AU2001267155B2 - Synthesis of cyclic compounds - Google Patents

Description

WO 02/00639 PCT/AU01/00781 1 Synthesis of cyclic compounds 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 cyclisation of substituted or unsubstituted halogenated 4-oxoalkanoic acids (halogenated levulinic acids) and to the products of such a method. The invention has particular application in the synthesis of furanones including fimbrolides (halogenated 5-methylene-2(5H)-furanones) and their synthetic analogues. The invention also relates to novel furnanone compounds and uses thereof.

Background Art 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 99/53915, the disclosures of which are incorporated herein by cross-reference). These metabolites can be isolated from red marine algae Delisea fimbriata, Delisea elegans and Deliseapulchra.

In spite of recently discovered biological significance offimbrolides, there is not at present a general method suitable for the large-scale synthesis of these metabolites.

The few reported syntheses of these metabolites utilise either the sulfuric acid-catalysed cyclisation ofbrominated levulinic acid at elevated temperatures (Tetrahedron 1997, 53: 15813-15826) or use (E)-P-bromo--lithioacrylate Org. Chem. 1985, 50, 2195- 2198) or allenes Org. Chem. 1995, 60, 1814-1822) as starting materials.

Known acids used in the cyclisation reaction include 100% or 98% sulfuric acid which causes a high degree of charring during the reaction thus producing large quantities of intractable materials. Furthermore copious amounts of water are required to quench these reactions, a process that generates a large quantity of aqueous acidic waste.

These reactions are non-selective, extremely difficult to control and lead to mixtures of different products due to scrambling of bromines atoms under these conditions. Exhaustive chromatography is required to separate the reaction products and this results in low yields of desired 4-bromo-5-(bromomethylene)-, (dibromomethylene)-, 4-bromo-5-(dibromomethylene)-2(5H)-furanones. The chromatography required is tedious and often impractical for large scale reactions.

The compounds 3-alkyl-4-bromo-5-(bromomethylene)-,and (dibromomethylene)-2(5H)-furanones are key intermediates in the synthesis of highly of active side chain functionalised furanones (see WO 99/54323, the disclosure of WO 02/00639 PCT/AU01/00781 2 which is incorporated herein by cross-reference). Accordingly, there is a need for more efficient and reliable syntheses of parent furanones.

We have found conditions that, surprisingly, enable the cyclisation of halogenated 4-oxoalkanoic acids under mild conditions. We have found this discovery to be particularly useful in cyclising brominated 4-oxopentanoic acids under mild conditions to afford high yields of brominated 2(5H)-furanones or furanones under mild conditions. Furthermore tetrahydrofuranones generated under these conditions can be dehydrobrominated to yield a range of 5-(methylene)-, (bromomethylene)-, 5-(dibromomethylene)- or 4-bromo-5-(bromomethylene)-2(5H)furanones. These furanones can be further functionalised to yield novel analogues of Delisea metabolites.

Disclosure of Invention In a first aspect, the present invention provides a method for the preparation of a compound of formula II RI

R

2 Re O R3 II R4 wherein R 1 and R 2 are independently H, alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic, hydrophobic or fluorophilic; R3, R 4

R

5 and R 6 are independently or all hydrogen or halogen; and provided that at least two of the R 3

R

4

R

5 and R 6 are halogens; the method comprising cyclising a compound of formula I WO 02/00639 PCT/AU01/00781 3 Rs

R

1

OOH

R

3

R

4 R2

I

wherein RI, R 2

R

3 R4, R5 and R 6 are as defined above, wherein the cyclisation is carried out in the presence of a mild acid catalyst or a dehydrating agent or a mixture thereof, optionally in the presence of solvent.

In formula II, a particular geometry is not to be taken as specified. For example, the formula covers both Z- and E- isomers.

The starting 4-oxoalkanoic acid of formula I and the tetrahydrofuranone of formula II preferably have the following substituents wherein: RI and R 2 are independently H, alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic, hydrophobic or fluorophilic;

R

3

R

4 R5 and R 6 are independently or all hydrogen or halogen; and provided that at least two of the R 3 R4, R 5 and R 6 are halogens Cl, Br, I); More preferably, the starting levulinic acid of formula I and the tetrahydrofuranone of formula II have the following substituents wherein:

R

1 and R2 are independently H, alkyl, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic, hydrophobic or fluorophilic; R3, R4, R 5 and R 6 are independently or all hydrogen or halogens; and provided that at least two of the R3, R4, R5 and R1 are halogens Br, I); Most preferably, the starting levulinic acid of formula I and the tetrahydrofuranone of formula II have the following substituents wherein: RI and R2 are independently H, alkyl, aryl or arylalkyl whether unsubstituted or substituted, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic, hydrophobic or fluorophilic;

R

3 R4, Rs and R6 are independently or all hydrogen or halogen; and provided that at least two of R3, R4, R5 and R6 are halogens Br, I); Most preferably, at least one of R5 and R6 is Br.

WO 02/00639 PCT/AU01/00781 4 The method of the present invention has particular application in the cyclisation of brominated 4-oxopentanoic acid compounds of formula I wherein at least two of R3, R4, R5 and R6 are halogens. Preferably, the compound of formula II produced by the method of the present invention is selected from halogenated 2-tetrahydrofuranones.

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 from alkyl, cycloalkyl, alkenyl, alkynyl, halo, haloalkyl, haloalkynyl, hydroxy, alkoxy, alkenyloxy, haloalkoxy, haloalkenyloxy, nitro, amino, nitroalkyl, nitroalkenyl, nitroalkynyl, nitroheterocyclyl, alkylamino, dialkylamino, alkenylamine, alkynylamino, acyl, alkenoyl, alkynoyl, acylamino, diacylamino, acyloxy, alkylsulfonyloxy, heterocyclyl, heterocycloxy, heterocyclamino, haloheterocyclyl, alkylsulfenyl, alkylcarbonyloxy, alkylthio, acylthio, phosphorus-containing groups such as phosphono and phosphinyl.

The term "alkoxy" denotes straight chain or branched alkyloxy, preferably C.-o 10 alkoxy. Examples include methoxy, ethoxy, n-propoxy, isopropoxy and the different butoxy isomers.

The term "alkenyl" denotes groups formed from straight chain, branched or mono- or polycyclic alkenes and polyene. Substituents include mono- or polyunsaturated alkyl or cycloalkyl groups as previously defined, preferably C 2 10 alkenyl.

Examples of alkenyl include vinyl, allyl, 1-methylvinyl, butenyl, iso-butenyl, 3-methyl- 2-butenyl, 1-pentenyl, cyclopentenyl, 1-methyl-cyclopentenyl, I1-hexenyl, 3-hexenyl, cyclohexenyl, 1-heptenyl, 3-heptenyl, 1-octenyl, cyclooctenyl, 1-nonenyl, 2-nonenyl, 3-nonenyl, 1-decenyl, 3-decenyl, 1,3-butadienyl, 1-4,pentadienyl, 1,3-cyclopentadienyl, 1,3-hexadienyl, 1,4-hexadienyl, 1,3-cyclohexadienyl, 1,4-cyclohexadienyl, 1,3cycloheptadienyl, 1,3,5-cycloheptatrienyl, or 1,3,5,7-cyclootatetraenyl.

The term "halogen" denotes fluorine, chlorine, bromine or iodine, preferably bromine or fluorine.

The term "heteroatoms" denotes O, N or S.

The term "acyl" used either alone or in compound words such as "acyloxy", "acylthio", "acylamino" or diacylamino" denotes an aliphatic acyl group and an acyl group containing a heterocyclic ring which is referred to as heterocyclic acyl, preferably a Cj.lo alkanoyl. Examples of acyl include carbamoyl; straight chain or branched alkanoyl, such as formyl, acetyl, propanoyl, butanoyl, 2-methylpropanoyl, pentanoyl, 2,2-dimethylpropanoyl, hexanoyl, heptanoyl, octanoyl, nonanoyl, decanoyl; alkoxycarbonyl, such as methoxycarbonyl, ethoxycarbonyl, t-butoxycarbonyl, t- WO 02/00639 PCT/AU01/00781 pentyloxycarbonyl or heptyloxycarbonyl; cycloalkanecarbonyl such as cyclopropanecarbonyl cyclobutanecarbonyl, cyclopentanecarbonyl or cyclohexanecarbonyl; alkanesulfonyl, such as methanesulfonyl or ethanesulfonyl; alkoxysulfonyl, such as methoxysulfonyl or ethoxysulfonyl; heterocycloalkanecarbonyl; heterocyclyoalkanoyl, such as pyrrolidinylacetyl, pyrrolidinylpropanoyl, pyrrolidinylbutanoyl, pyrrolidinylpentanoyl, pyrrolidinylhexanoyl or thiazolidinylacetyl; heterocyclylalkenoyl, such as heterocyclylpropenoyl, heterocyclylbutenoyl, heterocyclylpentenoyl or heterocyclylhexenoyl; or heterocyclylglyoxyloyl, such as, thiazolidinylglyoxyloyl or pyrrolidinylglyoxyloyl.

The term "fluorophilic" is used to indicate the highly attractive interactions between certain groups, such as highly fluorinated alkyl groups of C4-C10 chain length, have for perfluoroalkanes and perfluoroalkane polymers.

The mild acid catalysts may be selected from catalysts that are insoluble in the reaction medium or catalysts that are soluble in the reaction medium. Examples of insoluble acid catalysts include polyphosphoric acid, Eaton's reagent, acidic resins and polymers, Lewis acids, acidic metal salts.

Examples of soluble acid catalysts include chlorosulfonic acid, phosphoric acid, p-toluenesulfonic acid, trifluoromethanesulfonic acid, methanesulfonic acid, acetic acid, bromine, phosphorus tribromide and hydrobromic acid.

Examples of suitable dehydrating agents include phosphorus pentoxide, silica gel, molecular sieves, alumina, phosphorus oxychloride, acetic anhydride, N, N'dicyclohexyl-carbodiimide (DCC), trifluoroacetic anhydride, trifluorosulfonic acid anhydride (triflic anhydride).

Preferably cyclisation is carried out using phosphorus pentoxide or polyphosphoric acid by itself or mixed with a mineral acid. More preferably cyclisation is carried out using phosphorus pentoxide.

The cyclisation may be performed with a mild acid or a dehydrating agent 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, 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 trichloroethane.

The reaction is preferably carried out at mild temperatures. Preferably the cyclisation reaction is performed at a temperature in the range of from about 20-150 0

C.

WO 02/00639 PCT/AU01/00781 6 Where a solvent is present, the cyclisation may be performed at reflux temperature of the solvent, for example, at the reflux temperature of dichloromethane.

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.

In the case of halogenated 4-oxoalkanoic acids, whilst the parent levulinic acid is commercially available, the corresponding alkyl substituted 4-oxoalkanoic acids can be prepared from the condensation of ethyl acetoacetate with alkyl haloalkanoates followed by hydrolysis and decarboxylation of the keto ester (Tetrahedron, 1997, 53, 15813). Di-, tribromolevulinic acids are readily obtained by the bromination of the corresponding levulinic acids, for example with bromine and catalytic hydrobromic acid.

Alternatively the brominated 4-oxoalkanoic acid derivatives can be conveniently obtained by the addition of bromine or hydrogen bromide to the corresponding 4-oxo- 3-alkyl-2-pentenoic acids.

The present invention extends to these brominated intermediates including those novel compounds of formula I.

Representative examples of 3-alkyl-2,3-dibromo-4-oxoalkanoic acids (la-i) prepared for use in this invention are listed below.

WO 02/00639 PCT/AU01/00781 H COOH Br 0 la Br H COOH SBr 0 Id Br H COOH Br lb Br 0 Ic Br Br H OOH CH Br COOH Br 0 Br 0 0 1g 1 h The present inventors have found that with a judicious choice of acid catalysts and solvents, the brominated levulinic acids could be cyclised with few side products and in high yields to the corresponding tetrahydro-2(5H)-furanones. In particular the use of phosphorus pentoxide in dichloromethane provided very efficient cyclisation of the levulinic acids to The surprising results obtained by the present inventors are in sharp contrast to those reported in the literature for attempted cyclisation of brominated levulinic acid. It has been reported that when 5-bromolevulinic acid was treated with relatively mild dehydrating agents (trifluoroacetic acid or dicyclohexylcarbodiimide) the major product of the cyclisation reaction was a cyclic allyl bromide Am. Chem. Soc., 1981, 103, 5459).

No further reaction of the tetrahydrofuranone was observed even if the reaction was continued for a longer period of time. This reaction appears to be quite general and was repeated on a hundred gram scale.

Representative examples of tetrahydrofuranones (2a-i) that can be synthesised by this procedure arc listed below.

WO 02/00639 WO 0200639PCT/AU01/00781 BrBr Br 0:4H O::Br 0Br 2a Br 2b Br 2c Br H MeHH Br Br Br Br Br BrH 2d H 2e H 2f H H BrB Bt Br

B

0 0 2g H2h 2i in a second aspect, the present invention consists in a derivative of formula 11, wherein R, and R(2 are independently H, alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic, hydrophobic or fluorophilic; R(3, R 4

R

5 and R 6 are independently or all hydrogen or halogen; and provided that at least two of the R(3, 1(4, 1(5 and R 6 are halogens; with the proviso RI= R(2 R(3= R 4 Cl, R5= R 6 Br; RI,= R 2 R5,= R(6 H, KR3, R4 Cl.

The inventors have found the brominated tetrahydrofurariones of formula II can be dehydrobrominated to yield a range of 5-(methylene)-2(5H)-furanofles, (dibromnomethylene)-2(5H)-furanones or 4-bromo-5-(bromlornethy~le) 2 furanones.

We believe that the 2(5H)-furanones prepared in accordance with the present invention are novel compounds.

WO 02/00639 PCT/AU01/00781 9 In a third aspect, the present invention provides a method for the dehydrohalogenation of a compound of formula II above, provided that at least two of the R 3 R4, R5 and R 6 are halogens; to prepare a compound of formula IIIa or lIlb;

R

1

R

Ilia R 4 wherein R2 is an alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophobic, hydrophilic or fluorophilic; R4 is a halogen (X F, Cl, Br or I); R, and R3 are independently or both hydrogen or halogen; R R 2 O OY R 3 Ilb R4 wherein R2 is an alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophobic, hydrophilic or fluorophilic; R3 and R 4 are hydrogen and R, is a halogen; the method comprising contacting a compound of formula II with a base.

The compound of formula II used in the second aspect of the invention may be a compound of formula II produced by the method of the first aspect of the invention, although compounds of formula II produced by other methods may be used.

