sci, USA 95:7046-7050
Surette, M. G., M. B. Miller, and B. L. Bassler. 1999. Quorum sensing in Eschenchta coh, Salmonella typhimurium, and Vibrio han/eyi: a new family of genes responsible for autoinducer production. Proc, Natl. Acad. Sci., USA 96:1639-1644.
Any description of prior art documents herein is not to be taken as an admission that the documents form part of the common general knowledge of the relevant art.
It will be appreciated by persons skilled in the art that numerous variations and/or modifications may be made to the invention as shown in the specific embodiments without departing from the spirit or scope of the invention as broadly described. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive.
Any description of prior art documents herein is not to be taken as an admission that the documents form part of the common general knowledge of the relevant art.

53 WE CLAIM:
1 A method for the preparation of a compound of formula II

wherein R1 and R2 are independently selected from the group H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted substituted or unsubstituted oxoalkyl, substituted or unsubstituted substituted or unsubstituted alkenyl, substituted or unsubstituted substituted or unsubstituted aryl or arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophihc,
R3 and R4 are independently selected from the group H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl,
R5 is selected from the group consisting of H, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted alkylsilyl, substituted or unsubstituted substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophihc, or
forms part of an ammo acid, or
is a nucleoside, an ohgomer, a polymer, a dendnmer, a substrate or a surface,
the method comprising reacting a compound of formula I

54
wherein R1 and R2 are independently H, halogen, substituted or unsubstituted alkyl,
substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophihc,
R3 and R4 are independently H, halogen, substituted or unsubstituted alkyl, substituted or
unsubstituted alkoxy, substituted or unsubstituted aryl or arylalkyl, and R is hydroxy, halogen, and
"===" represents a double bond, in which case R is absent, or a single bond, provided that at
least one of R1, R2, R3 and R4 is halogen,
with a compound of formula R5NH2
wherein R5 is selected from the group consisting of H, substituted or unsubstituted alkyl, hydroxy, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophilic, or
forms part of an ammo acid, or
is a nucleoside, an oligomer, a polymer, a dendnmer, a substrate or a surface
2 A method as claimed in claim 1 wherein R5 is a residue of a naturally occurring compound
3 A method as claimed in claim 1 wherein R5 is a biomolecule
4 A method as claimed in claim 3, where R5 is a coenzyme or cofactor
5. A method as claimed in any one of claims 1 to 5, wherein R5 is an oligomer or a polymer and the oligomer or polymer is a biomolecule
6 A method as claimed in claim 5, wherein R5 is a peptide or polyamide
7 A method as claimed in any one of claims 1 to 5, wherein R5 is a protein residue
6
55
8 A method as claimed in claim 7, where R5 is an enzyme or a receptor
9. A method as claimed in any one of claims 1 to 5, wherein R5 is an ohgomer or polymer comprising nucleic acid residues.
10 A method as claimed in claim 9, wherein R5 is a polynucleotide
11 A method as claimed in claim 10, wherein the polynucleotide is DNA or RNA
12. A compound of formula II

wherein R1 and R2 are independently H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or iluorophilic;
R3 and R4 are independently H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl,
R5 is selected from the group consisting of H, hydroxy, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophilic, or
forms part of an amino acid, or
is a nucleoside, an ohgomer, a polymer, a dendnmer, a substrate or a surface

56
13 A compound as claimed in claim 12, wherein at least one of R1, R2, R3 and R4 is halogen
14 A method for preparing a compound of formula III, the method comprising dehydration a
compound of formula II as claimed in claim 12 or 13:

wherein R1 and R2 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophihc,
R3 and R4 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl or arylalkyl, and R5 is selected from the group consisting of H, substituted or unsubstituted alkyl, hydroxy, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophihc, or
forms part of an amino acid, or
is a nucleoside, an oligomer, a polymer, a dendrimer, a substrate or a surface
15 A method as claimed in claim 14, wherein at least one of R1, R2, R3 and R4 in formula III is
halogen
16 A method as claimed in claim 14 or 15, wherein the dehydration is carried out in the
presence of a dehydrating agent
15
57
17 A compound of formula III

wherein R1 and R2 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophihc,
R3 and R4 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl or arylalkyl, and
R5 is selected from the group consisting of H, substituted or unsubstituted alkyl, hydroxy, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophihc, or
forms part of an amino acid, or
is a nucleoside, an ohgomer, a polymer, a dendnmer, a substrate or a surface
18 A compound as claimed in claim 17, wherein at least one of R1, R2, R3 and R4 is halogen
19 A compound as claimed in claim 17 or 18, selected from the group consisting of

