AU2005291833A1 - Selective inhibitors - Google Patents

Description

WO 2006/037159 PCT/AU2005/001510 1 Selective Inhibitors Field of the invention. 5 The invention relates to a method of identifying compounds with selective biologically activities, and libraries of compounds. Background. Small molecules involved in molecular interactions with a biological target, be it 10 enzyme or receptor, are often described in terms of binding elements or pharmacophore groups which directly interact with the target, and non-binding components which form the framework of the bioactive molecule. In the case of peptide ligands or substrates for instance, a number of amino acid side chains usually form direct interactions with their receptor or enzyme, whereas specific 15 folds of the peptide backbone (and other amino acid residues) provide the structure or scaffold that controls the relative positioning of these side chains. In a peptidomimetic approach to drug discovery, the side chains of important amino acids may be systematically modulated to identify better binding interactions. This is referred to as a scanning approach. Unfortunately, the side chains of peptides 20 are rarely independent, such that each interaction cannot be optimised without consideration of the others. One way to overcome this problem is to construct diversity libraries. 25 So far, approaches for creating universal diversity have largely focused on the combination of substituents aspects. When it comes to creating diversity in WO 2006/037159 PCT/AU2005/001510 2 presentation of these substituents, pharmaceutical companies generally turn to the known heterocyclic scaffolds, with an emphasis on the so-called 'privileged structures'. Creating structural diversity in libraries has been highly desired but has been limited by the lack of structural diversity in the chemically useful scaffolds. 5 Monosaccharides provide an excellent sugar scaffold to design molecular diversity by appending desired substituents at selected positions around the sugar scaffold. The monosaccharide-based scaffold contains five chiral, functionalized positions, enabling attachment of various substituents at each position. This provides a 10 unique opportunity to create libraries of structurally diverse molecules, by varying the pharmacophoric groups, the scaffold and the positions of attachment of the pharmacophoric groups in a systematic manner. A pharmacophoric group in the context of these libraries is an appended group or substituent, or part thereof, which imparts pharmacological activity to the molecule. 15 Molecular diversity could be considered as consisting of diversity in pharmacophoric group combinations (diversity in substituents) and diversity in the way these pharmacophoric groups are presented (diversity in shape). Libraries of compounds in which either diversity of substituents, or diversity of shape, or both of 20 these parameters are varied systematically are said to scan molecular diversity. There is a need for methods to improve the development of drug candidates that purposely interact with selected targets, and not with other targets, in order to minimize side effects. Selectivity profiles are determined by biological assays, 25 either in vitro or in vivo, in which compounds exhibit a specific response in each WO 2006/037159 PCT/AU2005/001510 3 assay. The panel of specific responses represents the selectivity profile across the selected assays. The profile distinguishes actives against non-actives in each assay. Methods to improve the identification of selectivity profiles overcome or at least partially ameliorate this problem. 5 In previous applications (WO2004014929 and WO2003082846) we demonstrated that arrays of novel compounds could be synthesized in a combinatorial manner. The libraries of molecules described in these inventions were synthesized in a manner such that the position, orientation and chemical characteristics of 10 pharmacophoric groups around a range of chemical scaffolds, could be modified and/or controlled. In a later application (WO2004032940), we demonstrated that classes of molecules from the above cited applications exhibited biological activity when 15 screened against melanocortin and somatostatin GPCRs. Classes of molecules from the applications WO2004014929 and WO2003082846 were also tested against integrin receptors (Australian patent Application No. 2003900242). Selections of these molecules were also demonstrated to display activity against this class of receptors. 20 We have now found that libraries of molecules described in the applications WO2004014929 and WO2003082846 can be used to scan molecular diversity. This diversity approach provides an improved method, for effectively identifying selectivity profiles. 25 It will be clearly understood that, if a prior art publication is referred to herein, this reference does not constitute an admission that the publication forms part of the common general knowledge in the art in Australia or in any other country. 30 WO 2006/037159 PCT/AU2005/001510 4 Summary of the invention. In one aspect the invention provides a method of identifying biologically active 5 compounds with defined selectivity profile(s) comprising: (a) designing a library of compounds of formula 1 to scan molecular diversity; and (b) assaying the library of compounds in at least two different biological assays; wherein formula 1 represents: 0 ZR 1

R

5 X V X

R

4 X

XR

2 10

XR

3 Formula I wherein the ring may be of any configuration; Z is sulphur, oxygen, CH 2 , C(O), C(O)NRA, NH, NRA or hydrogen, in the case 15 where Z is hydrogen then R 1 is not present, RA is selected from the set defined for

R

1 to R 5 , or wherein Z and R1 together form a heterocycle, X is oxygen or nitrogen, when X is nitrogen, each X may combine independently with the corresponding R 2 to R 5 to form an azide, or wherein each X may also combine independently with any one of corresponding R 2

-R

5 to form a heterocycle; 20 R 1 to R 5 are independently selected from the group which includes but is not limited to H or an C1 to C20 alkyl or acyl; C2 to C20 alkenyl, alkynyl, heteroalkyl; C5 to C20 aryl, heteroaryl, arylalkyl or heteroarylalkyl, which is optionally substituted, and can be branched or linear. 25 In a preferred embodiment the invention relates to a library of compounds selected from compounds of formula 1 when used according to first said method. In a preferred embodiment, the invention relates to first said method wherein at least one X is nitrogen. 30 WO 2006/037159 PCT/AU2005/001510 5 In a preferred embodiment, the invention relates to first said method wherein two of X is nitrogen. In a preferred embodiment, the invention relates to first said method wherein X and 5 R 2 combine to form heterocycle. In a preferred embodiment, the invention relates to first said method wherein R 1 R 5 optional substituents are selected from OH, NO, NO 2 , NH 2 , N 3 , halogen, CF 3 ,

CHF

2 , CH 2 F, nitrile, alkoxy, aryloxy, amidine, guanidiniums, carboxylic acid, 10 carboxylic acid ester, carboxylic acid amide, aryl, cycloalkyl, heteroalkyl, heteroaryl, aminoalkyl, aminodialkyl, aminotrialkyl, aminoacyl, carbonyl, substituted or unsubstituted imine, sulfate, sulfonamide, phosphate, phosphoramide, hydrazide, hydroxamate, hydroxamic acid, heteroaryloxy, aminoaryl, aminoheteroaryl, thioalkyl, thioaryl or thioheteroaryl, which may optionally be 15 further substituted. The term "halogen" denotes fluorine, chlorine, bromine or iodine, preferably fluorine, chlorine or bromine. 20 The term "alkyl" used either alone or in compound words such as"optionally substituted alkyl","optionally substitutedcycloalkyl","arylalkyl"or"heteroarylalkyl", denotes straight chain, branched or cyclic alkyl, preferably C1-20 alkyl or cycloalkyl. Examples of straight chain and branched alkyl include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, amyl, isoamyl, sec-amyl, 1,2 25 dimethylpropyl,1, 1-dimethylpropyl, hexyl, 4-methylpentyl, 1-methylpentyl, 2 methylpentyl, 3methylpentyl,1, 1-dimethylbutyl, 2,2-dimethylbutyl, 3,3dimethylbutyl,1, 2-dimethylbutyl, 1,3-dimethylbutyl, 1,2,2trimethylpropyl,1, 1,2 trimethylpropyl, heptyl, 5methylbexyl, 1-methylhexyl, 2,2-dimethypentyl, 3,3 WO 2006/037159 PCT/AU2005/001510 6 dimethylpentyl, 4,4-dimethylpentyl, 1,2-dimethylpentyl, 1,3-dimethylpentyl, 1,4 dimethylpentyl, 1,2,3trimethylbutyl,1, 1,2-trimethylbutyl, 1,1,3-trimethylbutyl, octyl, 6-methylheptyl, 1-methylheptyl, 1,1,3,3 tetramethylbutyl, nonyl,1-, 2-, 3-, 4-, 5-, 6-or 7-methyloctyl,1-, 2-, 3-, 4-or 5-ethylheptyl, 1-, 2-or 3propylhexyl, decyl,1-, 2-, 3-, 4-, 5 5-, 6-, 7-or 8methylnonyl,1-, 2-, 3-, 4-, 5-or 6-ethyloctyl, 1-, 2-, 3or 4-propylheptyl, undecyll-, 2-, 3-, 4-, 5-, 6-, 7-, 8or 9-methyldecyl,1-, 2-, 3-, 4-, 5-, 6-or 7 ethylnonyl, 1-, 2-, 3-, 4-or 5-propyloctyl, 1-, 2-or 3-butylheptyl, 1-pentylhexyl, dodecyl,1-, 2-, 3-, 4-, 5-, 6-, 7-, 8-, 9or 10-methylundecyl,1-, 2-, 3-, 4-, 5-, 6-, 7-or 8ethyldecyl,l-, 2-, 3-, 4-, 5-or 6-propylnonyl, 1-, 2-, 3or 4-butyloctyl, 1-2 10 pentylheptyl and the like. Examples of cyclic alkyl include mono-or polycyclic alkyl groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl and the like. The term "alkylene" used either alone or in compound words such as "optionally 15 substituted alkylene" denotes the same groups as "alkyl" defined above except that an additional hydrogen has been removed to form a divalent radical. It will be understood that the optional substituent may be attached to or form part of the alkylene chain. 20 The term "alkenyl" used either alone or in compound words such as "optionally substituted alkenyl" denotes groups formed from straight chain, branched or cyclic alkenes including ethylenically mono-, di-or polyunsaturated alkyl or cycloalkyl groups as defined above, preferably C2-6 alkenyl. Examples of alkenyl include vinyl, allyl,l-methylvinyl, butenyl, iso-butenyl, 3-methyl-2butenyl, 1-pentenyl, WO 2006/037159 PCT/AU2005/001510 7 cyclopentenyl, 1-methyl-cyclopentenyl, 1-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, 1,3cyclohexadienyl, 1,4-cyclohexadienyl, 1,3 5 cycloheptadienyl, 1,3,5-cycloheptatrienyl and 1,3,5,7-cyclooctatetraenyl. The term "alkynyl" used either alone or in compound words, such as "optionally substituted alkynyl" denotes groups formed from straight chain, branched, or mono-or poly-or cyclic alkynes, preferably C2-6 alkynyl. 10 Examples of alkynyl include ethynyl,1-propynyl, 1-and 2butynyl, 2-methyl-2 propynyl, 2-pentynyl, 3-pentynyl, 4pentynyl, 2-hexynyl, 3-hexylnyl, 4-hexynyl, 5 hexynyl, 10undecynyl,4-ethyl-l-octyn-3-yl, 7-dodecynyl, 9-dodecynyl, 10 dodecynyl,3-methyl-I -dodecyn-3-yl, 2-tridecynyl, 11-tridecynyl, 3-tetradecynyl, 7 15 hexadecynyl, 3-octadecynyl and the like. The term "alkoxy" used either alone or in compound words such as "optionally substituted alkoxy" denotes straight chain or branched alkoxy, preferably C 1-7 alkoxy. Examples of alkoxy include methoxy, ethoxy, npropyloxy, isopropyloxy and 20 the different butoxy isomers. The term "aryloxy" used either alone or in compound words such as "optionally substituted aryloxy" denotes aromatic, heteroaromatic, arylalkoxy or heteroaryl WO 2006/037159 PCT/AU2005/001510 8 alkoxy, preferably C6-13 aryloxy. Examples of aryloxy include phenoxy, benzyloxy, l-napthyloxy, and 2-napthyloxy. The term "acyl" used either alone or in compound words such as "optionally 5 substituted acyl "or" heteroarylacyl" denotes carbamoyl, aliphatic acyl group and acyl group containing an aromatic ring, which is referred to as aromatic acyl or a heterocyclic ring which is referred to as heterocyclic acyl. Examples of acyl include carbamoyl; straight chain or branched alkanoyl such as formyl, acetyl, propanoyl, butanoyl, 2-methylpropanoyl, pentanoyl, 2,2-dimethylpropanoyl, hexanoyl, 10 heptanoyl, octanoyl, nonanoyl, decanoyl, undecanoyl, dodecanoyl, tridecanoyl, tetradecanoyl, pentadecanoyl, hexadecanoyl, heptadecanoyl, octadecanoyl, nonadecanoyl, and icosanoyl; alkoxycarbonyl such as methoxycarbonyl, ethoxycarbonyl, t butoxycarbonyl, t-pentyloxycarbonyl and heptyloxycarbonyl; cycloalkylcarbonyl such as cyclopropylcarbonyl cyclobutylcarbonyl, 15 cyclopentylcarbonyl and cyclohexylcarbonyl; alkylsulfonyl such as methylsulfonyl and ethylsulfonyl; alkoxysulfonyl such as methoxysulfonyl and ethoxysulfonyl; aroyl such as benzoyl, toluoyl and naphthoyl; aralkanoyl such as phenylalkanoyl (e. g. phenylacetyl, phenylpropanoyl, phenylbutanoyl, phenylisobutyl, phenylpentanoyl and phenylhexanoyl) and naphthylalkanoyl (e. g. naphthylacetyl, naphthlpropanoyl 20 and naphthylbutanoyl); aralkenoyl such as phenylalkenoyl (e. g. phenylpropenoyl, phenylbutenoyl, phenylmethacrylyl, phenylpentenoyl and phenylhexenoyl and naphthylalkenoyl (e. g. naphthylpropenoyl, naphthylbutenoyl and naphthylpentenoyl); aralkoxycarbonyl such as phenylalkoxycarbonyl (e. g. benzyloxycarbonyl); aryloxycarbonyl such as phenoxycarbonyl and WO 2006/037159 PCT/AU2005/001510 9 naphthyloxycarbonyl; aryloxyalkanoyl such as phenoxyacetyl and phenoxypropionyl; arylcarbamoyl such as phenylcarbamoyl; arylthiocarbamoyl such as phenylthiocarbamoyl; arylglyoxyloyl such as phenylglyoxyloyl and naphthylglyoxyloyl; arylsulfonyl such as phenylsulfonyl and naphthylsulfonyl; 5 heterocycliccarbonyl; heterocyclicalkanoyl such as thienylacetyl, thienylpropanoyl, thienylbutanoyl, thienylpentanoyl, thienylhexanoyl, thiazolylacetyl, thiadiazolylacetyl and tetrazolylacetyl; heterocyclicalkenoyl such as heterocyclicpropenoyl, heterocyclicbutenoyl, heterocyclicpentenoyl and heterocyclichexenoyl; and heterocyclicglyoxyloyl such as thiazolylglyoxyloyl and thienyglyoxyloyl. 10 The term "aryl" used either alone or in compound words such as "optionally substituted aryl", "arylalkyl "or "heteroaryl" denotes single, polynuclear, conjugated and fused residues of aromatic hydrocarbons or aromatic heterocyclic ring systems. Examples of aryl include phenyl, biphenyl, terphenyl, quaterphenyl, 15 phenoxyphenyl, naphthyl, tetrahydronaphthyl, anthracenyl, dihydroanthracenyl, benzanthracenyl, dibenzanthracenyl, phenanthrenyl, fluorenyl, pyrenyl, indenyl, azulenyl, chrysenyl, pyridyl, 4-phenylpyridyl, 3-phenylpyridyl, thienyl, furyl, pyrryl, pyrrolyl, furanyl, imadazolyl, pyrrolydinyl, pyridinyl, piperidinyl, indolyl, pyridazinyl, pyrazolyl, pyrazinyl, thiazolyl, pyrimidinyl, quinolinyl, isoquinolinyl, benzofuranyl, 20 benzothienyl, purinyl, quinazolinyl, phenazinyl, acridinyl, benzoxazolyl, benzothiazolyl and the like. Preferably, the aromatic heterocyclic ring system contains 1 to 4 heteroatoms independently selected from N, O and S and containing up to 9 carbon atoms in the ring.

