Multivalent Oligosaccharides Field of the Invention

[0001] The present invention relates to novel glycoclusters that are anti-microbial agents for use in the treatment of gastric and intestinal diseases. The present invention further provides for a method for the synthesis of these compounds and for the use of the compounds as therapeutic agents.

Background to the Invention

[0002] Oligosaccharides and proteins, heterogeneously coated on cell surfaces, provide a complex environment where a variety of functions take place. These include cell-cell interactions involved in the immune response, inflammation, cell signalling and infection by pathogens.

[0003] The design, synthesis, and development of carbohydrate- and glycoconjugate-based drug molecules that interfere with the binding events at cell surfaces is a potential emerging therapeutic area. Helicobacter pylori infects over half of the world's population and thought to be a leading cause of chronic gastritis, peptic ulcer, and gastric cancer. Campylobacter jejuni is another enteric and gastric bacteria which is the most common cause of bacterial diarrhea.

[0004] Both of these microbes as well as many other pathogens adhere to mucosa by binding blood group antigen-related and other carbohydrates expressed on epithelial cells. Breast feeding protects infants against bacterial infections from these microbes, which has been shown to be linked to the presence of large quantities of antigen oligosaccharides in human milk (Newburg, D. S.; Ruiz-Palacios, G. M.; Morrow, A. L. Annu. Rev. Nutr. 2005, 25, 37-58).

[0005] The glycans found in human milk function as soluble receptors that inhibit pathogens from adhering to their target receptors on the mucosal surface of the host gastrointestinal tract. However, monovalent carbohydrates typically bind to their putative receptors with low affinity. Multivalent oligosaccharides or glycoclusters have in some cases improved ability to inhibit protein-carbohydrate interactions because of glycoside cluster effect. There are a number of plausible mechanisms to explain why glycoclusters are more active, and this includes cases where cross-linking of glycan binding receptors can occur.

[0006] Notwithstanding the state of the art there remains a need for novel therapeutics that can be utilised in the treatment of gastrointestinal disorders.

Summary of the Invention

[0007] The present invention provides for novel multivalent oligosaccharides for use in the treatment of gastrointestinal (Gl) disorders. As used herein the term multivalent oligosaccharide refers to a glycoside (or glycosylamine) in which a plurality of oligosaccharides are bonded to a spacer or linker moiety.

[0008] The oligosaccharide of the present invention for use in the treatment of a Gl disorder may be of the general formula (la), a pharmaceutically acceptable salt thereof, an ester thereof, a solvate thereof or a hydrate thereof:

(la)

wherein:

each R ⁴ is independently selected from the group consisting of H, C-i-C-io aliphatic, CH ₂ C(0)NHCH ₂ C(0)OH, CH ₂ C(O)NHCH ₂ C(O)O(CrCi ₀ aliphatic), and CH ₂ C(0)NH(C Cio aliphatic);

each A is absent, or A, N and R ⁴ together define a C ₂ -C ₆ aliphatic or aromatic heterocycle;

B is selected from C=0, C=S, C(0)-C(0), C ₁ -C ₁ 0 aliphatic optionally interrupted by O, S, or N, C ₃ -C ₂₀ cycloaliphatic, C ₂ -C ₂₀ heterocyclic aliphatic, C ₅ -C ₂₀ aryl, C ₂ -C ₂₀ heteroaryl, and combinations thereof;

each D is an oligosaccharide moiety comprising lactose; and

n is 2 - 20 such that the value of n will not exceed the valence of moiety B.

[0009] n may be 2 - 6.