58
20 A compound of formula (VI)

wherein R1 and R2 are independently selected from H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophihc;
R3 and R4 are independently selected from H, halogen, substituted or unsubstituted alkyl,
substituted or unsubstituted aryl or arylalkyl,
R5 is selected from the group consisting of H, substituted or unsubstituted alkyl, hydroxy, substituted or unsubstituted alkoxy, substituted or unsubstituted oxoalkyl, substituted

59
or unsubstituted alkenyl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic or fluorophihc, or
forms part of an amino acid, or
is a nucleoside, an ohgomer, a polymer, a dendnmer, a substrate or a surface; and Z is selected from the group R2, halogen, OC(O)R2, =0, amine azide, thiol, R2, mercaptoaryl, arylalkoxy, mercaptoarylalkyl, SC(O)R2, OS(O)2R2, NHC(O)R2, =NR2 orNHR2
21 An ohgomer or a polymer formed by oligomensing or polymerising a compound of formula
II, III or VI directly or with one or more other monomers
22 An ohgomer or a polymer as claimed in claim 21, wherein the one or more other monomer is
selected from the group acrylate ester such as substituted or unsubstituted alkyl,
hydroxyalkyl, armnoalkyl, or substituted substituted or unsubstituted aryl acrylates or
methacrylates, crotonates, substituted or unsubstituted acrylonitnles, vinyl alcohols or
acetates, styrene and siloxanes.

23. A surface coating or polymer having incorporated therein a compound as claimed in claim
12 or claim 17
24. A composition comprising an effective amount of a compound as claimed in claim 12 or
claim 17 for treating an infection in a human or animal subject
25 A composition comprising an effective amount of a compound as claimed in claim 12 or claim 17 for treating an infection or condition in a subject that is characterised by biofilm formation.
26. A composition as claimed in claim 25, wherein the condition is cystic fibrosis
27. A composition as claimed in claim 25, wherein the condition is dental caries, periodontitis,
otitis media, muscular skeletal infections, necrotising fascitis, biliary tract infection,
osteomyelitis, bacterial prostatitis , native valve endocarditis, cystic fibrosis pneumonia,
26.
6o
meloidosis 28 A composition as claimed in claim 25, wherein the condition is nosocomial infection
29. A composition as claimed in claim 25, wherein the infection is ICU pneumonia or an
infection associated with sutures, exit sites, artenovenous sites, scleral buckles, contact
lenses, urinary catheter cystitis, peritoneal dialysis (CAPD) peritonitis, lUDs, endotracheal
tubes, Hickman catheters, central venous catheters, mechanical heart valves, vascular grafts,
biliary stent blockage, and orthopaedic devices, penile prostheses
30. A composition as claimed in claim 25, wherein the infection is selected from the group of a
skin infection, burn infection and wound infection
31 A composition as claimed in any one to claims 25 to 30, wherein the subject is an immunocompromised individual
32. A composition composing an effective amount of a compound as claimed in claim 12 or
claim 17 for treating or preventing biofilm formation on a surface
33. A composition as claimed in claim 32, wherein the surface is a biological surface.
34 A composition as claimed in claim 33, wherein the surface is a surface of a tissue, membrane
or skin
35 An implant device having at least one surface associated with a compound as claimed in
claim 12 or claim 17
36 An implant device as claimed in claim 35, wherein the device is an artificial heart valve or
hip joint, an indwelling catheter, pacemaker, surgical pin and the like
34
61
37 A biofilm removing or inhibiting composition comprising a compound as claimed in claim
12 or claim 17 and a vehicle or carrier, wherein the amount of the mixture is effective to
remove or disrupt a bacterial biofilm or inhibit normal biofilm formation
38 A composition composing a cleaning-effective amount of a compound as claimed in claim
12 or claim 17 for removing a biofilm from a biofilm-containing surface
39 A composition comprising an effective amount of a compound as claimed in claim 12 or
claim 17 wherein the amount is effective to prevent biofilm formation on a surface
40. A method of removing a biofilm from a non-biological surface comprising the step of administering a cleaning-effective amount of a compound as claimed in claim 12 or claim 17, to a biofilm-containing non-bio logical surface
41 A method of preventing biofilm formation on a non-biological surface comprising the step of administering an effective amount of a compound as claimed in claim 12 or claim 17 to a non-biological surface, wherein the amount is effective to prevent biofilm formation.
Novel synthesis methods to the products of such novel methods and to uses of these products. In particular, the present invention provides methods for the reactions of furanones, in particular fimbrolides, with amines. The invention has particular application in the synthesis of halogenated l,5-dthydro-pyrrol-2-one, 5-halomethylene substituted l,5-dihydropyrrol-2-ones (laetam analogues of fimbrolides), 5-amino substituted furanones and 5-ammomethylene-2(5H)-furanones and their synthetic analogues. The invention also relates to novel compounds and uses thereof.