WO 2006/037159 PCT/AU2005/001510 10 The term "heterocycle" used either alone or in compound words as "optionally substituted heterocycle" denotes monocyclic or polycyclic heterocyclyl groups containing at least one heteroatom atom selected from nitrogen, sulphur and oxygen. Suitable heterocyclyl groups include N-containing heterocyclic groups, 5 such as, unsaturated 3 to 6 membered heteromonocyclic groups containing I to 4 nitrogen atoms, for example, pyrrolyl, pyrrolinyl, imidazolyl, pyrazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazolyl or tetrazolyl; saturated to 3 to 6 membered heteromonocyclic groups containing 1 to 4 nitrogen atoms, such as, pyrrolidinyl,imidazolidinyl, piperidin or piperazinyl ; unsaturated condensed 10 heterocyclic groups containing 1 to 5 nitrogen atoms, such as, indolyl, isoindolyl, indolizinyl, benzimidazoyl, quinolyl, isoquinolyl, indazolyl, benzotriazolyl or tetrazolopyridazinyl; unsaturated 3 to 6-membered heteromonocyclic group containing an oxygen atom, such as, pyranyl or furyl; unsaturated 3 to 6-membered heteromonocyclic group containing 1 to 2 sulphur atoms, such as, thienyl; 15 unsaturated 3 to 6-membered heteromonocyclic group containing 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms, such as, oxazolyl, isoxazolyl or oxadiazolyl; saturated 3 to 6-membered heteromonocyclic group containing I to 2 oxygen atoms and 1 to 3 nitrogen atoms, such as, morpholinyl; unsaturated condensed heterocyclic group containing 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms, such 20 as, benzoxazolyl or benzoxadiazolyl; unsaturated 3 to 6-membered heteromonocyclic group containing 1 to 2 sulphur atoms and 1 to 3 nitrogen atoms, such as, thiazolyl or thiadiazolyl; saturated 3 to 6-membered heteromonocyclic group containing 1 to 2 sulphur atoms and 1 to 3 nitrogen atoms, such as thiazolidinyl; and unsaturated condensed heterocyclic group containing I to 2 WO 2006/037159 PCT/AU2005/001510 11 sulphur atoms and 1 to 3 nitrogen atoms, such as, benzothiazolyl or benzothiadiazolyl. In a preferred embodiment, the invention relates to first said method comprising a 5 library of compounds selected from compounds of formula II, O ZR 1

R

5 X HO-y "XR2

XR

3 Formula II 10 wherein R 1 , R 2 , R 3 , R 5 , Z and X are defined as in Formula I. In a preferred embodiment the invention relates to a library of compounds selected from compounds of formula 11. 15 In a preferred embodiment, the invention relates to first said method comprising a library of compounds selected from compounds of formula Ill, o A

R

5 X

R

4 X XR

XR

3 20 Formula Ill wherein A is defined as hydrogen, SR 1 , or OR 1 where Ri is defined as in Formula I, and 25 X and R 2 to R5 are defined as in Formula I.

WO 2006/037159 PCT/AU2005/001510 12 In a preferred embodiment the invention relates to a library of compounds selected from compounds of formula III. 5 In a preferred embodiment, the invention relates to first said method comprising a library of compounds selected from compounds of formula IV, RsOO

OR

1

R

5 0 R HO '

"NHR

2

OR

3 10 Formula IV wherein R 1 , R 2 , R 3 and R 5 are defined as in Formula I. In a preferred embodiment the invention relates to a library of compounds selected 15 from compounds of f formula IV. In a preferred embodiment, the invention relates to first said method comprising a library of compounds selected from compounds of formula V,

R

5 0O o OR 1 HO 9 "/NHR 2 20

OR

3 Formula V wherein R 1 , R 2 , R 3 and Rs are defined as in Formula I. 25 WO 2006/037159 PCT/AU2005/001510 13 In a preferred embodiment the invention relates to a library of compounds selected from compounds of formula V. In a preferred embodiment, the invention relates to first said method comprising a 5 library of compounds selected from compounds of formula VI, RSO O SR 1 RS0

HO

*

"

'

"'/NHR

2

OR

3 Formula VI 10 wherein R 1 , R 2 , R 3 and R 5 are defined as in Formula I. In a preferred embodiment the invention relates to a library of compounds selected from compounds of formula VI. 15 In a preferred embodiment, the invention relates to first said method comprising a library of compounds selected from compounds of formula VII, O5

SR

1 HO NHR 2

OR

3 20 Formula VII wherein R 1 , R 2 , R 3 and R 5 are defined as in Formula I. 25 In a preferred embodiment the invention relates to a library of compounds selected from compounds of formula VII.

WO 2006/037159 PCT/AU2005/001510 14 In a preferred embodiment, the invention relates to first said method comprising a library of compounds selected from compounds of formula VIII, R50 0

NHR

1 HO " "'NHR 2 5 OR 3 Formula VIII wherein R 1 , R 2 , R 3 and R 5 are defined as in Formula I. 10 In a preferred embodiment the invention relates to a library of compounds selected from compounds of formula VIII. In a preferred embodiment, the invention relates to first said method comprising a 15 library of compounds selected from compounds of formula IX,

R

5 0 Ho'" "'NHR 2

OR

3 Formula IX 20 wherein R 2 , R 3 and R 5 are defined as in Formula I. In a preferred embodiment the invention relates to a library of compounds selected from compounds of formula IX. 25 WO 2006/037159 PCT/AU2005/001510 15 In a preferred embodiment, the invention relates to said methods wherein biological assays involve Peptide Ligand class of GPCRs. In a preferred embodiment, the invention relates to first said method wherein 5 biological assays involve opioid, melanocortin, melanin-concentrating hormone, neurokinin, neuropeptide and urotensin receptors. In a preferred embodiment, the invention relates to first said method wherein biological assays involve 8-opioid (DOP), ic-Opioid (KOP), Melanocortin MC3, 10 Melanocortin MC4, Melanocortin MC5, Melanin-Concentrating Hormone (MCH1), pt-opioid (MOP), Neurokinin (NK1), Neuropeptide Y (NPY-Y1), Opioid (ORL1) and urotensin (UR2) receptors. In another aspect the invention provides a compound according to formula 1 in 15 which at least one X is nitrogen, and said X is combined with the corresponding R 2 R 5 to form a heterocycle. In a preferred embodiment, the invention provides a compound according to formula 1 wherein X and R 2 combine to form a heterocycle. 20 In a preferred embodiment, the invention provides a compound according to formula 1 wherein the heterocycle is heteroaryl, including triazoles, 25 benzimidazoles, benzimidazolone, benzimidazolothione, imidazole, hydantoine, thiohydantoine and purine 30 Detailed Description of the invention Embodiments of the invention will be described with reference to the following examples. Where appropriate, the following abbreviations are used. Ac Acetyl WO 2006/037159 PCT/AU2005/001510 16 DTPM 5-Acyl-1,3-dimethylbarbiturate Ph Phenyl TBDMS t-Butyldimethylsilyl TBDPS t-Butyldiphenylsilyl 5 Bn benzyl Bz benzoyl Me methyl DCE 1,2-dichloroethane DCM dichloromethane, methylene chloride 10 Tf trifluoromethanesulfonyl Ts 4-methylphenylsulfonyl, p-toluenesulfonyl DMF N,N-dimethylformamide DMAP N,N-dimethylaminopyridine oa,a-DMT a,a-dimethoxytoluene, benzaldehyde dimethyl acetal 15 DMSO dimethylsulfoxide DTT dithiothreitol DMTST Dimethyl(methylthio)sulphoniumtrifluoro- methanesulphonate TBAF tetra-n-butylammonium fluoride 20 Selectivity profiles are determined by biological assays, either in vitro or in vivo, in which compounds exhibit a specific response in each assay. The panel of specific responses represents the selectivity profile across the selected assays. The selectivity profile may be determined by testing compounds against (a) a series of commercially available assays, and/or (b) self-designed assays. The profile 25 distinguishes actives against non-actives in each assay, as indicated in Table 3 below.