[0010] The oligosaccharide may be of the general formula (lb), a pharmaceutically acceptable salt thereof, an ester thereof, a solvate thereof or a hydrate thereof:

(lb)

wherein:

each R ¹ is independently selected from the group consisting of OH, a hexose monosaccharide, a 6-deoxy-hexose monosaccharide, a pentose monosaccharide, and a methyl pentose monosaccharide;

each R ² is independently selected from the group consisting of OH, NHAc, and NHC(0)R ³ ; each R ³ is independently selected from C1-C10 aliphatic optionally interrupted by O, S or N, C3-C20 cycloaliphatic, C ₂ -C ₂ o heterocyclic aliphatic, C ₅ -C ₂ o aryl, C ₂ -C ₂ o heteroaryl, and combinations thereof;

each R ⁴ is independently selected from the group consisting of H, C1-C10 aliphatic, CH ₂ C(0)NHCH ₂ C(0)OH, CH ₂ C(0)N HCH ₂ C(0)0(C Cio aliphatic), and CH ₂ C(0)NH(C C10 aliphatic);

each A is absent, or A, N and R ⁴ together define a C ₂ -C ₆ aliphatic or aromatic heterocycle;

B is selected from C=0, C=S, C(0)-C(0), C1-C10 aliphatic optionally interrupted by O, S, or N, C ₃ -C ₂₀ cycloaliphatic, C ₂ -C ₂₀ heterocyclic aliphatic, C ₅ -C ₂₀ aryl, C ₂ -C ₂₀ heteroaryl, and combinations thereof;

each R ⁵ is H, or may define a C ₂ -C ₂₀ aliphatic bridge together with R ⁵ or R ⁶ of an adjacent oligosaccharide;

each R ⁶ is H, or may define a C ₂ -C ₂₀ aliphatic bridge together with R ⁵ or R ⁶ of an adjacent oligosaccharide; and

n is 2 - 20 such that the value of n will not exceed the valence of moiety B.

[0011] When R ⁶ defines a C ₂ -C ₂₀ aliphatic bridge together with R ⁵ or R ⁶ of an adjacent oligosaccharide it is to be construed as providing a structure as follows; for example, where n = 2 (a similar construction applies to the same definition for R ⁵ ):

[0012] n may be 2 - 6. B may be selected from C1-C10 aliphatic, C1-C10 aliphatic ether, C1-C5 aliphatic thioether, C ₆ -Ci ₀ aryl, and combinations thereof.

[0013] R ¹ may selected from the group consisting of OH , L-fucose, L-rhamnose, and L- arabinose. R ¹ may be OH or L-fucose. R ⁴ may be CH ₂ C(O)N H(Ci-Ci ₀ aliphatic).

[0014] In one embodiment, when each R ¹ and R ² are OH , each R ⁴ is not H .

[0015] An oligosaccharide may be of the general formula (Ic), a pharmaceutically acceptable salt thereof, an ester thereof, a solvate thereof or a hydrate thereof:

(lc)

wherein:

each R ¹ is independently selected from the group consisting of OH, a hexose monosaccharide, a 6-deoxy-hexose monosaccharide, a pentose monosaccharide, and a methyl pentose monosaccharide;

each R ² is independently selected from the group consisting of OH, NHAc, and NHC(0)R ³ ; each R ³ is independently selected from C1-C10 aliphatic optionally interrupted by O, S, or N, C3-C20 cycloaliphatic, C ₂ -C ₂ o heterocyclic aliphatic, C ₅ -C ₂ o aryl, C ₂ -C ₂ o heteroaryl, and combinations thereof;

each R ⁴ is independently selected from the group consisting of H, C1-C10 aliphatic, CH ₂ C(0)NHCH ₂ C(0)OH, CH ₂ C(0)NHCH ₂ C(0)0(C Cio aliphatic), and CH ₂ C(0)NH(C C10 aliphatic);

each A is absent, or A, N and R ⁴ together define a C ₂ -C ₆ aliphatic or aromatic heterocycle;

B is selected from C=0, C=S, C(0)-C(0), C1-C10 aliphatic optionally interrupted by O, S, or N, C ₃ -C ₂₀ cycloaliphatic, C ₂ -C ₂₀ heterocyclic aliphatic, C ₅ -C ₂₀ aryl, C ₂ -C ₂₀ heteroaryl, and combinations thereof; and

n is 2 - 20 such that the value of n will not exceed the valence of moiety B.

[0016] n may be 2 - 6. B may be selected from C1-C10 aliphatic, C1-C10 aliphatic ether, C1-C5 aliphatic thioether, C ₆ -Ci ₀ aryl, and combinations thereof.

