AU2003218916B2 - Disaccharides for drug discovery - Google Patents

Description

WO 03/093286 PCT/AU03/00494 1 DISACCHARIDES for drug discovery FIELD OF THE INVENTION This invention relates to methods for the preparation of combinatorial libraries of potentially biologically active disaccharide compounds. These compounds are variously functionalized, with a view to varying lipid solubility, size, function and other properties, with the particular aim of discovering novel drug or drug-like compounds, or compounds with useful properties. The invention provides intermediates, processes and synthetic strategies for the solution or solid phase synthesis of disaccharides, variously functionalised about the sugar ring, including the addition of aromaticity and charge, and the placement of pharmaceutically useful groups and isosteres.

BACKGROUND OF THE INVENTION From a drug discovery perspective, carbohydrate pyranose and furanose rings and their derivatives are well suited as templates. Each sugar represents a three-dimensional scaffold to which a variety of substituents can be attached, usually via a scaffold hydroxyl group, although occasionally a scaffold carboxyl or amino group may be present for substitution. By varying the substituents, their relative position on the sugar scaffold, and the type of sugar to which the substituents are coupled, numerous highly diverse structures are obtainable.

An important feature to note with carbohydrates, is that molecular diversity is achieved not only in the type of substituents, but also in the three dimensional presentation. The different stereoisomers of saccharides that occur naturally (examples include glucose, galactose, mannose etc,Fig offer the inherent structural advantage of providing alternative presentation of substituents.

WO 03/093286 PCT/AU03/00494 2 OH OH OH OH HOHO OI- OH OH O OH HOOH a,P-D-Galactose ac,P-D-Glucose a,p-D-Mannose Fig. 1 Although there are a number of examples of monosaccharides being used as scaffolds for drug discovery purposes' there are only a limited number of examples of disaccharides or higher saccharides being used as templates for the presentation of pharmaceutically useful functional groups.

Derivatised disaccharides and higher saccharides, represent a new class of compounds for drug discovery that are able to address a significant and different group of receptors from those addressed by monosaccharide scaffolds.

This group or receptors can be broadly described as those receptors in which the critical binding groups are distal to each other. In principle, monosaccharide scaffolds can be used to address up to five binding groups (more usually 3 binding groups would be chosen) the connection points on the scaffold are each separated by between 1 and 5 angstroms in space. Disaccharide scaffolds on the other hand can accommodate up to eight binding groups although more usually 3-4 binding groups would be chosen, the connection points for each of these groups being separated by as much as 10 angstroms in space. Obviously the appended functional groups may be separated by even greater distances in 3-dimensional space. The replacement of the glycosidic bond linking the two monosaccharide components with a spacer group can further increase the separation between binding groups of interest.

The ability to address more distally placed binding groups is an important feature for a number of biological receptor molecules including the G-protien coupled receptors, where at the extra-cellular opening to many of these receptors, the width of the binding channel is up to 14 angstroms. Additionally, disaccharide scaffolds can be used as probes of interactions which involve WO 03/093286 PCT/AU03/00494 3 large surface areas for example the protein-protien interaction of the CD4- GP120 system, an important interaction in the aetiology of the human immunodeficiency virus.

Through the development of a range of selectively protected and modified monosaccharide, cyclitols and tetrahydropyran building blocks, we have developed a system that allows the chemical synthesis of highly structurally and functionally diverse derivatised disaccharide and disaccharide analogue structures, of both natural and unnatural origin. The diversity accessible is particularly augmented by the juxtaposition of both structural and functional aspects of the molecules. In order to access a wide range of diverse structures, stereo-center inversion chemistry is required, so as to achieve nonnaturally occurring and hard to get sugars and sugar analogues in a facile manner. Other chemistries are also required that provide unnatural deoxy or deoxy amino derivative which impart greater structural stability to the drug-like target molecules. With a suite of reagents to effect a suitable range of chemistries on a solid support, allowing such things as; wide functional diversity, highly conserved intermediates, a limited number of common building block to be required, and with suitable chemistry to allow access to unusual carbohydrate stereo-representations and including access to deoxy and deoxy amino analogues, a methodology is then established that can create focused libraries for a known target, or alternatively diversity libraries for unknown targets for random screening.

It will be clearly understood that, although a number of prior art publications are referred to herein, this reference does not constitute an admission that any of these documents forms part of the common general knowledge in the art, in Australia or in any other country.

Many of the traditional methods of carbohydrate synthesis have proved to be unsuitable to a combinatorial approach, particularly because modern high-throughput synthetic systems require procedures to be readily WO 03/093286 PCT/AU03/00494 4 automatable. The compounds and processes described herein are particularly suited to the solid and solution phase combinatorial synthesis of carbohydratebased libraries, and are amenable to automation. The methods of the invention yield common intermediates that are suitably functionalized to provide diversity in the structure of the compounds so generated. Using the method described, it is possible to introduce varied functionality in order to modulate both the biological activity and pharmacological properties of the compounds generated.

Thus the compounds and methods disclosed herein provide the ability to produce random or focused combinatorial-type libraries for the discovery of other novel drug or drug-like compounds, or compounds with other useful properties in an industrially practical manner.

WO 03/093286 PCT/AU03/00494 SUMMARY OF THE INVENTION In a first aspect, the invention provides disaccharide compounds of formula I A-d-L-e-B formula I In which the groups A and B are independently chosen from R6 R6 R7 R4 R7 R4\ /,R7 T R1 T R R4 R2 R3 R2 R3 in which the ring may be of any configuration and the anomeric center where present may be of either the a or P configuration; Independently for each ring T may be O or CH 2 R6 and R7 are hydrogen, or together form a carbonyl oxygen; R1 may be hydrogen, OZ or SZ wherein; When R1 is N(Z)Y Y is selected from hydrogen, or the following; o o II -W

W

JNW

OH

Q

Z is selected from hydrogen or X1; Q is selected from hydrogen or W; WO 03/093286 PCT/AU03/00494 6 The groups Z and Y may be combined to form a monocyclic or bicyclic ring structure of 4 to 10 atoms. This ring structure may be further substituted with X1 groups; The groups W are independently selected from alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, arylalkyl or heteroarylalkyl of 1 to 20 atoms which is optionally substituted, branched and/or linear. Typical substituents include but are not limited to 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, substituted or unsubstituted imine, sulfate, sulfonamide, phosphate, phosphoramide, hydrazide, hydroxamate, hydroxamic acid; The groups XI are independently selected from alkyl, alkenyl, alkynyl, heteroalkyl, acyl, arylacyl, heteroarylacyl, aryl, heteroaryl, arylalkyl or heteroarylalkyl of 1 to 20 atoms which is optionally substituted, branched and/or linear. Typical substituents include but are not limited to 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, substituted or unsubstituted imine, sulfate, sulfonamide, phosphate, phosphoramide, hydrazide, hydroxamate, hydroxamic acid; Where RI is C(Z)Y; Y, where present, is selected from hydrogen, double bond oxygen to form a carbonyl, or triple bond nitrogen to form a nitrile.

Z may be optionally absent, or is selected from hydrogen or X2 Wherein X2 is independently selected from alkyl, alkenyl, alkynyl, heteroalkyl, aminoalkyl, aminoaryl, aryloxy, alkoxy, heteroaryloxy, aminoaryl, aminoheteroaryl, thioalkyl, thioaryl or thioheteroaryl, acyl, arylacyl, heteroarylacyl, aryl, heteroaryl, arylalkyl or heteroarylalkyl of 1 to 20 atoms WO 03/093286 PCT/AU03/00494 7 which is optionally substituted, branched and/or linear. Typical substituents include but are not limited to 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, substituted or unsubstituted imine, sulfate, sulfonamide, phosphate, phosphoramide, hydrazide, hydroxamate, hydroxamic acid, heteroaryloxy, aminoalkyl, aminoaryl, aminoheteroaryl, thioalkyl, thioaryl or thioheteroaryl, which may optionally be further substituted; The groups Z and Y may be combined to form a monocyclic or bicyclic ring structure of 4 to 10 atoms. This ring structure may be further substituted with X1 groups; Where R1 is OZ or SZ, Z is selected from hydrogen or X3, Wherein X3 is independently selected from alkyl, alkenyl, alkynyl, heteroalkyl, acyl, arylacyl, heteroarylacyl, aryl, heteroaryl, arylalkyl or heteroarylalkyl of 1 to 20 atoms which is optionally substituted, branched and/or linear. Typical substituents include but are not limited to 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, substituted or unsubstituted imine, sulfate, sulfonamide, phosphate, phosphoramide, hydrazide, hydroxamate, hydroxamic acid, heteroaryloxy, aminoalkyl, aminoaryl, aminoheteroaryl, thioalkyl, thioaryl or thioheteroaryl, which may optionally be further substituted; The groups R2, R3, R4 and R5 are independantly selected from the group consisting of hydrogen, N 3 OH, OX4, N(Z)Y, wherein N(Z)Y is as defined above or additionally Y is WO 03/093286 PCT/AU03/00494 8 S 0 N

Q

Q

Q

where Q and W are as defined above, and X4is independently selected from alkyl, alkenyl, alkynyl, heteroalkyl, aminoalkyl, aminoaryl, aryloxy, alkoxy, heteroaryloxy, aminoaryl, aminoheteroaryl, thioalkyl, thioaryl or thioheteroaryl, acyl, arylacyl, heteroarylacyl, aryl, heteroaryl, arylalkyl or heteroarylalkyl of 1 to atoms which is optionally substituted, branched and/or linear. Typical substituents include but are not limited to 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, substituted or unsubstituted imine, sulfate, sulfonamide, phosphate, phosphoramide, hydrazide, hydroxamate, hydroxamic acid; The groups Z and Y may be combined to form a monocyclic or bicyclic ring structure of 4 to 10 atoms. This ring structure may be further substituted with X1 groups; The groups A and B are linked together with a linking structure d-L-e, in which the groups d and e represent the connection points for A and B and replace one of the groups R1,R2,R3,R4,or R5 in each of the groups A and B and form the connection point for the linker L.

The groups d and e are independently chosen from a covalent bond or the following list: 0 z z I 0 WO 03/093286 PCT/AU03/00494

T

V O 11 9 z z 0 Z Z t~N z 0 0 ,IA z

II

0 Y L may be absent, or is selected from alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heteroalkyl, cycloheteroalkyl, aryl, heteroaryl, arylalkyl or heteroarylalkyl of 1 to 12 atoms which is optionally substituted, branched and/or linear, saturated or unsaturated. Typical substituents include but are not limited to 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, substituted or unsubstituted imine, sulfate, sulfonamide, phosphate, phosphoramide, hydrazide, hydroxamate, hydroxamic acid, heteroaryloxy, aminoalkyl, aminoaryl, aminoheteroaryl, thioalkyl, thioaryl or thioheteroaryl, which may optionally be further substituted; It is understood that the rules of molecular stoichiometry will be upheld by the default addition of hydrogens atoms as required.

A preferred embodiment of the first aspect provides for compounds of formula I in which in group A, T is oxygen, group A is a pyranose ring, WO 03/093286 PCT/AU03/00494 The linker, d-L-e, is a glycosidic linkage formed between the anomeric position R1 of group A, and any position R1 to R5 of group B, such that the d is L is absent, and e is a covalent bond Importantly, The R groups on each ring may be selected independently from each other. For example, R2 on ring A may be different from R2 on ring

B.

Exemplary structure of this embodiment include but are not limited to: R7 R6 R7 R6 R6 RR7 R7 R6 0 T RI R5 ,O 1 1R4 R2 R4 R2 R2 R4 R4R2 R3 R3 R3 R3 R6 R7 R7 R6 Ri T 0 R 5 X 0 R4 R4' R2 R3 R3 R7 R6 T R5 T R1 R7 R6 OR2 R3 R4 R2 R3 He R6 R7 R4

RI

R7 R6 o 0 R3 R6 R7 R6 R4 R2 R2 R3 R3 IIf R6 R4 R7 ^T RI WO 03/093286 PCT/AU03/00494 11 R7 R6 R6 0 0 R7

T

R4 R2 RI R3 R3 R2 Another preferred embodiment of the first aspect provides for compounds of formula I in which In group A, T is oxygen, group A is a pyranose ring, The linker, d-L-e, forms an amide linkage in which R6 and R7 of A is a C=O, R5 is d which is a covalent bond, L is absent, and any of R1, R2, R3, R4, R5 on B is e which is z Importantly, The R groups on each ring may be selected independently from each other. For example, R2 on ring A may be different from R2 on ring

B.

Exemplary structure of this embodiment include but are not limited to: 0 o

RI

R1 0 R6 R7 o T R6 N R5 RI 0 R7 R2 R4R1 R2 R3 R4 R2 R4 R3 R4 R3 IIIa R3 TrTT, WO 03/093286 WO 03/93286PCT/ATJO3/00494 R1 0 0 2

RI

Nj T R2' R4 I R6 R3 R5 R7 R6 0 Z R7 R4 RI 0 R2) R4N R3 R2 R3 111f 0 Ri 0 R6 N R7 R2:[ R4Z RI R3 R3 R2 ii Another preferred embodiment of the first aspect provides for compounds of formula I in which WO 03/093286 PCT/AU03/00494 13 in group A, T is oxygen, both groups A and B are pyranose rings, The linkage, d-L-e, is an ether type linkage in which any of R1 to R5 in group A and group B is d and e respectively and is and L must be present.

Importantly, The R groups on each ring may be selected independently from each other. For example, R2 on ring A may be different from R2 on ring

B.

Exemplary structure of this embodiment include but are not limited to: R7 R6 0 0 R6 R7 V R4 XJR2 R2 R4 R3 IVa R3 R6 R7 R1 R7 R6 0 R4 R2 R3 IVb R7 RG X R3 R3 IVd R7 R6 0 R1 R5 X 0 R7 R6 R4 Y O L O R3 LZ RV R4 R2 R3 R7 R6 R7 yR6 V R1 ZT L R4 R2 R4 R2 R3 R3 IVe WO 03/093286 WO 03/93286PCT/A1J03/00494 IVg Ilih R6 R7 R7 R6 RI 0 T Ri R2) R4 0 l,0R 4F R2 R3 R IVi R7 R6 R 0R6 R7 R4 R2 R3 IVk R7 R6 R6 R7 Rl 0 0 0R2 R2': R4

R

R3 IVj R6 R7 Rl RO R7 R, 0 Rq 0 0 R3R4 RR3 lvi R6 R7 R, 0R6 R7 R2: R4

ZR

R 2 R4 R3 IVm R3 R6 R7 R6 R7 R, 0 Ri T R2q R5 R5 R2 R4 R3 R3 R6 R7 R, 0 R6 R7 21q0 0 R24 R4 IVn R3 R6 R7 RI T Pl 0 6 R7 R5 R2 R4 R2 0 R3 IVo WO 03/093286 WO 03/93286PCT/A1J03/00494 IVq IVr R7 R6 R6 R7 0j R, RI R5 R4 R2 R2 R4 ivs R7 R6 R5 0 Rl R6 R7 RI T R 4 "R 2

R

R4 0 R4 L R 'Vt R7 R6 0

R

RB R7 R4 R3 0 0 T

R

R2R4 lWv R3 R6 R7 R7 R6 R6 R7 R7 R6 0 Rl Xi TX 0 R4R X X RR5 R5 0 R 2R R4 3 0 0 R R4 R4 03 0 ivw lvx R7 R6 R7 R6 0 R 1 R T Rl R4o 0~ R2 R3 R3 WO 03/093286 PCT/AU03/00494 16 Another preferred embodiment of the first aspect provides for compounds of formula I in which In group A, T is oxygen, The linkage, d-L-e, is a linkage in which R1 in group A is d, which is chosen from: a covalent bond; 0Z or o Z or or L must be present; and e is z J-0- I-

Z

0 0 0

Z

0 z s z z

S

z s Importantly, The R groups on each ring may be selected independently from each other. For example, R2 on ring A may be different from R2 on ring

B.