WO 2006/037159 PCT/AU2005/001510 17 The designing of libraries is based on methods known in the art, including designing to scan for molecular diversity using molecular modeling. The libraries may be designed by using molecular modeling techniques as described by Thanh Le et al (Drug Discovery Today 8, 701-709 (2003)). 5 Part A: Preparation of building blocks: In order to fully enable the invention, we detail below methods for the preparation of certain building blocks used in the preparation of the compounds of the invention. The building blocks described are suitable for both solution and solid 10 phase synthesis of the compounds of the invention. Compounds of the library as presented exhibit different selectivity profiles. It is also apparent from these relationships that new compounds with different selectivity profiles may be designed. 15 Example A: Synthesis of a 24 dinitrogen containing Galactopyranoside Building Block OH Ph Ph H (i) (ii)

S

0SMe 0 SMe WO SMe NHDTPM NHDTPM NHDTPM A-1 A-2 A-3 S(iii) N OTBDPS OTBDPS OH Mv -0 (iV) 0 BzO SMe C- zo SMe NHDTPM NHDTPM NHDTPM A-6 A-5 A-4 Conditions: (i) ca,a-dimethoxytoluene (a,ac-DMT), p-toluenesulphonic acid (TsOH), 20 acetonitrile (MeCN), 760C, 85%; (ii) Benzoylchloride (BzCI), triethylamine; DCM, WO 2006/037159 PCT/AU2005/001510 18 99%; (iii) methanol (MeOH)/MeCN/water, TsOH, 75 0 C, 98%; (iv) t butyldiphenylsilylchloride (TBDPS-CI), imidazole, pyridine, 120 0 C, 99% ; (v) Tf 2 O, pyridine, DCM, 0 0 C, 100%;(b) NaN 3 , DMF, 16hr, RT, 99%. 5 Example B: Synthesis of a 3-nitrogen containing Gulopyranoside Building Block Ph Ph OO H -\00 0 ~OH HO SMe (i) SMe (ii) SMe OBzCI N3 N 3 OBzCI B-1 B-2 B-3 (iii) O OTBDPS j SMe

N

3 OBzCI B-4 Conditions: (i) (a) trifluoromethanesulfonic anhydride (Tf 2 0), pyridine, -200C, dichloromethane (DCM), 1 hour, 100%, (b) sodium azide (NaN 3 ), NN 10 dimethylformamide (DMF), 50 0 C, 5 hours, quantitative; (ii) TsOH, MeCN/ MeOH/water (12:3:1), 90 0 C, 6 hours, 88%(iii) TBDPSCI, DMAP, pyridine, 120 0 C, 12 hours, 93% WO 2006/037159 PCT/AU2005/001510 19 Example C: Synthesis of a 2,6-dinitrogen substituted Glucopyranoside Building Block OH N 3 O (i) 0 zO SMe -zO SMe NHDTPM NHDTPM A-4 C-1 5 Conditions: (i) (a) Tosylchlodride, pyridine, RT, 24 hours, 33%(b) NaN 3 , DMF, RT, 168 hours. Example D; Synthesis of a 2-nitrogen containing Tallopyranoside Building Block 10 P h- P h P h - -0 000 HO SMe i TBDPS SMe TBDPSO SMe OBz OBz OH B-I D-1 D-2 (iii) OBz OH Ph---- O OO T (v) O(iV, 0 'NO TBDPSOL - .SMe TBDPSO SMe TBDPSO SMe D-5 D-4 D-3 Conditions: (i) TBDPSCI, imidazole, 1,2-DCE, reflux; (ii) NaOMe/MeOH; (iii) (a) Tf 2 0, pyridine, -200C, DCM, 1 hour, (b) NaN 3 , DMF, 500C, 5 hours; (iv) TsOH, 15 MeCN/MeOH/water; (v) benzoylchloride, DMAP, 1,2-DCE, -200C.

WO 2006/037159 PCT/AU2005/001510 20 Example E: Synthesis of two 3-nitrogen containing Altropyranoside Building Block HO OH O H 0 HO SMe OSMe O SMe E-1 E-2 E-3 PhOMe O OBn-OMe HO OBn-OMe 0 ~O\ oO ~ OBn-OMe HO SMe (iv) a HO SMe O S HSMe IN3 H ' N3 E-6 E-5 E-4 TBDPSO OBn-OMe TBDPSO OB HO HO S e TBDPSO OBz L SMe HO S (vr) HO SMe

N

3 I N 3

N

3 E-7 E-8 E-9 5 Conditions: (i) cyclohexanone dimethylacetal, TsOH, MeCN; (ii) p methoxybenzaldehyde dimethylacetal, TsOH, MeCN; (iii) DIBAL, -780C, diethyl ether; (iv) (a) Tf 2 0, pyridine, -200C, DCM, 1 hour, (b) NaN 3 , DMF, 5000, 5 hours; (v) TsOH, MeCN/MeOH/water; (vi) TBDPSCl, DMAP, 1,2-DCE; (vii) (a) CAN, (b) BzCI, DMAP, 1,2-DCE, (c) TsOH, MeCN/MeOH/water; (viii) TBDPSCI, DMAP, 1,2 10 DCE.

WO 2006/037159 PCT/AU2005/001510 21 Example F: Synthesis of a 2-nitrogen containing Glucopyranoside Buildinq Block Ph Ph HO 0-0 HO __CIS ) o-- qj (i) HO SMe 0 HO SMe SMe

N

3

N

3

N

3 F-I F-2 F-3 TROPSO -~HO (i()v) 0O HO < BzO SMe B O SMe

N

3 F-4N 3 F-5 Conditions: (i) a,a-DMT, TsOH, MeCN; (ii) 1,2-DCE, BzCl, DMAP; (iii) TsOH, 5 MeOH/MeCN; (iv) TBDPS-CI, DMAP, 1,2-DCE. HO 7 7 oTBDPSO HO SMe ) HO SMe

N

3 F-I F6 N 3 TBDPSO TBDPSOSe BzO7 + HO e HO S e Bzo

N

3

N

3 F-7 F-5 Conditions: (i) TBDPSCI, DMAP, pyridine, 120 0 C, 0.5 hours, 81%; (ii) a. (Bu)2SnO, MeOH; b. Benzoylchloride, RT, 24 hour; WO 2006/037159 PCT/AU2005/001510 22 Example G: Synthesis of a 2-nitrogen containing Allopyranoside Building Block Ph Ph Ph 00 O SMe () MsO~ SMe () O SMe

N

3 N 3 SMe F-2 G-1 OBz N 3 G-2 (lii) TBDPSO 0 V HO HO ----- " SMe , HHO SMe OBz 3 G-4 G-3 Conditions: (i) DCM/pyridine, MsCI, DMAP, 0 0 C; (ii) sodium benzoate, 5 dimethylsulphoxide (DMSO), 140 0 C; (iii) TsOH, MeOH/MeCN/water; (iv) TBDPS CI, imidazole, DCM, 1 hour, reflux. Part B: Bioloqical Assays Experimental Method 10 Cloned receptor membrane preparations from Perkin Elmer Biosignal T M were used in radioligand binding assays. Membranes (Al - All = Codes for Table 3: Results). Al Human 5-opioid (DOP), A2 Human ic-Opioid (KOP), A3 Human Melanocortin 15 (MC3), A4 Human Melanocortin (MC4), A5 Human melanocortin (MC5), A6 Human melanin-concentrating hormone (MCH1), A7 Human ip-opioid (MOP), A8 Human neurokinin (NK1), A9 Human neuropeptide Y (NPY-Y1), A10 Human opioid (ORL1) All Mouse urotensin (mUR2) WO 2006/037159 PCT/AU2005/001510 23 Materials and Methods Screening experiments were performed in either a 50 pl filtration or 25 pl FlashPlate assay format using the following protocol: 5 Table 1: Assay format, radioligands and reference ligands Assay Final conc. Reference Final Receptor format Radioligand (nM) ligand conc. (pM) 25 pl 125 MCH1 FlashPlate [2sl]-S36057 0.1 MCH 1 50 pl [12I]-NDP MC3 FlashPlate uMSH 0.25 NDP-uMSH 10 25 pl [ 125 1]-NDP MC4 FlashPlate aMSH 0.25 NDP-aMSH 10 25 pl [1 25 1]-NDP MC5 FlashPlate xMSH 0.25 NDP-aMSH 10 50 pl [1 25 ]-Substance NK1 Filtration P 0.1 L703,606 10 25 pl NPY-Y1 FlashPlate [ 125 1]-PYY 0.35 BIBP 10 25 pl ORL1 FlashPlate [1251]-Nociceptin 0.22 Nociceptin 1 25 pl p-opioid FlashPlate [ 3 H]-Naloxone 3 Naltrexone 10 50 pl [H] K-opioid Filtration Diprenorphine 1 nor-BNI 1 25 pl [3 H] 6-opioid FlashPlate Bremazocine 3 Naltrindole 1 25 pl [ 25 1]-Urotensin UR2 FlashPlate II 0.3 Urotensin II 10 WO 2006/037159 PCT/AU2005/001510 24 Table 2: Assay buffers Receptor Buffer MCH1 25 mM Hepes pH 7.0, 10 mM MgCI 2 , 1 mM EDTA and 0.5% BSA MC3 25 mrnM Tris-HCI pH 7.4, 1 mM MgCl 2 , 1.5 mM CaCI 2 , ImM NaCI and 0.2% BSA 25 mM Tris-HCI pH 7.4, 1 mM MgCI 2 , 1.5 mM CaCI2, 1 mM NaCI and 0.2% MC4 BSA 25 mM Tris-HCI pH 7.4, 1 mM MgCl 2 , 1.5 mM CaCI 2 , 1 mM NaCI and 0.2% MC5 BSA 40 mM Hepes pH 7.4, 5 mM MgCl 2 , 1 mM EDTA, 0.5% BSA, 0.025% NK1 bacitracin and 25 pM phosphoramidon 50 mM Tris-HCI pH 7.4,5 mM KCI, 1 mM MgCI 2 , 2 mM CaCI 2 , 120 mM NaCI, NPY-Y1 0.5% BSA and 50 pM thiorphan ORL1 50 mM Tris-HCI pH 7.4, 10 mM MgCI 2 , 1 mM EDTA and 0.5% BSA 50 mM Tris-HCI pH 7.4, 10 mM MgCI 2 , 1 mM EDTA, 0.5% BSA and 0.01% p-opioid bacitracin K-opioid 50 mM Tris-HCI pH 7.4 3-opioid 50 mM Tris-HCI pH 7.4, 10 mM MgCI 2 , 1 mM EDTA and 0.5% BSA UR2 50 mM Tris-HCI pH 7.4, 10 mM MgCI 2 , 1 mM EDTA and 0.5% BSA 5 Format 1: FlashPlate Assay Volumes 19.5 p1 buffer, 0.5 pl1 of compound diluted in DMSO, 5 pl of radioligand diluted in binding buffer. Format 2: Filtration Assay Volumes 10 44 p1 membranes diluted in buffer, 1 pAl of compound diluted in DMSO, 5 pl of radioligand diluted in binding buffer. Compound handling and dilutions The day prior to performing the experiment 50 pl DMSO was added to each well of 15 the compound plates to yield compounds at a final concentration of 10 mM. Daughter plates were then created by diluting the compounds further in DMSO to a concentration of 0.5 mM. The mother plates were frozen immediately.

WO 2006/037159 PCT/AU2005/001510 25 Protocols: Filtration Thaw membranes on ice then dilute membranes in binding buffer at a 5 concentration of 1 Unit per well. Dilute radio-ligand to 10 times the final concentration in binding buffer. Add 44 pl of diluted membranes to each well of the deep-well plate. Add 1 pl of DMSO (total value, 5 wells), reference ligand (non specific value, 3 wells) or compound to the corresponding wells in the deep-well plate. Initiate the reaction by adding 5 pl of radioligand to each well and vortex 10 gently. Incubate at room temperature for 1 hour. During incubation, pre-incubate the Multiscreen Harvest plates in 0.3% PEI. Filter over pre-soaked Multiscreen Harvest plates using a Tomtec Harvester. Wash 9 times with 500 gl of cold 50 mM Tris-HCI pH 7.4 at 40C and air-dry for 30 minutes at room temperature under a fume hood. Apply a bottom seal to the Multiscreen Harvest plates. Add 25 pl of 15 MicroScint-0 to each well. Apply TopSeal-A to the plate. Count for 30 seconds per well on TopCount Microplate Scintillation and Luminescence Counter (PerkinElmer) using a count delay of 60 seconds. FlashPlate 20 Immobilize membranes into FlashPlate microplates using PerkinElmer BioSignal's proprietary coating procedure. Dilute radioligand to 5x the final concentration in binding buffer. Add 19.5 pl buffer to each well of the FlashPlate. Add 0.5 pl of DMSO (total value, 5 wells), reference ligand (non-specific value, 3 wells) or compound to the corresponding wells in the FlashPlate microplate. Initiate the 25 reaction by adding 5 pl of radioligand to each well. Apply TopSeal-A onto FlashPlate microplates. Incubate at room temperature for 1 hour in the dark. Count for 30 seconds per well on TopCount Microplate Scintillation and Luminescence Counter (PerkinElmer) using a count delay of 60 seconds.