[0017] R ¹ may selected from the group consisting of OH, L-fucose, L-rhamnose, and L- arabinose. R ¹ may be OH or L-fucose. R ⁴ may be CH ₂ C(O)NH(Ci-Ci ₀ aliphatic).

[0018] In one embodiment, when each R ¹ and R ² are OH, each R ⁴ is not H.

[0019] Each R ¹ may be independently selected from the group consisting of OH, a hexose monosaccharide, a 6-deoxy-hexose monosaccharide, a pentose monosaccharide, and a methyl pentose monosaccharide; each R ² may be OH; each R ⁴ may be independently selected from the group consisting of H, CH ₂ C(0)NHCH ₂ C(0)0(C Cio aliphatic), and CH ₂ C(O)NH(C Ci ₀ aliphatic); each A may be absent, B may be selected from C(0)-C(0), C1-C10 aliphatic optionally interrupted by O, S, or N, C ₃ -C ₂₀ cycloaliphatic, C ₂ -C ₂₀ heterocyclic aliphatic, C ₅ -C ₂₀ aryl, C ₂ -C ₂₀ heteroaryl, and combinations thereof; and n is 2 - 6 such that the value of n will not exceed the valence of moiety B.

wherein linkage of the nitrogen of the oligosaccharide unit to B is illustrated by the squiggle bond.

[0021] The oligosaccharide may be of the general formula (Id), a pharmaceutically acceptable salt thereof, an ester thereof, a solvate thereof or a hydrate thereof:

(Id)

wherein:

each R ¹ is independently selected from the group consisting of OH , a hexose monosaccharide, a 6-deoxy-hexose monosaccharide, a pentose monosaccharide, and a methyl pentose monosaccharide;

each R ² is independently selected from the group consisting of OH, NHAc, and NHC(0)R ³ ; each R ³ is independently selected from C1-C1 ₀ aliphatic optionally interrupted by O, S or N , C3-C20 cycloaliphatic, C ₂ -C ₂ o heterocyclic aliphatic, C ₅ -C ₂ o aryl, C ₂ -C ₂ o heteroaryl, and combinations thereof;

each R ⁴ is independently selected from the group consisting of H, C1-C1 ₀ aliphatic, CH ₂ C(0)NHCH ₂ C(0)OH, CH ₂ C(0)NHCH ₂ C(0)0(C Cio aliphatic), and CH ₂ C(0)NH(C C10 aliphatic); and

R ⁷ is absent or is selected from C1-C1 ₀ aliphatic optionally interrupted by O, S, or N , C ₃ -C ₂₀ cycloaliphatic, C ₂ -C ₂₀ heterocyclic aliphatic, C ₅ -C ₂₀ aryl, C ₂ -C ₂₀ heteroaryl, and combinations thereof.

[0022] In one embodiment, when each R ¹ and R ² are OH, each R ⁴ is not H. [0023] Each R ¹ may be independently selected from the group consisting of OH , a hexose monosaccharide, a 6-deoxy-hexose monosaccharide, a pentose monosaccharide, and a methyl pentose monosaccharide; each R ² may be OH ; each R ⁴ may be independently selected from the group consisting of H , CH ₂ C(O)N HCH ₂ C(O)O(Ci-Ci ₀ aliphatic), and CH ₂ C(0)N H(C Cio aliphatic); R ⁷ may be absent or may be selected from C-I-C-IO aliphatic optionally interrupted by O, S, or N , C3-C20 cycloaliphatic, C ₂ -C ₂ o heterocyclic aliphatic, C ₅ -C ₂₀ aryl, C ₂ -C ₂₀ heteroaryl, and combinations thereof.

[0024] R ⁷ may be absent or selected from C1-C10 aliphatic, C1-C10 aliphatic ether, C1-C5 aliphatic thioether, C ₆ -Ci ₀ aryl, and combinations thereof.

[0025] R ¹ may selected from the group consisting of OH , L-fucose, L-rhamnose, and L- arabinose. R ¹ may be OH or L-fucose.

[0026] R ⁴ may be CH ₂ C(O)N H(C Ci ₀ aliphatic).