Exemplary structure of this embodiment include but are not limited to: WO 03/093286 PCT/AU03/00494 17 R6 R7 RI- r R 0 R7 ,R6 ROR4 R7 .R R6 R7 R7 R R3 R4 R2 R2 R4 R4 R2 R3 Va 3 R3 Vb R6 R7 RI

RS

R2 R4 0 R7 RB 0 O O2 0 R4 R2 R3 Vc R7 R6 T .T, N R3 R4 R2 Z R3 Vd Another preferred embodiment of the first aspect provides for compounds of formula I in which In group A, T is oxygen, The linkage, d-L-e, is a linkage in which R1 in group A is d, R1 in group B is e, and both d and e are independently chosen from: a covalent bond; o o Z or or L must be present; WO 03/093286 PCT/AU03/00494 18 Importantly, The R groups on each ring may be selected independently from each other. For example, R2 on ring A may be different from R2 on ring

B.

Exemplary structures of this embodiment include but are not limited to: Another preferred embodiment of the first aspect provides for compounds of formula I in which In group A, T is oxygen, The linker, d-L-e, is a linkage in which any R group Rito R5 in group A may be d and is selected from z 0 0 z z I I z

S

z z o S z z s And any R group R1 to R5 in group B may be e and e is WO 03/093286 PCT/AU03/00494 19 L must be present.

Importantly, The R groups on each ring may be selected independently from each other. For example, R2 on ring A may be different from R2 on ring

B.

Exemplary structures of this embodiment include but are not limited to the list below.

z 7 R7 R6R6R o L T R VR4 R2 0 R24 R Vila R3 R6 R7 Ri 1 z R7 R6 IR3) R4 0 NR 0 R4 R2 R3 Vi R6 R7 RI 0 0 R2

N-

0

R

VIRI

R3 R2 R7 R6 R7 R6B R R5

IB

RI0)R RB 0 R2 R4

N-

R3 0 z WO 03/093286 PCT/AU03/00494 Another preferred embodiment of the first aspect provides for compounds of formula I in which In group A, T is oxygen, The linkage, d-L-e, is a linkage in which any R group Rito R5 in groups A and B may be d and e respectively and d and e are independently selected from

Z

z z I I 0 z T Oz If i z I Ox z z 0 S Z Z s z

Z

N

s L must be present.

Importantly, The R groups on each ring may be selected independently from each other. For example, R2 on ring A may be different from R2 on ring

B.

Exemplary structures of this embodiment include but are not limited to the list below.

z z R7 R6 l R6 R7 R3 VIa R3 Villa WO 03/093286 PCT/AU03/00494 21 R6 R7 Z Z R1 R7 R6 I RI -r R4 R2 0

R

R3 VIIIb R6 R7 RI T Z R2 R4 R7 R6 L z R4 R2 0 R3 VIIIc R7 R6 R7 R6 T Ri 0 RI 0 0 '^uR2 R4 N L R3 R3 Z VIIId R7 R6 O R1 R4 R2 T R N L N T R1 0 0 Z R4 R2 0 R3 vine In a second aspect, the invention provides for a method of synthesis of compounds of formula I comprising the step of reacting two appropriately substituted and protected monosaccharide compounds A and B in solution.

In a third aspect, the invention provides for a method of combinatorial synthesis of compounds of the formula I comprising the step of immobilizing a compound of group B onto a support through any of the functionalized positions R1 to Said support may be soluble or insoluble. Non-limiting examples of insoluble supports include derivatised polystyrene, tentagel, wang resin, MBHA resin, WO 03/093286 PCT/AU03/00494 22 aminomethylpolystyrene, rink amide resin etc. Non-limiting examples of soluble supports include DOX-mpeg, polyethylene glycol etc.

Compounds of the invention are useful in screening for biological activity.

For the purposes of this specification it will be clearly understood that the word "comprising" means "including but not limited to", and that the word "comprises" has a corresponding meaning.

WO 03/093286 PCT/AU03/00494 23 General Method 1: Urea Formation To the diamine (1.05 mmol) dissolved in N,N-dimethylformamide (11 mL), was added isocyanate (1.99 mmol), and the solution stirred for 2.5 h. Toluene mL) was added and all solvent removed to leave an oil. This procedure was repeated twice more. This residue was then triturated with ether and the resulting solid filtered to afford the product.

General Method 2: Transesterification To a solution of the sugar (16.2 mmol) in a 1:1 mixture of methanol/dichloromethane (110 mL) was added sodium methoxide (6.5 mmol) and the whole stirred under nitrogen for 2 h. Amberlite IR 120 H was added until pH 5 was reached. The resin was filtered off and washed several times with methanol and the combined filtrates were then concentrated to dryness to leave a residue. The residue was either triturated with ether or purified by column chromatography to give the desired product.

General Method 3: Azide Reduction To a solution of the azido compound (0.30 mmol) in 4:1 N, N,dimethylformamide/methanol (5 mL) was added a solution of ammonium chloride (1.50 mmol) in water (0.5 mL). Activated zinc dust (8.98 mmol) was then added and the suspension stirred for 40 min. A second addition of ammonium chloride (0.50 mmol) in water (0.25 mL) and zinc dust (1.5 mmol) was made and the suspension stirred for a further 40 min. After this time chloroform (50 mL) was added and the suspension filtered through celite and washed with chloroform/NN,,-dimethylformamide These combined filtrates were then washed with brine, dried (MgSO 4 and all solvent removed in vacuo to typically leave solid.

General Method 4: HBTU Coupling To a solution of the acid (0.05 mmol) and HBTU (0.05 mmol) in dry N,dimethylformamide (0.2 mL) was added diisoproplyethylamine (0.03 mL, 0.17 WO 03/093286 PCT/AU03/00494 24 mmol) and the whole stirred for 10 min. A solution of the sugar amine (0.04 mmol) in dry N, N,-dimethylformamide (0.3 mL) was then added and the whole stirred for 16 h. Chloroform (15 mL) was then added and washed with water, citric acid, saturated sodium hydrogen carbonate, brine, dried (MgSC 4 and the solvents removed in vacuo to leave an oil. The crude was typically carried through to the next step without further purification.

General Method 5: Global deprotection for formation of the final product-1.

The sugar (0.04 mmol) was dissolved in a solution (3mL) of 93% dry dichloromethane, 5% triethylsilane, 2% trifluoroacetic acid and the reaction stirred at room temperature for 2 hours. The solvents were then removed in vacuo and the solution then freeze-dried to leave a white solid. This solid was then purified by prep HPLC.

General Method 6: Global deprotection for formation of the final product-2.

The sugar (0.0016mmol) was dissolved in a solution (0.1mL) of 83% dry dichloromethane, 15% p-thiocresol, 2% trifluoroacetic acid and the reaction stirred at room temperature for 0.5 hours. The solvents were then removed in vacuo to give the crude product.

General Method 7: DIC Coupling To the acid (52Limol) and HOBT (52gmol) in dry DCM (lmL) and dry DMF (1 drop) was added DIC (52pmol). The solution was stirred for 1 min then added to a solution of amine (35Lmol) in dry DCM (lmL). The reaction mixture was stirred at room temperature for 1h then diluted with DCM, washed with citric acid, saturated hydrogen carbonate, brine, dried over MgSO 4 and the solvents removed in vacuo.

General Method 8: Ester Hydrolysis To crude product 4 in dioxane (0.6mL) was added 1M aq. KOH (0.6mL). The reaction mixture was stirred at room temperature for 1h then concentrated in vacuo. The residue was dissolved in CH 3 CN (2mL) and DCM (0.5mL) and WO 03/093286 PCT/AU03/00494 stirred with Amberlite for Ih. The solution was filtered and concentrated in vacuo to yield a residue which was subsequently purified by prep HPLC.

General Method 9: Fmoc Cleavage followed by DIC coupling Fmoc protected amino compound (-50pmol) was dissolved in acetonitrile (2.4mL) and piperidine (60pL,0.60pmol) was added. The mixture was stirred over night, the solvents evaporated in vacuo and the residue azeotroped with toluene to afford a residue. The residue was taken up in dry dichloromethane (2mL) and a solution of octanoic acid (12pL, 75pmol) and DIC (12pL, 75pmol) in dry dichloromethane (2mL) (stirred for 5 min at room temperature prior to addition) was added. Stirring was continued for 1h and the mixture was diluted with dichloromethane (50mL), washed with 10% citric acid, satd. sodium bicarbonate solution, filtered over a pad of cotton, and the solvents removed in vacuo to afford the product as a crude mixture.

General Method 10: Amine Deprotection The fully protected block [2 mmol] was suspended in butanol (15 ml) and ethylene diamine (15 ml) and the mixture heated at reflux for 20 h. The solvents were evaporated, the residue taken up in chloroform, washed with dilute brine, dried (MgSO 4 and evaporated. The compound was loaded onto a pad of silica with chloroform and eluted with 9% methanol in chloroform to yield the pure diamine quantitatively.

General Method 11: Diamine Coupling The diamine (17 mg, 0.023 mmol) was dissolved in dry chloroform (0.5 ml), DIPEA (3 mg, 4 pi, 1 equiv) added and the solution cooled to -78 0 C. A solution of FMOC-CI (4.2 mg, 0.7 equiv) in chloroform (0.2 ml) was added dropwise and allowed to warm to rt slowly before stirring for 16 h. The mixture was partitioned between chloroform and water, the organic layer washed with NaHCO 3 brine, dried (MgSO4) and evaporated to dryness. This gave the monoprotected amine as the major product.

WO 03/093286 PCT/AU03/00494 26 General Method 12: Thiourea Formation To a solution of the sugar (0.48 mmol) in N,N-dimethylformamide (4.8 mL), was added 4-fluorophenylisothiocyanate (0.48 mmol), and the solution stirred at room temperature for 4 h. Toluene (5 mL) was added and all solvent removed to leave an oil.

General Method 13 CBz formation To a solution of the sugar (0.48 mmol) in chloroform (4.8 mL), was added diisopropylethylamine (0.52 mmol). After 5 minutes benzyl chloroformate (0.52 mmol) was added and the solution stirred at room temperature for 2 h.

Chloroform (5 mL) was then added and washed with water, 10% citric acid, saturated sodium hydrogen carbonate, brine, dried (MgSO 4 and the solvents removed in vacuo to leave an oil.

General Method 14: Sulphonamide formation To a solution of the sugar amine (0.0267 mmol) and diisopropyethylamine (0.014 ml, 0.08mmol) in dry dichloromethane (0.25 mL), was added p-toluenesulfonyl chloride (10.2 mg, 0.054 mmol) and the solution stirred for 72 hrs. The reaction mixture was diluted with dry dichloromethane (2mL), washed with water (2ml), dried (MgSO 4 and the solvents removed in vacuo to afford the product as a crude mixture.

General Method 15: Acylation with acetic anhydride To a solution of the sugar amine (0.04 mmol) and diisopropyethylamine (0.02 ml, 0.12mmol) in dry dichloromethane (0.4 mL) was added dropwise acetic anhydride (0.015 mL, 0.12 mmol) and the solution stirred for 1 h. The solution was concentrated in vacuo to yield the crude product.

General Method 16: Fmoc Cleavage Fmoc protected amino compound (0.48mmol) was dissolved in chloroform (4mL) and piperidine (Iml). The mixture was stirred for 1 hr, evaporarted to dryness. The residue was taken up in acetonitrile (4ml) and the solid was filtered off and the filter cake washed with acetonitrile (1ml). The acetonitrile WO 03/093286 PCT/AU03/00494 27 solutions were combined and evaporated to dryness. The residue was purified by column chromatography (dichloromethane:methanol 20:1) to afford the desired product.

General Method 17: Hydrolysis of triflouroacetate ester and purification of the final products.

The sugar (0.04 mmol) was dissolved in a solution of methanol (2mL) and concentrated aqueous ammonium hydroxide (2ml); the reaction stirred at room temperature for 2 hours. The solvents were then removed in vacuo and the solution then freeze-dried to leave a white solid. This solid was then purified by prep HPLC.

WO 03/093286 PCT/AU03/00494 28 Example 1: Preparation of a compound with a glycosidic linkacge as described by compounds of formula 11 O 0h~O Ph 0O~ DdeHN BrDdH DdeHN 3 1I -d 1-a 1

N

3 0-oj me+HO AcO-'

N

3 DdeHN OMe 2 I6 0 J 1 -e 0 OMe 7 DdeHN 0 HO--0 BnO\ 0 1-h/O Conditions: Methyl bromoacetate, NaH, DMF, 80%; Bu 3 75%; (1-c) Ac 2 O, Pyridine, 95%; (1-dl) NaCNBH 3 MeQTf, DCM, 70%; (i) dithiothreitol, (Hi) Ac 2 O, Pyridine, 80% overall; NaOMe/MeOH, (Hi) TFA, (mi) NH 3

.H

2 0; 4-chloroindole-2-carboxylate, HBTU, DMF, DIPEA, (Hi) NaCH.

WO 03/093286 PCT/AU03/00494 29 Example 2: Preparation of a compound with an amide linkage as described by compounds of formula Ill Ph-'-O 0 0 HO \S O l e 11

OH

AcO7-k 14 OAc Conditions: AliyI-Br, DMF, NaH, 85%; 9-BBN, THF, NaQAc, H 2 0 2 (ii) Ph 3 P, CBr 4 (iii) NaN 3 DMF, (iv) DTT, Boc 2 O, NH 3

.H

2 0; HBTU, DIPEA, DMF; DTT, (ii) HBTU, DIPEA, isobutyric acid; (i) NaOMe/MeOH, (ii) TFA.

WO 03/093286 PCT/AU03/00494 Example 3: Preparation of a compound with an ether linkage as described by compounds of formula IV

O

0 12 NHDde Me Ac Oc 3-a 3-b 0c QCOOc H OA c Ac 0 NH 2 NHA 18 /1 3-c AcO OBn OH OBn NHAc OIe HAC 720 3-d 741 AcO QAc OHH 21 0 0 0 N OZ H OH o

NH

Ob

NH

2

NH

2 Conditions: NaCNBH 3 (Ri) NH- 3

.H-

2 0, (iii) Ac 2 O, pyridine; NBS, acetone, (ii) TFA, (iii) Ac 2 O, pyridine, (iv) NH- 3

H-

2 O, HBTU, DIPEA, 2amninobenzoic acid; grubbs catalyst; NaCMe/MeCH.

WO 03/093286 PCT/AU03/00494 31 Example 4: Preparation of a compound with an ether linkage as described by compounds of formula IV 'OAc 4-c 4-e Conditions: Vinylbenzyl chloride, NaH, DMF; 9-131N, NaDAc, H 2 0 2 Chloroacetyichloride, (ii) NBS, acetone, TFA, (iii) Ac 2 O, pyridine, (iv)

NH

3

.H

2 0, HBTU, DIPEA, indole-2-carboxylate, (vi) thiourea; MeQTf; (4-e) NaCNBH 3 (ii) NaOH.