WO 2006/037159 PCT/AU2005/001510 26 Data Analysis Percentage inhibition was calculated using the following formula: % inhibition = (compound - Total) x 100 5 Non Specific - Total Key to Blocks for Table 3: Results. H R6 OR6 OR6 OR6 HO \ HO 0 HO ~ HO 0 R3O- NH ORI H ORI R3O N S RI NHSRI R3~ 0 7 R30 N R2 R2 O R2 O R2 A B C D OR6 OR6 H N0 R1 HO H R3 N N0 H H RI R30 NH R2 R2 E

F

WO 2006/037159 PCT/AU2005/001510 27 Table 3: Radioligand Binding Results NO BLOCKRI R2 R3 R6 Al A2 A3 A4 AS A6 A7 A8 A9 A10 AlI I A X1 X14 X1 X24 -+ 4++ + -2 - -.... ..... .. .... ........... ..... .% 2 .- - ..... 2 A X1 X15 X1 X24 - + + + ... 4 .. .... ...... . .... ..- k ... -.- .. ............... 3 A X1 X14 X2 X24 - - - + + + 4 _A X1 X16 X2 X24 -- ++ + ......... ~~~ ~~ ...... ....... ........ .. 1 - 8 ... .......... ........ ............ . ......................... ........... . ... ,.. ..... ..... . .......... . 5 A X1 X15 X2 X24 - - - .- + + ... i ... .... .. ....... .... x 2 ) Y -. 2 ...... ... ....... ....... ... .......... . ...... ... .. .2 .. Y ... .. .. ......... ... ...... ......... 7 A X1 X16 X8 X24 - + + + .... .2 .. .... .... .... .. x .... .! .X . _ ... . .. ................ 8 XA 1 X17 X8 X24 -- + -+ ....... i...... ... . ......... .... .. .. ....... . ...... ...-....... 9 A X1 X17 X8 X24 -- ----- +--- ..... .4 ....... _A ........ X .. ..! ... ... . ... . ....... ...... .... .... .... ... ..... ......... . .... ................ 10 A XI X143 X + + + - - 11 A XI X16 X3 X24++ + + + + + 12 A X1 X151X3 X24-- ---------- ---- 13 A XI X17 X3 X24 - + + + 14 A X2_ X14 X1 X24 - + + + + -- ..2 .. ... ... ............. ...... _. . 1. ..... !. X _-- . ...... ... . ***...... . ................. .... .... ... .. 15 A X2 X14 XI X24 - - + + + + - - - ... 3 ...... A .... ..6 ... ........ . ........ ...... I....._.......... 16 A X2 X16 XI X24 + + + + + + + + - - .. ........ . .. A ............ ...... .... . . ... ........ .x .......... ........... .... ...... ........ ....... .. .... .. -.- .... ....... 17 A X2 X14 2 X24 - + + + + + 18 A X2 X16X2 X24- - - - + + + + - - 19 A X2 X16 X8 X24 - ----- + + - - 20 A X2 X15 X8 X24 - + - - 21 A X2 X17 8 X24 - - 22 A X2 X14X3 X24-- + 24 A X8 X16X X24- - - - + + - + - - 25 A X8 X17 XI X24- - - + + + - + - 26 A X8 X15 X2 X24 - +-- 27 A X8 X14 X3 X24- - - - + 28 A X3 X14 XI X24 - - + + + - + - ...... .... .... .. ... .. . .. ...... . _.. .. ! ... ..... .......... 29 A X3 X16 XI X24 - + + + + - + - - 30 A X3 X17 X1 X24 - - + + + + + + - - 31 A X3 X14 X2 X24 - - + - + - + - - - 32 A X3 X6 X2 X24 + - - - + - - + - - 33 A_ X3 X17 X2 X24--- --- --- + - + - - 34 A X3 X16 X8 X24------ - - + - - 35 A X3 X15 X8 X24 - +-- 36 A X3 X17 X8 X24- ------ + + - - 37 A X3 X14 X3 X24- - + + + + 38 A X3 X16X3 X24- - - - + + - + - - 39 A X3 X17 X3 X24-- ---- + - + - - 40 A X7 X14 X1 X24- - + + + + 41 A X7 X16 X1 X24 - - - + + + - + - - 42 A X7 X15 X1 X24 - - -+ - - - - 43 A X7 X X24- + + + + + + + - - WO 2006/037159 PCT/AU2005/001510 28 44A X7 _[)( 4 X 2 - X24- - +1+1 45 _A X7 X6 X2 X24V -f 46 A 7X16 X8 X24 -- -. j4 47 A4 X7 X15 X8 X24- - - I 48 A X7 X14-.X,3 X24,- - - + +I 49 A X7 -X16 X3 X24 50 A X7 X17 X3 "**,,X24------- 51 _A_ XI X14 X24X 1.. . - +.... ... .... .. . ... .... + ..... .. .. 52 A Xl X16X24X1 - -++--++ 53.....-.A Xl .... X15.X24..X1--..-....-----....-..-- .... 55 A XI X14 X24 X2 - , -+++++-+ 57 A lXI X17 X24X2 .... . .. .+ + t it . ..... ..... . ...... 58! j X4 X24 X -+ + ++ 59 A lXI X16 X24 X8-+ + + --. 60 AXi X15 X24 X8... - - - +4 61 A Xi _X17 X4 X8- + ~ ~ - -- 1 62.- . A.... .i .. X2 X 3 .. .... .t ...... 64 A Xl X15 X24 X3- -I - - + - - 6*5 AX1 17 X24 ;X + + - + + - ..-..--....- ....- - -... .... ..... -. -....- .. . . ...... .. . 68 A X2 X145X24 X1 - -+ + + + + + - 69 A X2 X 624 X1 + - + + + + + + -+- 680AX2 X15X24 Xl - + + + +-+ +--..

72 A X2 X175X24 X2 -- 4- -+ + 73AX2 X16X24 X2 +- + +- + - + 72 A X2 XI65X24 X2-------- + - 74]A X2 X1 4X24 X8- + + ++ .. A X2 ..... X 3 -1 ..... .+ ... . + ....- . ..... 70 A62 1 X24 X8 + + - - - - 83 A X2 X156X24 X8 + -- 4 - 85 A AX 7 X 24XI .- - +........ ... ... .... .. ............ 78 A X2 X14 X24 X ---. +- 88 A -X2 IXI6 X24 X2 ------- +------ 80 A X2 X157X24X2-------------+++-- WO 2006/037159 PCT/AU2005/001510 29 90 A X8 X14 X24 X3- -- - + 91 A X8 16X24X3- -- - + 92 A X8 X15 X24 X3 - - - + - - 93 A X8 X17X24X3 ---- + - + + 94 A X3 X16 X24 X1 -- + + - + + 95 A X3 X15 X24 X1 - - -+ - + - + 96 A X3 X17 X24 X1 -+++ + - +++ 97 A X3 X14 X24 X2 - - + + - - + - 98 A X3 X16 X24 X2 - - -+ + + - + - - 99 A X3 X15 X24 X2 - - - + - - + - - ....... . ........ .....-.. ,- * ... ............ ..... .... . .... . . ...... ...... ..... .. ........... _ _ ........ .. . .. . ..... 100 A X3 X17X24 X2 + + + + + 101 A X3 X14 X24 X8 - - -- +-- --- 102 A X3 X16 X24 X8 - - - - .+ -- -- ..... .... ..... ... ................... .. ... .. ...... ... .... I.. .......... ... I. ............. I 11. - I. ... ........ . .... . ........ ... ........ . ..... .. . .. .. .. 103 A X3 X15 X24 X8 +-- - - + - 104 A X3 X17 X24 X8 - -- + - + - - 05 A X3 X14 X24 X3 - -- ++ - - + - - 106 A X3 X16 X24 X3 - + + - - + - - 107 A X3 X15 X24 X3 - - + - - + 108 A X3 X17 X24 X3 - - +++- - - 109 A X7 X14 X24 X1 - - + . 110 A X7 X16 X24 X1 - - - .-.---- +.-- 11 A X7 X17 X24 X1 - + - - -+ - - 113 A X7 9X14 X24 X2 - - -+ - - + 114 A X7 X16 X24 X2 - - - - + - 115 A X7 IX15 X24 X2- + 17 A X7 X176 X24 X8 - - - - + - - 18 A X7 X17 X24 X8 - -- . 119 A X7 X14 X24 X3 - - - -1. - 120 A X7 _16 X24.X3 - +- - - + - - 120 A X7 X165 X24 X3- - + -- ....... ...... ... . ... ..... ................ .... .. ... .... ..... ....... ... .. .. ..... .... . ....... ..... .I ...... .......... I ~ .. _ _ _ ..... . ...... 121 ~A X7 IS X24 X3 .....- 122 A X7 X17 X24 X3- - - ++ - - + - 123 A X16 X25 X2 X24- - - - + - - + - - 124 A X16 X25 X24 X2- - - - + - - + - - 125 A X16 X23 X24 X8 - ...----- - 126 A X16 N3 X8 X2- - + - +- 127 A X16 N3 X2 X8 - - + + - ....- + - 128 A X24 X11 X8 X26- - ... - + - 129 A X1 X14 X4 X24 - ...---- - + - - 130 A XI X4 X24--- -- - + - - 131 A X1 X15 X4 X24-- ---- + P - - - 132 A X X17 X4 X24 + - - + - 133 A X2 X14X4 X24 - - - - - 135 A X2 X17 X4 X24 -- +- - - _La.4~~~~~~ ........ ............ ±x_~ x4 .. 5 .. . .... ... ..... _.!_ x _ _ x 4 ...... .. =.. ..

WO 2006/037159 PCT/AU2005/001510 30 136 A iX8 X15 X4 X24 - - - + - 137 A X3 X14X4X24 - - + - + 138 A X3 X16 X4 X24 +-- --- +- 139 A X3 X17'X4 X24 - .4 -++- 140 A X7 X15 X4X24 141 A X4 X14 Xl X24- -- - + + - - - - 142 A X4 X16 X X24 - - - - - -+ - - - - 143 A 4 X14 X 24----- - -- --- +[+ - . . . . . .. ..-. .. .. 144 A X4-" X17"' X1 X24--*"" -* - +. +..+ 145 A X4 XI6 X3 X24 - + 146 AXI X14 X24X4 + -+ - + 147 X 6 X24X4 + - + + + + 148 A I X15 X2 X4 + 150 A X2 X14 X24X4 +-.- +. +.--. --. 151 A X2 X6X2414 ------- - - - + 152 A X2 X171X24X-- - - - - 153 A X8 X15 X24 X4 + 15 A X8 . ... ......................... 155 A X3 XI4X24 X4- - - 156 A X3 X15 X24 X4 - - - + - - + - - 157 A X3 X14X24 X4 - . - + .- . 158 A X7 X14 X24 X4 - * .- .-..- +. - - 159 A X7 X16 X24 X4 - - + 160 A X7 X15 X24X4 161 A X7 X17 X:4 4 ....... ....... .............. 162 A X4 X14 X24 X1 163 A X4 X16 X24X1 - - + + + - - - - 164 A X4 X15 X24X1 - - + - - - + - - 165 A X4 X14X24X2--------- --- + - - 16 A X4 X16 X24 --- ++ 167 A X4 X15 X24 X2.- -- - - -- + - 168 A X4 X17 X24 X + 169 A X4 X15 X24 X3 - --- 171 B X8 X14 X - + - + 172 B IX8 X20X8 X24 73 B X8 X16X8 X24-- +- 174 B X8 X15X8 4X4- -+- 175 B X8 X17X8 X24 + 176 B X8 X19X3 X24...... ... ..... ......... 177 B X8 X14 X3 X24 - +++ - - + 178 B 8 X2 X X3 X24 - - - + - + 179 B 8 X16X3 X24+ - + + + - + 180 B 8 X15X3 X24 - + 181 B X8 X17X3 X24 - - + + - + . . ... ... . . .. .. .I . - , . - . - . . .. ......... .... ...