[0027] The Gl disorder may be associated with Campylobacter jejuni or Helicobacter pylori.

[0028] The Gl disorder may be selected from the group consisting of gastric cancer, mucosa- associated lymphoid tissue (MALT) lymphoma, peptic ulcers, duodenal ulcers, chronic gastritis, non-ulcer dyspepsia, lymphocytic gastritis, granulomatous gastritis, gastric outlet obstruction, gastric hyperplastic polyps, Menetrier's disease, watery or bloody diarrhoea, abdominal cramps, traveller's diarrhoea, bacteraemia, immunoproliferative small intestinal disease, and

combinations thereof.

[0029] The oligosaccharides of the present invention may also find use in the treatment of extra-gastric disorders caused by bacterial gastrointestinal pathogens, for example

Campylobacter jejuni or Helicobacter pylori. Such extra-gastric disorders may be selected from idiopathic thrombocytopenic purpura, iron deficiency anemia, chronic fatigue syndrome, chronic idiopathic urticaria, reactive arthritis, typhoid-like syndrome, Guillain-Barre syndrome, meningitis, myocarditis, myopericarditis, and acute atrial fibrillation.

[0030] The present invention also provides for a method of treating a gastrointestinal disorder in a patient in need thereof, comprising administering a pharmaceutically acceptable amount of an oligosaccharide according to the present invention to the patient.

[0031 ] In a further aspect, the multivalent oligosaccharide of the present invention as defined supra may find use in the reduction and/or control of bacterial gastrointestinal pathogen carriage in animals. The multivalent oligosaccharide of the present invention as defined supra may find use in the reduction and/or control of Campylobacter jejuni or Helicobacter pylori carriage in animals. The animal may be a farm animal. The farm animal may be poultry. The poultry may be chicken.

[0032] This may be particularly advantageous in preventing human infection with such bacteria through the consumption of contaminated food. [0033] In yet a further aspect, the present invention provides for an animal feed composition comprising a multivalent oligosaccharide of the present invention as defined supra. The animal feed composition may be chicken feed.

[0034] In yet a further aspect, the present invention provides for a compound of the general formula (lb):

(lb)

wherein:

each R ¹ is independently selected from the group consisting of OH, a hexose monosaccharide, a 6-deoxy-hexose monosaccharide, a pentose monosaccharide, and a methyl pentose monosaccharide;

each R ² is independently selected from the group consisting of OH, NHAc, and NHC(0)R ³ ; each R ³ is independently selected from C-I-C-IO aliphatic optionally interrupted by O, S or N , C3-C20 cycloaliphatic, C ₂ -C ₂ o heterocyclic aliphatic, C ₅ -C ₂ o aryl, C ₂ -C ₂ o heteroaryl, and combinations thereof;

each R ⁴ is independently selected from the group consisting of H , C1-C1 ₀ aliphatic, CH ₂ C(0)NHCH ₂ C(0)OH, CH ₂ C(0)NHCH ₂ C(0)0(C Cio aliphatic), and CH ₂ C(0)NH(C C10 aliphatic);

each A is absent, or A, N and R ⁴ together define a C ₂ -C ₆ aliphatic or aromatic heterocycle;

B is selected from C=0, C=S, C(0)-C(0), C1-C1 ₀ aliphatic optionally interrupted by O, S, or N, C ₃ -C ₂₀ cycloaliphatic, C ₂ -C ₂₀ heterocyclic aliphatic, C ₅ -C ₂₀ aryl, C ₂ -C ₂₀ heteroaryl, and combinations thereof;

each R ⁵ is H, or may define a C ₂ -Ci ₀ aliphatic bridge together with R ⁵ or R ⁶ of an adjacent oligosaccharide;

each R ⁶ is H, or may define a C ₂ -Ci ₀ aliphatic bridge together with R ⁵ or R ⁶ of an adjacent oligosaccharide; and

n is 2 - 20 such that the value of n will not exceed the valence of moiety B,

provided that:

i) when n = 2, each R ¹ = R ² = OH, each R ⁵ = R ⁶ = H , each A is absent, and each R ⁴ is H or CH ₂ C(0)NHCH ₂ C0 ₂ Me ii) when n = 2, each R ¹ = R ² = OH, each R ⁵ = R ⁶ = H, and each A, N and R ⁴ together define a triazole

[0035] n may be 2 - 6. B may be selected from C1-C10 aliphatic, C1-C10 aliphatic ether, C1-C5 aliphatic thioether, C ₆ -Ci ₀ aryl, and combinations thereof.