WO 03/093286 PCT/AU03/00494 32 Example 5: Preparation of a compound with an ether linkage as described by compounds of formula V AcO Oc AcO 27 NHDde AcO H 5-b Ac O Z- N3 ACO Ac.

28NDde 29 j NHDde 31 'N 3 I 32 NHDde PhIV 0 HO

K

N

3 H AcO/ 33 0 0 NHDde Conditions: NaOMe, -40'C- 3-azidopropyl bromide, NaH, DMF; DTT; NaOH; HBTU, DIPEA, DMF; OTT, (ii) HBTU, Boc- Glycine, (iii) NaCNBH 3 TFA, (iv) I H-pyrazole-I -carboxamidine hydrochiorie, (v)

NH

3

H

2 0, (vi) mono-methyl malonate, DCC, (vii) NaOMe.

WO 03/093286 PCT/AU03/00494 33 Example 6: Preparation of a compound with a linkage as described by compounds of formula V and VI

N

3

OTBOPS

HO

0 0 36A

R

2 ANH OR 3 R"O2 HO 4

OH

OR no 43 R 3 .0 Conditions: wang OTCA resin, BF 3 .Et 2 O, (ii) NaOMe/MeOH, (iii) R 1 -Br, base, (iv) TBAF, MeOH, THF, R 3 -Br, base, (vi) DT, (vii) HBTU, DIPEA, DMF,

R

2

-CO

2 H; m-CPBA; amine 38, DMF; TFA cleavage; (6-e) grubbs cleavage; TEA cleavage.

WO 03/093286 PCT/AU03/00494 34 Example 7: Preparation of a compound with a linkage as described by compounds of formula VI Ph~VOPh---O 01 Aco 7- a

LW

0

C

N3 N3 Me 0 4 Conditions: NaOMe/MeOH, (ii) Methyl bromoacetate, NaH, DMF; (7-b) DTT, be nzoylchlo ride, DMF; SP-nitrilase immobilised enzyme, acetonitrile, water; HBTU, DIPEA, 1,5-diaminopentane (ii) TEA, acetonitrile, H 2 0.

WO 03/093286 PCT/AU03/00494 Example 8: Preparation of a compound with a linkage as described by compounds of formula VII

O

Ac 3 AcO I -NH 2 0- NHBoc NHBoc 49 so

-O

8-b 0- SMe MeO 2 OH OH 0 OH OH a HN0

H

2 51 ci Condition: DTT; DTT; (ii) 3,4-dichlorobenzoyl chloride, (iii) NaOH; HBTU, DIPEA, DMVF, (ii) NaOMe/MeOH, (iii) TEA, acetonitrile, water.

WO 03/093286 PCT/AU03/00494 36 Example 9: Preparation of. a compound with a linkagie as described by compounds of formula ViII Ph-'-OPh-\JO

O

0h~ 0 0 0 0 39a H AcO 0.&NH2 NHDcde H N, 0 2 MeO 5 1 CICI 19bNHBoc OH OH 0HO

OH

K

0

H

AcO~. 0 0 NH c: _OMea 2 54 CI Conditions: NH 3

.H

2 0, (ii) 3,4-dichlorobenzoyl chloride, (iii) DTT; (i) succinic anhydride, (ii) HBTU, DIPEA, (iii) NaCH, (iv) TFA, acetonitrile, water.

WO 03/093286 PCT/AU03/00494 37 Example 10: Synthesis of 1,5-Anhydroqalactitol Acceptor

N

3 PMB N OPMB N 3

OPMB

1-a -b CIBzO A HO.- b HO N 1-c 1 1 N3PMB

N

3 OPMB NS OPMB HO O 1-e AO-' 1-d HO NH OH 0 O O 59 58 10-a. Synthesis of 1, 5-anhydro-4-azido-2,4-dideoxy-2-DTPM-6-O-(4methoxybenzyl)-D-galactitol (56) Compound 56 was prepared according to the procedure described in General Method 2, 6 H (400 MHz; CDCI 3 3.25 (3 H, 3.26 (3 H, 3.65 (5 H, 3.80 (3 H, 4.09 (3 H, 4.50 (2 H, q, J 9.5 Hz and J 3.6 Hz), 6.89 (2 H, d, J 8.8 Hz), 7.26 (2 H, d, J 8.8 Hz), 8.21 (1 H, d, J 13.6 Hz), and 10.15 (1 H, brt, J 11.4 Hz); LCMS [M+H]+=475.

Synthesis of 2-amino-1,5-anhydro-4-azido-2, 4-dideoxy-6-O-(4methoxybenzyl)-D-galactitol (57) To a solution of the sugar 56 (16.0 mmol) in a 3:1 mixture of dry methanol/N, N,-dimethylformamide (120 mL) was added hydrazine monohydrate (86.3 mmol) and the resulting reaction mixture was stirred for 3 h. The resulting precipitate was removed by filtration and the filtrate concentrated in vacuo. The residue was then redissolved in dichloromethane, washed with saturated sodium chloride, dried (MgSO 4 and all solvent removed under reduced pressure to leave a solid 57. The solid was used directly in the next step.

WO 03/093286 PCT/AU03/00494 38 Synthesis of 1,5-anhydro-4-azido-2-(3-carboxybenzyl)-2,4-dideoxy-6-O-(4methoxybenzyl)-D-galactitol (58) To a solution of the amine 57 (16.2 mmol) in methanol (55 ml), was added phthalic anhydride (216.2 mmol), and'the whole stirred for 2 h. The mixture was then evaporated to dryness under reduced pressure and the residue azeotroped with toluene to leave a cream solid 58.

Synthesis of 3-O-acetyl- 1, 5-anhydro-4-azido-2, 4-dideoxy-6-O-(4methoxybenzyl)-2-phthallimido-D-galactitol (59) The acid 4 (16.3 mol) was suspended in dry pyridine (19 ml), cooled to 0 C, and acetic anhydride (48.7 mmol) added. The suspension was then stirred for 1 h at 0°C followed by 18 h at room temperature. The solvent was then removed in vacuo and the resulting residue azeotroped with toluene, redissolved in chloroform and washed with water, 10% citric acid, saturated sodium hydrogen carbonate, brine, dried (MgSO 4 and the solvent removed in vacuo to leave a yellow solid 59.

Synthesis of 1,5-Anhydro-4-azido-2,4-dideoxy-6-O-(4-methoxybenzyl)-2phthallimido-D-galactitol Compound 60 was prepared according to the procedure described in General Method 2, yield (77% from 55), 6H (400 MHz; CDCI 3 3.58 (1 H, dd, J 14.3 Hz and J 7.3 Hz), 3.66 3.74 (1 H, 3.82 (3 H, 3.82 3.93 (1 H, 4.03 (1 H, t, J 11.4 Hz), 4.10 (1 H, d, J 3.7 Hz), 4.42 4.52 (1 H, 4.52 (2 H, 4.66 (1 H, dd, J 3.7 Hz and J 10.7 Hz), 6.90 (2 H, d, J 8.9 Hz), 7.28 (2 H, d, J 8.9 Hz), 7.74 (2 H, m) and 7.67 (2 H, [M+NH 4 456.

WO 03/093286 PCT/AU03/00494 39 Example 11: Synthesis of a Trichloroacetimidate Donor.

OH OPMB OPMB o pMBO HO- PSMe MBO PMBO OHH O cl M OMBO PMB 11-c 6 66 65 64 NH 2

CF

3

CF

3 11-a. Thiomethyl 2-azido-2-deoxy-3,4,6-tri-O-(4-methoxybenzyl)-f-Dglucopyranoside (62) The thioglycoside 61 (21.2 mmol) was added in portions to a suspension of sodium hydride (84.8mmol) in DMF (106 ml) at 0°C. After 20 min, the mixture was allowed to return to room temperature and stirred for 30 min prior to recooling to 0°C. To the suspension was then added 4-methoxybenzyl chloride (11.5 ml) over 20mins. The reaction mixture was then allowed to return to room temperature and stirred for 16 h. The resulting solution was cooled to 0 C and quenched with ammonium chloride solution. The reaction mixture was partitioned between water and chloroform, and the organic layer subsequently washed with brine, dried (MgSO 4 and evaporated. Residue was purified by column chromatography to give the desired product 62 as a solid in quantitative yield, 8H (400 MHz; CDCI 3 2.22 (3 H, 3.32 3.48 (3 H, 3.54 3.70 (3 H, 3.80 (3 H, 3.81 (3 H, 3.82 (3 H, 4.17 (1 H, d, J 9.5 Hz), 4.46 (1 H, d, J 9.3 Hz), 4.49 (1 H, d, J 10.3 Hz), 4.55 (1 H, d, J 11.6 Hz), 4.63 (1 H, d, J 5.2 Hz), 4.82 (2 H, 6.75 6.95 (6 H, 7.09 (2 H, d, J 8.9 Hz), 7.24 (2 H, d, J 8.9 Hz) and 7.30 (2 H, d, J 10.3 Hz); LCMS [M+Na]f 618.2.

11-b. Synthesis of 2-Azido-2-deoxy-3,4,6-tri-O-(4-methoxybenzyl)-Dglucopyranose (63) WO 03/093286 PCT/AU03/00494 To a solution of thioglycoside 62 (112.9 mmol) in acetone (450 mL) at 0°C, shielded from light, was added water (277.8 mmol), N-iodosuccinimide (134.9 mmol), followed by TMSOTf (11.2 mmol), and the solution stirred for 90 min.

Chloroform (400 mL) was added and the chloroform layer separated and washed with saturated sodium hydrogen carbonate solution, saturated Na 2

S

2 03 solution, brine, dried (MgSO 4 and concentrated in vacuo to leave a solid. The solid was triturated with ether to give the desired product 63 as a cream coloured solid SH (400 MHz; CDC13) 3.42 (1 H, dd, J 3.0 Hz and J 10.0 Hz), 3.52 3.66 (3 H, 3.78 (3 H, 3.81 (3 H, 3.82 (3 H, 3.97 (1 H, t, J 9.8 Hz), 4.03 (1 H, dq, J 2 Hz, J 4.9 Hz and J 9.8 Hz), 4.42 (2 H, t, J 10.3 Hz), 4.53 (1 H, d, J 11.8 Hz), 4.62 (1 H, br d, J4.9 Hz), 4.73 (1 H, d, J 10.6 Hz), 4.82 (2 H, 6.81 6.93 (6 H, 7.05 (2 H, d, J 8.8 Hz), 7.24 (2 H, d, J 11.7 Hz) and 7.30 (2 H, d, J8.8 Hz); LCMS [M+Na] 588.3.

11-c. Synthesis of 2-Amino-2-deoxy-3,4,6-tri-O-(4-methoxybenzyl)-Dglucopyranose (64) To a solution of the azide 63 (110.2 mmol) in dry DMF (275 mL) was added dithiothreitol (220.4 mmol). The reaction mixture was then degassed with a stream of nitrogen, cooled to 0 C, and triethylamine (220.4 mmol) added. The solution was then allowed to return to room temperature and subsequently stirred for 24 h. The solution was then diluted with ethyl acetate and washed with water, brine, dried (MgSO 4 the solvents removed in vacuo, and resulting residue treated with diethyl ether (-250 mL) to give the desired product 64 as a white solid LCMS [M+H]f 540.25.

11-d. Synthesis of 2-Deoxy-3,4, 6 -tri-O-(4-methoxybenzyl)-2-trifluoroacetamido- D-glucopyranose To a solution of the amine 64 (75.6 mmol) in dry chloroform (400 ml) at 0°C was added diisoproplyethylamine (113.4 mmol) and trifluoroacetic acid (16.0 mL, 113.4 mmol). The resulting reaction mixture was then stirred for 30 min after which time the reaction was allowed to return to room temperature and stir for a further lh. At this time the reaction was cooled to 0°C and further WO 03/093286 PCT/AU03/00494 41 diisoproplyethylamine (113.4 mmol) and trifluoroacetic acid (113.4 mmol) added, the reaction was allowed to return to room temperature and stirred for 3h. The reaction mixture was then poured into dilute sodium hydrogen carbonate and the mixture stirred for 30 min. The solid was washed with water, transferred to a flask and dried by co-evaporation with acetonitrile to give the crude product. Ethyl acetate was then added to the residue and the resulting suspension refluxed for 2h. Petrol ether (-300mL) was then added to this, the mixture cooled to room temperature, and the resulting solid filtered and dried under high vacuum to give the desired product 65 as a solid 6 H (400 MHz; CDC13) 3.04 3.17 (1 H, 3.54 3.60 (2 H, 3.61 3.71 (2 H, m), 3.79 (3 H, 3.80 (3 H, 3.81 (3 H, 4.01 [dt, J 3.0 Hz and J 10.4 Hz] and 4.16 [dt, J3.7 Hz and J 10.1 Hz] (1 4.43 (2 H, t, J 10.4 Hz), 4.50 4.60 (2 H, 4.66 4.78 (2 H, 5.23 (1 H, d, J 4.0 Hz), 6.11 (1 H, br d, J 2.9 Hz), 6.80 6.90 (6 H, 7.06 (2 H, d, J 11.1 Hz), 7.18 (2 H, d, J 12.6 Hz) and 7.24 (2 H, d, J 11.1 Hz); LCMS [M+Na] 658.2.

11-f. Formation of Imidate To a solution of the lactol 11 (8.7 mmol) in tetrahydofuran (540 mL) was added trichloroacetonitrile (172.9 mmol), followed by potassium carbonate (103.9 mmol), and the suspension stirred for 8 days. After 8 days the suspension was filtered through celite, washed with tetrahydrofuran and all the solvent removed in vacuo to leave a brown oil. "The oil was then purified by column chromatography (eluent toluene/acetone; 20:1) to give the desired product 12 as a brown semi-solid WO 03/093286 PCT/AU03/00494 42 Example 12: Synthesis of Disaccharide and Functional isation 12-c 12e PMB( 0 OHl HN OH HO O0 NH NH NH q NH 72 CF3 CF, 12-a. Glycosylation with a Trichioroacetimidate Donor to Afford Disaccharide (67) WO 03/093286 PCT/AU03/00494 43 To a solution of the acceptor 60 (1.8 mmol) and the donor 66 (2.7 mmol) in dry 1,2-dichloroethane (84 mL) was added 3A acid-washed molecular pellets (4 g) and the resulting mixture stirred for 20 min. To the mixture was then added TMSOTf (1.8 mL of a 0.1M solution in dry 1,2-dichloroethane, 0.18 mmol) and the reaction then stirred for 15 min. After this time triethylamine (6 mL) was added and the suspension filtered, washed with dichloromethane and all solvent removed in vacuo to leave a yellow solid. This resulting solid was triturated with ether and filtered to give the disaccharide 67 as a cream solid 6H (400 MHz; CDCl3) 3.26 3.35 (1 H, 3.42 3.66 (5 H, 3.68 3.72 (1 H, m), 3.74 (3 H, 3.76 (3 H, 3.76 3.82 (4 H, 3.78 (6 H, 3.87 3.96 (1 H, 4.32 4.54 (6 H, 4.62 4.79 (3 H, m, [4.66, dd, J 3.4 Hz and J 13.3 Hz]), 5.02 (1 H, dd, J 3.8 Hz and J 12.6 Hz), 5.04 (1 H, d, J 10.7 Hz), 6.34 (1 H, d, J 10.3 Hz), 6.76 6.92 (8 H, 7.07 (4 H, dd, J 1.6 Hz and J 9.0 Hz), 7.20 7.24 (4 H, 7.69 7.74 (2 H, m) and 7.77 7.85 (2 H, LCMS [M+H+Na] 1078.2.