Dehydrobromination of the tetrahydrofuranones to 5-(methylene)-, (bromomethylene)- and 5-(dibromomethylene)-2(5H)-furanones may be accomplished by treatment of the tetrahydrofuranones with a base e.g. 1,4-diazabicyclo[2.2.2]octane (DABCO), 4-(dimethylamino)pyridine (DMAP), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), triethylamine, sodium or potassium carbonate, sodium or potassium acetate and N,N'-diisopropylethyl amine (Hunig's base).

WO 02/00639 WO 0200639PCT/AU01/00781 Representative examples of furanones (3a-n) that can be synthesised by this procedure are listed below.

0 -Zz H 0 3a Br

H

0 Br 3d Br

H

Br 0 0 Br 3b Br 3c B

H

SBr '0 3e (88) Br

SH

0 M 3f

H

Br

SH

0 70 3g

H

Br

SH

00 3h

H

SH

0 0B Br 3j (63)

B

H

0 0

H

3k Me H 0 Br 3m (64) Br In a fourth aspect, the present invention consists in a 2(5H)-furanone derivative having formula 11la or M~b; WO 02/00639 PCT/AU01/00781 11 R1 R2 O 0o R 3 IlIa R4 wherein R2 is an alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophobic, hydrophilic or fluorophilic;

R

4 is a halogen (X F, Cl, Br or I); R, and R 3 are independently or both hydrogen or halogen; with the proviso that RI H, R 2 Me or Ph, R 3 I, R4 H; and R1 H, R 2 OMe, R3 Cl, R 4

C,

R1 R2 0 'NR3 O \.R3 Illb R4 wherein R2 is an alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophobic, hydrophilic or fluorophilic; and

R

3 and R 4 are hydrogen and Ri is a halogen.

In a fifth aspect, the present invention provides a method for the halogenation of a compound of formula HI, Ia or IlIb above to prepare a compound of formula IV

R

1 R2 O R

IV

wherein R 2 is independently H, halogen, alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophilic or fluorophilic;

R

4 and Rs are halogen (X F, Cl, Br or R 4 can also be OH or alkoxy; R, and R 3 are independently or both hydrogen or halogen; and WO 02/00639 PCT/AU01/00781 12 is a single bond or double bond, the method comprising contacting a compound of formula III with a halogenating agent in the absence of solvent or in the presence of an unreactive or reactive solvent or reagent..

Halogenation of the 2(5H)-furanones of formula (III) to or 5-halo-5-dihalomethyl-2(5H)-furanones of formula (IV) may achieved by treatment of the 2(5H)-furanone with an halogenating agent e.g. bromine, chlorine, iodine, Nbromosuccinimide, N-chlorosuccinimde, iodine monochloride, phenyltrimethylammonium bromide perbromide, pyridinium tribromide and cupric bromide.

Unreactive solvents and reagents are non-nucleophilic organic solvents or ionic liquids, including dichloromethane, chloroform, toluene, diethyl ether, N,Ndimethylformamide, N-methylpyrrolidinone, butylmethylimidazolium tetrafluoroborate.

Reactive solvents and reagents are nuceophilic organic or inorganic substances such as water, methanol, acetic acid, lithium chloride, benzylamine and silver nitrate.

Representative examples offuranones (4a-p) that can be synthesised by this procedure are listed below.

WO 02/00639 WO 0200639PCT/AU01/00781 Br Me 0 0 Br Br 4a 4b 4c 4e (120) 0 o BrB r 4f Br Br 0 Br B 4i Me 0 4g 4h 4i 4U 1(2)4m 4n 41(72) WO 02/00639 PCT/AU01/00781 14 In a sixth aspect, the present invention consists in a 2(5H)-furanone of formula

IV,

RO R

IV

wherein R2 is independently alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophilic or fluorophilic;

R

4 and R5 are halogen (X F, Cl, Br or R4 can also be OH or alkoxy; Ri and R 3 are independently or both hydrogen or halogen; and is a single bond or double bond.

In yet a seventh aspect, the present invention provides a method for the dehydrohalogenation of a compound of formula IV above, to prepare a compound of formula V

R

1 R2 V R4 wherein R2 is a H, alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophobic, hydrophilic or fluorophilic; R4 is a halogen (X F, Cl, Br or I); R, and R 3 are independently or both hydrogen or halogen; the method comprising contacting a compound of formula IV with a base.

The compound of formula IV used in the seventh aspect of the invention may be a compound of formula IV produced by the method of the fifth aspect of the invention, although compounds of formula IV produced by other methods may be used.

Dehydrohalogenation of the to 5-halo-5-halomethyl- or 2(5H)-furanones of formula (IV) may be accomplished by treatment of the furanones WO 02/00639 PCT/AU01/00781 with a base e.g. 1,4-diazabicyclo[2.2.2]octane (DABCO), 4-(dimethylamino)pyridine (DMAP), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), triethylamine, sodium or potassium carbonate, sodium or potassium acetate and N,N'-diisopropylethyl amine (Hunig's base).

Surprisingly the present inventors have found that the dehydrohalogenation of compounds of formula (IV) where R1 is a halogen cannot be achieved satisfactorily by the use of the above mentioned reagents. Pleasingly the present inventors have found that dehydrohalogenation of these compounds can be achieved successfully by the use of N,N-diisopropylethyl amine.

Representative examples of furanones (5a-m) that can be synthesised by this procedure are listed below.

WO 02/00639 WO 0200639PCT/AU01/00781 Br Me Br Ph oj0 Br

B

0:

H

5f (105) Br Br r 0 Sr ~Br BD; e 0

B

5j Br Br 0 SBr Sk Br

B

Br 61 Br Br 0 4 E3 5m Br WO 02/00639 PCT/AU01/00781 17 In an eighth aspect, the present invention consists in a 2(5H)-furanone of derivative of formula V, wherein R2 is an alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophobic, hydrophilic or fluorophilic; R4 is a halogen (X F, C1, Br or I); R, and R 3 are independently or both hydrogen or halogen; with the proviso that R H, R2 Me or Ph, R3 I, R4 H; and

R

1 H, R 2 OMe, R 3 Cl, R4 C1.

Furthermore the present inventors have also found that halogenated 4oxoalkanoic acid substrates of formula where R5 and R6 are halogens, when treated with sulfuric acid undergo lactonisation with concomitant dehalogenation rather than oxidation as reported in the literature. The halogen produced in this reaction is consumed in situ to produce compounds of formula (VI).

For example when 3-alkyl-2,3-dibromo-4-oxopentanoic acid was treated with sulfuric acid it underwent clean cyclisation with concomitant debromination and utilisation of the liberated bromine through a bromination-dehydrobromination sequence to yield 4alkyl-5-(bromomethylene)-2(5H)-furanone in good yields. Similarly 2,3,5-tribromo-4oxopentanoic acid and 2,3-dibromo-4-oxopentanoic acid gave (bromomethylene)-2(5H)-furanone in high yields.

Furthermore compounds of formula (VI) can also be prepared by the treatment of the starting 3-alkyl-4-oxo-2-pentenoic acids with sulfuric acid followed by the addition of bromine.

Accordingly, in a ninth aspect, the present invention provides a method of concomitant cyclisation and dehalogenation of compounds of formula I as defined above to form a compound of formula the method comprising contacting the compound with sulfuric acid or other strong acid.

ORO: R3 VI R4 wherein R2 is a H, halogen, alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophobic, hydrophilic or fluorophilic; WO 02/00639 WO 0200639PCT/AU01/00781 R(4 is a halogen (X Cl, Br or 1); Ri is hydrogen; and 1R3 is a hydrogen or halogen; The sulfuric acid type reagent may be, for example, concentrated sulfuric acid, oleum, chlorosulfonic acid, or a mixture of sulfuric acid with one or more other like agents.

Examples of compounds of formula I that may be used in the method of the seventh aspect include 3-ly-,-irmo4ooetni acid, 2,3, 5-tribromo-4oxopentanoic acid, 2,3 -dibromo-4-oxopeltaloic acid, 2,5-dibromo-4-oxopentaloic acid, 2,3,5,5-tetrabromo-4-oxopeltanoic acid, 2,3,3 -tribromo-4-oxopentaloic acid and 2,3,3,5-tetrabromo-4-oxopentanoic acid.

Representative examples of furanones (6a-g) that can be synthesised by this procedure are listed below.

0 05

HO:

6a (73) Br 6b Br 6c (103)B h 0

H

0 H 0 69 Br WO 02/00639 PCT/AU01/00781 19 In a tenth aspect, the present invention consists in a 2(5H)-furanone of derivative of formula VI, wherein R2 is an alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophobic, hydrophilic or fluorophilic; R4 is a halogen (X F, Cl, Br or I); R1 is hydrogen; and R3 is a hydrogen or halogen; with the proviso that R 1 H, R 2 Me or Ph, R 3 I, R4 H; and

R

1 H, R 2 OMe, R 3 Cl, R4 Cl.

In an eleventh aspect the present invention consists in a fimbrolide derivative, having a formula (VII), wherein R2 is a H, alkyl, alkoxy, polyethyleneglycyl, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophobic, hydrophilic or fluorophilic;

R

4 is a hydrogen, halogen (X F, Cl, Br or I);

R

1 and R 3 are independently or both hydrogen or halogen;

Z

R1 IR2 O Ra VII R4 Z is independently selected from the group R2, halogen, OC(0)R 2 amine azide, thiol, R 2 mercaptoaryl, arylalkoxy, mercaptoarylalkyl, SC(0)R 2 OS(0) 2

R

2 NHC(0)R 2 =NR2 or NHR 2 prepared by functionalizing a fimbrolide of formula (VIII) wherein, R 1

R

2

R

3 and R 4 are as defined above, with reagent according to our complete specification. (see WO 99/54323, the disclosures of which are incorporated herein by cross-reference).

Reagents including halogenating and oxidising agents (N-halosuccinimide, lead tetraacetate, selenium dioxide, Jones reagent), nucleophiles including (organic metal carboxylates, organic alcohols, dimethyl sulfoxide and organonitriles) and electrophiles including (organic acids, isocyanates, carboxylic or sulfonic acid halides and diethylaminosulfur trifluoride).

WO 02/00639 PCT/AU01/00781 R1 0Q r'NR3 VIII R4 In a twelfth aspect the present invention provides an oligomer or polymer formed by oligomerising or polymerising a compound of formula III- VII, described in the in the present invention directly or with one or more monomers.

The one or more other monomer may be any suitable polymerisable copolymer e.g. acrylate ester such as alkyl, hydroxyalkyl, aminoalkyl, or substituted aryl acrylates or methacrylates, crotonates, substituted or unsubstituted acrylonitriles, vinyl alcohols or acetates, styrene and siloxanes.

In a thirteenth aspect, the present invention consists in incorporation of fimbrolides either in surface coatings or polymers through the newly introduced functionality on the alkyl chain or the alkyl chain itself via direct polymerisation or copolymerisation with suitable monomers.

In an fourteenth aspect, the present invention consists in a fimbrolide derivative produced by the method according to the first, third, fifth, seventh, ninth, or eleventh aspects of the present invention.

In a fifteenth aspect, the present invention consists in the use of a fimbrolide derivative according to the present invention. The present inventors have found that many of the fimbrolide derivatives having the formula (III), (VI) and (VII) have antimicrobial and/or antifouling properties. Accordingly, the fimbrolide derivatives are suitable for use as antimicrobial and/or antifouling agents.

In a sixteenth aspect, the present invention provides methods of use of fimbrolides of formula II in medical, scientific and/or biological applications.

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.

Brief description of the drawings Figure 1 is a graph showing growth of Staphylococcus aureus against compound 120; Figure 2 is a graph showing growth of Staphylococcus aureus against compound 63; WO 02/00639 PCT/AU01/00781 21 Figure 3 is a graph showing growth of Staphylococcus aureus against compound 105; Figure 4 is a graph showing growth of Candida albicans against compound 73; Figure 5 is a graph showing Growth of Candida albicans against compound 113.

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

General. Melting points are uncorrected. Microanalyses were performed by Dr H.P. Pham of The University of New South Wales Microanalytical Laboratory. 1

H

NMR spectra were obtained in CDC1 3 on a Bruker AC300F (300 MHz) or a Bruker DMX500 (500 MHz) spectrometer. 13 C 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 5 scale internally referenced to the solvent peaks: CDC13 (6 7.26, 6 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-Elmer 298 or a Perkin-Elmer 580B spectrophotometer and refer to paraffin mulls. The electron impact mass spectra were recorded on an VG Quattro mass spectrometer at 70eV ionisation voltage and 200 0 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 Ssilica gel 60GF 2 5 4 (Art. 7730).

General method A:-for the synthesis of 4-oxo-2-alkenoic acids Orthophosphoric acid (30 ml) was added to a mixture of glyoxylic acid (0.21 mol) and an alkanone (0.63 mol). The mixture was heated in an oil bath maintained at 80-85 0 C for 4h and then stirred at room temperature overnight. The mixture was extracted with dichloromethane/diethyl ether 3 x 100 ml). The combined organic phase was washed with brine (3 x 75 ml), dried over sodium sulfate and evaporated to yield a dark brown oil. The crude product was chromatographed on a silica gel WO 02/00639 WO 0200639PCT/AU01/00781 22 column using initially dichioromethafle/light petroleum 1) to remove the unreacted alkanone followed by ethyl acetate/dichloronlethafle to yield the 4-oxo-2-alkefloic acid as a pale yellow oil that solidified on keeping at room temperature.

The following compounds were prepared according to method A.

4-Oxo-2-penteloic acid Prepared from glyoxylic acid (40 g, 0.44 mol), acetone (60 g, 1.00 mol) and orthophosphoric acid 60 ml) Pale yellow prisms (10.5 g, m.p. 120-121 0

C

(lit m.p. 121-122C).

3-Metl-4-oxo-2-peflteUoic acid Prepared from glyoxylic acid (16.1g, 0.18 mol), 2-butanone (45.6g, 0.63 mol) and orthophosphoric acid (30 ml). 1H n.m.r. 5 (CDCI 3 2.07, s, 3K, CH3; 2.40, s, CH 3 6.57, s, 1H1, H2. 13 C n.m.r. 5 (CDCI3): 13.2, CU 3 26.2, CU 3 125.2, C2; 152.6, C3; 171.2, Cl; 199.8, C4.

3-Propy-4-oxo-2-pelteloic acid Prepared from glyoxylic, acid (I15.2g, 0. 17 mol), 2-hexanone (30Og, 0.3 mol) and oithophosphoric acid (30 ml). 1H1 n.m.r. 5 (CDCl3) 0.88, t J 7.2 Hz, 3K1 CH3; 1.44, m, 211, CU 2 ;2.41, s, 311 CU 3 2.78, tJ 7.5 Hz, 211,C2; 6.51, s, CU. 13 C n.m.r. (CDCl3): 14.0, CH3; 22.5, 26.5, CU 2 28.6, CU 3 124.5, C2; 157.3, C3; 170.4, COOH; 199.8 C2.