WO 2006/037159 PCT/AU2005/001510 31 182 B X8 X19 X1 X24 - - - 183 B X8 X14 XI X24 --. + +-.. 184 B X8 X20 X1 X24 -- - + - - +- - 185 B X8 X6X X24 -- - + + + - + - - ... .2 .... .2 ....... .......... . 186 B X8 X15 X1 X24 - - - + - - - 187 B X8 X17 X1 X24 - - - + + + - + - .-... . ......... 188 B X8 X19 X2 X24 -- ------ -- 189 B X8 X14X2 X24 - + ....... + + - + - .- .. 190 B X8 X20 X2 X24 - - - + - - + - I -. . . .. ......... ...... X '' -;= .. . . .... .............. ........ ... . ..... ..... ........ .. ...... ......... ... ................. 2 + -- -- .. . 191 B X8 X16 X2 X24 - - + + +- + - . ......... ... .. X 3 ...... i .......... ..... ....... ...... ........... .... ........... ...... ..... .... ........... . ............ 2 . . . . .. . 192 B X8 X15 X2 X24 - - - - + - - + - . x..... .x.....4. .... .......... ............................ ................... .... .. ... 193 B X8 X17 X2 X24 - - + + + + - - ......... 1..5 ............. - ..- .. -.-.... .... ... -..... - .. ".... .. -- - .. ..... ...... ......... .. . ... .......... .... .... -... ........ ... ... ... ... ..... - ....... . .... ..... ... . ............ .... ...... .. -. . ... ........... . .......... . i. ....... .. . ....... ............ , ... ..... ...... 194 B X3 X14 X8 X24 - - + + - - + - ' . ......... ...... .... ..... ...... ............ ...... . , ...................... +..... .... .. ....... . - . ... ........ .... ... .......... .+ . ... . 196 B X3 X16 X8 X24 + - ----- + - - 198 x 21 - - -+ -- + - 2004 B X3_ Xi4~3X4 - + + 4- + - 201 B X3 X19 X3X24 - - -+ + - - + - 26 B X3 X16 X1X24 - -_ - + + + - + - - ... ............. ...... ..... .

. ....... .. ... ............... .. ++... .... . ... ....... 4 ..... .............. .+ .... .... .. 207 B X3 X15 X3 X24 - + - + - + - - 204 B X3 X16 X2 X24 - + -- 4-++ - 4 - 205 B X3 X15 X X24- - - + + - - + - - 210 B X3 X17 X2 X24 - - + + + - + - 24 B X3 X1 ~XX24- - + - + - - + - - 2 B X3 X1 X2 X24 - - - ..... - - -- -... ..... ............... .... ....... 4... ....... ..... .................. ... .... . ........ . ...... . . i....... .. ... . ....... . ....... .... ............ 209 B X3 X1 4IX2 X24 - -+ + + -- + - 21 B X3 X1 IX 2 X4 - - + -... -....... .. 212 B X3 X19 X2 X24 - - - + - - + - 213 B X73X14 X X24 + -+ + 4- - - + - - ........ ... ..... ......... x .. i ... .. . ... ......................... ...... .......... .... .......... ................ .. .... ... . . .. . -.-............... I. .... ....... ..... ....... _ .._ -_ . ........ ... .. ...... .... 215 B X7 X15 X3 X24 - - - - - + - 216 B X7 X17 X3 X24 - + + + - + - - 2 B X7 X19 X3 X24 - - - - - - + - -........ ... 225 B X7 X20 XI X24 - - + - + - - ... ... .. .. .... ..... ..... ...... ... .. ............. .. ........... ....... -.... ...... ...... .... .. ..... ..... .. . ......... ......................... ...... .... ...... . . . ... ...... . .... ... .. .... .... - i ..... ..... . . . .. ....... ..... . .. . .. . . . . . . . 22 B X7 X15 JXI X24 - - + - - + - 22 . ........ ........ .................... B X 7 X 20 X X - . -.. .... . + . - -........ - ......... .. ......... ....... 223 B X7 X19X2 X24 .- - + - + - i'- -*- '- -i ... .. ..... ..... . ... F . ...... ...... . .... .... .. .. ... . ..... ... ... .... ...... .. .... ... ......... ... . ....... ... ......... .. .. .. .. ... . ............... ..... . ......... .. .. .. 2 X7 X14 XI2 X24 -........... I ~ ~ ~ _ .x2.......... ------ .......I.........o_ ! 2.... .. .. 4 ._ : .. _.-_.,..._,, .............._ 225 B X7 X209X2 X24-------- ------- 226, B X7 X16 X2 X24-_ + + + + - - 227 B X7 X0 2 X X24------------+- -- WO 2006/037159 PCT/AU2005/001510 32 228 B X7 X15 X2 X24 - - -F +- - + .. . ... I ... .. .. .. . .... ... . .. . . .. . .. . .... . ......... . . 229 B X7 X17 X2 X24- - - + + - + - - 230 B X2 X14X8 X24- - - - + - - + - - 231 B X2 X20 X8 X24 ----. -- +-- 232 B X2 X16 X8 X24--- --- - +- 233 B X2 X15 X8 X24 --- ---- +-- 234 B X2 X17X8 X24--- --- - +-- 235 B X2 X19 X3 X24, +--- --- +-- 236 B X2 X14 X3 X24- - + + + + + 237 B X2 X20 X3 X24 - + + 238 B X2 X16 X3 X24+ - + + + + + + 239 B X2 X15 X3 X24 - - - - + + - - 240 B X2 X17X3 X2 - + + + + + + + + 241 B X2 X19 X1 X24 - - + + - + - 242 B X2 X14 XI X24- - + + + + - - 243 B X2 X20 XI X24-- + + - + 244 B X2 X16 X1 X24 - -+ + + + + + 245 B X2 X15 X- X24 - + 246 B X2 X19 X2 X24 - -- - - + - - 247 B X2 X14 X2 X24 - + + +- - + - - 248 B X2 X20 X2 X24 - - + - - + 249 B X2 X16 X2 X24- - + + - - 250 B X2 X17 X2 X24 - -- + + + +- + --. .......... ..... .... .... ..... .. ............ 251 D X24 X16 X8 X8 - +- 252 D X24 X17 X8 X8 -- - + 253 D X24 X14 X8 X3 - - + - - + 254 D X24 X16 X8 X3- ----- - + -- -- - 255 D X24 X14 X8 X1 - - - + - - + - 256 D X24 X20 X8 X + - - 257 D X24 X16 X8 X1 - -+ 258 D X24 X15 tX8 IX1 - ------ + - - 259 D X24 X17 X8 X1 - - - +--- + - - 260 D X24 X14 X8 X2- - - +- ----- 261 D X24 X20 X8 X2 - - - - +-- --- 262 D X24 X16 X8 X2 - - - - + - - + - - 263 D X24 X15 X8 X2 - ---- - + - - 264 D X24 X17 X8 X2 - -- - - + -- 265 D X24 X14 X3 X8 - - + -- + - - 266 D X24 X20 X3 X8- ----- - +- - .......... ..... .. . . .... .. ..... ... . .... ....... ....... ...... . .... - .. ..... ..... ..... ... ............ .. .......... .......... 267 D X24 X16 X3 X8 - - - + -- + 68 . . . 2 x ........ .... 269 D X24 X179 X3X - - - - - - + - - 270 D X24 X14 X3 X3 - - + + + - - + - - 2Z 0 _ _ ... ..... ....... i _.. [. .. . ....... ... .. .... ....... 271 D X24 X20 X3 X3 - ----- + .. ...-- 273 .D X24 X15X3 X3.--... -.. ---- ..- - - WO 2006/037159 PCT/AU2005/001510 33 274 D X24 X7 X3 X3- 275 D X24 X14 X3 X1 - - - + + - - + - - 276 D X24 X20 X3 X1------+-- 277D X24 X16 X3 X1 278 D X24 X5 X3 X1 --- - - + - 279 D X24 X17 X3 XI - -- - - - + 280 D X24 X19 X3 X2 - - 1 + 281 D X24 X14 X3 X2 - - + + + - - + - 282 D X24 X20 X3X2- - - +- - + - 28 D 2 D X16 X3 - - ++ - + - 284 D X24 X14 X1 X8 - - - + - - + - 285 D X24 X20 Xi Xs - X- 286 D X24 X16 X1 X8 - - -4+- -. + 287 DX24X7iX---------- +---- . . . . . . . ..........-.......... .. . .. 288 D X24 X19 Xl X3-- - - -+ 289 D X24 X14 X1 X3 -+.+ -.- + 290 D X24 X20 Xl X3 - - - 291 D X24 X 6X1 X3 - - - +-.-.. ............ . .... .. - . ... . ..... - ... .... .. .... . . ...~ 292 D X24 X167X1 X3- - - - + - - +- -- 293 D X24 X15 X1 XI 294 D X24 X1 XI Xl - - - - - - - + - - 296 D X24 X14 X1 X2 + 297 D X24 X20 X1 X2 - - - - - - - + - - 298 D X24 X16 X1 X2 - - - + -.- + -. 299 D X24 X15X1 X2 - - - + ------ 300 D X24 X19 X2 X2 - - - + - + - -. 302 D X24 X14 X2 X8- - -+ - - +- 303 D X24 X16 X2 X8 - - - + + - - +- 304 D X24 X15 X2 X8 - - - 305 D X24X 17 X2 X8 - - - - + - - + - - 306 D X24 X19 X2 X3 307 D _X24-X1"4X"2 X3----"_ * . + .. . . + . -.. 308 D 24 X20 X2 X3 + - 309 D X24 X16 X2 X3- -i-- + - - + - 310 D X24 X15X2 X--. + 311 D X24 X17 X23 + 312 D X24 X14 X2 XI + + + + 313 D X24 . 20X2 X1 -.- -. + - - + 314 D 2 X - -+ 315 D X24X15 X2 X 316 D (24 X19 X2 X2 - 317 D X24 X14 X2 X2- - - + 318 D X24 X20 X2 X2- - .- .- + 319 D X24X16X2X2 - - ++ -+ WO 2006/037159 PCT/AU2005/001510 34 320 D X24 XI5 X2 X2 - - + 321 D X24 XI7 X2 X2 + + + + 322 D X24 X22 X5 X8---- -- +-- 323 D X24 X23 X5 X3- ------ i ]~~.. .... . ...... .2- . .. 5- -1= ....... . ..... ... ..... ... ........ ........ 324 C X24 X21 X5 X8 -- .-. - +- -- - 325 D X24 X8 X8 X5 - - - - 326 D X24 X23 X8 X5-- -- --- .- - + -- - .