12-b. Amine Deprotection to Afford the Diamino Derivative (68) The protected disaccharide 67 (0.95 mmol) was suspended in nbutanol/ethylenediamine 14 mL) and heated at reflux for 16 h. The solvent was then removed in vacuo, and the residue taken up in chloroform, washed with dilute brine, dried (MgSO4), and solvent removed in vacuo to leave an oil.

The oil was purified by column chromatography (eluent 10:1, chloroform/methanol) to give the desired product 68 as a gummy solid [72%, used directly in next step[, SH (400 MHz; CDCI 3 2.91 (1 H, dd, J 8.2 and J 10.3 Hz), 2.96 3.08 (1 H, 3.21 3.29 (1 H, 3.35 3.50 (4 H, 3.52 3.68 H, 3.74 3.82 (12 H, singlets), 3.91 3.98 (1 H, 4.16 {1 H, J 3.6 Hz) and 4.32 J 7.8 4.35 4.52 (7 H, 4.58 4.64 (1 H, 4.71 {1 H, J 10.3 Hz) and 4.91 J 10.9 6.80 6.92 (8 H, 7.11 (2 H, d, J 8.8 Hz), 7.2 7.32 (6 H, LCMS 830.35.

12-c. Urea Formation (69).

WO 03/093286 PCT/AU03/00494 44 Compound 69 was prepared according to the procedure described in General Method 1, yield used directly in next step], LCMS 1204.53.

12-d. Azide Reduction Compound 70 was prepared according to the procedure described in General Method 3 used directly in next step], LCMS [M+H]f 1178.65.

12-e. Lipid HBTU Coupling (71) Compound 71 was prepared according to the procedure described in General Method 4, LCMS [M+H] 1374.83.

12-f. Global Deprotection (72) Compound 72 was prepared according to the procedure described in General Method 5, yield over two steps from compound 70] as a white solid; LCMS 894.22.

WO 03/093286 PCT/AU03/00494 Example 13: Synthesis of an Aminoacid Linked Lipidic Side Arm o

HN-<

0_ 0 OH

OH

0 N =<NH 13-d 7 75. R=cyclohex 76. R=H

F

3 76 CF 3 13-c

YI

13-a. DIC Coupling of a Aminoacid Side Arm (73) Compound 73 (using 0.034 mmo! of 70) was prepared according to the procedure described in General Method 7; HPLC Method A, [M+HI+=1475.

13-b. Deprotection of the Boc Protected Amine and 4-Methoxybenzyl Groups (74) Compound 74 was prepared according to the procedure described in General Method 5; HPLC Method A, Rt=4.82min, [M+H]'=895.

WO 03/093286 PCT/AU03/00494 46 13-c. DIC Coupling of a Lipidic Side Arm Compound 75 was prepared according to the procedure described in General Method 7; HPLC-Method A, Rt=6.18 mins, [M+H]*=1117; RT=7.16min, [M+H]+=1147.

13-d. Ester hydrolysis to provide the free acid (76) Compound 75 was prepared according to the procedure described in General Method 8, yield 25.7% from 70; HPLC Method A, Rt=4.75min, [M+H]*=939.

WO 03/093286 PCT/AU03/00494 47 Example 14: Synthesis of an Aminoacid Linked Lipidic Side Arm-2 0

,PBH

2 N OPMB 0 1 OPMB HN OPM PMO LOB0 14-aH PMBO 0 0=<14a 0 NH NI NH NH oW O= H

NI

7

CF

3

CF

3

CF

3

F

C02BZ

F

OH HN H OPMB H N' PI1 HOO 14-c PMBOX-jCXROO HO _NH-OI PMBO-~ NHNH N H q NH NH NH

N

79q

CF

3

CF

3

CF

3

CF

3 14-a. DIC Coupling of a Aminoacid Side Arm (77) WO 03/093286 PCT/AU03/00494 48 Compound 77 was prepared (using 0.034 mmol of 70) according to the procedure described in General Method 7, HPLC Method A, Rt=7.3 mins, =1497.

14-b. Deprotection of the Boc Protected Amine and 4-Methoxybenzyl Groups (78) Crude 77 was stirred at room temperature in DCM (2mL), TFA (40p1L) and TES (100L) for 2 hrs. The reaction mixture was concentrated in vacuo, HPLC Method A, Rt=4.5 mins, [M+H]*=917.

14-c. DIC Coupling of a Lipidic Side Arm (79) Compound 79 was prepared according to the procedure described in General Method 7; HPLC Method A, Rt=5.15mins, [M+H]+=1014.

14-d. Ester hydrolysis to provide the free acid Compound 79 was prepared according to the procedure described in General Method 8, yield (from 70) 24.5%; HPLC-Method A, Rt=4.50 mins, [M+H]+=925.

WO 03/093286 PCT/AU03/00494 49 Example 15: Synthesis of 2-deox-2-(3-trifluoromethl)-ureido-3-Dglucoipyranosyl 1 .5-anhvdro-2 ,4-d ideoxv-4-(1 -decanesulphonamido)-2-(3trifluoromethyl)-ureido-D-cialactito 4 with a Suiphonamide Lipidic Side Arm OP'MB H2N OPMB PMBO 0 PMBO NH

N

0<NH 04NH

CF

3 70 CF 3 CI 0 O=S-O /N6 OPMB

HNOPMBI

15-a )PPMBO-- L 02 SPMBO MH NH N= H 04NH 81 CF 3 82 CF 3

OH

,-HO 0 0

NH

NH

NH

CF

3 83 C F, Formation of a Sulphonamide Linked Lipidic Side Arm (82) To a solution of compound 70 (0.034 mmol) and pyridine (36.4 equiv) in dry dichioromethane (imi) was added 81 (20 equiv) in several portions. The resulting reaction mixture was stirred under nitrogen atmosphere for 2hr and saturated sodium bicarbonate solution (20m1) added. After 30 min stirring, the aqueous phase was extracted with dichloromethane; the combined organic solutions were dried over MgSO 4 and evaporated in vacuo to dryness. The residue was purified by preparative TLC (eluent: neat EtOAc) to furnish the crude 2-deoxy-3 ,4,6-tri-O-(4-methoxybenzyl)-2-(3-trifluoromethyl)-ureido-P-Dglucopyranosyl I ,5-anhyd ro-2,4-dideoxy-6-(4-methoxybenzyl)-4-( 1decanesulphonamido)-2-(3-trifluoromethyl)-ureido-D-galactito 82 (HPLC WO 03/093286 PCT/AU03/00494 Method A, Rt=7.71 mins, The product was used for the next step without further purification.

Global Deprotection to provide the Final Product (83) Compound 83 was prepared according to the procedure described in General Method 5, (10.8% from 70) HPLC Method A, Rt=11.85 mins, [M+H]+=902.53) WO 03/093286 PCT/AU03/00494 51 Example 16: Synthesis of 2-deoxv-2-(3-trifluoromethyl)-u reido-13-Dglucopyranosyl I .5-anhydro-2,4-dideoxy-4-( 1 -decvlphosphonamido)-2-(3trifluoromethvl)-ureidc-D-aalactito 7 ,OPMB H 2 N /OPMB PMBO-S 0 PMB0:-- NH

N

NH

NH

CF

3 70

CF

3 CI 0

O=P-CP

o=P-c[HN

OH

,OPMB

OPMB

16-a PMBO- 0 PMBO NH N

NHH

NH

NH

84

CF

3 85 ~CF 3 16-a. Formation of a Phosphonamide Linked Lipidic Side Arm To a solution of 1-decyiphosphonic acid (I mmol) and dimethylformamide (1.7iil) in dry dichioromethane (4ml) was added drop-wise oxalyichloride (261 .2 d, 3mmol). The resultant solution was stirred at room temperature under nitrogen atmosphere for 1 hr and evaporated to dryness in vacuo. The residue was dried under high vacuum for 1 hr to afford compound 84 as brownish liquid. To a 84 (Immol) in DCM (4m1) was added triethylamine (6951, 5mmol) followed by compound 70 [0.034 mmol]. The resultant reaction mixture was stirred under nitrogen atmosphere for lhr and 1N hydrochloride solution (20ml) added; the aqueous phase was extracted with dichloromethane (3x20m1); the combined WO 03/093286 PCT/AU03/00494 52 organic solutions were dried over MgSO4 and evaporated in vacuo to dryness.

The product was used for the next step without further purification. (HPLC Method A, Rt=7.67min, [M+H]+=1382.82).

16-b. Global Deprotection to provide the Final Product (86) Compound 86 was prepared according to the procedure described in General Method 5, yield 8% (from 70); HPLC Method A, Rt=5.36 mins, [M+H]t=902.5) WO 03/093286 PCT/AU03/00494 53 Example 17: Synthesis of 2-deoxv-2-(3-trifluoromethyl)-u reido-f3-D- -ilucopyrariosyl 1. 5-anhvdro-2,4-dideoxy-4-(N-octanoylcilycylam ido)-2-(3trifluoromethyl)-ureido-D-galactitoI OPMB H2N OPMB PMBO) O PMBD- NH N NH NH q CF 3 CF 3

HO

-O

HN'

NH- OPMB 01 16-a pmBOX

PMBO

NH

NH

87 CF 3 88

OH

16-b

H-

HO 0

NH

NH

CF

3 17-a. DIC Coupling to form an Amide Linked Lipidic Side Arm (88) Compound 88 was prepared (from 0.034 mmol of 70) according to the procedure described in General Method 7. The crude 88 was used for the next step without further purification; HPLC Method A, Rt=7.l7min; 361.80.

17-b. Global Deprotection to provide the Final Product (89) Compound 89 was prepared according to the procedure described in General Method 5, yield 24% (from 70); HPLC Method A, Rt=4.86 mins; fM+H]+=881 .36.

WO 03/093286 PCT/AU03/00494 54 Example 18: Synthesis of 2-deoxv-2-(3-trifl uoromethvl)-u reido-13-Dalucopyranosyl I .5-anhyd ro-2.4-d ideoxy-4-(N-octanoyl-N'-yI -3,4-dioxocvclob utene- 1.2-diamine)-2-(3-trifluoromethvl)-ureido-D-galactitoI 92.

0 0 OPMB H2N OPMB

OR

o 0OPMB HN OPMB NH18-a 18< PMO H NH PMBO o 18- O(N =HEtO OEt PMBO- NH N q 3NH

NH

F3 3

C

3 70

CF

3

CF

0 0 0 0 N N OPMB HN OFMB /OH HN OH PMBO a 18-C PMBON NH

N

NH oKNH NH NH NH NH NH CF CIF 3 C F, F C,91 92G 1 8-a. Coupling of Diethyl Squarate to form a Vinylogous Amide Amine 70 (0.034 mmol) was dissolved in Ethanol and DIPEA (35mmol) and diethyisquarate (70pmol) was added. After stirring at room temperature a complete conversion of the starting material to 5 was observed; HPLC Method A, Rt=6.70 mins; 302.47.

18-b. Coupling of an Alkylamine Side Chain to the Squarate (91) Decylamine (lO0pmoI) was added in the mixture (from 18-a) and heated to 50'C over night. The mixture was diluted with ethyl acetate, washed twice with citric acid, satd. sodium bicarbonate solution, and the solvents removed in vacuo. The crude 91 was used without further purification in the next step; HPLC Method A, Rt=7.66 mins, [M+H]+=141 3.15.

WO 03/093286 PCT/AU03/00494 18-c. Global Deprotection to Provide the Final Product (92) Compound 86 was prepared according to the procedure described in General Method 5 to give pure 92, yield 26.2% (from 70); HPLC Method A, Rt=5.61mi; =933.32.

WO 03/093286 PCT/AU03/00494 56 Exam pie 19: Synthesis of 2-deoxy-2-(3-trifluoromethyl)-u reido-13-Dg lucopyranosyl 1, 5-an hyd ro-2 ,4-dideoxy-4-(( N-hexanovl)-4-am ino-butyrovl)-2- (3-trifluoromethyl)-u reido-D-cialactitol OPMB H 2 N OPMB 0 PMBO 0 PMB O N NH N 0= H 0 NH+ 0 NH

CF

3

CF,

93 0

NF-

OH NH OH 19-b LQ HO NHO H_ NH

N

O<NH 04NH

CF

3

CF

3 0 0=NH DH OPMB NH OPMB 19-a PMBO LO PMBO

N

NH N NH NH

CF

3

CF

3 94 19-a. DIC Coupling to form an Amide Linked Lipidic Side Arm (94) Compound 94 was prepared (from 0.034 mmoi of 70) according to the procedure described in General Method 7; HPLC Method A, Rt=6.85min; 1361.47. The crude product was directly used for the next step.

19-b. Global Deprotection to Provide the Final Product Compound 95 was prepared according to the procedure described in General Method 5, to give pure 10, yield 42.1% (from 70); HPLC Method A, Rt=4.60min; [M+H]+=881.39.

WO 03/093286 PCT/AU03/00494 57 Example 20: Synthesis of a Disaccharide with a Lipidic Side Chain and Acid Function.

NHFmoc 0 PMOH0 2 C NHFmoc OPMB HN OPMB PMOS NH_ 20 MB H N H PMBO 07

CF

3

CF

3 96

CF

3

CF

3 NH 2 0

CO

2 tBu 0HN OPMB HN OP -MB Y COAtU PMBO LO OPMB HN DPMB 2- PMB O 0 NH 20-c PMB NHB- NH=NHH NH NH qF, qF3 ~97 0 09 CFF3 CF 9 0

HN

002 C0 2 OH HN OH HO- C0 Hoj NH

N

NH

NH

q

F

3

F

3 9 NB. Reaction series was completed separately for both the R and the S isomers.

WO 03/093286 PCT/AU03/00494 58 DIC Coupling to form an Amide Linked Lipidic Side Arm (96).

Compound 96 was prepared according to the procedure described in General Method 7, the crude product was directly used for the next step; HPLC Method A, Rt=7.70 mins; [M+H]+=1585.45.

and 20-c. Fmoc Deprotection Followed by Amide Formation (98) Compound 98 was prepared according to the procedure described in General Method 9, the crude product was directly used for the general deprotection, (HPLC Method B, Rt=18.51 mins, =1489.77).

global Cleavage to Afford the Final Product.

Compound 99 was prepared according to the procedure described in General Method 5, yield (R isomer) 27.5% from 70. yield (S isomer) 10.5% from HPLC Method A, Rt=4.88 mins; [M+H]+=953.46 WO 03/093286 PCT/AU03/00494 59 Example 21: Synthesis of a Disaccharide with a PEG Side Chain.

OPMB H2N OPMB PMBO n BO- NH NH NH

NH

CF

3 70 CF 3 o 0, O O H/-O SPMB NH NH NH NH 0 0 100

CF

3

CF

3 21-b 0 0 OPMB HN OPMB

PMBO

PMBO NH NH /NH NH NH CFa 101 CF, 21-a. DIC Coupling to form an Amide Linked PEG Side Arm (100) Compound 100 was prepared according to the procedure described in General Method 7; HPLC Method A, Rt=6.81 mins; [M+H]*=1338.49. The crude product was directly used for the next step.