3-Buty-4-oxo-2-peflteloic acid Prepared from glyoxylic acid (15.2g, 0.17 mol), 2-heptanone (34.2g, 0.3 mol) and orthophosphoric acid (30 ml). 1H n.m.r. 6 (CDCl3) 0.91, t J 7.2 Hz, 3K1 CH3; 1.38, m, 411, CU 2 2.39, s, 3K1 CU- 3 2.77, t J 7.2 Hz, 2H, CH2; 6.49, s, CH.

3-HexyI-4-oxo-2-pelteloic acid Prepared from glyoxylic acid (5.35g, 7.0 mmol), 2-nonanone (14.2g, 1.0 mol) and orthophosphoric acid (20 ml). vmax 2925, 2850, 1700, 1450, 1410, 1350, 1220, 1120, 870, 770, 720 cm- 1 IH n.m.r. 6 (CDCl3) 0.87, t J 7.2 Hz, 311 CU3; 1.29, m, 811, CR2; 2.39, s, 3K4 CU 3 2.77, t J 6.8 Hz, 2H1, CH2;5 6.50, s, CII WO 02/00639 WO 0200639PCT/AU01/00781 23 3-lleptyI-4-oxo-2-pelteloic acid Prepared from glyoxylic acid (5.10g, 6.7 mmol), 2-decanone (15.0g, 9.6 mmol) and orthophosphoric acid (20 ml). vmax 2925, 2820, 1690, 1460, 1380, 1240, 1120, 880, 720 cm- 1 IH n.m.r. 8 (CDC13) 0.87, t J 7.2 Hz, 3K1 CR 3 1.29, m, 1011 CR2; 2,39, s, 3H1, CR 3 2.77, t J 6.8 Hz, 2K, CR 2 6.50, s, CH. 13 C n.m.r. 8 (CDCl3): 13.9, CH3; 22.5, 26.6, 26.9, 29.1, 29.6, 31.6 CR 2 28.8, CR3; 124.3, C2; 157.9, C3; 170.9, COOR; 199.9 C2.

3-DecyI-4-oxo-2-peflteloic acid Prepared from glyoxylic acid (4.94g, 6.5 mmol), 2-tridecanone (12.9g, mmol) and orthophosphoric acid (20 ml). 111 n.m.r. 8 (CDCl 3 0.87, t J 7.2 Hz, 3K1 GCl 3 1.30, m, 16H, CR2; 2.39, s, 3K1 CR 3 2.77, t 1 7.0 Hz, 211, CR 2 6.50, s, CR.

3-Methy-4-oxo-2-hexeloic acid Prepared from glyoxylic acid (16.1g, 0.18 mol), 3-pentanone (45.6g, 0.63 mol) and orthophosphoric acid (30 ml). 111 n,m.r. 8 (CDC1 3 1.23, t J 7.2 Hz, 311 CR3; 2.56, s, 3H, CR3; 2.76, q J 7.2 Hz, CR2; 6.55, s, CH.

2-Methyl-4-oxo-2-pelteloic acid and 3,5-dimethy-5-hydroxy-2-(5H)-furaflone Orthophosphoric acid (30 ml) was added dropwise to a mixture of pyruvic acid (I12.7g, 0. 14 mol) and dry AR. grade acetone (25g, 0. 54 mol). The mixture was heated under reflux in an oil bath maintained at 80-85'C for 5h and then stirred at room temperature for 72 h. The mixture was extracted with dichloromethale/diethyl ether 1, 3 x 100 ml). The combined organic phase was washed with brine (3 x 75 ml), dried over sodium sulfate and evaporated to yield a dark brown oil (12.7g). The crude product was chromatographed on a silica gel column using initially dichioromethane as the eluent to yield 3,-iehl5hdox--5-)frnn (3.2g, 25%) (111 n~m.r. 6 (CDCl3) 1.67, s, 311, CR3; 1.91, d J 1.5 Hz, CR 3 3.41, bs, 111, OH; 6.85, q J 1.5 Rz, 114. 13C n.m.r. 8 (CDCI3): 10.2, CR3; 24.5, CR3; 104.6, C5; 131.3, C3; 148.0, C4; 172.3, C2.) followed by ethyl acetateldichioromethane 1) to yield the 2-miethyl-4oxo-2-pentenoic acid (2.6g, 20%) as a pale yellow oil which solidified on keeping at room temperature.

4-Oxo-3-phenyl-2-pelteloic acid and 5-hydroxy-5-methy1-4-phefl-l 4 5

H)-

furanone WO 02/00639 PCT/AU01/00781 24 Orthophosphoric acid (30 ml) was added dropwise to a mixture of glyoxylic acid (15.2g, 0.20 mol) and benzyl methyl ketone (40.2g, 0.30 mol). The mixture was heated in an oil bath maintained at 80-85 0 C for 4h and then stirred at room temperature for 24 h. The mixture was poured onto brine (100 ml) and extracted with dichloromethane/diethyl ether 2 x 150 ml). The combined organic phase was washed with brine (3 x 75 ml), dried over sodium sulfate and evaporated to yield a brown viscous oil (30g). Light petroleum (100 ml, 60-80 0 C) was added to the viscous oil and the mixture cooled in an ice bath for 2h. The resulting solid was collected, washed with saturated sodium bicarbonate solution (75 ml), water and recrystallised form dichloromethane/light petroleum to yield 5-hydroxy-5-methyl-4-phenyl- 2 furanone (8.9g, 21%) as a colourless needles. 1 H n.m.r. 6 (CDCl3) 1.81, s, 3H, CH3; 6.25, s, H3; 7.47, m, 3H, ArH; 7.80, m, 2H, ArH. 1 3 C n.m.r. 8 (CDC13): 25.1, CH3; 107.3, C5; 114.3, C4; 128.2, 128.9, 129.0, 131.3, ArH; 166.1, C3; 171.0, C2. The bicarbonate extract was acidified with hydrochloric acid (6N) and extracted with dichloromethane (3 x 75 ml). The combined extract was washed with brine, dried over anhydrous sodium sulfate and evaporated to yield 2-methyl-4-oxo-2-pentenoic acid (8.7g, 23%) as a pale yellow oil that solidified on keeping at room temperature. 1H n.m.r. 6 (CDC13) 2.27, s, CH3; 6.70, s, 1H, H4, 7.18, m, 2H, ArH; 7.47, m, 3H, ArH.

13C n.m.r. 8 (CDC13): 28.0, CH 3 125.2, C2; 128.9, 129.0, 129.4, 131.3, ArH; 152.5, C3; 169.7, C1; 199.3, C4.

GENERAL METHODS FOR THE SYNTHESIS OF HALOGENATED 4- OXOALKANOIC

ACIDS

Examples of the preparartion of 2,3-dibromo-4-oxoalkanoic acids, and dibromo- and 3,5,5-tribromolevulinic acids are provided below.

General method for the synthesis of 2,3-dibromo-4-oxoalkanoic acids A solution of bromine 0.045 mol) in dry dichloromethane (8 ml) was added slowly to an ice-cooled solution of 4-oxo-2-alkenoic acid (0.03 mol) in dry dichloromethane (30 ml). The mixture was stirred in an ice-bath for 0.5h and then at room temperature for 0.5 h. The resulting solution was washed with aqueous sodium metabisulfite (0.5 M, 30 ml) and brine (30 ml). The solution was dried over sodium sulfate and evaporated to dryness to yield the crude 2,3-dibromo-4-oxoalkanoic acid as a pale brown oil (60-65%).

The crude product was used for the lactonisation step without further purification.

WO 02/00639 WO 0200639PCT/AU01/00781 2,3-Dibromo-4-oxopentaloic acid 4-Oxo-2-pontenoic acid afforded a pale yellow oil (0.93 g, 39%) as a mixture of diastereoisomers, IH n.m.r. 6 (CDC1 3 2.42, s, 311, CR 3 2.44, s, 311, CH 3 4.65-4.74, m, 112 and 113; 6.32, broad s, COOR. 13 C n.m.r. 5 (CDCl 3 27.2, 27.5, CR 3 40.0, 1, C2; 46.6, 52. 0, C3; 171.2, 172.3, Cl1; 196.9, 200.3, C4.

2,3-Dibromo-3-mety-4-oxopeltaloic acid 3-Methyl-4-oxo-2-pentenoic acid (5g, 39 mmol) and bromine (7.5g, 46.9 mmol) in dichloromethane (100 ml) gave a mixture of diastereoisomnersas a pale yellow oil (8.2g, 1 Hn.m.r. S(CDC1 3 2.18, bs, 311,CR3; 2.20, bs, 311,CR 3 5.01,s, 111 112; 5.08, s, 11,112; 6.00, bs, COOFI. 13 C n.m.r. 5 (CDC13): 22,5, 24.0, 24..2, 25.1,

CH

3 52.3, 53.4, C2, 61,8, 65.3, C3; 171.8, 172.8, Cl1; 198.9, 201.4, C4.

2,3-Dibromo-3-buty-4-oxopeltaloic acid 3-Butyl-4-oxo-2-pefltenoic acid (2.O0g, 13.0 mmol) and bromine (3.2g, mmol) in dichloromethane (35 ml) gave a mixtu~re of diastereoisomers as a colourless oil (3.7g, IH n.m.r. 5 (CDC1 3 0.93, m, 311, CR 3 0.95, m, 311, CH 3 1.42, broad m. CH2; 1,87, broad m, CH 2 2.18, bs, 311, CR 3 2.54, broad m, CR2; 4.87, bs, 111, 112; 5.00, s, 111, 112; 7.4, bs, COOR.

2,3-Dibromo-3-hexyI-4-oxopeltaloic acid 3-Hexyl-4-oxo-2-penteloic acid 12g, 13.4 mmol) and bromine (2.4g, mmol) in dichloromethane, (3 5 ml) gave a mixture of diastereoisomers as a colourless oil (3.4g, 111 n.m.r. 5 (CDCI 3 0.8k,~ t, J 7.1 Hz, 311 CR3; 0.90, t, J 7.1 Hz, 311,

CH

3 1,33, broad mn, CR2; 1.50-1.67, broad m, CR 2 2.00-2.30, m, CR2; 2.54, s, CR 3 2.57, s, CR 3 4.83, s, 11-L 112; 5.00, s,1H) 112.

2,3-Dibromo-3-hepty-4-oxopeltaloic acid 3-Reptyl-4-oxo-2-pentenoic acid ig, 5.0 mmol) and bromine (1 .2g, mmol) in dichloromethane (35 ml) gave a mixture of diastereoisomers as a colourless oil (1.6g, IH n.m.r. 8 (CDCl 3 0.88, broad t, 311 C113; 1.28, broad m, CR2; 1.50- 1.67, broad m, CR2; 2.00-2.3 0, m, CR2; 2.54, s, CH3; 2.57, s, CR 3 4.87, s, 111 H12; 4.99, s, 111, 112.

2,3-Dibromo-3-methy-4-oxohexaloic acid WO 02/00639 PCT/AU01/00781 26 3-Methyl-4-oxo-2-hexenoic acid (1.01g, 7.1 mmol) and bromine (1.7g, 10.7 mmol) in dichloromethane (35 ml) gave a mixture of diastereoisomers as a pale yellow oil (1.7g, 1 H n.m.r. 5 (CDC13) 1.16, t J 7.1 Hz, H6; 1.18, t J 7.1 Hz, H6, 2.17, q J 7.1 Hz, H5; 2.18, q J 7.1 Hz, 115; 2.19, bs, 3H, CH3; 2.20, bs, 3H, CH 3 5.03, s, 1H, H2; 5.14, s, 1H, H2.

2,3,5-Tribromo-4-oxopentanoic acid A solution of bromine (10 g, 0.06 mol) in dry dichloromethane (8 ml) was added slowly to an ice-cooled solution of 4-oxo-2-pentenoic acid (3.6 g, 0.03 mol) in dry dichloromethane (30 ml). The mixture was stirred at.room temperature for 0.5 h and then at reflux for 1 h. After cooling to room temperature, the resulting solution was washed with aqueous sodium metabisulfite (0.5 M, 30 ml) and brine (30 ml). The solution was dried over sodium sulfate and evaporated to dryness to yield the crude 2,3,5-tribromo-4-oxopentanoic acid a pale brown oil (7.2 g, 1 H n.m.r. 5 (CDC1 3 4.06, d, J 12.4 Hz, Ha 5 4.27, d, J 12.4 Hz, Hb 5 4.67, d, J 11.3 Hz, H2; 5.20, d, J 11.3 Hz, H3.

Synthesis of 3,5-dibromo-4-oxopentanoic acid A solution of bromine (70 g, 0.44 mol) in dry dichloromethane (80 ml) was added slowly to a solution of 4-oxopentanoic acid (23.2 g, 0.2 mol) in dry dichloromethane (700 ml) containing hydrobromic acid (33% in acetic acid, 12 drops).

The mixture was warmed at 50 0 C for 0.5 h, and then at reflux for 1 h. It was cooled to room temperature and the resulting solution was washed successively with water (100 ml), aqueous sodium metabisulfite (0.5 M, 100 ml) and brine (100 ml). The organic phase was dried over sodium sulfate, and evaporated to dryness to yield the crude dibromo-4-oxopentanoic acid as a white solid. The crude product was recrystallised from chloroform to yield the acid as colourless needles (44 g, 76%) m.p. 112-113 0

C

(lit. m.p. 113 0

C).

Synthesis of 3,5,5-tribromo-4-oxopentanoic acid A solution of bromine (43 g, 0.27 mol) in dry dichloromethane (80 ml) was added slowly to a solution of 4-oxopentanoic acid (10 g, 0.90 mol) in dry dichloromethane (70 ml) containing hydrobromic acid (33% in acetic acid, 12 drops).

The mixture was warmed at 50 0 C for 0.5 h, and then at reflux for 1 h. It was cooled to room temperature and the resulting solution was washed successively with water (100 ml), aqueous sodium metabisulfite (0.5 M, 100 ml) and brine (100 ml). The solution WO 02/00639 WO 0200639PCT/AU01/00781 27 was dried over sodium sulfate and evaporated to dryness to yield the crude 3,5,5tribromo-4-oxoPefltafloic acid as a yellow oil. 1H1 n.m.r. 8 (CDCl3) 3.13, dd, J16.8 Hz 17.7 Hz, Ha 2 3.4 1, dd, J16.8 Hz 17.7 Hz, J 1 b 2 5. 18, dd, 1 6.8, 6.8 Hz, 113; 6.47, s, The corresponding alkyl substituted di- and tri-brominated 4-oxopentanoic acids were similarly prepared.

GENERAL METHODS FOR THE SYNTHESIS OF HALOGENATED 2-

TETRAHYDROFURANONES

Examples of the preparation of 3 ,4-dibromo-4-alkyb5-(methylene)-, (bromomethylene)-, 3-akl4bom--dboomtyee- and 3 -alkyl-3,4dibromo-5-(alkylidene)-2(5H-tetrahydrof.1ranone are provided below.