327 X24 X25X3 X5 328 D.... X.... 24,x8 X3... X--- - ---........--....... ---.......-- ......... ... ...... ...... .. . . .. i- i ...... 8 ...... ... ....... .. . ....... ..... .............. ...... ......... ... ....... 329 D X24 X21 X3 X5 - .. ..... ... ... ... .... .-x i ...... .....-. ........ ..... .... ... . ........... .. .............. . I.-.......- ..... .... 330 C 24 X2. .. X8 X8 X5-- - - --...... .-- --..... 331 C X24 X23 X8 X5 - - I- - - - - +--- -.- -.......... . ...... ... .. ...... .... x : i < ; .... .-, ...... .... ., ... .. .... .... .. .... .: .... .. .. . .. 332 C X24 X25 X3 X5 ... ...... ... ..... .. 2-.:.i .: ..... .. .. .... ......... ......... .. ... ... ......... .. .............. ... .. 333 C X24 X8 X3 X5 - - + - - ~ ~ ~ ~ ~ . ................... x, - ,2 ...... : .. .... :........ ....... .......... 34 C X24 X23 X3 X5 - + 336 A X1 X17 X1 X24 + + + + + + + + - - . ...... A ....... , x ] ::: ,: ::..... I: -- ......... .............. ...... .... ........ ......... .: ....... .. I , .. ... ... ...... J -: :........_... ........ ...... ... .. .... ............. ... ....... ..-...... . 337 A X2 X15 X1 X24 - - - + + + + - : : ::::::::::: ::::::::::::::: ::::::::::. ._ ... 2 .. ..................................... 338 A X2 X15 X2 X24-------++-- i..4: ......... A ......... X3 .. _X.......,.L .. :2 _:........*_ ...... ............... . ... ...... :... ...... 339 A X2 X15 X2 X24 - - + + + + + - -........ ................ .... .. ............... .......... I... . .......... . .... ...... ....... . . .................. . ..... ..... ... . 340 A X2 4 XI X3X4-------------+

--

341 A X2 X17 X3 X24 - -- -- + + + - .344 .. A .. X3 X. IS .2 X24 - - - -.. - - - +.... - .....- -............. 345 A X7 X15 X2 X24 --. ............. ....... .. ... ..................................... . . .. .............. ... ....... ... ....... .. .......... . .. . ....... .... ..... .................... .. ....... ........ 346 A X7 X15 2 X24 + + + + + +- + + " , -/~~~~ ~~~ ......... .. ........ ..-. - 2 ...................... . ..... ... .. ....... . .... .... ...... . ...... ...... .. . ............. ..... ..... ..... ..........2... .. , .. ....... .. ... .... ........ ... .. ... .......... . ... ...... .......... . ...... 348 B X3 X28 X24X2 - - - - + I.. .. .... .. I..... ...... ..... I.. ... .. _. ... .......... .... ... ....... ........ ..... ... ... .+ .......: ..... ....... ...... 349 B X3 X18 X24 X2 - - - -I- + 350 B X3 X14 X24 X2 - - - + + - - I...5 ......... ........ ........ X ... 2 . . ..... ... ... .................. ... * ... .... ........... .. ............. .... ............ ....... ....... 352 ..... A X3 X28 X24 2 ......... ......... .......... - - -..... - .- - -......... ...... - .-...........-- 343 A X3 X29 X24 X2---------------- - 356 E X20 X24 X9 X30 - - - - - - - - -- + - ..... ..................... 357 E X20 X25 X6 X30 -------- +------- ---- 358 EB X20 X2 "

"

X9X30 - -..--.... - 3 . ......... . I ....... .... I.... .............. !x 6 ........... .... ... ........ . ..... .... ..... ......... .........,.....: - . ...... . .... ...... ... ...... ....... 359 E X13 X24X8 X30- --- - - + - 361 E X12 X25 X3 X30 - - +------ - 362 E X 12.. *2.- .... * . .X30- ...-- - -..- -............... ............ .3 .. .. . . .. ... ... .............. ...... ..... ..... ..... .-...... ...... .... ...... ... ........ ......... = .......... .......... ... 363 E X10 X25 X9 X30 - ------ +- - 364 B X8 X20X3X2 - + + + + 365 B X8 X20 X3 X8 - - - - - - - - - - - WO 2006/037159 PCT/AU2005/001510 35 367 BX8X20 X21X Ix ~ + +- 368 B X8 X20 X2 Xk3-- +--+ 369 _B X8 X20X8 X2-I . =-1-- 370 B X8 X20 X8 X8---------------------...... 371 B X8 X20~7 X8~ 372 B X8 Xl 5 X3 IX2 -- ++--+ 373 _B X8 X 5 X3 X8 - -++ B7 X8 X 5 X3 X3 -. . + ................ 375 B X8_ X15 X2 X28 + + -- + - - B7 X8 X15 X8X2- + +- - +-I - . -..... 37...... B ... 1 X8 X8. - ... .. .... . ...-..... 380 B~ X8 XI X8 X3 + 3 81 B X 2 X 20 X 3 2 - .. ...-.. .. - [.. . . .. . ........ .... 382 B X2 X20 X3 8-- + + 385 B X2 X20 X2 X8- - + + + + -+ -U 386 B X2 X20 X2 3-- + + +- ~ 387 B X2 X20 X8 X2- + + +- +- . 3 8 .. ... ..... ....... ~~~~~~~~~~~...........*........................ ... ... .. ........ ... . . . . . . 390 B X2 X 5 X3 X2- + ++-+ 391 B X2 X 5X3 X8- - +-- - - 392 B X2 X1 5X3 X3- + -- + - - 393 B X2 X15 X2 X2- +- + + 394 B X2 X 5 X2 X8- - + + +- - +-- 395 B IX2 X 5 X2 X3- + + - - + - + 397 B X2 X1 5X8 X2- + + + - - + - + 398 B X2 X15 X8X3- +4-- -+ 399 B XI X14 X3X2- + + +4 - - + - 400 B XI~i X2 X8 + + ++ + - + -+ 4068 1 14 X X3 + + + + +- + 408 B Xl X14X23X2- -- -~4. 49B_ XI X14 X2X8- tt~. .~-t~ 408 XI X146X2X3 + ++ + - + 4118B Xl X146X2X2- - ++ +- - +- WO 2006/037159 PCT/AU2005/001510 36 412 _ 1 XIi6 X2 X8+- -~ + + + 413 B XI X16X2 X3- +~+ + + + 414 _B XI- X6 X1X2- -+-- -- 415 B'- X Xi i -6X X8- - +'-," . . .-... 416 B XI X6 X X3- + + + ... . ..... ... ... 417 B X3 X14 X3X2- + + + 418 _B X3 X14 X3 X8- + + +++-+ 419 B X3X4 X3 X3- +I+ ++ 420 B X3 X14 X2X2-- +I+ +- -+ 421 B X3 X14 X2 X8 ++++ .42'2 .. .... -X .3 X .14 ...X.2 -X 3 - . .. ....... ....... -t......t..... 424 _B X3 X14XI X8 + + + - + - - 425 B X3 X4 XX3- -++ + - + -+ 426 B X3 X16 X3X2- + + +++ + ++ 427 B X3 X16 X3X8--+ +- + 428 B X3X16 X3 X3- jK 429 B X 3 X 16 X2 X2. . ..... ... . .. .... ... ..... .... 430 BX3 X16 X2 X8- +>"+ - + 432 B X3 X16 Xi X2 - - + + + - + - - 4373 X XIX8X--+-+ ~~~~~~. . ... . ............ .... ... . . . .... 44 B X3 X16 X3 1X3 + T- .t4. ...... 442 A X2 X20 X2 X2-- + + + + .t - 443- -. 1---. 12X ~ 2 . -I. 1.. 1 1....... 44537 X X20 X2 X2 + ++ + 1.. + 487 X2 X20XX3 + + + +- - 448 AX2 X20 X8X + + 449 A3 X 30X 8 X.-... + .. ..... .... 450 A X2 X20 X8X + + - + 453 A X2 X20 XX + + + + - +- - + 454 A X2 X205X2- + + +- + 45 A X2 X20X X + + ++ + + - + 446 A02X1X X3 -, -. + -- + -- 457 A X2 X20X2X38----------+ +- - - WO 2006/037159 PCT/AU2005/001510 37 458 A X2 X15 X 3 X3 - + + + + + +- 459 A X2 X15 X3 X3 - - + 460 A X2 X15 X2 X2 461 A X2 X15 X2 X2 - + + + 462 A X2 X15 X2X8- - -+ - - 463 AX X1 2 463 A X2 X15 X2 X3- ++ + - + - 464- 2 2 X5X2X3 - +- + 465 A X2 X15 X8 X2 - - + - - + - 466 A X2 X15 X8 X2 - - + + + - + - 467 A X2 X15 X8 X8 -- -+ A468 A X2 X15 X - + + - + 469A X2 X15X8 X3 --- + + - - + - - 1470 A 2 X15s 2X 471 A X3 X20 X3 - - - + - - + - 472 A X3 X20X3 X2 - - + + + - + 473 AX3 X20 X3 X8 --- + + + + - 47 A X3 X20 X3 X8 -- + 475 A X3 X20 X3 X3 - + + - + - - 476 A X3 X20 X3 X3 - - + + + + + + - 477 A X3 X20 X2 2 - - 478 A X 20X2X2 + + + + 479 A X3 X20 X2 X8 480 A X3 X20 X2 X3 - + + + + - S4811 A X3X 0X X +1 ++- +- 482 A X3 X20 X X - + + + + 483 A 3 X20 X8 X + . 484 A X3X X8 X2 486 A X3 X20X8 X8 - - - - + - 4876 A X3 X20 X8 X8 - - - - - - - + - - 488 A X 488 A X3 X20 X8 X3 - - - + + - - + 4891 A X3X1X X + + 490 A X3 X15X3X2 + + + + + + 491 A X3 X15X3X8- - - + 493 A X3X15X3X3 - + + + + 494 A X3 15X3 X3 - + + + + - + - + 4961 A X2++++3 497J A X3 1 2X 4985 A X3 15X2 X2 - + + + + - + 4996 AX3 15 X2 X2 - - + + - 497 A X3 X15 X2 X8 - - + + + + + + 498i A - X3 Xk15 X2.X.8 - .. .... ..... .+... .... ..... . .....- + o X3X15 2 X - + + + + 500 A X3 X15 X2X3 + + + + - - 501 A X3 X15X88 -- +- - - 5021A X3X5X8X2 503 _A X 3X+ WO 2006/037159 PCT/AU2005/001510 38 504 A X3 X15 X8 X8 - - - + - + 505 . A X3 X15 X8-X3- - - - ++- - 506 A X3 X15 X8 X3- -- L- - - + - + 507 A X3 X14 X3 X2- - - - + - + -.. 508 A X3 1X14 X3 X2 - + - +- + 509 A X3 X14 X3 X8-- + +++ ++- - ..... - --- . .l ....... . .. . ........ . . - . .. ._ -_ .... .... .. .. .. .... 510 A X3 XI4 X3 X8- ++. - +- + I 3 ...... ......... ........... .- - .3 ._ 4 - k ... , -, -; : ............ .... : -: .. ... ......... 511 A X3 X14 X3 X3- - +++ + ++- + .5.:!.4 A -X-3-~ ~ ~..X .4..... " - -.. ..... ...... :......... :" ... + . .... .....'.'. .. ." --...-... L" .... .:....... i ...... ...... .: ........ .... ... £ x . . .. . .. ... . 512 A X3 X14 X2-X2- - + +++ ++- + 513 A X3 X14 X2 X2 - + + + + + + + + 514 A X3 X14 X2 X8 - - +++ - + .. -........ 515 A X3 X14 X2 X3 - - + + - + - + ........ .... .. X 3 .... . .. ........ I........ ..... .. ... 516 A X3 X14X3 +... + - - 517 A X3 X14 XI X2 - + + ...... + + - + - - -.... .518 A X3 X1X X. ... ........ ..... +. 519 A X3 X14 X1 X8 - - + - - ...-.... 520 A X3 X14 X X3 - - +++ + - - -......... ........ ..... .... .1 6 X 2 ......X......8..... .. ... ... ....... .. ... .. . .. ... ....... 521 A X3 X14 X1 X3- - + + - + + 5223 A X3 X16 X3X2- + . .. ........ _ .... .......... ... . .. ...... .. . . _ x .

_ .........- . _ . _. ........ , .......... .s . .... ....... ... . ... ... ..... ... .... _...._....... I......... .... . _, ... :... :.._ . .... . :...... . 522 A X3 X16 X3 X2 - + + + + + + + .. 3 ........ A ... ... , _ . .. ._ .. x

....