21-b. global Cleavage to Afford the Final Product (101).

Compound 101 was prepared according to the procedure described in General Method 5 (101 purified by preparative chromatography on a C18 column), yield 21.8% (from 70); HPLC Method A, Rt=4.27min; [M+H]+=958.52.

WO 03/093286 PCT/AU03/00494 Example 22: Synthesis of a Range of Lipid Conjugates OPMB H 2 N OPMB PMBO R1HN OPMB PMBO O 0 PMBO NH O=NH 22-a O NH NH NH NH or 22-b N 0 H 102a-e

CF

3 C F 3

FCF

22-c HO R1HN OH HH

NH

NH 0 NH 103a-e

CF

3

CF

3 22-a. Preparation of Amido Derivatives at C-4 (102a, 102b, 102c, 102e) Compounds 102a, 102b, 102c, 102e were prepared according to the procedure provided in General Method 4.

22-b. Reaction with Acetic Anhydride (102d) Added dropwise to a solution of the sugar amine (0.04 mmol) in dry dichloromethane (0.4 mL) was acetic anhydride (0.12 mmol) and the solution stirred for 16 h. Chloroform (15 mL) was then added and washed with water, citric acid, saturated sodium hydrogen carbonate, brine, dried (MgSO 4 and the solvent removed under reduced pressure to give the title compound quantitatively as an oil, LCMS [M+H-]=1220.5.

22-c. global Cleavage to Afford the Final Product (103a-103e).

WO 03/093286 PCT/AU03/00494 61 Compound 103a-e were prepared according to the procedure described in General Method Table 1: Derivatives prepared in Example 22.

Comp. No. RI R2 Molecular Ion Yield 102a Rla R2a [M+H]'1276.2 87% 102b Rib R2a [M+H]+=1374.2 Quant.

102c Ric R2a [M+H]+=1358.3 Quant.

102d Rid R2a [M+H]+=1220.5 Quant.

102e Rie R2a- [M+Hr= 1346.5 Quant.

103a Ria R2b [M+H]+=796.3 103b Rib R2b [M+H]+=894.3 49% 103c Ric R2b 103d Rid R2b [M+H]+=740.1 103e Rie R2b [M+H] 4 =866.2 41% Side Arms for Table 1.

WO 03/093286 0 Rla PCT/AJO3/10494 0 SRid Rib Ric 0 Rle OMe R2a Example 23: Synthesis of a Further Disacoharidic Library Part 1: Preparation of Compounds 111-01 to 111-30 yH R2b WO 03/093286 PCT/AU03/00494 N3 OR4

R

4 0

RO

NH N C)=A 0

CF

3 67

KJ

23-a

R

4 0

NH

2

OR

4 NH NH 2 104

R

4

N

3

OR

4

H

2 NH 2 23-b R 4 O R 4

R

4 23-c NH NHFmoc 105

R

4 0, NH OR 4

R

4

R

4 0

NHR

2 NHFmoc 107-01 to 107-05

NI

3

OR

4 0R OO 23-d NHRz NHFmoc 106-01 to 106-07

R

4 0 R O R 4 23-e R 4

Q..

0 23

NHR

2 NHFmoc 108-01 to 108-10

R

4 0. OR 4

NR

4 0H2

NHR

2

NH

2 23-g

R

4 0 R 3

OR

R

4 O-7-Q

Y

R

4 0 0

NHR

2

NHR

1 110-01 to 110-31 109-01 to 109-05 23-h HO R 3

OH

HO 0 Oj NHR NHR, 111-01 to 111-30 Conditions: (23-a) General Method 10; (23-b) General Method 11; (23-c) For General Methods used for specific compounds see Table 2; (23-d) General Method 3; (23-e) For General Methods used for specific compounds see Table 2; (23-f) and General Method 9; (23-h) For General Methods used for specific compounds see Table 2.

WO 03/093286 64 Part 2: Preparation of Compound 113-01 to 113-03 PCT/AU03/00494

R

4 0

NH

2

OR

4 O -O 23-i

R

4 0

NH

2

NH

2 104

HO,

NHR

2

NHR

1 113-01 to 113-03

R

4 0 R OR4

R

4 0 HR 2 NHR 1 112-01 to 112-03 O R4

K

Conditions: For General Methods for specific compounds see Table 2; (b) General Method Part 3: Preparation of Compounds 114-01-114-03 for the Synthesis of Alternate Urea Building Blocks For the Preparation of Further Derivatives at R 3 as Exemplified by Structures 117-01-117-15 (by employing 5 different lipidic side chains).

WO 03/093286

R

4 0 N3 O R4 R4 O 23-k

R

4 H

NH

2 68

R

4 0 O4NH 2OR 4 R4? I R4 0

NHR

2

NHR

1 115-01 to 115-03 PCT/AU03/00494

R

4 0

R

R

4 0-

NHR

2 NHRi 114-01 to 114-03 R OR4

R

4 0 1HR01 NHR 1 116-01 to 116-15 HO R I R4

NHR

2

NHR

1

K

117-01 to 117-15 Conditions: General Method 1; General Method 3; General Methods 1,4,7,14 and 15 as appropriate depending upon lipidic side chain to be coupled General Method 5 6 and 17.

Table 2: Experimental Results, Intermediates and Products for Example 23

R

4 0 OR 4 0 R 3

R

4 0 O

NHR

2

NHR

1 Product SM* M R1 R2 R3 R4 Rt (min) Yield 68 67 10 I I L K 830.5 6.07 74.8 104 67 10 I I M K [M+H 804.5 5.36 18 105 68 11 H I L K 1052.52 8.0-12.0 32 106-01 105 15 H N L K [M+H 1094.6 6.90 100 106-02 68 11 H H L K 1274.7 7.66 19.5 106-03 105 4 H C L K M+H 1224.7 7.32 84 106-04 105 14 H D L K [M+Na= 1228.6 7.45 100 106-05 [M+Na] 1261.7 7.38 WO 03/093286 PCT/AU03/00494 106-06 105 14 H F L K 1205.7 7.35 98 106-07 105 13 H G L K [M+Naf 1208.8 7.43 82 107-01 106-03 3 H C M K Detected in 108-01 form 100 107-02 106-04 3 H D M K Detected in 108-02 form 96 107-03 106-05 3 H E M K Detected in 108-03 form 100 107-04 106-06 3 H F M K Detected in 108-04 form 100 107-05 106-07 3 H G M K Detected in 108-05 form 100 108-01 107-01 15 H C B K 1240.7 6.99 100 108-02 107-02 15 H D B K 1222.7 7.07 108-03 107-03 15 H E B K 1255.5 6.99 100 108-04 107-04 15 H F B K 1221.3 7.42 86 108-05 107-05 15 H G B K [M+Na] 1224.8 7.05 108-06 107-01 4 H C A K 1366.9 8.09 108-07 107-02 4 H D A K 1348.9 8.01 67.7 108-08 107-03 4 H E A K 1381.9 8.33 88 108-09 107-04 4 H F A K 1347.4 7.86 100 108-10 107-05 4 H G A K 1328.6 8.18 100 109-01 108-06 16 I C A K Detected in 110-02 form 67 109-02 108-07 16 I D A K Detected in 110-03 form 61 109-03 108-08 16 I E A K Detected in 110-04 form 109-04 108-09 16 I F A K Detected in 110-05 form 58 109-05 108-10 16 I G A K Detected in 110-06 form 51 110-01 68 15 N N L K 914.5 5.71 100 110-02 109-01 15 N C A K 1186.76 7.50 100 110-03 109-02 15 N D A K 1168.77 7.54 100 110-04 109-03 15 N E A K 1201.8 7.33 100 110-05 109-04 15 N F A K 1167.5 7.30 100 110-06 109-05 15 N G A K 1148.5 7.51 100 110-07 109-01 4 C C A K 1316.7 7.59 86.8 110-08 109-02 4 C D A K 1298.6 7.94 100 110-09 109-03 4 C E A K 1331.71 7.90 100 110-10 109-04 4 C F A K 1297.5 7.58 100 110-11 109-05 4 C G A K 1278.7 7.82 100 110-12 109-01 14 D C A K 1298.5 7.68 84.7 110-13 109-02 14 D D A K 1280.4 7.92 100 110-14 109-03 14 D E A K 1313.83 8.03 100 110-15 109-05 14 D G A K [M+H 1260.5 7.89 28.2 110-16 109-01 1 E C A K [M+Na] =1353.8 7.86 100 110-17 109-02 1 E D A K 1313.81 7.81 100 110-18 109-04 1 E F A K 1312.8 7.64 100 110-19 109-05 1 E G A K 12 93 8 7.88 100 110-20 109-01 12 F C A K [M+H] 1297.9 7.60 110-21 109-02 12 F D A K 1279.8 7.92 77 110-22 109-03 12 F E A K 1312.84 7.65 100 110-23 109-04 12 F F A K 1278.8 7.73 73 110-24 109-05 12 F G A K 1259.9 7.64 110-25 109-01 13 G C A K 1278.8 7.64 66 110-26 109-02 13 G D A K 1260.

8 7.91 110-27 109-03 13 G E A K 1293.87 7.96 100 110-28 109-04 13 G F A K 1259.8 7.75 72 110-29 109-05 13 G G A K 1240.8 7.67 110-30 109-03 1 0 E A K Compound fragments to 110- 100 31 under LCMS conditions 110-31 P E A K 1202.81 7.26 111-01 110-07 5 C C A I 836.4 5.33 100 WO 03/093286 PCT/AU03/00494 111-02 110-08 5 C D A I 818.4 5.57 100 111-03 110-09 5 C E A I [M+H] 851.5 5.43 100 111-04 110-10 6 C F A I 817.4 5.44 18 111-05 110-11 5 C G A I 798.5 5.54 100 111-06 110-12 5 D C A I 818.4 5.56 100 111-07 110-13 4 D D A I 800.4 5.47 100 111-08 110-14 5 1 D E A I M+H] 833.4 5.83 100 111-09 110-15 5 D G A I 780.4 5.41 100 111-10 110-16 5 E C A I 851.3 5.64 100 111-11 110-17 5 E D IA I M+H] 833.3 5.75 100 111-12 110-18 5 E F A I 832.5 5.53 16 111-13 110-19 5 E G A I M+H] =813.3 5.69 100 111-14 110-20 6 1 F C A I M+H] 817.4 5.49 51 1 111-15 110-21 6 F D A I [M+H 799.4 5.46 48 111-16 110-22 5 F E A I M+Hl 832.5 5.65 2 111-17 110-23 6 F F A I M+H] 798.4 5.36 111-18 110-24 6 F G A I [M+H 779.5 5.58 53 111-19 110-25 5 G C A I 798.5 5.30 100 111-20 110-26 5 1 G D A I 780.5 5.57 100 111-21 110-27 5 G E A I 813.4 5.69 100 111-22 110-28 6 G F A I [M+H] 779.5 5.08 111-23 110-29 5 G G A I [M+H 1 760.5 5.28 100 111-24 108-06 5 H C A I 886.55 6.07 100 111-25 108-07 5 H D A I [M+H] 868.58 5.99 100 111-26 108-08 5 H E A I 901.4 6.00 100 111-27 108-09 6 H F A I [M+Hl] 867.6 6.03 111-28 108-10 5 H G A I M+H 848.6 5.91 100 111-29 110-30 5 P E A I M+H] 722.4 4.63 100 111-30 109-03 5 I E A I 679.5 4.62 100 112-01 104 15 N N B K [M+Na 952.7 5.01 47 112-02 104 4 C C Q K M+H] 1320.5 6.99 76 112-03 104 1 E E R K M H] 1365.4 7.21 92 113-01 112-01 5 1 N N B I 450.26 0.69 100 113-02 112-02 5 C C Q I [M+H 840.4 4.53 100 113-03 104 5 I I M I 324.2 0.62 100 114-01 68 1 S S L K 1204.9 7.02 100 114-02 68 1 T T L K [M+H 1096.5 6.82 100 115-03 68 1 U U L K [M+HW] 1204.3 7.25 100 111-01-1 111-01 17 C C A I As Start Material N/A 43 111-02-1 111-02 17 C D IA I As Start Material N/A 19.2 111-06-1 111-06 17 D C A I As Start Material N/A 34.5 111-07-1 111-07 17 D D A I As Start Material N/A 51 111-08-1 111-08 17 D E A I As Start Material N/A 27 111-10-1 111-10 17 E C A I As Start Material N/A 16 111-11-1 111-11 17 E D A I As Start Material N/A 111-29-1 111-29 17 P E IA I As Start Material N/A SM*=Starting Material M=Method of Synthesis (General Method) Rt All compounds in Table 2 were analysed by HPLC Method A.

Note: Under the employed analytical conditions, compounds containing amino group (compounds classed as 68, 104, 105, 107, 109) elute in unusual broad WO 03/093286 PCT/AU03/00494 68 peaks, sometimes several minutes wide; therefore, most of the time they are detected as acetamide derivatives obtained via acylating with acetic anhydride.] Substituents for Table 2 H H A 0 0

B

F

F

F

0 0= =0

D

F

HN

S

F

0

G

Cc 0

H

I K N H 2

N

N

SI

0

CF

3

HN

S

H

2 N 0

P

HN

T

c1 HN -e0

U

F

w WO 03/093286 WO 03/93286PCT/ATJO3/00494 HPLC Methods.

HPLC Method A Time H 2 0% MeCN% Flow Rate mL/min 0 95 5 2 1 95 5 2 7 0 100 2 12 0 1002 Agilent SB Zorbax C1 8 4.6 x 50mm (5pmr, LC Mobile Phase: Acetonitrile Water 0.1/ %formic acid HPLC Method B Agilent SB Zorbax Cl8 4.6 x50mm (5gm,80A) LC Mobile Phase: Acetonitrile Water 0.1% formic acid WO 03/093286 PCT/AU03/00494 Example 24: Synthesis of an Alpha linked Disaccharidic compound Selective Removal of Protecting Groups for Diversity-Part I Ph 0 N 3 OBnOMe 0 0 24-a OCIBz Phth 118 60 Ph TBDPSO <i 19Nhth 24-b

TB[

24-d

H

24-e2 24-el I 23a 24-a: glycosylation Compound 118 [1.Ommol] and 60 [l.5mmol] were dissolved in dry dichloromethane [l6mLJ and stirred with molecular sieves [4A, acid washed] at room temperature for 1h. To the mixture was then added 2,6-di-tert.butylpyridine [I .6mmol] and DMTST [I .6mmol], and the reaction stirred at room temperature. After 2.5h further 60 [0.2mmol], 2 ,6-di-tert.-butylpyridine [.2mmol] WO 03/093286 PCT/AU03/00494 71 and DMTST [0.2mmol] were added and the reaction stirred at room temperature for 1h. The reaction was then quenched with triethylamine, the solvents were removed in vacuo and the product purified by column chromatography (silica, petrolether/ethyl acetate 2:1) to give 119 as a colourless foam HPLC Method A, Rt=8.09 mins, [M+Na]+=1087.56; 1H-NMR (CDCI 3 8.00 2H, Ar), 7.82-7.65 2H, Ar), 7.95-7.14 19H, Ar), 6.90 2H, Ar), 5.52 (dd, 1H, H- J1'-2'=3.4Hz, J2'-3'=10.1Hz), 5.24 1H, 4.82 1H, CHPh), 4.72- 4.63 2H), 4.54 (dd, 1H, J3'-4'=3.6Hz), 4.38 (AB, 2H, CH 2 Ar, Jgem=11.7Hz), 3.95-3.86 2H), 3.83 3H, OCH 3 3.83-3.76 1H), 3.62- 3.56 1H), 3.49 (dd, 1H, J=5.9Hz, J=9.0Hz), 3.38 (dd, 1H, J=7.7Hz, J=9.2Hz), 3.37 (dd, 1H, H4',J4'-5'<IHz), 3.30 (dd, 1H, H1-a, Jgem=12.5Hz, J1a,2=1.9Hz), 3.25 1H), 2.92 (dd, 1H, H-1b, Jlb,2<1), 0.88 9H, tBu).