General method for the synthesis of 3,4-dibromo-4-alkyF$5(methylefle)-, 2tetrahydrofuraloeS 3,-irm--ly--mtyln)2ttayrfrnn Phosphorus pentoxide (11.4 g) was added with stirring to a solution of 2,3 dibromo-3-alkyl-4-oxopentaloic acid (4.6 g, 16.0 mmol) in dry dichloromethane ml). The mixture was heated at reflux with stirring for 2 h, and cooled to room temperature. The resulting mixture was filtered through a pad of celite, washed with brine, dried over sodium sulfate and evaporated to yield the tetrahydrofuranone as a pale yellow oil (60-80%).

This product was used in the next step without further purification.

Method C was used to prepare the following compounds.

3,-irm--ehl5(ehln)2ttayrfrnn Colourless oil as a mixture of diastereoisomers. 1H1 n.m~r. 8 (CDC1 3 2.10, s, 3H, CH 3 2.17, s, 3HL CH3; 4.48, s, 113; 4.77, s, 113; 4.89, d, J 3,8 Hz, CHa; 4.92, d, J 3.8HRz, CHb; 5.02, d, J3.8 Hz, CHa; 5.08, d, J3.8 Hz, CHb.

3,-irm--rpl5(ehln)2ttayrfrnn Pale yellow oil as a mixture of diastereoisomers. 111 n.m.r. 6 (CDCl3) 1.01, t, 3K-1 CH3; 1.05, m, 311 CH3; 1.62, m, 211 C112; 1.65, m, 211, CH2; 2.45, mn, 211, CH2; 2.47, mn, 211, CH2; 4.66, s, H13; 4.71, s, 113; 4.90, d, J 3.3 Hz, CHa; 4.91, d, J 3.7 Hz, CHb; 4,91, d, J 3.3 Hz, CHa; 5.04, d, J 3.7HI-z, CHb.

WO 02/00639 WO 0200639PCT/AU01/00781 28 3,-irm--uy--mtyln)2ttayrfrnn Pale yellow oil as mixture of diastereoisomers. IH n.m.r. 5 (CDCl 3 0.98, t, 3H, C113; 1.00, t, 311, CH 3 142, mn, 2H, CH 2 1.69, m, 211, GIl 2 2.28, mn, 211, CH 2 4.66, s, H3; 4.71, s, 113; 4.86, d, J 3.8 Hz, CHa; 4.88, d, J 3.7 Hz, Glib; 5.03, d, J 3.7 Hz, GHa; 5.05, d, J 3,7 Hz, GHb.

3,4-Dibromo-4-hexyI-5(methy~le)2-etrahydrofuranone Pale yellow oil as a mixture of diastereoisomers. 1 H n.m.r. 5 (CDCl 3 0.92, t, 311 CH3; 1.35, mn, 611K GH2,) 1.76, mn, 211, GH 2 2.23, m, 2H1, GH2; 4.62, s, 113; 4.66, s, 113; 4.87, d, 1 3.2 Hz, Gila; 4.88, d, 1 3.2 Hz, Glib; 5.03, d, J 3.2 Hz, CHa; 5.05, d, J 3.2 Hz, Glib.

3,-irm--etl5(ehln)2ttayrfrnn Pale yellow oil as mixture of diastereoisomers. 111 n.m.r. 8 (CDCl3) 0.90, t, 311, GH-3; 1.32, mn, 811, CH 2 1.76, m, 211, GCl 2 2,30, m, 2H1, GH2; 4.63, s, H13; 4.66, s, 113; 4.86, d, J 3.8 Hz, CHa; 4.87, d, 1 3.8 Hz, GHb; 5.02, d, J 3.8 Hz, CHa; 5.04, d, J 3.8 Hz, CHb.

Synthesis of 3,-irm--ty--ehldn)2ttayrfrnn Pale yellow oil (3.68 g, 84%) as a mixture of diastereoisomers. 1H1 n.m.r. 5 (CDGI3) 1.91, d, J7,1 Hz, 5-(GHM~e); 1.92, d, 1 7.1 H~z, 2.03, s, 311, C113; 2.12, s, 311, GCl 3 4.48, s, 114; 4.76, s H4; 5.34, q, 1H, 5-(CEHNe); 5.36, q, Ili Synthesis of 4-bromo-5-(bromomethy~le)-2-tetrahlydrofuranone Phosphorus pentoxide (22.5 g) was added with stirring to a solution of dibromo-4-oxopentanoic acid (30.4 g, 0.11 mol) in dry dichloromethane (500 ml). The mixture was heated at reflux with stirring for 2 hi, and cooled to room temperature. The resulting mixture was filtered through a pad of filter aid, washed with brine, dried over sodium sulfate and evaporated to yield the tetrahydrofuranone (mixture of Z- and Eisomers in 4: 1 ratio) as a pale yellow oil (23.4 g, The oil solidified on standing at room temperature overnight. Crystallisation from dichioromethane/light petroleum (60-80 0 C) gave an analytically pure sample of the Z-isomer. m.p. 79'C. vmnax 3094, 3027, 2947, 1799, 1634, 1393, 1289, 1114, 949, 838, 747, 718, 660 cnr'. kmax 2 79 nm (s 4938). 111 n.m.r. 6 (CDGI3) 3. 10, dd, J2.3 Hz, 18.8 Hz, Ha 3 3.45, dd, J 7.9 Hz, WO 02/00639 WO 0200639PCT/AU01/00781 29 18.8 Hz, Hb 3 5.07, m, H4; 5.90, s, 5-CHBr. 13C n.m.r. 8 (CDC1 3 37.2, C3; 40.5, C4; 86.9, 5-CHBr; 152.3, C5; 169.5, 02.

E-isomer: 1H1 n.m.r. 5 (0DCl 3 3.09, dd, J2.3 Hz, 18.8 Hz, Ha 3 3.38, dd, J7.9 Hz, 18.8 Hz, Hb 3 5.15, m, H4; 6.19, s, 5-CHBr, 13 C n.m.r. 8 (CD13): 37.0, C3; 39.8, C4; 90.1, 5-CHBr; 151.9, C5; 170.4, C2.

Synthesis of 4-rio3btl5(irmmthln)2ttayrfrnn Phosphorus pentoxide (10.0 g) was added with stirring to a solution of 2-(1,3,3tribromo-2-oxopropyl)hexafloic acid (7.7 g, 0, 11 mot) in dry dichloromethane (150 ml).

The mixture was heated at reflux with stirring for 2 h, and cooled to room temperature.

The resulting mixture was filtered through a pad of filter aid, washed with brine, dried over sodium sulfate and evaporated to yield the tetrahydrofuranone as a pale brown oil.

g, 82%) that solidified on standing at 40C overnight. vmax 2958, 2930, 2871, 1820, 1782, 1638, 1455, 1127, 1086, 965, 849, 757, 718 cnr 1 1 11n.m.r. 5 (CDCl3) 0. 93, t, J 6.7 Hz, (H14)3; 1 3 7- 1. 81, mn, (H13')2- (111)2; 3,22, t, J 7.5 Hz, H13; 4.79, s, H4.

13C n.m.r. 8 (CDCl 3 14.1, C4'; 22.5, 03'; 28.9, Cl'; 31.6, 02T; 43.6, 04; 53.7, C3; 76.5, 5-C~r 2 149.8, CS; 172.5, 02.

GENERAL METHODS FOR THE SYNTHESIS OF HALO GENATED

FURANONES

Examples of the preparation of 3-boo4akl5(ehyee-(H-uaoe 5-(bromomethylene)-2(SH)-, 5-(dibromomethylene)-2(5H)- and 3 are provided below.

General method Part for the synthesis of 3-bromo-4-alky1-5-(mlcthylefle)- 1,8-Diazabicyclo[5.4.0]unrdec-7-ene (DBU/DABCO or DMAP) (14.8 mmol) was added dropwise to a cooled stirred solution of 3,4-dibromo-4-alkyl-5-(aethylenC)- 2-tetrahydrofuranone (0.0 16 mol) in dichloromethane (30 ml). The mixture was stirred at room temperature for 0. 5 h, washed with dilute hydrochloric acid (2N, 20 ml), brine ml), dried over sodium sulfate, and evaporated to dryness. The crude product was passed through a short plug of silica gel to yield the 3-bromo-4-methyl-5-(methylene)- 2(511)-furanone (50-83%) as a colourless oil.

3-rm--ehl5(ehln)25)frnn WO 02/00639 WO 0200639PCT/AU01/00781 Pale brown oil 111 n.m.r. 6 (CDCl 3 2.17, s, 3H1, CH 3 5.01, d, J 3.0 Hz, CHa; 5,22, d,1J3.0HI-z, CHb.

3 -Ilromo-4-propyl-5-(methylene)-2(51u)-furanone Pale brown oil IH n.m.r. 5 (CDC13) 1.02, t, J 7.1 Hz, 3H, CH 3 1.68, m, 2H1, 0112; 2,52, t, J17.1 Hz, 2H1, CH 2 5.01, d, 3.1 Hz, CHa; 5.24, d, J13.1 Hz, CHb.

3 -Bromo-4-butyl-5-(methylene)-2(5HJ-furanone Pale brown oil IH n.m.r. 6 (CDCI 3 0.96, t, J 7.2 Hz, 3H, OH 3 1.37, m, 2H, 0112; 1.64, m, 211, CH 2 2.52, t, J 7.3 Hz, 211, OH 2 5.02, d, J 3,1 Hz, CHa; 5.24, d, J 3.1 Hz, C 0 b.

3-.Bromo-4-hexyl-5-(methylene)-2(51J).furanone Pale brown oil 1 H n.m.r. 5 (ODC1 3 0.93, t, 1 7.1 Hz, 3H1, 0113; 1.35, m, 6H, CH 2 1. 62, m, 2H,0112; 2.53, in, 2H, 0112; 5. 01, d, J13. 0Hz, CHa; 5.24, di, J13.0 Hz, 011 b.

3 -Bromo- 4 -heptyl-5-(rnethylene)-2(5H)-furanone Pale brown oil 111 n.m.r. 5 (CDCd 3 0.89, t, J 7.3 H, 3H1, GCl 3 1.29, m, 811, 0112; 1. 61, mn, 211, CH 2 2.5 3, t, J 7.5 Hz, 2H1, 0112; 5.02, d, J13.2 Hz, CHa; 5.24, d, 1 3.2 Hiz, C14b. 130C n.mi. r. 5 (CDC1 3 13.9, 0113; 22.5, 26.2, 28.1, 28.8, 29.1, 3 1. 0112; 95.3, 5-(CH 2 112.2, C3; 154.3, C5; 154.6, C4; 164.6, C2.

Synthesis of 5-(bromomethylene)-2(5H)-furanone 1,4-Diazabicyclo[2.2.2]octane (DABCO) (2.8 g, 0.025 mol) was added to a stirred solution of a mixture of and (Z)-isomers of tetrahydro-2-fiuranone (4.2 g, 0.0 16 mol) in dichioromethane (20 ml). The mixture was stirred at room temperature for 2 h and filtered. The filtrate was washed with dilute hydrochloric acid (2N, 10 ml), brine (20 ml), dried over sodium sulfate, and evaporated to dryness. The crude product was recrystallised from light petroleum (60-800C) to yield S-(bromoinethylene)-2(5H)-furanone as colourless needles (2.2 g, 76%) m.p. 820C. vmnax 29 05, 2840, 1770, 1740, 1630, 1540, 1450, 1370, 1290, 1160, 1100, 1070, 915, 880, 815, 770, 720 cm- 1 Xmax 284 nm (s 13 53 1H n.mn.r. 6 (CDC1 3 6.12, s, CHBr; 6.32, d, 1 5.1 Hz, 113; 7.40, d, 1 5.1 Hz, 114. 13C n.m.r. 5 (CDCl 3 92.5, CHrBr; 120.7, 03; 141.8, 04; 152.4, C5; 168.3, C2. Mass spectrum: m/z 176 (M 81 Br), 100%); 174 (M 79 Br), 100); 148 142 122 120 95 WO 02/00639 PCT/AU01/00781 31 Synthesis of 3-butyl-5-(dibromomethylene)-2(5H)-furanone 1,4-Diazabicyclo[2.2.2]octane (DABCO) (1.2 g, 10.4 mmol) was added to a stirred solution of 4-bromo-3-butyl-5-(dibromomethylene)-tetrahydro-2(5H)-furanone (2.0 g, 6.9 mmol) in dichloromethane (20 ml). The mixture was stirred at room temperature for 2 h and filtered. The filtrate was washed successively with dilute hydrochloric acid (2N, 10 ml), brine (20 ml). It was dried over sodium sulfate and evaporated to dryness to yield the furanone as pale brown solid. The crude product was recrystallised from light petroleum (60-80 0 C) containing a small amount of dichloromethane to yield 3-butyl-5-(dibromomethylene)-2(5H)-furanone as a white solid (1.0 g, m.p. 48-49 0 C. Vmax 3080, 2900, 2840, 1740, 1590, 1445, 1330, 1255, 1040, 960, 890, 840, 820, 705 cm- 1 Xmax 30 3 nm 13682). 1 H n.m.r. 8 (CDC13) 0.92, t, J7.2 Hz, 1.32, m, 1.56, m, 2.32, t, J7.3 Hz, (H1') 2 7.27, br s, H4. 13C n.m.r. 8 (CDCl 3 13.7, C4'; 22.3, C3'; 25.4, Cl'; 29.3, C2'; 78.8, 5-CBr 2 134.0, C4; 138.0, C3, 149.7, C5; 166.7, C2. Mass spectrum: m/z 312 (M 81 Br2), 310 (M 81 Br, 79 Br), 14); 308 283 281 279 270 268 266 231 229 202 200 198 189 187 172 161 159 149 (28).

One pot syntheses of 3-bromo-4-alkyl-5-(methylene)-2(5H)-furanone, 5-(dibromomethylene)-2(5H)- and The general methods C and D were carried out in one sequence without the isolation and purification of intermediates.

Synthesis of 3-Bromo-4-methyl-5-(methylene)-2(5H)furanone Phosphorus pentoxide (11.5 g) was added with stirring to a solution of 2,3dibromo-3-methyl-4-oxopentanoic acid (4.6g, 16.0 mmol) in dry dichloromethane ml). The mixture was heated at reflux with stirring for 2 h. It was cooled to room temperature and the resulting mixture was filtered through a pad of filter aid and treated with 1,4-diazabicyclo[2.2.2]octane (DABCO) (2.24 g, 14.8 mmol). The mixture was stirred at room temperature for 1 h, washed with dilute hydrochloric acid (2N, 10 ml), brine (20 ml), dried over sodium sulfate, and chromatographed on a short plug of silica gel to yield the 3-bromo-4-methyl-5-(methylene)-2(5H)furanone as a colourless oil (2.14 g, 83%).