Documents:

00460-kolnp-2005-abstract.pdf

00460-kolnp-2005-assignment.pdf

00460-kolnp-2005-claims.pdf

00460-kolnp-2005-correspondence.pdf

00460-kolnp-2005-correspondence_1.1.pdf

00460-kolnp-2005-correspondence_1.2.pdf

00460-kolnp-2005-correspondence_1.3.pdf

00460-kolnp-2005-description(complete).pdf

00460-kolnp-2005-form-1.pdf

00460-kolnp-2005-form-13.pdf

00460-kolnp-2005-form-13_1.1.pdf

00460-kolnp-2005-form-18.pdf

00460-kolnp-2005-form-3.pdf

00460-kolnp-2005-form-3_1.1.pdf

00460-kolnp-2005-form-3_1.2.pdf

00460-kolnp-2005-form-5.pdf

00460-kolnp-2005-international publication.pdf

00460-kolnp-2005-international search authority report.pdf

00460-kolnp-2005-others document.pdf

00460-kolnp-2005-pct others.pdf

00460-kolnp-2005-power of authority.pdf

460-kolnp-2005-granted-abstract.pdf

460-kolnp-2005-granted-assignment.pdf

460-kolnp-2005-granted-claims.pdf

460-kolnp-2005-granted-correspondence.pdf

460-kolnp-2005-granted-description (complete).pdf

460-kolnp-2005-granted-examination report.pdf

460-kolnp-2005-granted-form 1.pdf

460-kolnp-2005-granted-form 13.pdf

460-kolnp-2005-granted-form 18.pdf

460-kolnp-2005-granted-form 3.pdf

460-kolnp-2005-granted-form 5.pdf

460-kolnp-2005-granted-letter patent.pdf

460-kolnp-2005-granted-pa.pdf

460-kolnp-2005-granted-reply to examination report.pdf

460-kolnp-2005-granted-specification.pdf

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Patent Number

215543

Indian Patent Application Number

460/KOLNP/2005

PG Journal Number

09/2008

Publication Date

29-Feb-2008

Grant Date

27-Feb-2008

Date of Filing

18-Mar-2005

Name of Patentee

BIOSIGNAL LIMITED

Applicant Address

RUPERT MYERS BUILDING, GATE 14, BARKER STREET, UNSW SYDNEY, NEW SOUTH WALES 2052

Inventors:

#	Inventor's Name	Inventor's Address
1	KUMAR, NARESH	33 WHITE AVEUE, MAROUBRA, NEW SOUTH WALES 2035

PCT International Classification Number

C07D 207/36

PCT International Application Number

PCT/AU2003/001053

PCT International Filing date

2003-08-19

PCT Conventions:

#	PCT Application Number	Date of Convention	Priority Country
1	2002950862	2002-08-19	Australia