24-b: azide reduction Compound 119 [0.164mmol] was treated according to the procedure described in General Method 3 (NB: compound smears over several minutes on HPLC column (HPLC Method =1039.53. The solution of the crude product 120 was directly used for the next conversion.

24-c: amide coupling Crude 120 was treated with undecanoic acid 120mg [0.65mmol] according to the procudure described by General Method 7. 1 The residue containing 121 was purified on a silica column (gradient petrolether/ethyl acetate 2:1 to petrolether/ethyl acetate 1:1 with 2% triethylamine) to give 121 HPLC Method A, Rt=9.14 mins; [M+Na] =1229.69, and unreacted 120 24-d: silylether cleavage To a solution of 121 in DMF [3mL], was added a 1 molar solution of TBAF in THF [0.5mL] and acetic acid [30pL], and the mixture heated to 65°C for 6h. The reaction mixture was diluted with ethyl acetate and the solution washed with saturated sodium bicarbonate solution and water, the dried over magnesium WO 03/093286 PCT/AU03/00494 72 sulfate and the solvents removed in vacuo to give crude 122 (100% conversion by ELSD); HPLC Method A, Rt=7.17 mins; [M+H]+=969.55.

24-el to e2: removal of acid labile protecting groups (24-el) Crude 122 was dissolved in dry dichloromethane [5mL] and triethylsilane [0.5mL] and trifluoroacetic acid [0.lmL] were added. After stirring at room temperature for 10min conversion to 123a was complete (HPLC Method A, Rt=6.80min, (24-e2). Further stirring at room temperature for 3h gave 123 (100% conversion by ELSD); HPLC Method A, Rt=6.71 mins; [M+H]r=761.43.

Selective Removal of Protecting Groups for Diversity-Part 2 24-fl

TBDF

124a 24-f2 24-f to f2: removal of acid labile protecting groups (24-fl). Compound 121 [8pmol] was dissolved in dry dichloromethane [1mL] and triethylsilane [0.1mL] and trifluoroacetic acid [0.02mL] were added. After stirring at room temperature for 2min conversion to 124a was complete (HPLC Method A, Rt=8.54 mins, [M+H]+=1187.49). (24-f2). Further stirring at room temperature for 3 hrs gave 124 (100% conversion by ELSD); HPLC Method A, Rt=7.87 mins, [M+H]+=999.56.

WO 03/093286 73 Selective Removal of Protecting Groups for Diversity-Part 3 PCT/AU03/00494 24-g

TBDI

TBDI

121 125 24-h

TBDI

24-g: phthalimido cleavage To a solution of 121 [O.10mmol] in ethanol [5mL] was added hydrazine hydrate [0.05mL], and the solution refluxed for 20h. The solvents were removed in vacuo and the residue co-evaporated with toluene to give crude 125 (100% conversion by ELSD). Product smears over several minutes on HPLC (HPLC Method [M+H]+=1077.43.

24-g: sulfonamide formation Compound 125 [1.8pmol] was reacted with tosylchloride [5mg] according to General Method 14 to give 126 (100% conversion by ELSD), HPLC Method A, Rt=9.25 mins; [M+H]*=1231.65, [M+Na] =1253.63.

WO 03/093286 PCT/AU03100494 74 Selective Removal of Protecting Groups for Diversity-Part 4 Ph Ph 0 K~ HN 24-i I HN TBDPSO 0TBDPSO BZCIO L. BzCIO 125 NH 2 127 'F 3 Ph 0P 0 24-jH 0M 24- 0H TBDPSO

K

HO

TBDPSO

128 0CF 3 NH NH O=NH

C

HN Id-HN-

F

3 C 129 H Ph 0Ph 0

HNH

240 H0 ~Oe0NO 24- qMe 24-mi A- HO 0 10 HO 0 NH- H CF 3 NHN CF 3

F

3 C 130 HNF 3 C 131a H

OH

H

24-m 0

,H

~NH 0=NH CF3

F

3 CP 131

H

WO 03/093286 PCT/AU03/00494 24-i: Urea Formation Compound 125 [0.10mmol] was was reacted with 3-trifluoromethylphenyl isocyanate [0.4mmol] according to General Method 1 to afford 127 (73% purity by ELSD); HPLC Method A, Rt=9.04 mins; [M+H]+=1264.75.

24-j: Ester Cleavage Compound 127 [0.1Ommol] was treated according to the procedure described in General Method 2 (with the exception that only MeOH was used as solvent) to afford crude 128; HPLC Method A, Rt=8.60 mins; [M+H]+=1126.48, =1148.46.

24-k: Carbamate Formation To a solution of 128 [0.10mmol] in dry DMF [5mL], was added 3trifluoromethylphenyl isocyanate [0.4mmol] and DBU [45pL], and the solution heated to 80'C for 20h. The reaction mixture was diluted with ethyl acetate, washed with saturated sodium bicarbonate solution and water, dried over magnesium sulfate, and the solvents removed in vacuo to give 129.

24-1: Silylether Cleavage To a solution of 129 in DMF [3mL], was added a 1M solution of TBAF in THF and acetic acid [30pL], and the reaction mixture heated to 65 0 C for 6h.

The reaction mixture was diluted with ethyl acetate and the solution washed with saturated sodium bicarbonate solution, water, dried over magnesium sulfate, and the solvents removed in vacuo to give crude 130.

24-ml to 24-m2: Removal of Acid Labile Protecting Groups (24-ml). To a solution of compound 130 in dichloromethane [5mL] was added triethylsilane [0.5mL] and trifluoroacetic acid [0.1mL]. After stirring at room temperature for 2min conversion to 131a was complete. (24-m2). Further stirring at room temperature for 3 hrs gave 131 quantitatively.

WO 03/093286 76 Selective Removal of Protecting Groups for Diversity-Part PCT/AU03/00494 ,24-qi 24-n: Silylether Cleavage To a solution of 127 in DMF [3mL], was added a 1M solution of TBAF in THF and acetic acid [30pL], and the mixture heated to 65°C for 6h. The reaction mixture was then diluted with ethylacetate and the solution washed with saturated sodium bicarbonate solution, water, dried over magnesium sulfate, and the solvents removed in vacuo to give crude 132.

WO 03/093286 PCT/AU03/00494 77 24-o: Carbamate Formation To a solution of 132 [O.10mmol] in DMF [5mL] was added 3trifluoromethylphenyl isocyanate [0.4mmol] and DBU [45pL], and the solution heated to 80°C for 20h. The reaction mixture was diluted with ethyl acetate, washed with saturated sodium bicarbonate solution, water, dried over magnesium sulfate, and the solvents removed in vacuo to give 133.

24-p: Ester Cleavage Compound 133 [0.10mmol] was treated according to General Method 2 (with the exception that only MeOH was used as solvent) to provide crude 134.

24-ql to 24-q2: Removal of Acid Labile Protecting Groups (24-q1). Compound 134 was dissolved in dry dichloromethane [5mL] and triethylsilane [0.5mL] and trifluoroacetic acid [0.1mL] were added. After stirring at room temperature for 2min conversion to 135a was complete. (24-q2).

Further stirring at room temperature for 3 hrs gave 135.

Example 25: Synthesis of a Beta 1--6 Linked Disaccharidic Compound For Drug Discovery-1 OAc A OAc N3-OBnOMe O 0 A SM O 25-a AcO N AcO-A Me+ HO NPhth _O NPhth NPhth HO 136 60 137 NPhth OAc AcO 0 N3 AcO 0 -b NPhth

O

AcO- N o IQ NPhth WO 03/093286 PCT/AU03/00494 78 Glycosylation To a solution of 60 [0.48 mmol] and 136 [0.70 mmol] in dichloroethane [8mL] was added DMTST [0.48 mmol], and the mixture stirred for 45 mins at room temperature. At that time further DMTST [0.21 mmol] was added, and after stirring for 20 mins the reaction was quenched by the addition of triethylamine mL]. The reaction mixture was then diluted with dichloromethane, washed with 10% citric acid, saturated sodium bicarbonate solution, dried over magnesium sulfate, the solvents evaporated in vacuo and the product purified by column chromatography (silica, petrol ether/ethyl acetate 1:1) to give 137 as a colorless foam HPLC Method A, Rt=5.06 mins; [M+Na]+=758.49.

Acetylation To a solution of 137 [2mg] in pyridine [0.2ml] was added acetic anhydride and the ensuing reaction mixture stirred at room temperature for 2 hrs.

The reaction mixture was then diluted with dichloromethane and washed with citric acid, saturated sodium bicarbonate solution, dried over magnesium sulfate and the solvents evaporated in vacuo to give 138; 1 H-NMR (CDCIs): 7.85-7.62 8H, Ar), 5.75 (dd, 1H, J2', 3 '=10.4Hz, J 3 4 5.70 (dd, 1H, H-3, J 2 3 =11.0Hz, J 3 4 =3.4Hz), 5.39 1H, Jil, 2 5.12 (dd, 1H,

J

4 5 4.64 (ddd, 1H, 4.25 (dd, 1H, 4.00 (dd, 1H, H-4), 3.88-3.76 3H, H-la, H-5, 3.70-3.52 5H, H-lb, H-6a, H-6b, H-6a', 2.07, 1.98, 1.87, 1.71 (each s, 3H, Ac).

Example 26: Synthesis of a Beta 1--6 Linked Disaccharidic Compound For Drug Discovery-2

OPMB

PMB BnO PMH BO'MB BnOOH PMBO O II 26-a PMBO- H BnO TBDPSO SMe NH CCI BzCI OHO TBDPSO SMe 3BzCCF 66 baz 139 140 WO 03/093286 PCT/AU03/00494 79 26-a. Formation of a Beta 1-,6 Linked Disaccharide A solution of thioglycoside 139 [0.154mmol], trichloroacetimidate 66 [1.5 eq.

with respect to thioglycoside] and 4 angstrom molecular sieves [0.26g] in 1,2- DCE [2.6mL] was stirred at room temperature for 15 mins. At this time TMSOTf [0.3 eq.] was added. The reaction was allowed to stir for 30mins at which time it was quenched by the addition of triethylamine [2mL]. The reaction mixture was diluted with DCM, filtered and the resulting filtrate concentrated in vacuo to afford a residue. The residue was purified by column chromatography [toluene/acetone, 20:1] to provide the product as a colourless foam HPLC Method A, Rt=8.02 mins; [M+Na] =1316; 'H-NMR, CDC1 3 8 4.07 1-H, H-la J 12 =9.2 Hz), 4.42 1-H, H-lb, J 1 2 =8.2 Hz) indicating two beta linkages.

WO 03/093286 PCT/AU03/00494 Example 27: Synthesis of a Methyl Glycoside Disaccharide for Drug Discovery

OH

O 'u27-a Se 27-zHO 0 SMe HO-VO

SM

e Oz OSMe BzHB 141 NHDTPM 142 NHDTPM 143 NHDTPM 127-c N -OTBDPS

OTBDPS

S27-e Tfo 0 27-d Bo SMe BzO SM HDTPM 145 NHDTPM 27-f 146 N OTBDPSTBDPS 27-g N 3

OTBDPS

HO-- 0 sM 27g 0 ASMe 27-h NHDTPM HO

H

147 148 N

OTBDPS

-BzO 4 SMe 144 NHDTPM

OPMB

PMBO 0 NH PMBO- NHO -CC 3 O/ 66

CF

3 OPMB N TBDPS PMBO 0 0 O\OMe

PMBO

NH

N

CF

3 151 27-a. Benzoylation of the 3-OH Position.

To a solution of 141 [37.2 mmol] in 1,2-dichloromethane [140 mL] at 0°C was added DMAP [2 eq.] and benzoyl chloride [1.5 The reaction mixture was allowed to return to room temperature, and stirred for 2 hrs. Methanol was added and the reaction mixture was stirred for a further 15 mins. The reaction mixture was then diluted with CHCI 3 washed with 10% citric acid solution, WO 03/093286 PCT/AU03/00494 81 saturated NaHCO 3 solution, saturated brine solution, dried (MgSO 4 concentrated in vacuo. Compound was passed through a plug of silica to give 142 and used directly in the next step without further purifucation.

27-b. Benzylidene Cleavage To a solution of thioglycoside 142 [36.7 mmol] in a mixture of MeCN/MeOH/H 2 0 155mL] was added p-toluenesulphonic acid [200mg]. The resulting reaction mixture was stirred at 75 0 C for 2 hrs. The reaction was allowed to cool, to room temperature, water was added [50 mL] and the volatile solvents [MeCN and MeOH] removed in vacuo. The resulting suspension was filtered and the collected solid washed further with water followed by petroleum ether and then dried under vacuum to afford pure 143 (99.8% pure by ELSD); HPLC Method A, Rt=3.80 mins.

27-c. Sialyl Protection of a 6-OH Group.

To a suspension of the diol 143 [10mmol] in pyridine [20 mL] was added imidazole [1 mmol] and the resulting reaction mixture was then heated to 120 0 C. At this time TBDPS-CI [12mmol] was added in portions and the reaction was stirred for 1 hr at 120 0 C. After this time further TBDPS-CI [0.4 eq.] was added and the reaction was allowed to stir for a further hour. The reaction mixture was then cooled, annd the volatiles removed in vacuo. The residue was taken up in DCM and washed with 1 molar HCI solution, dried (MgSO 4 and the solvent removed in vacuo. The residue was washed with petroleum ether to afford pure 144 as a white solid HPLC Method A Rt=6.66 mins (100% purity by ELSD); [M+H]+=718.57.

27-d. Formation of a 4-O-Triflate.

To a solution of 144 [2 mmol] in DCM [20 mL] was added pyridine [4mmol] and the resulting mixture cooled to 0°C. At this time triflic anhydride [3.2mmol] was slowly added and the reaction mixture was then allowed to return to room temperature. The reaction was allowed to stir for one hour at room temperature at which time it was diluted with DCM and washed with a solution of 0.5 molar WO 03/093286 PCT/AU03/00494 82 HCI, dried (MgSO 4 and the solvent removed in vacuo to afford pure 145 HPLC Method A, Rt=7.63 mins; [M+H]+=850.66.

27-e. Formation of an Axial Azido Derivative.

To a solution of triflate 145 [1mmol] in DMF was added NaN 3 [3 mmol] and the resulting reaction mixture was allowed to stir at room temperature for 10 hrs.

The reaction mixture was concentrated in vacuo, and the residue washed with water followed by petroleum ether. The solid was then dried to provide the product 146 HPLC Method A, Rt=7.23 mins; =743.5.

27-f. Removal of a Benzoyl Group by Transesterification.

Compound 147 was prepared according to the procedure described in General Method 2 and purified by column chromatography [30% ethyl acetate/petroleum ethers] to afford a white solid HPLC Method A, RT=6.53 min; [M+H]*=639.2.