WO 02/00639 PCT/AU01/00781 32 Synthesis of 5-(bromomethylene)-2(5H)-furanone Phosphorus pentoxide (22.5 g) was added with stirring to a solution of dibromo-4-oxopentanoic acid (30.4g, 0.11 mol) in dry dichloromethane (500 ml). The mixture was heated at reflux with stirring for 2 h. It was cooled to room temperature and the resulting mixture was filtered through a pad of filter aid and treated with 1,4diazabicyclo[2.2.2]octane (DABCO) (2.8 g, 0.025 mol). The mixture was stirred at room temperature for 2 h and filtered. The filtrate was washed with dilute hydrochloric acid (2N, 10 ml), brine (20 ml), dried over sodium sulfate, and evaporated to dryness.

The crude product was recrystallised from light petroleum (60-80 0 C) to yield the as colourless needles (2.1 g, 73%) m.p. 82-83 0

C.

Synthesis of 5-(dibromomethylene)-2(5H)-furanone Phosphorus pentoxide (20 g) was added with stirring to a solution of 3,5,5tribromo-4-oxopentanoic acid (14.5 g, 4.1 mmol) in dry dichloromethane (200 ml).

The mixture was heated at reflux with stirring for 2 h, and cooled to room temperature.

The resulting mixture was filtered through a pad of filter aid and the filtrate treated with a solution of4-(dimethylamino)pyridine (DMAP) (5.8 g, 4.7 mmol) in dichloromethane ml). The mixture was stirred at room temperature for 2 h and filtered. The filtrate was washed with dilute hydrochloric acid (2N, 10 ml), brine (20 ml), dried over sodium sulfate, and evaporated to dryness. The crude product was chromatographed on silica using light petroleum (60-80 0 C) as the eluent to yield the 5-(dibromomethylene)-2(5H)furanone as pale yellow prisms. (4.2 g, 40%) m.p. 135 0 C. vmax 2905, 2840, 1805, 1783, 1558, 1257, 1102, 1067, 960, 887, 826 cm- 1 Xmax 306 nm (e 4300). 1 H n.m.r. 6 (CDCl3) 6.40, d, J5.3 Hz, H3; 7.66, d, J5.3 Hz, H4. 13 C nm.r. 8 (CDC13): 81.7, CBr2; 122.3, C3; 140.6, C4; 150.7, C5; 167.6, C2. Mass spectrum: m/z 176 (M 8 1 Br), 100%); 174 (M 79 Br), 100).

GENERAL METHODS FOR THE SYNTHESIS OF HALOGENATED Examples of the preparation of 5-(bromomethyl)-3,5-dibromo- and (dibromomethyl)-5-bromo-2(5H)-furanones are provided below.

General method for the synthesis of 4-alkyl-5-(bromomethyl)- 3 4-alkyl-3,5-dibromo-5-(dibromomethyl)-, 4-alkyl-5-bromo-(bromomethyl)-5- and 4alkyl-5-bromo-5-(dibromomethyl)-2(5H)-furanones WO 02/00639 WO 0200639PCT/AU01/00781 33 A solution of bromine (17 mmol) in dichioromethane (10 ml) was added dropwise to an ice-cooled solution of 3-rm--ly--mthln)25)frnn (18.7 mmol) in dichioromethane (20 ml). The mixture was stirred in ice for 0. 5h and then at room temperature for further I h. It was washed with saturated sodium metabisulfite solution (30 ml) followed by brine (40 ml). The resulting solution was dried over anhydrous sodium sulfate and evaporated to yield the 3,5-dibromo-5bromomethyl-4-alkyl-2(5H)-furanofle (65-85%) as a colourless oil.

5-Booehl-,-irm--ehl25)frnn Pale yellow *solid vmax 2925, 2850, 1780, 1650, 1430, 13 80, 1295, 1260, 1160, 1130, 970, 850, 740 cm-1. 'IIIn~m.r. 8 (CDCl 3 2.19, s, 311, CU 3 3.97, d, J 11.7 Hz, CHaBr; 4.26, d,J1 1.7 Hz, CHbBr. 13 C n.m~r.58(CDC13): 12.9, CU 3 33.8, CH2B3r; 90.9, C5; 112.3, C3; 161.5, C4; 163.6, C2. Mass spectrum: m/z 352 (N4 81 Br 3 350 (M 8 lBr 2 79 Br), 348 (M (SIBr), 79 Br 2 346 79 Br 3 271 269 265 213 190 188 162 158 (20; 122 120 3,5-Dibromo-5(dibromomthy)-4methylI4( H).furanone Colourless prisms IH n.m.r. 5 (CDCI 3 2.26, s, 3H, CU 3 5.91, s, (CHBr 2 13 C n.m.r. 5 (CDCl3): 13.7, CH3; 43.1, 5-(CIIBr2), 93.8, C5; 112.9, C3; 143.3, C4; 161.4, C2.

5-(Bromomethy)-3,5-dibromo-4-butyI-2(5H)-furaflone Pale yellow oil 111 n.m.r. 5 (CDCI 3 1.00, t, J 7.3 Hz, 3H, CH3; 1.45, m, 2H, CH2; 1.80, m, 2H, CU 2 2.45, t, J17.1 Hz, 2H, 4.00, d, J 10.7 Hz, CHaBr; 4.28, d, J 11.7 Hz, CHbBr. 13 C n.m~r. 8 (CDCI 3 13.5, CH3; 23.0, 27.5, 28.3, CH2; 34.2, CH 2 Br; 91.3, CS; 112.4, C3; 163.8, C4; 164.2, C2.

5-Booehl-,-irmo4hxl25)frnn Pale yellow oil 111 n.m.r. 5 (CDCl3) 0.92, t J 7.1 Hz, 31-L CH3; 1.35, m, 411, CU 2 1.46, m, 211, CU 2 1.82, m, 211, CH2; 2,47, t, J 8.1 Hz, 2K1 CH2; 4.00, d, J 11.7 Hz, CHaBr; 4.27, d, J 11.7 Hz, CHjBr.

5-(Bromomethy)-3,5-dibromfo-4-hepty1-2(5H) 4 furalofe Pale yellow oil vmax 3038, 2953, 2927, 2855, 1793, 1635, 1463, 1415, 1377, 1265, 1227, 1165, 1129, 973, 881, 743 cm- 1 X,,nax 244 nm(s 4330). 1 11n.m.r.

WO 02/00639 WO 0200639PCT/AU01/00781 34 (CDC1 3 0.90, t, J 7.1 Hz, 3H, GCl 3 1.32, m, 81H, CH 2 1.82, m, 2H1, GH 2 2.47, t, J 8.1 Hz, 2H, CH2; 4.00, d, J 11.7 Hz, CHaBr; 4.27, d, J 11.7 Hz, CH 13 Br. 13 C n.m.r. (CDCl 3 13.9, C11 3 22.5, 26.3, 27.8, 28.6, 29.8, 31.5, CH 2 34.2, CH 2 Br; 91.3, 112.4, C3) 163.8, C4) 164.3, C2.

5-(Dibromomethyl)-3,5-dibromo-4-hepty-2(5H)furalofe Pale yellow oil vmax 3005, 2953, 2926, 2855, 1796, 1630, 1464, 1377, 1260, 1165, 1143, 968, 885, 799, 746, 720 cur 1 2..max 242 nm(0 312). lHn.m.r. S (CDCl 3 0.90, t, J 7.2 Hz, 3K1 CH 3 1.32, m, 8H1, C112; 1.86, m, 211, C11 2 2.50, m, 2H1,

CH

2 5.93,) s, 5-GH~r 2 1 3 C n.m.r. 5 (CDCI3): 13.9, CH3; 22.5, 26.5, 28.2, 28.6, 29.8, 3 1.5, GCl 2 43.4, 5-CHBr2; 94.0, C5; 112.6, C3; 163.3, C4; 164.4, C2.

5-(Bromomethy)-5-bromO-4-phenl2(5H)furalofl Colourless prisms 1 11n.m.r. 5 (CDCl 3 4.11, d 11.3 Hz, CHaBr; 4.3 8, d, J 11.3 Hz, CHbBr. 13 C n.m.r. 5 (CDCl 3 34.8, GH 2 Br; 89.4, C5; 116.9, C3; 128.0, 129.2, 131.8, Ph; 165.9, C4; 167.2, C2. Mass spectrum: mlz 334 (M (SIBr 2 332 (M 8 1Br), 79 Br), 330 (N4 79 Br 2 253 251 172 116 115 102 (100).

GENERAL METHODS FOR THE SYNTHESIS OF HALOGENATED Examples of'the preparation of 3-bromo-4-alky1-5-(bromomethylefle)-2(5H)furanone produced are provided below.

General method for the synthesis of 3-bromo-4-alkyl-5-(brolomlethylefle)- A solution of N,N' -diisopropylethyl amine (Hunig' s base) (36 mmol) in dichioromethane (10 ml) was added dropwise with stirring to an ice cooled solution of 3, 5-dibromo-5 -bromomethyl-4-alkyl-2(5H)-furarlone (7.2 mmol) in dichloromethane (40 ml). The mixture was allowed to warm to room temperature and further stirred at room temperature for 60h, The resulting mixture was washed with aqueous hydrochloric acid (100 ml, 2M), dried over anhydrous sodium sulfate and evaporated to yield 3-rm--ly--booehln)25)frnn as a dark brown oil. The crude product was cliromatographed on a silica gel column using dichioromethane/light petroleum (60-80 0 C) as eluent to yield the pure furanone, as a pale yellow oil.

WO 02/00639 PCT/AU01/00781 3-Bromo-4-methyl-5-(bromnomethylene)-2(5H)-furanone Pale yellow oil vmax 2905, 2850, 1760, 1630, 1580, 1450, 1380, 1270, 1230, 1105, 980, 900, 720 cm-l. max 290 nm (a 20570). 1 Hn.m.r. 8 (CDC13) 2.16, s, 3H, CH3; 6.22, s, 5-(CHBr). 13 C n.m.r. 6 (CDCl3): 11.5, CH3; 90.4, 5-(CHBr); 112.2, C3; 149.3, CS; 152.0, C4; 163.4, C2. Mass spectrum: m/z 270 (M 8 1 Br2), 268 (M (lBr, 79 Br), 80); 266 8 1 Br2), 242 240 238 189 187 161 159 133 131 122 120 3-Bromo-5-(dibromomethylene)-4-methyl-2(5H)-furanone Colourless prisms vmax 2951, 2924, 2853, 1771, 1592, 1576, 1462, 1428, 1382, 1232, 1020, 906, 847, 746, 718 cm-1. Xmax 307 nm (s 24576). 1 H n.m.r. 6 (CDCI3) 2.50, s, 3H, CH 3 13 C n.m.r. S (CDCl 3 16.4, CH3; 81.6, 5-(CBr2); 115.9, C3; 148.8, CS; 149.5, C4; 162.2, C2. Mass spectrum: m/z 350 (M 8 1 Br 3 348 (M 8 1Br 2 79 Br), 100%); 346 (M4 8 1 Br) 7 9 Br 2 100%); 344 (M 7 9 Br 3 269 267 265 241 239 237 213 211 209 202 200 198 3-Bromo-5-(bromomethylene)-4-propyl-2(5H)-furanone Pale yellow oil vmax 2936, 2843, 1760, 1595, 1440, 1380, 1270, 1190, 1100, 1010, 950, 845, 760, 710 cm-1. m,,ax 287 nm (8 13757). 11 n.m.r. 6 (CDCl 3 1.01, t, J 7.5 Hz, 3, CH 3 1.67, m, 2H, CH2;2.52, t, J 7.2 Hz, t, 21H, CH2; 6.23, s, (CHBr). 1 3 C n.m.r. 8 (CDCl 3 13.9, CH 3 21.8, 28.0, CH 2 90.8, 5-(CHBr); 112.0, C3; 151.5, C5; 153.3, C4; 163.6, C2. Mass spectrum: m/z 298 (M 8 1 Br2), 296 (M 8 1 Br, 7 9 Br), 40); 294 8 1 Br2), 217 (100); 215 189 187 159 161 136 (100).