. ... ... . ... .. .......... ....... .. ----- _ ... _ ; _ . . ....... -, ...... ........ 523 A X3 X16 X3 X2 - + + + + + + 524 A X3 X16 X3 X8 - - - + + - + 525 A X3 XI X3 X8 - + + + + + - +-.- ...... .F. ......- . ..... ...... .. . ... ...... .. 2 .. .. .. .. ..... ..... 526 A X3 X16 X3 X3- - - - + + - 5 43.. ... ........... .E ......... .... . ..... -1 -l . _) .. .. 2 . 2 . . .... .......... ...... ... .... . = _ _ .. _ ,..._ : ; ....... ...... 527 A X3 X6 X3 X3 - + + + + + - + 528 A X3 X16 X2 X2 - - . .. + 529 A X3 X16 X2 X2 - + .+ + + - + - .. 530 A X3 X16 X2 X8 - + j+ +- + + + + -+ .. 5_4,,......,..B................_.,..... ..... x...x2.,.. . .. ....... ... .,.. _ - _ - . . ......... 531 A X3 X16 X2 X8 + + + + 532 A X3 X16 X2 X3 - i- + + + + + + -+ 533 A X3 X6 XX2 - - +)+ + + - +- 53 A 63 X 8+IX- ++44+ -+ 535 A X3_ X16 X1 X8 - + + + + + + + - - 536 A X3 X16 X1 X3 - + + + + - + 5374 A X 3 X I6 X I X .-.. + 4 - + .- ..... .... . ..... 538 A X3 X16 X1 X3 + + + + + - + + + + 539 F - X14 X3 X3 - + - - - + + +-- 540 F - X14 X3 X2 - + - 541 F...................... - . .,....j. .. .... ..................................... 542 F - +1XX-------------- 543 E X8 X15 X2 X24 - -. - - - + - - 544 F X20 X3 XI - + 545 X2X20 X2 X24 - .... ----- ------------- 546 B X2 X20 X2 X24 +----- 547 B X2 X16 X2X24 . -+ - + -..

548 B X7 X14X1 X24 + + 549 B X7 X14X1 X24 -+- ....- -- -- - WO 2006/037159 PCT/AU2005/001510 39 Key to Table 3: Results "+" indicates greater than 50% inhibition at 10 pLM, "-" indicates less than 50% 5 inhibition at 10 tM. "P" indicates precipitation XI - X30 are sidearms selected from the figure below. Figure 1: Sidearms for Table 3. © N N 'N' KC C cl OH NH 2 H X1 X2 X3 X4 X5 X6 X7

NH

2 Ho H 10 X8 X9 X1O X11 X12 X13 NH H H NkNH 2 A N NH N NH H NH2 A f ,- N NH v

NH

2

NH

2

NH

2 X14 X15 X16 X17 NH 2 X18 NH N NH2 CH3

A.NH

2 P <cH X19 X20 NH NH X21 X22 X23 X24 X25 NH O 2NH 2 H N NH 2 H 10 6 X2 HX2 X30 10 X26 X27 X8X29 X3 WO 2006/037159 PCT/AU2005/001510 40 Throughout the specification and the claims (if present), unless the context requires otherwise, the term "comprise", or variations such as "comprises" or "comprising", will be understood to apply the inclusion of the stated integer or group of integers but not the exclusion of any other integer or group of integers. 5 Throughout the specification and claims (if present), unless the context requires otherwise, the term "substantially" or "about" will be understood to not be limited to the value for the range qualified by the terms. 10 It should be appreciated that various other changes and modifications can be made to any embodiment described without departing from the spirit and scope of the invention.

Claims (30)

1. A method of identifying biologically active compounds with defined selectivity profile comprising: 5 (a) designing a library of compounds of formula 1 to scan molecular diversity; and (b) assaying the library of compounds in at least two different biological assays; wherein formula 1 represents: R5X 0 ZR 1 R 5 X/' O R R 4 X XR 2 XR 3 10 Formula I wherein the ring may be of any configuration; Z is sulphur, oxygen, CH 2 , C(O), C(O)NRA, NH, NRA or hydrogen, in the case where Z is hydrogen then R 1 is not present, RA is selected from the set 15 defined for R 1 to Rs, or wherein Z and R1 together form a heterocycle; X is oxygen or nitrogen, when X is nitrogen, each X may combine independently with the corresponding R 2 to R 5 to form an azide, or wherein each X may also combine independently with any one of corresponding R 2 20 R 5 to form a heterocycle; R 1 to R 5 are independently selected from the group which includes but is not limited to H or an C1 to C20 alkyl or acyl; C2 to C20 alkenyl, alkynyl, heteroalkyl; C5 to C20 aryl, heteroaryl, arylalkyl or heteroarylalkyl, which is 25 optionally substituted, and can be branched or linear.

2. The method according to claim 1 wherein at least one X is nitrogen.

3. The method according to claim 1 wherein two of X is nitrogen. WO 2006/037159 PCT/AU2005/001510 42

4. The method according to claim 1 wherein X and R 2 combine to form a heterocycle.

5 5. The method of claim 1 wherein R 1 -R 5 optional substituents are selected from the group consisting of OH, NO, NO 2 , NH 2 , N 3 , halogen, CF 3 , CHF 2 , CH 2 F, nitrile, alkoxy, aryloxy, amidine, guanidiniums, carboxylic acid, carboxylic acid ester, carboxylic acid amide, aryl, cycloalkyl, heteroalkyl, heteroaryl, aminoalkyl, aminodialkyl, aminotrialkyl, aminoacyl, carbonyl, 10 substituted or unsubstituted imine, sulfate, sulfonamide, phosphate, phosphoramide, hydrazide, hydroxamate, hydroxamic acid, heteroaryloxy, aminoaryl, aminoheteroaryl, thioalkyl, thioaryl or thioheteroaryl, which may be further substituted. 15

6. The method according to claim 1, wherein the library of compounds is selected from compounds of formula II, R5X" 0 ZR 1 R 5 XH HO "XR2 XR 3 Formula II 20 wherein Z is sulphur, oxygen, CH 2 , C(0), C(O)NRA, NH, NRA or hydrogen, in the case where Z is hydrogen then R 1 is not present, RA is selected from the set defined for R 1 to Rs, or wherein Z and R1 together form a heterocycle; 25 X is oxygen or nitrogen, when X is nitrogen, each X may combine independently with the corresponding R 2 to R 5 to form an azide, or wherein each X may also combine independently with any one of corresponding R 2 R 5 to form a heterocycle; WO 2006/037159 PCT/AU2005/001510 43 R 1 to R 5 are independently selected from the group which includes H or an C1 to C20 alkyl or acyl; C2 to C20 alkenyl, alkynyl, heteroalkyl; C5 to C20 aryl, heteroaryl, arylalkyl or heteroarylalkyl, which may be substituted, and can be branched or linear. 5

7. The method according to claim 1, wherein the library of compounds is selected from compounds of formula III, 0 A R 6 X R 4 X XR 2 XR 3 10 Formula I1I wherein A is defined as hydrogen, SR 1 , or OR 1 , 15 R 1 to R 5 are independently selected from the group which includes H or an C1 to C20 alkyl or acyl; C2 to C20 alkenyl, alkynyl, heteroalkyl; C5 to C20 aryl, heteroaryl, arylalkyl or heteroarylalkyl, which may be substituted, and can be branched or linear, 20 X is oxygen or nitrogen, when X is nitrogen, each X may combine independently with the corresponding R 2 to R 5 to form an azide, or wherein each X may also combine independently with any one of corresponding R 2 R 5 to form a heterocycle. 25 WO 2006/037159 PCT/AU2005/001510 44

8. The method according to claim 1, wherein the library of compounds is selected from compounds of formula IV, R 5 0 0 OR, HR "'N H R 2 OR 3 5 Formula IV wherein R 1 , R 2 , R 3 and R 5 are independently selected from the group which includes H or an C1 to C20 alkyl or acyl; C2 to C20 alkenyl, alkynyl, 10 heteroalkyl; C5 to C20 aryl, heteroaryl, arylalkyl or heteroarylalkyl, which may be substituted, and can be branched or linear.

9. The method according to claim 1, wherein the library of compounds 15 is selected from compounds of formula V, R5O 1*,O OR, HO" " '"NHR 2 OR3 Formula V 20 wherein R 1 , R 2 , R 3 and R 5 are independently selected from the group which includes H or an C1 to C20 alkyl or acyl; C2 to C20 alkenyl, alkynyl, heteroalkyl; C5 to C20 aryl, heteroaryl, arylalkyl or heteroarylalkyl, which may be substituted, and can be branched or linear. 25 WO 2006/037159 PCT/AU2005/001510 45

10. The method according to claim 1, wherein the library of compounds is selected from compounds of formula VI, R5O SR 1 R 5 0 " R HO" ' "'NHR 2 OR3 5 Formula VI wherein R 1 , R 2 , R 3 and R 5 are independently selected from the group which includes H or an C1 to C20 alkyl or acyl; C2 to C20 alkenyl, alkynyl, 10 heteroalkyl; C5 to C20 aryl, heteroaryl, arylalkyl or heteroarylalkyl, which may be substituted, and can be branched or linear.

11. The method according to claim 1, wherein the library of compounds is selected from compounds of formula VII, 15 RsO SR 1 R 5 0 HO \ ' " "NHR 2 OR 3 Formula VII 20 wherein R, R 2 , R 3 and R 5 are independently selected from the group which includes H or an C1 to C20 alkyl or acyl; C2 to C20 alkenyl, alkynyl, heteroalkyl; C5 to C20 aryl, heteroaryl, arylalkyl or heteroarylalkyl, which may be substituted, and can be branched or linear. 25 WO 2006/037159 PCT/AU2005/001510 46

12. A method according to claim 1 wherein the library of compounds is selected from compounds of formula VIII, RO 0 NHR, HO"" "'NHR 2 OR 3 5 Formula VIII wherein R 1 , R 2 , R 3 and R 5 are independently selected from the group which includes H or an C1 to C20 alkyl or acyl; C2 to C20 alkenyl, alkynyl, 10 heteroalkyl; C5 to C20 aryl, heteroaryl, arylalkyl or heteroarylalkyl, which may be substituted, and can be branched or linear.

13. The method according to claim 1, wherein the library of compounds 15 is selected from compounds of formula IX, R 5 0 HO" ""'NHR 2 OR 3 Formula IX 20 wherein R 2 , R 3 and R 5 are independently selected from the group which includes H or an C1 to C20 alkyl or acyl; C2 to C20 alkenyl, alkynyl, heteroalkyl; C5 to C20 aryl, heteroaryl, arylalkyl or heteroarylalkyl, which may be substituted, and can be branched or linear. 25

14. The method according to claim 1 wherein the biological assays involve Peptide Ligand class of GPCRs. WO 2006/037159 PCT/AU2005/001510 47

15. The method according to claim 14 wherein biological assays involve opioid, melanocortin, melanin-concentrating hormone, neurokinin, neuropeptide and urotensin receptors. 5

16. The method according to claim 15 wherein biological assays involve 8-opioid (DOP), ic-Opioid (KOP), Melanocortin MC3, Melanocortin MC4, Melanocortin MC5, Melanin-Concentrating Hormone (MCH1), lt-opioid (MOP), Neurokinin (NK1), Neuropeptide Y (NPY-Y1), Opioid (ORL1) and 10 urotensin (UR2) receptors.

17. A library of compounds selected from compounds of formula 1, when used according to claim 1. 15

18. A library of compounds selected from compounds of formula II, when used according to claim 6.

19. A library of compounds selected from compounds of formula Ill, when used according to claim 7. 20

20. A library of compounds selected from compounds of formula IV, when used according to claim 8.

21. A library of compounds selected from compounds of formula V, 25 when used according to claim 9.

22. A library of compounds selected from compounds of formula VI, when used according to claim 10. 30

23. A library of compounds selected from compounds of formula VII, when used according to claim 11. WO 2006/037159 PCT/AU2005/001510 48

24. A library of compounds selected from compounds of formula VIII, when used according to claim 12. 5

25. A library of compounds selected from compounds of formula IX, when used according to claim 13.

26. A biologically active compound identified by the method of claim 1, 10

27. A compound according to formula 1 in which at least one X is nitrogen, and said X is combined with the corresponding R 2 -R 5 to form a heterocycle.