27-g. Deprotection of the 2-Amino Group.

To a solution of the sugar 147 [4.29mmol] in DMFIMeOH 45 mL] at room temperature was added hydrazine hydrate [0.52 mL]. The reaction mixture was stirred for two hours at which time it was filtered and the filtered solid washed with methanol. The filtrates were combined, the solvents removed in vacuo, residue taken up in CHC13, washed with saturated brine, dried (MgSO4), and the solvent again removed in vacuo to provide a white solid 148 HPLC Method A, Rt=5.96 mins; [M+H] =473.3.

27-h. Reprotection of the 2-Amino Group.

To a solution of the sugar 148 [0.22 mmol] in MeOH [1.25 mL] was added phthalic anhydride [0.4mmol] and triethylamine [1 drop] and the solution was allowed to stir overnight. The reaction mixture was then concentrated in vacuo.

The residue was the dissolved in dry pyridine [0.25 mL], cooled to 0°C, and acetic anhydride [60 pL] added dropwise. The reaction was allowed to stir overnight. The reaction was then concentrated, the residue taken up in CHCIs WO 03/093286 PCT/AU03/00494 83 and washed with 10% citric acid solution, saturated sodium bicarbonate solution, saturated brine solution, dried (MgSO4), the solvent removed in vacuo, and the residue purified by column chromatography [20% ethyl acetate/petroleum ethers] to afford the product as a white solid 149 HPLC Method A, Rt=7.29 mins; [M+H]+=645.35.

27-i. Glycosylation to Form the O-Methyl Glycoside To a solution of the sugar 149 [0.775mmol] in DCM [5 mL] was added 3 angstrom molecular sieves, MeOH [12 mmol] and finally DMTST [2.32 mmol].

The reaction mixture was allowed to stir for 30 mins at which time the reaction was quenched with triethylamine [2.37 mmol], filtered and the filtrate concentrated in vacuo. The residue was taken up in DCM and washed with water, 10% citric acid solution, saturated sodium hydrogen carbonate solution, saturated brine, dried (MgSO 4 and the solvent removed in vacuo to provide a yellow oil. The oil was purified by column chromatography [20% ethyl acetate/petroleum ethers] to provide the product as a yellow oil HPLC Method A, Rt=7.20 mins; [M+Na]'=651.3.

27-j. Zemplen Deprotection.

Compound 150 was prepared according to the procedure described in General Method 2 and was purified by column chromatography [25% ethyl acetate/petroleum ethers] as a white solid HPLC Method A, Rt=6.88 mins; [M+Na] =609.7.

27-k. Formation of an O-Me Glycoside, Beta 1-3 Linked Disaccharide.

Donor 60 [0.128 mmol] and acceptor [85.2 mmol] were dissolved 1,2-DCE mL]. 4 Angstrom molecular sieves were added and the mixture was stirred for mins. TMSOTf [2.8 pimol] was then added and the reaction left to stir for mins. The reaction mixture was then quenched with triethylamine, diluted with

CHCI

3 washed with saturated NaHCO 3 solution, dried (MgSO 4 and the solvents removed in vacuo. The residue was purified by column WO 03/093286 PCT/AU03/00494 84 chromatography to afford 151 HPLC Method A, Rt=7.60 mins; [M+Na]+=1226.67 Example 28: Formation of Alternatively Linked Disaccharide Scaffolds.

0 9 SMe 28-a 0 9 O 28-a OORacceptor 0 OMe 2 0 OMe 153a-153h 28-a. Glycosylation with a Trichloroacetimidate Donor to Afford Disaccharides (153-a to 153-h) To solutions of the acceptor molecules 152a-152h (1.8 mmol) and the donor 2 (2.7 mmol) in dry 1,2-dichloroethane (84 mL) is added 3A acid-washed molecular pellets (4 g) and the resulting mixture stirred for 20 min. To the mixture was then added Methyl Triflate (1.8 mL of a 0.1M solution in dry 1,2dichloroethane, 0.18 mmol) and the reaction then stirred for 30 mins. After this time triethylamine (6 mL) was added and the suspension filtered, washed with dichloromethane and all solvent removed in vacuo. This residue was purified by column chromatography to yield the title compounds as indicated in table 3 below.

Table 3: Disaccharide Products From Glycosylation with Donor 2 WO 03/093286 WO 03/93286PCT/A1J03/00494 1 52c 153c1, 153c2 1 63d 1 53e 1653f "II 563g P h 7 0 0 M e O 2 C 4 o00 0 153h WO 03/093286 PCT/AU03/00494 86 28-b. Azide Reduction and deprotection (154a-154h) Compounds 153a-153h are hydrogenolysed at 60 psi for 1 hour with catalytic palladium on activated charcoal in ethanol, to yield upon filtration and evaporation, the corresponding diamines in which the benzylidene ring has also been cleaved as indicated in the table below. These diamines may be used in their crude form for further reactions.

IP-O 0 12

NH

2 0a 154a

IDPS

Me0 2 C C4 WO 03/093286 PCT/AU03/00494 87 28-c. Amide formation- HBTU Coupling (155a-155h) Compounds 155a(i)-155a(iv) to 155h(i)-155h(iv) are prepared by reaction of the diamines 164a to 164h with carboxylic acids according to the procedure described in General Method 4 in a combinatorial manner. This every diamine may be reacted with an excess of every carboxylic acid to produce the bisamide products.

Table 4: Acids i-v Shown Below Are Reacted with Diamnines to Give Products as Listed

H

0 ci ~BocHN- O0H iii iv

S~T

3 i Acids Diamnine ii iii iv 154a 155a(i) 155a(ii) 155a(iii) 155a(vi) 154b 155b(i) 155b(ii) 155b(iii) 155b(vi) 154c1 155c1(i) 155c1 (ii) 155cl (iii) 155c (Ai) 154c2 155c2(i) 155c2(ii) 155c2(iii) 155c2(vi) 154d 155d(i) 155d(ii) 155d(iii) 155d(vi) 154e 155e(i) 155e(ii) 155e(iii) 155e(vi) 145f 155f(i) 155f(ii) 155f(iii) 155f(vi) 154g 155g(i) 155g(ii) 155g(iii) 155g(vi) 154h 155h(i) 155h(ii) 155h(iii) 155h(vi) WO 03/093286 PCT/AU03/00494 88 Examples of Table 4 0

H

N HON HO HHO- N

O

H

H

HO 'OTBDPS Me O

CF

3 HN 0 HS O 0 0 OMe 55b(ii) 155e(i) 15 5b i i

CF

28-d. Silyl Deprotection The t-butyldiphenylsilyl groups are removed from Compounds 155 (as appropriate) by treatment of the silylated compound in N,N-dimethylformamide with tetrabutylammonium fluoride, followed by removal of the solvents in vacuo and purification by mass based fractionation on a C18 HPLC column.

Example 29: Formation of Alternatively Amide Linked Disaccharide Scaffolds Reduction of an azide to an amine Amino sugars can be obtained by reduction of corresponding azido sugars according to the procedure described in General Method 3. Alternatively, the azide may be reduced selectively by hydrogenolysis at atmospheric pressure over 5% palladium on charcoal in methanol for 30 minutes. This latter method is suitable for the reduction of azides in the presence of benzyl ethers. Filtration of the solution and removal of the solvents in vacuo yields the crude aminosugar suitable for further reaction. Hydrogenolysis is also employed to remove the carbobenzyloxy group from compound 152d (see Amines Used in Example 28).

Formation of an anhydride and reaction with an amine Anhydrides are formed according to the following general method. Azaleic acid monomethyl ester, suberic acid monomethyl ester or fumaric acid monoethyl ester [2 equivalents] are dissolved in anhydrous dichloromethane to form a WO 03/093286 PCT/AU03/00494 89 millilolar solution. To this solution is added diisopropylcarbodiimide [1 equivalent] and triethylamine [1 equivelant] and the solution stirred at room temperature for 45 minutes. After this time, the solution of acid anhydride is evaporated, redissolved in N,N-dimethylformamide to form a 10 millimolar solution and added to a solution of the crude sugar amine (selected from Amines Used in Example 28) in N,N-dimethylformamide. The reaction mixture is stirred for 1 hour. The reaction mixture is quenched with water, acidified to pH 4 and extracted with ethyl acetate and back extracted with 10% sodium hydrogen carbonate solution to yield the crude sugar amide. The solvents are removed in vacuo and the esters hydrolysed by the addition of 5 equivelant of lithium hydroxide to an wet methanolic solution of the crude ester. Acidification of this mixture followed by removal of the solvents in vacuo yields the crude half acid amide which is partially purified by passing through a short bed of silica gel.

This crude material in which all other protecting esters have been cleaved is suitable for further reaction. Compounds formed by this process are displayed in Half Acid Amides formed in Example 29 below.

Formation of a Dimeric Derivative The crude sugar half acid amide, is then dissolved in N,N-dimethylformamide and treated with 1 equivalent of ethyl diisopropylamine, 1 equivelant of HBTU and finally 1.3 equivalents of the crude sugar amine. The reaction mixture is stirred for 30 to 60 minutes at room temperature, quenched by the addition of water and solvents removed in vacuo. The crude residue is finally purified by mass based fractionation to furnish the desired bis amide linked scaffolds. A combinatoral matrix of acid and amine results in a wide diversity of bis amide linked scaffolds as exemplified in Table WO 03/093286 WO 03/93286PCT/AU03/00494 Amines used in Example 29: Ph -'-OH 2

N

NH

2 -GOMe BocHN-f 13 156

HH

1 52d Ph -O HBoc

H

2 N OPMB 0 59 Half Acid Amides formed in Example 29: 157A-C 0 -k OPMB

N-

0 160A-C HN 0,

H

I 58A-C

H

RYN,

0 IBDPSO Bz IGIA-C

OH

-O Fumaric Where R= A 0 B OHSuberic 0 CH Azeleic WO 03/093286 PCT/AU03/00494 91 Table 5: Products Resulting From Reaction of Compounds From "Amines used in Example 29", with "Half Acid Amides formed in Example 29" Amine Acid 13 156 50 59 152d 157A 157A1 3 157B 157133 157C 157C13 158A 158A1 3 158AI 6 168B 1681313 168B166 158C 158C13 I158016 169A 159AI 3 159A1 56 159A50 169B 1691313 159B156 1591350 159C 159C13 15SC156 159C50 160A 160A1 3 160A156 160A50 160A59 160B 160B13 16OB156 160B50 160B59 1600 160C1 3 160C156 160C50 160C59 161A 161 Al3 161A156 161 A50 161A59 161 Al52d 16113B 1611313 161B156 161 B50 161 B59 161Bi52d 61 C 161013 1610156 16105 16105 161152dl oi Example structures from Table 158B1 56 161A59 WO 03/093286 PCT/AU03/00494 92 Example 30: Formation of Amide Linked Disaccharide Scaffolds Glucuronic acid 14, is dissolved in N,N-dimethylformamide to form a millimolar solution. To this solution is added triethylamine [1.1 equivalents] followed by HBTU [1.05 equivalents. The mixture is stirred for 3 minutes at room temperature, after which time a concentrated solution of the amine [20-30 millimolar; 1 equivalent], as prepared in Example 29 [amines 13, 156, 152d, and 59] is rapidly added. The reaction mixture is stirred for a further 45 minutes, then quenched with an equal volume of 10% citric acid in water, and extracted with ethyl acetate. The organic layers are dried over magnesium sulfate and solvents removed in vacuo to yield the crude product which is further purified by column chromatography, to yield the desired product.

Reaction products: Ph-O S1 NH OMe BocHN- AcO N 3 OAc 162 Ph--O O H AcO A

-N

AcO N AcO

N

3 HBoc 162 NH OPMB AcO O0 AcAO cO-Ac 165 HN"' H

OBZ

0

OTBDPS

Ac166O AcO-- N 3 OAc 166 164 164 The Boc, isopropylidene and benzylidene protecting groups may be removed by treatment with TFA according to general procedure 5, acetate and benzoate protecting groups are removed according to general procedure 2.

WO 03/093286 PCT/AU03/00494 93 Example 31: Synthesis of an Alkylated 2-Deoxy-2-Amino Disacoharidic Compound 0 0 OPM H OMBOPMB HN OPMB PMBO\~~ 31 -a PBO- NH NH 2

PMBOHN

O<N H 109-03 N= H16

F

3 C F 3 Cd 1 31-b H HN

O

HO

F

3 0 168,169 3-1-a and 31-b. N-A Ikylation To a solution of the sugar 109-03 [7.4mg] in THF/MeOH [84 1 tL/9.4 tIL] was added benzaldehyde [0.7lgtL] and the mixture stirred for 2 hrs at room temperature. To the mixture was then added acetic acid [0.5liL] and NaCNBH3 [0.8mg] and the reaction was allowed to stir overnight at room temperature. The reaction was neutralised and concentrated in vacuo. The residue was taken up in DCM and washed with a saturated brine solution, dried (MgSO 4 and the solvent removed in vacuo. The residue was treated with AC 2 O/pyridine solution for two hours for the purpose of analysis; HPLC Method A, Rt (168 monobenzylated-monoacetylated) =7.56 mins, (169 bis-benzylated) =8.77 mins; WO 03/093286 PCT/AU03/00494 bis-benzylated mono-benzylated-mono-acetylated [M+H]+=1339.9.

[M+H]'=1391.9, Example 32: Synthesis of Benzimidazole Compounds OPMB N OPMB

PV

PMBo o 32-a P M B 0

NH

2

NH

2 68 32-b 32-a. Fluorine Displacement A solution of 3-fluoro-2-nitro-trifluoromethylbenzene [0.0715 mmol], triethylamine [0.0861 mmol] in DMF [250iL] was added to a flask containing diamine 68 [0.0241 mmol]. The resulting reaction mixture was then stirred at 0 C for 16 hrs. At this time the reaction was allowed to cool and the product was purified by preparative TLC [mobile phase ethyl acetate]. The product was collected in quantitative yield; HPLC Method A, Rt=7.51mins; [M+Na]'=1230.6 32-b. Formation of Benzimidazole A solution of SnCl2 [300pL of SnCI 2

.H

2 0 at 0.32 molar in DMF] was added to a flask containing compound 170 [2.48 Rmol]. The reaction mixture was then stirred at 80 0 C for 16 hrs. The reaction mixture was diluted with EtOAc/H 2 0 WO 03/093286 PCT/AU03/00494 5mLI, filtered through a pad of celite. The filtrated was separated into aqueous and organic layers and the organic layer washed with H 2 0, dreid (MgSO 4 and the solvent removed in vacuo to afford a mixture of products 171 and 172; HPLC Method A, Rt (171) 7.18 mins, Rt (172) 7.41 mins; (17 1) 1186.8, (172) 11. 58.8.

Example 33: Synthesis of a N-Acetyl-lactosamine Based Library of 6'-Hvdroxy Phosphonates

OR

3

OR

HO 0BnO OTBOPS Oi

R

BflO 0 1 R O~v HHN R2 BnO .~SMe 0 BnO HN N 0174 OPiv K0 /ODR 0-D O 33-a

O

4

OBM

173-1,2 133-b

OR

3

OR

3 0 -7 0 0 -Ti- 0

R

5 0 I R, 33-5o B0nO R Z% BnOS K9 B HN HJ 176-1,2 0 (BnO) 2 FP R 177-1,2 0 33-d O 3O

OR

3 v OH OH

R

5 0- O0iv R, 33-e HO0O 0 HO cHN3- .H R AcHN0 R 2 OH 0CH79 178- atol 179 (BnO 2 R 178-2a to 21(BOP\ R 0 0 For compounds 173-1, 175-1, 176-1, 177-1 and 178-1, RI= OBn and R 2

=H

For compounds 173-2, 175-2, 176-2, 177-2 and 178-2, R1= H and R2=OMe Reactions were carried out in identical series for resins 174-1 and series 174-2.