3-Bromo-4-butyl-5-(bromomethylene)-2(5H)-furanone Pale yellow oil vmax 2950, 2850, 1750, 1590, 1450, 1380, 1310, 1270, 1195, 1100, 1050, 1010, 950, 890, 840, 760, 720 cm-1. Am,, 295 nm (s 9827). 1H n.m.r. 6 (CDCl3) 0.96, t, J 7.2 Hz, 3H, CH3; 1.42, m, 21H1, CH2; 1.57, m, 2H, CH2; 2.53, t, J 7.5 Hz, t, 2H, CH2; 6.23, s, 5-(CHBr). 1 3 C n.m.r. 6 (CDCl 3 13.5, CH3; 22.6, 25.8, 30.3, CH 2 90.7, 5-(CHBr); 111.6, C3; 151.3, C5; 153.5, C4; 163.6, C2. Mass spectrum: m/z 312 (M 81 Br2), 310 (M 81 Br, 79 Br), 16); 306 8 1Br 2 270 268 266 231 229 217 215 189 187 159 161 WO 02/00639 PCT/AU01/00781 36 3-Bromo-5-(dibromomethlylene)-4-butyl--2( H)-furanone Pale yellow needles vmax 2980, 2952, 2850, 1766, 1615, 1575, 1462, 1380, 1150, 1009, 910, 830, 760, 720 cmnr 1 X,,ax 310 nm (8 2350). 1 H n.m.r. 6 (CDC13) 0.97, t, J 7.2 Hz, 3, CH 3 1.45, m, 2H, CH2; 1.59, m, 2H, CH 2 2.88, t, J Hz, 2H, CH2. 13C n.m.r. 8 (CDC1 3 13.6, CH 3 22.6, 28.0, 29.9 CH2; 81.2, 5-(CBr2); 115.4, C3; 148.4, C5; 153.4, C4; 162.2, C2. Mass spectrum: m/z 392 (M 8 1 Br 3 390 (M 8 lBr 2 7 9 Br), 388 (M 8 Br), 79 Brz), 386 7 9 Br 3 350 348 346 342 311 309 (100); 307 269 267 (100); 265 239 227 3-Bromo-4-hexyl-5-(bromomethylene)-2(5H)-furanone Pale yellow oil vmax 3092, 2955, 2928, 2857, 1786, 1679, 1636, 1595, 1465, 1364, 1301, 1219, 1183, 1050, 982, 914, 840, 765, 750 cm 1 ,max 290 nm (8 5647). 1H n.m.r. 6 (CDCl 3 0.90, t, J 7.2 Hz, 3H, CH3; 1.33, m, 6H, CH2; 1.61, m, 2H, CHi 2 2.52, t, J 7.9 Hz, t, 2H, CH 2 6.22, s, 5-(CHBr). 13C n.m.r. 6 (CDCl3): 13.9, CH3; 22.3, 26.1, 28.2, 29.0, 31.2, CH2; 90.5, 5-(CHBr); 111.7, C3; 151.4, C5; 153.5, C4; 163.5, C2. Mass spectrum: m/z 340 (M 8 1 Br2), 338 (M 8 1 Br, 7 9 Br), 40); 336 8 1 Br2), 270 268(40); 266 259 257 215 213 189 (100); 187 (100); 178 176 159 930); 149 135 121 3-Bromo-4-heptyl-5-(bromomethylene)-2(5H)-furanone Pale yellow oil vmax 3094, 2953, 2928, 2856, 1789, 1636, 1596, 1464, 1377, 1268, 1183, 1046, 981, 891, 840, 764, 749 cm-1. Xmax 293 nm (8 14610). IH n.m.r. 6 (CDC13) 0.89, t, J 7.2 Hz, 3, CH3; 1.33, m, 8H, CH2; 1.59, m, 2H, CH2; 2.52, t, J 7.5 Hz, t, 2H, CH2; 6.22, s, 5-(CHBr). 13C n.m.r. 6 (CDCI3): 13.9, CH3; 22.5, 26.1, 28.2, 28.7, 29.3, 31.5, CH 2 90.5, 5-(CHBr); 111.6, C3; 151.4, CS; 153.5, C4; 163.5, C2. Mass spectrum: m/z 354 (M 8 1 Br2), 352 (M 8 1 Br, 7 9 Br), 30); 350 (M, 8 tBr 2 270 268(50); 266 231 257 215 213 189 (100); 187 (100); 173 (100); 145 135 121 3-Bromo-4-decyl-5-(bromomethylene)-2(5H)-furanone Pale yellow oil vmax 2925, 2850, 1789, 1636, 1590, 1450, 1380, 1100, 980, 890, 750, 720 cm- 1 1 1H n.m.r. 6 (CDC13) 0.89, t J 7.0 Hz, 3, CH3; 1.32, m, 14H, CH2; 1.58, m, 21H1, CH2; 2.52, t J 7.5 Hz, t, 2H, CH2; 6.23, s, 5-(CHBr) 13C n.m.r. 8 (CDCl 3 14.0, CH3; 22.5, 26.1, 28.2, 29.0, 29.1, 29.3, 29.4, 31.7, CH2; 90.7, (CBHBr); 111.6, C3; 151.3, C5; 153.6, C4; 163.5, C2. Mass spectrum: m/z 396 (M WO 02/00639 WO 0200639PCT/AU01/00781 37 81 Br 2 394 (M 81 Br, 79 Br), 392 8 lBr 2 315 313 270 268 266 257 231 215 189 187 3 -Bromo- 4 -methyl-5-(ethylidene)2(5H)-furanone Pale yellow solid vmax 2952, 2850, 1778, 1630, 1430, 1380, 1270, 123 8, 1105, 1047, 912, 854, 720 cm- 1 1H n.m~lr. 5 (CDCI 3 1.93, d, J 7.2 Hz, 311, GCl 3 2.12, s, 31-, C14 3 5.47, q, J 7.2 Hz, 5-(CHMe). 13 C n~m.r. 6 (CDCl 3 11.5, C11 3 11.6, CH 3 109.2, 5-(CH-Me); 110.0, C3; 149.9, CS; 150,8, C4; 165.0, C2. Mass spectrum: mlz 204 (M 81 1Br), 202 79 13r), 192 190 178 176 148 146 GENERAL METHODS FOR THlE SYNTHESIS OF 4-ALKYL-, AND 4- BROMO-5-(HALOMETYLEN)2(5II)-FURANONES Examples of the preparation of 4 -alkyl-5-(bromomethylene)- and (bromomethylene)-2(5H-)ffranone are provided below.

Synthesis of 4 -alkyl-S5-(bromomethylene). and 4 2(511)-furanone General method G:- Synthesis of 4 -alkyl-5-(bromonlethylene)-2(5J1)-uranones 2 3 -Dibromo-3-alkyl-4-oxopentanoic acid (5 g) was added with stirring to hot concentrated sulfuric acid (30 ml) held at 100 0 C. The mixture was further heated with stirring at this temperature for 20 minutes, cooled to ambient temperature and treated slowly with ice water. The resulting oily product was extracted with dichioromethane (3 x 75 ml). The combined extracts were washed with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate and evaporated to yield the as a Pale yellow oil. The crude product was chromatographed on a silica gel column using dichioromethane/light petroleum 1) as eluent to yield pure 4 -alkyl-5-(bromomethylene)-2(5g-.}.furanone (50-60%) as a pale yellow oil. Further elution of the column with dichioromethane yielded 4 -alkyl-5-bromo-5-dibromomethyl- and (dibromomethylene)-2(51)-fjuranones General method H:- WO 02/00639 WO 0200639PCT/AU01/00781 38 3 -alkyl-4-oxo-2-pentenoic acid (7.8 mmol) was added with stirring to hot concentrated sulfuric acid (10 ml) held at 100C The mixture was further heated with stirring at this temperature for 10 minutes, cooled to ambient temperature and treated slowly with bromine (11.7 mmol). The mixture was stirred at room temperature for minutes and then at 1 00 0 C for fulrther 15 minutes. The mixture was cooled to room temperature and poured over crushed ice. The resulting oily product was extracted with dichloromethane (3 x 50 ml). The combined extract was washed with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate and evaporated to yield a pale yellow oil. The crude product was chromatographed on a silica gel column using dichioromethane/light petroleum 1) as eluent to yield (40-60%) as a pale yellow oil. Further elution of the column with dichioromethane yielded mixtures of 4-alkyl-5-bromo-5bromomethyl-, 4 -alkyl-5-bromo-5-dibromomethyl1 and 4 2 (5J1)-furanones (20-3 some of which were purified.

The following 4 -alkyl-5-(bromomethylene).2(51)fujranones were prepared according to methods G and H.

-(Bromomethylene)4methyl12(5H)-furanone Colourless prisms (Found: (HRESMS) m/z 210.9370. C6H 5 BrO 2 Na 79 Br) requires m/z 210.9365). vmax 2920, 2850, 1765, 1590, 1460, 1380, 1230, 1020, 910, 850, 750, 720 cm- 1 Xac281 n 20675). 1H n.m.r. 5 (CDC1 3 2. 11, d J 1.3 H1z, 31L CR3; 5.98, q J 1.3 Hz, 113; 6.02, s, 5-(CHBr). 13C n.m.r. 5 (CDC1 3 12.1, CH 3 89.8, 117.9, C3; 153.6, CS; 154.0, C4; 168.0, 02. High resolution mass: observed 210.9370 C6H5iBrO2Na requires: Mass spectrum: m/z 190 (M 81 lBr), 188 (M, 79 Br), 162 160 122 120 5-Bromo-5-(dibromomethyI)..4.methyl2(5R)furanone Colourless needles (Found: (IHIRSMS) m/z 368.7749. C6HSBr 3

O

2 Na 79 Br) requires m/z 368.7732). vmax 3006, 2923, 2853, 1779, 1642, 1595, 1460, 1380, 1330, 1290, 1182, 1140, 1085, 911, 868, 750, 720 cm- 1 X .,212 nm (s 8401). IH n.m.r. 5 (CDC1 3 2.28, d J 1.5 Hz, 3K1 0113; 5.89 s, 5-(CI{IBr 2 6.,10, q J 1.5 Hz, H3.

13C n.m.r. 5 (CDC 3 13.5, CH; 43.1, 5-(CHBr 2 94.5, C5; 118.5, C3; 166.6, C4; 166,9, C2. Mass spectrum: m/z 352 (Mv 8 1IBr 3 350 (M 8 IBr 2 79 348 (M 1 Br) (1 9 B3r 2 346 (Mt 79 Br 3 269 267 265 241 239 237 915); 213 211 209 202 204 206 (12).

WO 02/00639 PCT/AU01/00781 39 5-(Bromomethylene)-4-propyl-2(5H)-furanone Pale yellow oil vmax 3080, 2950, 2850, 1780, 1630, 1595, 1460, 1380, 1330, 1290, 1150, 1085, 1020, 920, 830, 750, 720 cm 1 l. 1H n.m.r. 8 (CDC13) 0.90, t, J 7.5 Hz, 3H, CH3; 1.65, m, 2H, CH2; 2.44, t, J 7.5 Hz, 2H, CH2; 6.03, s, H3; 6.10, s, (CHBr). 1 3 C n.m.r. 5 (CDCI3): 13.5, CH3; 21.2, 27.9, CH2; 89.2, 5-(CHBr); 116.2, C3; 153.0, C5; 157.7, C4; 167.7, C2. Mass spectrum: m/z 218 (M 8 1Br), 216 (M, 79 Br), 203 201 190 188 137 (100); 122 118 109 5-(Dibromomethylene)-4-propyl-2(5H)-furanone Colourless prisms vmax 3070, 2950, 2850, 1775, 1630, 1590, 1460, 1380, 1150, 1090, 1020, 920, 830, 760 cm- 1 1 H n.m.r. 8 (CDCI3) 1.04, t, J 7.2 Hz, 3H,

CH

3 1.70, m, 2H, CH2; 2.77, t, J 8.4 Hz, 2, CH 2 6.18, t, J 1.5 Hz, H3. 1 3 C n.m.r. 6 (CDCI3): 13.6, CH3; 21.2, 31.7, CH2; 80.2, 5-(CBr2); 119.4, C3; 149.8, C5; 158.2, C4; 165.9, C2. Mass spectrum: m/z 298 (M (81Br 2 296 (M 81 Br), 79 Br) 294 (M 79 Br 2 283 281 279 268 254 217 (100); 215 (100); 202 200 198 189 187 174 172 170 5-(Bromomethylene)-4-butyl-2(5H)-furanone Pale yellow oil vmax2995, 2 9 50, 28 50, 1780, 1610, 1595, 1460, 1380, 1350, 1290, 1150, 1090, 1020, 920, 840, 750, 720 cm-1. ma,,x 281 nm 10540). 1H n.m.r. 6 (CDCl3) 0.90, t, J 7.5 Hz, 3, CH 3 1.41, m, 2H, CH2; 1.62, m, 2H, CH2; 2.42, t, J 8.2 Hz, 21, CH 2 6.02, s, H3; 6.10, s, 5-(CBBr). 13 C n.m.r. 6 (CDC3): 13.9, CH 3 22.4, 25.9, 27.7, 28.7, 31.3, CH 2 89.3, 5-(CHBr); 116.1, C3; 152.9, C5; 158.1, C4; 167.8, C2. Mass spectrum: m/z 232 (M 8 1Br), 230 79 Br), 203 201 190 189 151 123 109 (100).

5-(Bromomethylene)-4-hexyl-2( H)-furanone Pale yellow oil vmax 3095, 2930, 2850, 1778, 1638, 1600, 1450, 1160, 1028, 920, 758 cm- 1 ax 283 nm (s 5093). IH n.m.r. 6 (CDCl3) 0.90, t, J 7.1 Hz, 31g

CH

3 1.32, m, 8H11, CH2; 1.62, m, 2H11, CH2; 2.42, t, J 6.8 Hz, 2, CH 2 6.02, s H3; 6.09, s, 5-(CHBr) 13 C n.m.r. 5 (CDC3): 13.6, CH3; 22.3, 25.7, 29.9, CH 2 89.4, 116.1, C3; 153.0, CS; 158.3, C4; 167.8, C2. Mass spectrum: m/z 260 (M( 81 Br), 258 (Mv, 79 Br), 190 (40);188 179 151 137 109 WO 02/00639 PCT/AU01/00781 5-Bromo-5-(Bromomethyl)-4-hexyl-2( H)-furanone Pale yellow oil vmax 3111, 3039, 2954, 2929, 2854, 1793, 1637, 1600, 1465, 1414, 1377, 1263, 1237, 1178, 1151, 1124, 992, 910, 858 cm-1. max,, 214 nm (8 1054). 1 H n.m.r. 8 (CDC 3 0.91, t, J 7.1 Hz, 3, CH 3 1.35, m, 4H, CH2; 1.47, m, 2H,

CH

2 1.71, m, 2H, CH 2 2.51, m, 2H, CH2; 3.94, d, J 11.7 Hz, CHaBr; 4.26, d, J 11.7 Hz, CH 1 Br; 6.03, bs, 113. 13 C n.m.r. S (CDCI3): 13.9, CH 3 26.2, 26.8, 28.6, 31.3, CH2; 33.8, 5-(CH 2 Br); 91.8, C5; 116.8, C3; 167.8, C4; 171.3, C2. Mass spectrum: m/z 342 (M 8 1 Br 2 340 (M 8 1 Br), 79 338 7 9 Br 2 325 314 261 259 191 189 152 915); 137 110 (Bromomethylene)-4-heptyl-2( H)-furanone Pale yellow oil vmax 3095, 2920, 2850, 1770, 1630, 1595, 1450, 1360, 1280, 1150, 1100, 1020, 910, 840, 750, 720 cm- 1 X.max 214 nm (g 9590). 1H n.m.r. 6 (CDCl3) 0.89, t, J 7.1 Hz, 3, CH1 3 1.32, m, 10H, CH2; 1.62, m, 21H, CH2; 2.44, t, J 8.4 Hz, 2H, CH1 2 6.01, sH3; 6.09, s, 5-(CHBr). 13 C n.m.r. 5 (CDCI3): 13.9, CH 3 22.5, 25.9, 27.8, 28.7, 29.0, 31.5, CH2; 89.2, 5-(CHBr); 116.1, C3; 153.0, CS; 158.1, C4; 167.7, C2. Mass spectrum: miz 274 (M 81 Br), 272 79 Br), 193 (100); 190 188 165 151 123 109 (100).