28. A compound according to claim 27 wherein X and R 2 combine to 15 form a heterocycle.

29. A compound according to claim 28, wherein the heterocycle is heteroaryl. 20 30. A compound according to claim 29, wherein the heteroaryl is selected from triazoles, benzimidazoles, benzimidazolone, benzimidazolothione, imidazole, hydantoine, thiohydantoine and purine. WO 2006/037159 PCT/AU2005/001510 49 AMENDED CLAIMS received by the International Bureau on 17 January 2006 (17.01.06) 1. A method of identifying biologically active compounds with defined selectivity profile comprising: 5 (a) designing a library of compounds of formula 1 to scan molecular diversity; and (b) assaying the library of compounds in at least two different biological targets; wherein formula 1 represents: Rex ZR 1 R 4 X . XR 2 10 XR 3 Formula I wherein the ring may be of any configuration; Z is sulphur, oxygen, CH2, C(0), C(O)NRA, NH, NRA or hydrogen, in the 15 case where Z is hydrogen then R 1 is not present, RA is selected from the set defined for R to R! 5 , or wherein Z and RI together form a heterocycle; When X is oxygen, R 1 to R 5 are independently selected from the group which includes but is not limited to H or an Cl to C20 alkyl or acyl; C2 to 20 020 alkenyl, alkynyl, heteroalkyl; C5 to C20 aryl, heteroaryl, arylalkyl or heteroarylalkyl, which is optionally substituted, and can be branched or linear wherein Ri-R 5 optional substituents are selected from the group consisting of OH, NO, NO2, NH 2 , Na, halogen, CF 3 , CHF 2 , CH 2 F, nitrile, alkoxy, aryloxy, amidine, guanidinlums, carboxylic acid ester, aryl, 25 cyoloalkyl, heteroalkyl, heteroaryl, aminoalkyl, aminodialkyl, aminotrialkyl, aminoacyl, carbonyl, substituted or unsubstituted imine, sulfate, sulfonamide, phosphate, phosphoramide, hydrazide, hydroxamate, hydroxamic acid, heteroaryloxy, aminoaryl, aminoheteroaryl, thioalkyl, thioaryl or thioheteroaryl, which may be further substituted; or AMENDED SHEET (ARTICLE 19) WO 2006/037159 PCT/AU2005/001510 50 when X is nitrogen, each X may combine independently with the corresponding R 2 to R 5 to form an azide, or wherein each X may also combine independently with any one of corresponding R 2 -R6 to form a heterocycle, wherein R 1 to R 5 are independently selected from the group 5 which includes but is not limited to H or an C1 to C20 alkyl or acyl; C2 to C20 alkenyl, alkynyl, heteroalkyl; 05 to C20 aryl, heteroaryl, arylalkyl or heteroarylalkyl, which is optionally substituted, and can be branched or linear wherein R 1 -Rs optional substituents are selected from the group consisting of OH, NO, NO 2 , NH 2 , N 3 , halogen, CF 3 , CHF 2 , CH2F, nitrile, 10 alkoxy, aryloxy, amidine, guanidiniums, carboxylic acid, carboxylic acid ester, carboxylic acid amide, aryl, cycloalkyl, heteroalkyl, heteroaryl, aminoalkyl, aminodialkyl, aminotrialkyl, aminoacyl, carbonyl, substituted or unsubstituted imine, sulfate, sulfonamide, phosphate, phosphoramide, hydrazide, hydroxamate, hydroxamic acid, heteroaryloxy, aminoaryl, 15 aminoheteroaryl, thioalkyl, thioaryl or thioheteroaryl, which may be further substituted. 2. The method according to claim 1 wherein at least one X is nitrogen. 20 3. The method according to claim 1 wherein two of X is nitrogen. 4. The method according to claim 1 wherein X and R 2 combine to form a heterocycle. 25 5. The method of claim 1 wherein R 1 -Rs optional substituents are selected from the group consisting of OH, NO, NO 2 , NH 2 , N 3 , halogen, CFa, CHF 2 , CH 2 F, nitrile, alkoxy, aryloxy, amidine, guanidiniums, aryl, cycloalkyl, heteroalkyl, heteroaryl, aminoalkyl, aminodialkyl, aminotrialkyl, aminoacyl, carbonyl, substituted or unsubstituted imine, sulfate, 30 sulfonamide, phosphate, phosphoramide, hydrazide, hydroxamate, AMENDED SHEET (ARTICLE 19) WO 2006/037159 PCT/AU2005/001510 51 hydroxamic acid, heteroaryloxy, aminoaryl, aminoheteroaryl, thioalkyl, thioaryl or thioheteroaryl, which may be further substituted. 6. The method according to claim 1, wherein the library of compounds is selected from compounds of formula II, R 5X0 ZR 1 ReXH HO1-1 "I XR2 XR3 Formula II wherein Z is sulphur, oxygen, CH 2 , C(O), C(O)NRA, NH, NR ^ or hydrogen, in the case where Z is hydrogen then R 1 is not present, RA is selected 10 from the set defined for R, to R6, or wherein Z and R1 together form a heterocycle; X is oxygen or nitrogen, when X is nitrogen, each X may combine independently with the corresponding R 2 to Rs to form an azide, or 15 wherein each X may also combine independently with any one of corresponding R 2 -R 5 to form a heterocycle; R, to R 5 are independently selected from the group which includes H or an 01 to C20 alkyl or acyl; C2 to C20 alkenyl, alkynyl, heteroalkyl; C5 to 20 C20 aryl, heteroaryl, arylalkyl or heteroarylalkyl, which may be substituted, and can be branched or linear. 7. The method according to claim 1, wherein the library of compounds is selected from compounds of formula III, 25 0 A R 5 X R 4 X XR 2 AMENDED SHEET (ARTICLE 19) AMENDED SHEET (ARTICLE 19) WO 2006/037159 PCT/AU2005/001510 52 Formula III wherein A is defined as hydrogen, SR1, or OR 1 , 5 R 1 to R 5 are independently selected from the group which includes H or an C1 to C20 alkyl or acyl; C2 to C20 alkenyl, alkynyl, heteroalkyl; C5 to C20 aryl, heteroaryl, arylalkyl or heteroarylalkyl, which may be substituted, and can be branched or linear, 10 X is oxygen or nitrogen, when X is nitrogen, each X may combine independently with the corresponding R 2 to R5 to form an azide, or wherein each X may also combine independently with any one of corresponding R 2 -R 5 to form a heterocycle. 15 20 8. The method according to claim 1, wherein the library of compounds is selected from compounds of formula IV, R 5 0- 0 R HRsO, 'OINHR 2 ORa 25 Formula IV wherein R 1 , R 2 , R 3 and R, are independently selected from the group which includes H or an C1 to 020 alkyl or acyl; C2 to C20 alkenyl, alkynyl, AMENDED SHEET (ARTICLE 19) WO 2006/037159 PCT/AU2005/001510 53 heteroalkyl; 05 to C20 aryl, heteroaryl, arylalkyl or heteroarylalkyl, which may be substituted, and can be branched or linear. 5 9. The method according to claim 1, wherein the library of compounds is selected from compounds of formula V, HO\P"' Y 'rNHR 2 O)R3 10 Formula V wherein R 1 , R 2 , R3 and R6 are independently selected from the group which includes H or an Cl to C20 alkyl or acyl; C2 to C20 alkenyl, alkynyl, heteroalkyl; C5 to C20 aryl, heteroaryl, arylalkyl or heteroarylalkyl, which 15 may be substituted, and can be branched or linear. 10. The method according to claim 1, wherein the library of compounds is selected from compounds of formula VI, sO O O SR I H O''"'NH R 2 20 OR3 Formula VI wherein R 1 , R 2 , Rs and R 5 are independently selected from the group 25 which includes H or an Cl to 020 alkyl or acyl; C2 to C20 alkenyl, alkynyl, AMENDED SHEET (ARTICLE 19) WO 2006/037159 PCT/AU2005/001510 54 heteroalkyl; C5 to C20 aryl, heteroaryl, arylalkyl or heteroarylalkyl, which may be substituted, and can be branched or linear. 11. The method according to claim 1, wherein the library of 5 compounds is selected from compounds of formula VII, R 5 0" 0 SR 1 HO ' "'NHR 2 OR 3 Formula VII 10 wherein R 1 , R 2 , R 3 and R 5 are independently selected from the group which includes H or an C1 to C20 alkyl or acyl; C2 to C20 alkenyl, alkynyl, heteroalkyl; CS to C20 aryl, heteroaryl, arylalkyl or heteroarylakyl, which may be substituted, and can be branched or linear. 15 12. A method according to claim I wherein the library of compounds is selected from compounds of formula VIII, RSO , NHR 1 HO\'"' "NHR 2 20 OR 3 Formula VIII wherein R 1 , R 2 , Rs and Rs are independently selected from the group 25 which includes H or an Cl to 020 alkyl or acyl; 02 to C20 alkenyl, alkynyl, AMENDED SHEET (ARTICLE 19) WO 2006/037159 PCT/AU2005/001510 55 heteroalkyl; C5 to C20 aryl, heteroaryl, arylalkyl or heteroarylalkyl, which may be substituted, and can be branched or linear. 5 13. The method according to claim 1, wherein the library of compounds is selected from compounds of formula IX, RsO NR, R5 01~~ HO"rJ ' INHIR 2 ORa Formula IX 10 wherein R 2 , R 3 and Rs are independently selected from the group which includes H or an C1 to C20 alkyl or acyl; C2 to C20 alkenyl, alkynyl, heteroalkyl; 05 to C20 aryl, heteroaryl, arylalkyl or heteroarylakyl, which may be substituted, and can be branched or linear. 15 14. The method according to claim 1 wherein the biological assays involve Peptide Ligand class of GPCRs. 15. The method according to claim 14 wherein biological assays 20 involve opicid, melanocortin, melanin-concentrating hormone, neurokinin, neuropeptide and urotensin receptors. 16. The method according to claim 15 wherein biological assays involve 8-opioid (DOP), iK-Opioid (KOP), Melanocortin MC3, Melanocortin 25 MC4, Melanocortin MC5, Melanin-Concentrating Hormone (MCHI1), opiold (MOP), Neurokinin (NKI), Neuropeptide Y (NPY-Y1), Opioid (ORLI) and urotensin (UR2) receptors. AMENDED SHEET (ARTICLE 19) WO 2006/037159 PCT/AU2005/001510 56 17. A library of compounds selected from compounds of formula 1, when used according to claim 1. 18. A library of compounds selected from compounds of formula II, 5 when used according to claim 6. 19. A library of compounds selected from compounds of formula Ill, when used according to claim 7. 10 20. A library of compounds selected from compounds of formula IV, when used according to claim 8. 21. A library of compounds selected from compounds of formula V, when used according to claim 9. 15 22, A library of compounds selected from compounds of formula VI, when used according to claim 10. 23. A library of compounds selected from compounds of formula VII, 20 when used according to claim 11, 24. A library of compounds selected from compounds of formula VIII, when used according to claim 12. 25 25. A library of compounds selected from compounds of formula IX, when used according to claim 13. 26. A biologically active compound identified by the method of claim 1. AMENDED SHEET (ARTICLE 19) WO 2006/037159 PCT/AU2005/001510 57 27. A compound according to formula 1 in which at least one X is nitrogen, and said X is combined with the corresponding R 2 -R 5 to form a heterocycle. 5 28. A compound according to claim 27 wherein X and R 2 combine to form a heterocycle. 29. A compound according to claim 28, wherein the heterocycle is heteroaryl. 10

30. A compound according to claim 29, wherein the heteroaryl is selected from triazoles, benzimidazoles, benzimidazolone, benzimidazolothione, imidazole, hydantoine, thiohydantoine and purine. AMENDED SHEET (ARTICLE 19)