After step 33). After step 33-b each series was divided into 12 portions for the individual alkylations.

WO 03/093286 PCT/AU03/00494 96 Alkylating Agents for Example 33, where R= Sulphonate or R=Halide.

I N1 C1 N NN N RR R 0'" R RR Bn OBn Bn OBn Bn OBn P BnOOn O n BnOp\ OBn OBn NO2 AcHN NNO R R

R

BnO OBn B OBn B O Bn OBn OBn Bnd \O\ OBn

F

F N F H F FO R 0N R R :CC F 0 Bn0 P BnO' O OBn OBn BnObn 33-a. Glycosylation Resin [0.47 mmol] was weighed into a reactor and molecular sieves [200 mg], thioglycoside donor sugar 174 [2.35 mmol] and dichloromethane mL] was added. To the mixture was then added DMTST [2.35 mmol]. The reaction vessel was sealed, shaken and reacted for 5 hours. At this time the reaction was then quenched by the addition of triethylamine, and the molecular sieves removed from the resin. The resin was then washed with DMF, MeOH/CHC13 and dichloromethane. The resin was then dried under vacuum.

33-b. Solid Phase Silylether Deprotection A solution of PSHF (proton sponge hydrogen fluoride) (0.5 Molar in DMF/Acetic Acid, 95:5) was prepared. The resins [1.41 mmol] was added to the solution and the reaction was stirred at 65 0 C for 24 hours. The resin was then washed with WO 03/093286 PCT/AU03/00494 97 DMF, MeOH/CH 3 COOH/THF, 1:1:8, THF and DCM, and then dried under high vacuum.

33-c. Solid Phase Alkylation Resins 176 [0.047 mmol] were individually reacted with a 0.25 molar solution of tert-butoxide in DMF (5 min) and then an alkylating agent (see above), [0.25 molar of alkylating agent in DMF, 20 min] was reacted with the resin. The resins were washed with DMF and again treated with the two solutions, this procedure was repeated a further four times. The final wash of the resins was performed as above; with DMF, THF/MeOH/ CH 3

CO

2 H THF, DCM and MeOH. The resins were then dried overnight.

33-d. Cleavage of Disaccharide from, Resin The resins 177 [0.047 mmol] were separately treated with a 7% hydrazine hydrate/DMF solution [2 mL] overnight. The resin was filtered and the resin washed with DMF. The filtrates were combined and the solvent removed in vacuo. The residue was taken up in DCM and washed with water and saturated brine solution, dried (MgSO 4 and the solvent removed in vacuo. The residue was then treated with a solution of Ac20/pyridine [1 mL, 1:3] for three hours.

The Solvents were removed in vacuo and the product purified by column chromatography.

33-e. Removal of the Pivaloyl Protecting Group To a solution of NaOMe/MeOH/THF [2 mL, ~2 molar] was added the pivaloyl protected disaccharide 178 [0.03 mmol]. The reaction mixture was heated at reflux until TLC indicated the reaction was complete. At completion, reaction mixture pH was reduced to ~5 with amberlite IR-120-H* resin. The reaction was filtered, and the solvent concentrated in vacuo.

33-f. Deprotection of hydroxyphosphonates and Benzyl Ether Cleavage Compound 178 (after 33-e) [0.0193mmol] was dissolved in dry dichloromethane [2 mL] under a nitrogen atmosphere, the solution cooled to 0 C and WO 03/093286 PCT/AU03/00494 98 trimethylsilyl bromide [0.Q97mmol] was added. After stirring at 0 0 C for 30 mins a solution of ammonia in methanol [12ltL of 28%aq ammonia in 2Oml- methanol] was added. The solvents were removed to give the crude free hydoxyphosphonate as an ammonium salt. Final products were purified by mass fractioning H PLC.

Table 6: Final products Synthesised in Example 33.

Comp. No. R RI R2 Note 179a Ra H OMe 179b Rb H OMe 179c Re H OMe 179d Rd H OMe 179e Re H OMe 179f Rf H OMe 179 -BR H OMe 179h Rh H OMe 179i Ri H OMe 179j RiH OMe 179k Rk H OMe 1791 RI H OMe 179m Ra HOH OH,H Anomeric lactol 179n Rb H,OH OH,H Anomeric lactol 179o Rc H,OH OH,H Anomeric lactol 17p Rd H,OH OH,H Anomeric Iactol 179q Re H,OH OH,H Anomeric lactol 179r Rf H,OH OH,H Anomeric lactol 179s Rg H,OH OH,H Anomeric lactol 179t Rh H,OH OH,H Anomeric lactol 179u Ri H,OH OH,H Anomeric lactol 179v Rj H,OH OH,H Anomeric lactol 179w IRk KOH OHH Anomeric lactol 179x RI H,OH QH,H tAnomeric l'actol WO 03/093286 WO 03/93286PCT/AU03OO-194 Side Arms for Table 6 Ra Rb Rc

CI

Rd AcHN Rf N0 2 Rg Re

H

N

0 Rk

RI

WO 03/093286 100 Example 34: Preparation of Guanidine PCT/AU03OO-194

R

4 0> R4

R

4 0 R0 NH

,NH

NH

CF

3

F

110-20 34-a HOR3

O

NH

,NH

0

S=NH

CF

3

F

111-14 R

OH

HO

HO 0 NH NH

CF

3

F

180 43-b 111-14 180 HO

R

3

OH

HO 0 HO O

Z

R

5

N

CF

3 F 1 Scheme 4: general method 12 NH 4 0H MeOH (1:1) WO 03/093286 PCT/AU03/00494 101 34-a. Reaction Conditions to form a Thiourea Compounds 111-14 and 180 were prepared as a mixture (unpurified) by reaction of compound 110-20 according to the procedure described in General Method 12. The mixture was used directly in the next step.

34-b. Formation of a Guanidine The sugar mixture (111-14 and 180) (0.025 mmol) was dissolved in methanol and concentrated aqueous ammonium hydroxide (0.5ml) was added.

The reaction was stirred at room temperature for 4 hours. The solvents were then removed in vacuo and the residue purified by LCMS.

In a cognate manner, benzylamine, ethylamine, and other primary or secondary amines can be substituted for ammonia to yield the corresponding substituted guanidiniums. Products are shown in table 7.

Table 7. Guanidinium products Product Starting Synth. R5* R2 R3 R4 M+H Rt Yield material method (mins) 180 110-20 12 H C A I 937.5 6.31 49 181 111-31 17 H C A I 800.37 5.23 182 110-20 34-b Bn C A I ND ND ND 183 110-20 34-b Et C A I ND ND ND 184 110-20 34-b Me C A I ND ND ND substituents are Hydrogen Benzyl Ethyl or Methyl (Me): Substituents R2-R4 are as found in Table 2, Example 24.

Claims (19)

1. A disaccharide compound of formula I A-d-L-e-B formula I wherein A and B are independently chosen from R6 R6 R7 R4 R7 T R 1 T R1 R4 R2 WR2 R3 R2 R3 T is O; R6 and R7 are hydrogen, or together form a carbonyl oxygen; R1 is hydrogen, -N(Z)Y OZ or SZ wherein; when R1 is N(Z)Y Y is selected from the group consisting of hydrogen or the following; 0 0 0 0 0 II II 0 NIIW OH wherein; Z is selected from hydrogen or X1, Q is selected from hydrogen or W, AMENDED SHEE Ie-lchi6iM PCT/AU2003/000494 Received 05 August 2004 103 W is selected from the group consisting of alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, arylalkyl or heteroarylalkyl of 1 to 20 non- hydrogen atoms, X1 is selected from the group consisting of alkyl, alkenyl, alkynyl, heteroalkyl, acyl, arylacyl, heteroarylacyl, aryl, heteroaryl, arylalkyl or heteroarylalkyl of 1 to 20 non-hydrogen atoms, where R1 is C(Z)Y; Y, is absent or is selected from hydrogen, double bond oxygen to form a carbonyl, or triple bond nitrogen to form a nitrile, Z is absent or is selected from hydrogen or X2, wherein X2 is selected from the group consisting of alkyl, alkenyl, alkynyl, heteroalkyl, aminoalkyl, aminoaryl, aryloxy, alkoxy, heteroaryloxy, aminoaryl, aminoheteroaryl, thioalkyl, thioaryl or thioheteroaryl, acyl, arylacyl, heteroarylacyl, aryl, heteroaryl, arylalkyl or heteroarylalkyl of 1 to 20 non-hydrogen atoms where R1 is OZ or SZ, Z is selected from hydrogen or X3, wherein X3 is selected from the group consisting of alkyl, alkenyl, alkynyl, heteroalkyl, acyl, arylacyl, heteroarylacyl, aryl, heteroaryl, arylalkyl or heteroarylalkyl of 1 to 20 non-hydrogen atoms, The groups R2, R3, R4 and R5 are selected from the group consisting of hydrogen, N 3 OH, OX4, N(Z)Y, wherein N(Z)Y is as defined above or Y is Q S N Q Q Q where Q and W are as defined above, IAMENDED SHE'T lra- qM 104 and X4 is independently selected from the group consisting of alkyl, alkenyl, alkynyl, heteroalkyl, aminoalkyl, aminoaryl, aryloxy, alkoxy, heteroaryloxy, aminoaryl, aminoheteroaryl, alkylcarbamoyl, arylcarbamoyl or cN heteroarylcarbamoyl, acyl, arylacyl, heteroarylacyl, aryl, heteroaryl, arylalkyl or heteroarylalkyl of 1 to 20 non-hydrogen atoms, Sd and e represent the connection points for A and B and replace one 0o of the groups R1, R2, R3, R4, or R5 in each of the groups A and B and form c the connection point for the linker L, d is a covalent bond, e is and L is 0 absent, or Z and Y combine to form a ring of 4 to 10 non-hydrogen atoms; c 10 and with the provisos that: a) any of the groups R1 to R5 on either ring may not combine together to form a cycle, b) when a group is OX4, X4 may not be an acetyl, chloroacetyl, dichloroacetyl, trichloroacetyl, trifluoroacetyl, benzoyl, pivaloyl, or levulinoyl protecting group commonly used in carbohydrate synthesis, c) X4 may not be another carbohydrate ring, a cyclitol ring or contain another carbohydrate ring, d) at least one of R2 to R5 on each ring must be OX4 or N(Z)Y, e) all of the X4 substituents may not be the same, f) when R2 on both rings is N(Z)Y, R1 may not be O-Allyl, and g) the compound of formula I must contain at least 2 and not more than 7 substituents selected from OX4 or N(Z)Y.

2. The compound of claim 1, wherein when R1 is N(Z)Y, W is substituted with at least one moiety 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, substituted or unsubstituted imine, sulfate, sulfonamide, phosphate, phosphoramide, hydrazide, hydroxamate, and hydroxamic acid. PCT/AU2003/000494 Received 05 August 2004 105

3. The compound of claim 1, wherein when R1 is N(Z)Y, Z and Y are combined to form a monocyclic or bicyclic ring structure of 4 to 10 non-hydrogen atoms.

4. The compound of claim 1, wherein X1 is substituted with at least one moiety 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, substituted or unsubstituted imine, sulfate, sulfonamide, phosphate, phosphoramide, hydrazide, hydroxamate, and hydroxamicacid.

5. The compound of claim 1, wherein; when R1 is C(Z)Y; X2 is substituted with at least one moiety 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, substituted or unsubstituted imine, sulfate, sulfonamide, phosphate, phosphoramide, hydrazide, hydroxamate, hydroxamic acid, heteroaryloxy, aminoalkyl, aminoaryl, aminoheteroaryl, thioalkyl, thioaryl and thioheteroaryl.

6. The compound of claim 1, wherein; when R1 is C(Z)Y; Z and Y form a ring structure of 4 to 10 non-hydrogen atoms.

7. The compound of claim 6, wherein the ring structure is substituted by X1 groups.

8. The compound of claim 1, wherein; r- VENDED SHEE IrStMj PCT/AU2003/000494 Received 05 August 2004 106 when R1 is OZ or SZ; X3 is substituted with at least one moiety 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, substituted or unsubstituted imine, sulfate, sulfonamide, phosphate, phosphoramide, hydrazide, hydroxamate, hydroxamic acid, heteroaryloxy, aminoalkyl, aminoaryl, aminoheteroaryl, thioalkyl, thioaryl and thioheteroaryl.

9. The compound of claim 1, wherein; when R1 is OZ or SZ; X4 is substituted with at least one moiety 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, substituted or unsubstituted imine, sulfate, sulfonamide, phosphate, phosphoramide, hydrazide, hydroxamate, and hydroxamic acid. The compound of claim 1, wherein Z and Y are combined to form a ring structure of 4 to 10 non-hydrogen atoms.

11. The compound of claim 10, wherein the ring structure is substituted with X1 groups.

12. The compound of claim 1, wherein in group A, T is oxygen, group A is a pyranose ring, and the linker, d-L-e, is a glycosidic linkage formed between the anomeric position R1 of group A, and any position R1 to R5 of group B, such that the d is L is absent, and e is a covalent bond AMENDED SH EET Irs-wV-t PCT/AU2003/000494 Received 05 August 2004 107

13. The compound of claim 12, wherein the structure of formula 1 is R7 R6 R6 R7 R4 2R2 R2 R4 R3 R3 IIa

14. The compound of claim 12, wherein the structure of formula 1 is R7 R6 R7 R6 T R1 S R4 R2 R4 R2 R3 R3 R 3 Ib The compound of claim 12, wherein the structure of formula 1 is R6 R7 R7 R6 R1 T 0 O R4 R4' Y R2 R3 R3

16. The compound of claim 12, wherein the structure of formula 1 is R6 R7 R1 T R2 R4 R7 R6 R4 R2 R3 IId -lreT~M 8 PCT/AU2003/000494 Received 05 August 2004 108

17. The compound of claim 12, wherein the structure of formula 1 is R7 R6 T R1 R7 R6 O R2 R3 R4 R2 R3 IIe

18. The compound of claim 12, wherein the structure of formula 1 is R6 R7 R4 R7 R6T T R4 R2 R4 R2 R3 R3 HIf The compound of claim 12, wherein the structure of formula 1 is The compound of claim 12, wherein the structure of formula 1 is R6 R4-- R7 TRI A~FNFDSHF-e lra -IjelDWEw 109 The compound of claim 12, wherein the structure of formula 1 is

22. A method of synthesizing a disaccharide compound of claim 1 comprising reacting compound A and compound B in solution.

23. A method of combinatorial synthesis of compounds of claim 1 comprising the step of immobilizing a compound of group B onto a support through at least one of the functionalized positions R1 to

24. The method of claim 23, wherein the support is selected from the group consisting of derivatised polystyrene, tentagel, wang resin, MBHA resin, aminomethylpolystyrene, rink amide resin DOX-mpeg, and polyethylene glycol. ALCHEMIA LTD April 2007