5 -(Dibromomethylene)-4-heptyl-2( H)-furanone Pale yellow solid vmax 3085, 2925, 2850, 1759, 1586, 1463, 1360, 1280, 1175, 1068, 959, 836, 720 cm 1 ,max 296 nm (8 8857). 1H n.m.r. 5 (CDC1 3 0.89, t, J 7.1 Hz, 3H, CH3; 1.36, m, 10H, CH2; 1.66, m, 2H, CH2; 2.76, t, J 7.0 Hz, 21, CH2; 6.17, t, J 1.5 Hz, H3. 13C n.m.r. 5 (CDCl3): 13.9, CH3; 22.5, 27.8, 28.8, 29.0, 29.8, 31.5, CH2; 80.2, 5-(CBr2); 119.3, C3; 149.8, CS; 158.5, C4; 166.0, C2. Mass spectrum: m/z 354 (M (81Br 2 352 (M (3lBr, 79 Br), 10); 350 79 Br2), 273 270 268 266 240 211 200 189 187 5-Bromo-5-(bromomethyl)-4-heptyl-2( H)-furanone Pale yellow oil vmax 3111, 3039, 2954, 2929, 2856, 1794, 1638, 1465, 1416, 1378, 1264, 1237, 1178, 1152, 1124, 990, 910, 858, 710 cm-1. 296 nm (6 8857). 1 H n.m.r. 6 (CDC13) 0.91, t, J 7.1 Hz, 3H11, CH 3 1.31-1.45, m, 8H, CH 2 1.73, m, 2H, CH2; 2.32, m, 1H, CH2; 2.53, m, 1, CH2; 3.93, d, J 11.7 Hz, CHaBr; 4.26, d, J 11.7 Hz, CHbBr; 6.03, bs, H3. 13 C n.m.r. S (CDC13): 13.9, CH 3 22.5,26.2, 26.8, 28.8, 28.9, 31.5, CH2; 33.8, 5-(CHBr); 91.7, CS; 116.8, C3; 167.8, C4; 171.2, C2. Mass spectrum: m/z 356 (M (81Br 2 354 (M (81lBr), 79 Br), 352 79 Br 2 WO 02/00639 WO 0200639PCT/AU01/00781 41 31(5), 314 281 250 232 194 189 166 151 137 110 -(Bromonlethylef)4deCY- 2 Pale yellow oil vmax 3 11 8 3088, 2923, 2852, 1781, 1761, 1602, 1465, 1160, 1100, 923, 890, 760 cm- 1 1H1 n.m.r. 5 (CDCl3) 0.88, t, J 7.2 Hz, 311, CH 3 1.32, mn, 1411, CH2; 1.62, m, 2H1, GCl 2 2.44, t, J18.2 Hz, 2K1 CH2; 6.02, s, H3; 6.09, s, (CIJBr). 1 3 C n.m.r. 5(CDCI3): 14.1, CH3; 22.7, 26.0, 27.9, 29.2, 29.2, 29.3, 29.4, 29.5, 31.9, CH2; 89.3, 5-(CI-JBr); 111.6, C3; 153. 1, C5; 158.2, C4; 167.8, C2. Mass spectrum: mlz 316 (N4( 8 1Br), 314 79 Br), 282 267 253 235 217 207 190 188 175 165 910); 151 133 123 915); 109 920).

-(Bromomethy~le)4phenylI2(Sl)-furanone Colourless prisms vmjax 3090, 2920, 2850, 1760, 1630, 1605, 1590, 1440, 1385, 1340, 1260, 1160, 1080, 950, 900, 840, 760, 710 cm1. X 251 nm (F 2263).

In.m.r. 6(CDCl3) 6.25, s, 5-(CIIBr); 6.30, s,H13. 13 C n.m.r.5S(CDCl3): 93.1 116.1, C3; 128.2, 129.2, 130.9, Ph; 151.8, C5; 155.7, C4; 167.0, C2. Mass spectrum: m/z 252 (M 81 Br), 250 (Ki 79 11r), 172 144 116 114 102 (100).

Synthesis of 4 -bromo-5-(bromfomethylene)2(5H)furanone Wi Crude 2,3,5-tribromo-4-oxopefltanoic acid (1.0 g) was treated according to method G to yield 4 -bromo-5-bromomethylene2(5HA-franone (0.6g, 83%) as colourless needles m.p. 97-98 0 C. vmax 3 1 60 3120, 1805, 1783, 1644, 1565, 1314, 1244, 1156,. 1101, 976, 892, 831 cur 1 X.max 290 n (6 8230). 1 H n.m.r. 6 (CDCI3) 6.42, s, 5-CHBr; 6.50, s, H3. 13 C n.m.r. 5 (CDCl3): 93.8, 5-CHBr; 121.0, C3; 135.3, C4; 151.5, C5; 165.5, C2.

(ii) 2,3-Dibromo-4-oxopefltafloic acid (1 g) was treated according to method

G

to yield 4 -bromo-5-bomomethylene2(5H)-furanone (0.4g, 50%) as colourless needles m.p. 97-98 0

C.

GENERAL METHODS FOR THE SYNTHESIS OF 4-(1'-HALOALKYL)-5 (HALOMETHYLENE)-, AND 3-RM WO 02/00639 PCT/AU01/00781 42 An example of 3-bromo-4-(l-bromoalkyl)-5-(halomethylene)-2(H)-furanone produced is provided below.

General method for the synthesis of 3-bromo-4-(1-bromoalkyl)- (halomethylene)-, and 4 -(l-bromoalkyl)-5-(halomethylene)-2(5H)-furanone N-Halosuccinimide (2.43 mmol) was added to a solution of 3-bromo-4or 4-alkyl-5-(halomethylene)-2(5H)-furanone (2.43 mmol) in carbon tetrachloride (15 ml) containing a few crystals ofbenzoyl peroxide. The mixture was irradiated with a 250 W lamp and refluxed in an oil bath for 18h. After cooling the mixture to room temperature it was filtered and the precipitate washed with carbon tetrachloride (50 ml). The combined filtrate and washings were evaporated under reduced pressure and the crude product was purified by silica gel chromatography using dichloromethane light petroleum (1 2) as the eluent to yield the 1-haloalkyl compounds (45-55%).

3-Bromo-4-(l'-bromoheptyl)-5-(bromomethylene)-2(5H)-furanone Colourless oil vmax 3101, 2954, 2927, 2866, 1790, 1632, 1585, 1463, 1377, 1270, 1177, 992, 897, 774, 748 cm-1. lmax 305 nm (e 11284). 1H n.m.r. (CDCl3) 0.88, t, J 7.2 Hz, 3, CH3; 1.29, m, 6H, CH 2 1.48, m, 2H, CH 2 2.25, m, 2H, CH2; 4.87, t, J 7.9 Hz, 2H, CH2; 6.65, s, 5-(CHBr). 13C n.m.r. 6 (CDC13): 13.9, CH 3 22.3, 27.7, 28.2, 31.3, 37.7, 40.4, CH2; 93.5, 5-(CHBr); 112.6, C3; 148.5, C5; 150.6, C4; 162.3, C2. Mass spectrum: m/z 434 (M (81Br 3 432 (M 1 Br 2 79 Br), 430 (M (81Br) 79 Br 2 428 (M 79 Br 3 408 391 354 352 350 328 (10);326 270 268 (100); 266 229 213 915); 189 173 Biological activity of furanones Growth of Staphylococcus aureus and Candida albicans against furanones (Figures 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, Nutrient Broth, containing furanones at the concentrations 1-50 Pg/ml. The bacteria were incubated at 37C and growth was measured at 610 nm.

WO 02/00639 PCT/AU01/00781 43 Results The results demonstrated that furanone 63 prolonged the lag phase of growth at and 50 pg/ml. The effect of prolonged lag phase of growth was also demonstrated with compound 105. Ten pg/ml inhibited the growth of S. aureus for 72 hrs. No growth was detected with compound 120 at 1 lg/ml.

Candida albicans were grown in Sabouraud dextrose media at 37C. One percent of overnight culture was inoculated to 10 ml of growth media containing furanones at concentrations 1-50 Rg/ml. The growth was measured at 610 nm for 72 hrs for compound 73 and 24 hrs for compound 113. Ten .g/ml of compound 73 completely inhibited the growth of C. albicans while 5 pg/ml gave a prolonged lag phase of 8 hrs.

Compound 113 completely inhibited the growth for at least 24 hrs at 10 .g/ml.

Effect of furanones as inhibitor of AHL-mediated quorum sensing, attachment/biofilm formation, two-component signal transduction assay and AI-2 activity 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 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.

Attachment/Biofilm 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. 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 aeruginosa, is added to each well and incubated for 24h. Following incubation, the wells are rinsed 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/biofilm formation in the WO 02/00639 PCT/AU01/00781 44 presence of the furanones is calculated as the percentage of the controls, which are not exposed to the furanones.

Two-Component signal transduction 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 grown overnight in M9 medium at 37 0 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 0 C and shaken at 180 rpm. The OD61o of the growing cultures was monitored regularly and when the OD 610 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 0 C incubator and shaken for 4 hours (OD61o approximately 0.7) before being removed and put on ice.

Samples were then removed for Deta-galactosidase assays carried out according to the method of Miller (1972).

V. harveyi 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 0

C

with shaking in AB medium. Cells were diluted 1:5,000 into 30 0 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 sterile medium to 100 ul. Ten ul of V.

han'eyi BB152 AI-2+) supernatant was used as a positive control and 10 ul of E. coli DH5o supernatant or sterile media was used as a negative control. This strain of E. coli has previously been shown to harbor a mutation in the AI-2 synthase gene, ygaG, which results in a truncated protein with no AI-2 activity (Surette et al. 1998).

The microtiter plates were incubated at 30 0 C with shaking at 175 rpm. Hourly determinations of the total luminescence were quantified using the chemiluminescent setting on a Wallac (Gaithersburg, MD) model 1450 Microbeta Plus liquid scintillation counter. The V harveyi cell density was monitored by the use of a microplate reader WO 02/00639 PCT/AU01/00781 (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.

The results of these experiments are summarised in the table 1.

Table 1. Effect of furanones as inhibitor of AHL-mediated quorum sensing, attachment/biofilm formation, two-component signal transduction assay and AI-2 activity WO 02/00639 PCT/AU01/00781 46 Christensen, G. W. A. Simpson, J. J. Younger, L. M. Baddour, F. F. Barrett, D. M.

Melton, and E. H. Beachey. 1985. Adherence of coagulase-negative staphylococci to plastic tissue culture plates: a quantitative model for the adherence of staphylococci to medical devices. J. Clin. Microbiol. 22(6):996-1006.

Andersen, J. C. Sternberg, L. K. Poulsen, S. P. Bjorn, M. Givskov, and S. Molin.

1998. New unstable variants of green fluorescent protein for studies of transient gene expression in bacteria. Appl.,Environ. Microbiol. 64(6):2240-2246.

Jin, 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 Escherichia coli transmembrane receptor, Tazl. 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. and B. L. Bassler. 1998. Quorum sensing in Escherichia coli and Salmonella typhimurium. Proc. Natl. Acad. Sci., USA 95:7046-7050.

Surette, M. M. B. Miller, and B. L. Bassler. 1999. Quorum sensing in Escherichia coli, Salmonella typhimurium, and Vibrio harveyi: 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.

Claims (56)

1. A method for the preparation of a compound of formula II R1 R2 .R 6 0 0 wherein R 1 and R2 are independently H, alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic, hydrophobic or fluorophilic; R3, R 4 R 5 and R6 are independently or all hydrogen or halogen; the method comprising cyclising a compound of formula I R 1 ISOOH R 3 R4 R 2 wherein R 1 R2, R3, R4, R 5 and R6 are as defined above, wherein the cyclisation is carried out in the presence of a mild acid catalyst or a dehydrating agent or a mixture thereof, optionally in the presence of solvent.

2. A method according to claim 1 wherein at least two of the R3, R4, Rs and R6 are halogens;

3. A method according to claim 1 or claim 2 wherein in the compound of formula I and the compound of formula II: R1 and R2 are independently H, alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic, hydrophobic or fluorophilic; WO 02/00639 PCT/AU01/00781 48 R 3 R4, R 5 and R6 are independently or all hydrogen or halogen; and provided that at least two of the R 3 R4, R 5 and R6, which may be the same or different, are halogens.

4. A method according to claim 1 wherein in the compound of formula I and the compound of formula II: R 1 and R 2 are independently H, alkyl, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic, hydrophobic or fluorophilic; R3, R4, R5 and R6 are independently or all hydrogen or halogens; and provided that at least two of the R 3 R4, R5 and R6, which may be the same or different, are halogens.

A method of claim 1 wherein in the compound of formula I and the compound of formula II: R1 and R2 are independently H, alkyl, aryl or arylalkyl whether unsubstituted or substituted, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic, hydrophobic or fluorophilic; R3, R4, R 5 and R6 are independently or all hydrogen or halogen; and provided that at least two of R 3 R 4 R 5 and R, which may be the same or different are halogens.

6. A method according to any one of the preceding claims wherein at least one of and R 6 is Br.

7. A method according to claim 1, wherein the compound of formula I is a brominated 4-oxopentanoic acid, wherein at least two of R3, R4, R5 and R6, which may be the same or different are halogens.

8. A method according to claim 1 wherein the compound of formula II is selected from halogenated 2-tetrahydrofuranones.

9. A method according to any one of the preceding claims, wherein the reaction is carried out in the presence of a mild acid catalyst(s).

10. A method according to claim 8. wherein the catalyst(s) is/are insoluble in the reaction medium WO 02/00639 PCT/AU01/00781 49

11. A method according to claim 10, wherein that catalyst is selected from the group consisting of polyphosphoric acid, Eaton's reagent, acidic resins and polymers, Lewis acids and acidic metal salts.

12. A method according to claim 9, wherein the catalyst is/are soluble in the reaction medium.

13. A method according to claim 12, wherein the catalyst is/are selected from the group consisting of chlorosulfonic acid, phosphoric acid, p-toluenesulfonic acid, trifluoromethanesulfonic acid, methanesulfonic acid, acetic acid, bromine, phosphorus tribromide and hydrobromic acid.

14. A method according to any one of the preceding claims which is carried out in the presence of a dehydrating agent.

A method according to claim 14, wherein the drying agent is selected from the group consisting of phosphorus pentoxide, silica gel, molecular sieves, alumina, phosphorus oxychloride, acetic anhydride, N, N'-dicyclohexyl-carbodiimide (DCC), trifluoroacetic anhydride and trifluorosulfonic acid anhydride (triflic anhydride).

16. A method according to claim 15, wherein the dehydrating agent is phosphorus pentoxide or polyphosphoric acid optionally mixed with a mineral acid.

17. A method according to claim 16 wherein the dehydrating agent is phosphorus pentoxide.

18. A method according to any one of the preceding claims wherein the cyclisation is carried out in the presence of at least one solvent.

19. A method according to claim 18, wherein the solvent is selected from the group consisting of alkyl acetates, aromatic hydrocarbons, chlorinated alkanes, tetrahydrofuran, diethyl ether, dioxane and C1-C3 acids.

20. A method according to claim 19 wherein the solvent is selected from aromatic hydrocarbons and chlorinated alkanes. WO 02/00639 PCT/AU01/00781

21. A method according to claim 20, wherein the solvent is selected from the group consisting of dichloromethane, dichloroethane and trichloroethane and mixtures of two or more thereof.

22. A method according to any one of the preceding claims wherein the cyclisation reaction is performed at a temperature in the range of from about 20-150 0 C.

23. A method according to any one of claims 18 to 22, wherein at least one solvent is present and the cyclisation is performed at reflux temperature of the solvent.

24. A method according to any one of the preceding claims, wherein the cyclisation reaction is carried out for at least 2 hours.

25. A compound of formula II produced by a method in accordance with any one of the preceding claims.

26. A compound of formula I RI COOH 2R2 I wherein RI and R 2 are independently H, alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic, hydrophobic or fluorophilic; R3, R4, Rs and R 6 are independently or all hydrogen or halogen; and provided that at least two of the R3, R4, R5 and R6 are halogens;

27. A compound according to claim 26 selected from the group consisting of compounds of formulae la-i: PCT/AOb1//00781 Received 8 May 2002 51 Br Br Sr H H H Ho H' JOOH la lb 1c Br Br Br H OOH H H H BBr Id e f H -OO 1O H Br-r Br O r Br B B r- Ig 1h li

28. A tetrahydro-2(5H)-furanone derivative of formula II, Ri R 3 II wherein R 1 and R2 are independently H, alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic, hydrophobic or fluorophilic; R 3 R 4 Rs and Re are independently or all hydrogen or halogen; and provided that at least two of the R 3 R 4 R 5 and Re are halogens; excluding compounds in which: R,=R 2 H, R 3 =R4=C and R.=R=Br; and R 1 ,=R2=R 5 =R6=H and R 3 R 4 =CI AMENDED SHEET IPEA/AU WO 02/00639 PCT/AU01/00781 52

29. A compound according to claim 28, which is selected from the group consisting of compounds of formula 2a-i Br Br Br B 2a Br 2b Br 2c Br H Me H H Br Br/ B Br Br Br H H 0 f H 2d H 2e H 2f B Br B B Br H H H 2g 2h 2i

30. A method for the dehydrohalogenation of a compound of formula II R 1 R 2 R 5 R 6 11 R4 wherein R 1 and R 2 are independently H, alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic, hydrophobic or fluorophilic; R 3 R, R5 and R6 are independently or all hydrogen or halogen, provided that at least two of the R3, R 4 R5 and R6 are halogens; to prepare a compound of formula IIIa or Ib; WO 02/00639 PCT/AU01/00781 53 OR R3 Ilia R4 wherein R2 is an alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophobic, hydrophilic or fluorophilic; R 4 is a halogen RI and R 3 are independently or both hydrogen or halogen; RI R2 Illb R4 wherein R2 is an alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophobic, hydrophilic or fluorophilic; R 3 and R 4 are hydrogen and Ri is a halogen; the method comprising contacting a compound of formula II with a base.

31. A method according to claim 30, wherein the base is selected from the group consisting of 1,4-diazabicyclo[2.2.2]octane (DABCO), 4-(dimethylamino)pyridine (DMAP), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), triethylamine, alkali metal carbonate, alkali metal acetate and N,N'-diisopropylethyl amine (Hunig's base).

32. A method according to claim 30 or claim 31, wherein the compound of formula III is selected from the group consisting of furanones of formula 3a-n: WO 02/00639 WO 0200639PCT/AU01/00781 H 0: 3a Br H SBr 3b Br 0 ~Br 3d Br H SBr '0 3e (88) Br BE Me SH W H Br SH 0 0 3g H Br 0: 3h H M1 Br SH 0 Br 3j (63) Me Br 3 m 6 4 Br 3k H

33. A compound according to formula Mia or ifb. Ri R 2 0 0 ~R 3 ilia R PCT/AUOI//00781 Received 8 May 2002 56 wherein in figure Ila R 2 is an alkyl, alkoxy, oxoalkyl, alkenyl, Mry or arlalky whether unsubstituted or substituted, straight chain or branched chain, hydrophobic, hydrophilic or fluorophilic; 1 4 is a halogen; Ri and R 3 are independently or both hydrogen or halogen', excluding compounds in which:- R 1 =Ht R 2 =Me or Ph, RZ3-1, R4=H; and R 1 1 2 =O!Ve, R13=Cl, R.=Cl; 0 R wherein in formula tulb R7 is an alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophobic, hydrophilic or fluorophilic; R 3 and R 4 are hydrogen; and R, is a halogen,

34, A method for the halogenation of a compound of formula 11, Ilila or Illb R, Rp wherein R, and R 2 are independently H, alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic, hydrophobic or fluorophilic; R 4 R 5 and R 6 are independently or all hydrogen or halogen; and provided that least two of the R 3 R 4 Rs arnd R 6 are halogens; AMENDED GHEE7 IPEA/AU WO 02/00639 PCT/AU01/00781 56 R 1 R 2 Ilia R wherein R2 is an alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophobic, hydrophilic or fluorophilic; R4 is a halogen Ri and R3 are independently or both hydrogen or halogen; R1 R2 OR R3 Illb R4 wherein R2 is an alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophobic, hydrophilic or fluorophilic; R3 and R4 are hydrogen and R, is a halogen; to prepare a compound of formula IV RI R2 0 R 3 IV wherein R2 is independently H, halogen, alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophilic or fluorophilic; R 4 is selected from halogen, OH and alkoxy; R5 are halogen; R, and R3 are independently or both hydrogen or halogen; and is a single bond or double bond, WO 02/00639 PCT/AU01/00781 57 the method comprising contacting a compound of formula II, IIIa or Ifb with a halogenating agent optionally in the presence of at least one unreactive or reactive solvent or reagent.

A method according to claim 34, wherein the halogenating agent is selected from the group consisting of bromine, chlorine, iodine, N-bromosuccinimide, N- chlorosuccinimde, iodine monochloride, phenyltrimethylammonium bromide perbromide, pyridinium tribromide and cupric bromide.

36. A method according to claim 34 or 35, wherein the unreactive solvent or reagent is selected from non-nucleophilic organic solvents or ionic liquids.

37. A method according to claim 36, wherein the solvent or reagent is selected from the group consisting ofdichloromethane, chloroform, toluene, diethyl ether, N,N- dimethylformamide, N-methylpyrrolidinone and butylmethylimidazolium tetrafluoroborate.

38. A method according to claim 34 or 35 wherein the at least one reactive solvent or reagent is a nuceophilic organic or inorganic substance.

39. A method according to claim 38, wherein the at least one reactive solvent or reagent is selected from the group consisting of water, methanol, acetic acid, lithium chloride, benzylamine and silver nitrate.

A method according to any one of claims 34 to 39 wherein the compound of formula IV is selected from the group consisting of compounds of formula 4a-p: WO 02/00639 WO 0200639PCT/AU01/00781 58 Br Me Br 0 Brr0 BroBr 4a 4b Br E o0 Br 0 4d Ph Vf 4g B r Me B Br 0 B Br 0 0 Br Br 4i 4j Me B Br H B r 0 0 Br Br 0 Br Br 41(72) 4m Brr 0 4c 4e (120) 4h 4k 4n WO 02/00639 PCT/AU01/00781 59

41. A compound of formula IV, R 1 R2 0 R 4 IV wherein R 2 is independently alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophilic or fluorophilic; R4 is selected from halogen, OH and Oalkyl; Rs are halogen; R 1 and R 3 are independently or both hydrogen or halogen; and is a single bond or double bond.

42. A method for the dehydrohalogenation of a compound of formula IV: RI R2 0 R 4 R3 IV wherein R 2 is independently H, halogen, alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophilic or fluorophilic; R 4 is selected from halogen, OH and Oalkyl; Rs are halogen; R I and R 3 are independently or both hydrogen or halogen; and is a single bond or double bond. to prepare a compound of formula V WO 02/00639 PCT/AU01/00781 R1 R 2 O -O R 3 V R4 wherein R2 is a H, alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophobic, hydrophilic or fluorophilic; R4 is a halogen; R 1 and R 3 are independently or both hydrogen or halogen; the method comprising contacting a compound of formula IV with a base.

43. A method according to claim 42 wherein the base is selected from the group consisting of 1,4-diazabicyclo[2.2.2]octane (DABCO), 4-(dimethylamino)pyridine (DMAP), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), triethylamine, alkali metal carbonate, alkali metal acetate and N,N'-diisopropylethyl amine (Hunig's base).

44. A method according to claim 42 wherein Ri of formula (IV) is a halogen and the base is N,N-diisopropylethyl amine.

A method according to any one of claims 42 to 44, wherein the compound of formula V is selected from the group consisting of a compounds WO 02/00639 WO 0200639PCT/AU01/00781 Br Me 00 6a Br Br 0 0 H 5b (99) Br Ph 0 0 Se Br B 0: H Sf (105) E3r Brr 0 ~Br 0 ~Br i Br 5 Br 0 0 5 r Br Br 0~ 0 k Br 44. A compound of formula V: PCT/AU01//00781 Received 8 May 2002 62 RI R V R4 wherein R is an alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophobic, hydrophilic or fluorophilic; R4 is a halogen; Ri and R are independently or both hydrogen or halogen; excluding compounds in which: Ri=H, R 2 Me or Ph, R I, R 4 H; and R, H, R2 OMe, R CI, R4 CI.

46. A method of tandem cyclisation, dehalogenation, rehalogenation and dehydrohalogenation of compounds of formula: R4 2 I wherein Ri and R 2 are independently H, alkyl, alkoxy, oxoalkyl, alkenyl, aryl, or arylalkyl whether unsubstituted or substituted, optionally interrupted by one or more hetero atoms, straight chain or branched chain, hydrophilic, hydrophobic or fluorophilic; R 8 RF, R 5 and Re are independently or all hydrogen or halogen; and provided that at least two of the Re, R4, R 6 and Re are halogens; to form a compound of formula the method comprising contacting the compound with a strong acid RI R2 O 0 R3 VI R AMENDED SHEET IPEA/AU WO 02/00639 PCT/AU01/00781 63 wherein R2 is a H, halogen, alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophobic, hydrophilic or fluorophilic; R4 is a halogen; RI is hydrogen; and R3 is a hydrogen or halogen.

47. A method according to claim 46, wherein the strong acid is a sulfuric acid type reagent.

48. A method according to claim 47, wherein the sulfuric acid type reagent is selected from the group consisting of concentrated sulfuric acid, oleum, chlorosulfonic acid, and a mixture of two or more thereof with one or more other like agents.

49. A method according to any one of claims 46 to 48, wherein the compound of formula I is selected from the group consisting of 3-alkyl-2,3-dibromo-4-oxopentanoic acid, 2,3,5-tribromo-4-oxopentanoic acid, 2,3-dibromo-4-oxopentanoic acid, dibromo-4-oxopentanoic acid, 2,3,5,5-tetrabromo-4-oxopentanoic acid, 2,3,3-tribromo- 4-oxopentanoic acid and 2,3,3,5-tetrabromo-4-oxopentanoic acid.

A method according to claim 49, wherein the compound of formula VI is selected from the group consisting of compounds of formula 6a-g: PCT/AU01//00781 Received 8 May 2002 6a (73) o s Br 6g Br

51. A compound of formula VI: R1 R VI 4 wherein R 2 is an alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophobic, hydrophilic or fluorophilic; R 4 is a halogen; R 1 is a hydrogen; and R 3 is a hydrogen or halogen; excluding a compounds in which RI=H, R 2 =Me or Ph, R3=I,R 4 and Ri=H, R 2 =OMe, R=CI, R 4 =CI.

52. A compound of formula (VII): AMENDED SHEET IPEA/AU WO 02/00639 PCT/AU01/00781 z RI R2 O O R 3 VII R4 wherein R2 is a H, alkyl, alkoxy, polyethyleneglycyl, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophobic, hydrophilic or fluorophilic; R 4 is a hydrogen, halogen; R, and R 3 are independently or both hydrogen or halogen; Z is independently selected from the group R 2 halogen, OC(0)R 2 amine azide, thiol, R 2 mercaptoaryl, arylalkoxy, mercaptoarylalkyl, SC(0)R 2 OS(0) 2 R 2 NHC(0)R, =NR 2 or NHR 2 prepared by functionalizing a fimbrolide of formula (VIII) wherein, R 1 R 2 R 3 and R 4 are as defined above, with reagents selected from the group consisting of halogenating, oxidising agents, nucleophiles and electrophiles. RI 0 R3 VIII R4

53. A compound according to claim 52 wherein the halogenation and oxidising agents are selected from N-halosuccinimide, lead tetraacetate, selenium dioxide, Jones reagent, organic metal carboxylates, organic alcohols, dimethyl sulfoxide, organonitriles, organic acids, isocyanates, carboxylic or sulfonic acid halides and diethylaminosulfur trifluoride.

54. An oligomer or polymer formed by oligomerising or polymerising a compound selected from the group consisting of compounds of formula IIIa, lb VII, directly or with one or more monomers WO 02/00639 PCT/AU01/00781 66 wherein R2 is an alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophobic, hydrophilic or fluorophilic; R 4 is a halogen R, and R3 are independently or both hydrogen or halogen; Ri R2 Illb R4 wherein R2 is an alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophobic, hydrophilic or fluorophilic; R3 and R4 are hydrogen; and R, is a halogen. R1 R2 R5 O R IV wherein R2 is independently H, halogen, alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophilic or fluorophilic; R4 is selected from halogen, OH and Oalkyl; are halogen; RI and R3 are independently or both hydrogen or halogen; and is a single bond or double bond. RI ,R2Z V R4 WO 02/00639 PCT/AU01/00781 67 wherein R2 is a H, alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophobic, hydrophilic or fluorophilic; R4 is a halogen; R t and R3 are independently or both hydrogen or halogen; R1 R2 oo R3 VI R4 wherein R2 is a H, halogen, alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophobic, hydrophilic or fluorophilic; R4 is a halogen; RI is hydrogen; and R3 is a hydrogen or halogen; R2 O R3 VII R4 wherein R2 is a H, alkyl, alkoxy, oxoalkyl, alkenyl, aryl or arylalkyl whether unsubstituted or substituted, straight chain or branched chain, hydrophobic, hydrophilic or fluorophilic; R4 is a hydrogen, halogen; R, and R3 are independently or both hydrogen or halogen; Z is independently selected from the group R2, halogen, OC(O)R 2 amine azide, thiol, R2, mercaptoaryl, arylalkoxy, mercaptoarylalkyl, SC(O)R 2 OS(0)2R2, NHC(O)R2, =NR 2 or NHR2, An oligomer or polymer according to claim 54, where the at least one monomer is selected from the group consisting of acrylate ester such as alkyl, hydroxyalkyl, aminoalkyl, or substituted aryl acrylates or methacrylates, crotonates, substituted or unsubstituted acrylonitriles, vinyl alcohols or acetates, styrene and siloxanes.

WO 02/00639 PCT/AU01/00781

56. An article incorporating at least one compound of formula II to VII in a surface coating(s) or polymer(s) through the newly introduced functionality on the alkyl chain or the alkyl chain itself via direct polymerisation or copolymerisation with suitable monomers.