[0036] R ¹ may selected from the group consisting of OH , L-fucose, L-rhamnose, and L- arabinose. R ¹ may be OH or L-fucose. R ⁴ may be CH ₂ C(O)N H(Ci-Ci ₀ aliphatic).

[0037] In one embodiment, when each R ¹ and R ² are OH , each R ⁴ is not H .

[0038] The compound of the present invention may be of the general formula (lc), a

pharmaceutically acceptable salt thereof, an ester thereof, a solvate thereof or a hydrate thereof:

(lc)

wherein:

each R ¹ is independently selected from the group consisting of OH, a hexose monosaccharide, a 6-deoxy-hexose monosaccharide, a pentose monosaccharide, and a methyl pentose monosaccharide;

each R ² is independently selected from the group consisting of OH, NHAc, and NHC(0)R ³ ; each R ³ is independently selected from C1-C10 aliphatic optionally interrupted by O, S, or N, C3-C20 cycloaliphatic, C ₂ -C ₂ o heterocyclic aliphatic, C ₅ -C ₂ o aryl, C ₂ -C ₂ o heteroaryl, and combinations thereof;

each R ⁴ is independently selected from the group consisting of H, C1-C10 aliphatic, CH ₂ C(0)NHCH ₂ C(0)OH, CH ₂ C(0)NHCH ₂ C(0)0(C Cio aliphatic), and CH ₂ C(0)NH(C C10 aliphatic);

each A is absent, or A, N and R ⁴ together define a C ₂ -C ₆ aliphatic or aromatic heterocycle; B is selected from C=0, C=S, C(0)-C(0), C1-C10 aliphatic optionally interrupted by O, S, or N, C3-C20 cycloaliphatic, C ₂ -C ₂ o heterocyclic aliphatic, C ₅ -C ₂ o aryl, C ₂ -C ₂ o heteroaryl, and combinations thereof; and

n is 2 - 20 such that the value of n will not exceed the valence of moiety B,

provided that:

i) when n = 2, each R ¹ = R ² = OH, each A is absent, and each R ⁴ is H or

CH ₂ C(0)NHCH ₂ C0 ₂ Me

B is not = 2, each R ¹ R ² = OH, and each A, N and R ⁴ together define a triazole

[0039] n may be 2 - 6. B may be selected from C1-C10 aliphatic, C1-C10 aliphatic ether, C1-C5 aliphatic thioether, C ₆ -Ci ₀ aryl, and combinations thereof.

[0040] R ¹ may selected from the group consisting of OH , L-fucose, L-rhamnose, and L- arabinose. R ¹ may be OH or L-fucose. R ⁴ may be CH ₂ C(O)N H(Ci-Ci ₀ aliphatic).

[0041] In one embodiment, when each R ¹ and R ² are OH , each R ⁴ is not H .

[0042] Each R ¹ may be independently selected from the group consisting of OH , a hexose monosaccharide, a 6-deoxy-hexose monosaccharide, a pentose monosaccharide, and a methyl pentose monosaccharide; each R ² may be OH ; each R ⁴ may be independently selected from the group consisting of H , CH ₂ C(0)N HCH ₂ C(0)0(C Cio aliphatic), and CH ₂ C(O)N H(C Ci ₀ aliphatic); each A may be absent, B may be selected from C(0)-C(0), C1-C10 aliphatic optionally interrupted by O, S, or N , C ₃ -C ₂₀ cycloaliphatic, C ₂ -C ₂₀ heterocyclic aliphatic, C ₅ -C ₂₀ aryl, C ₂ -C ₂₀ heteroaryl, and combinations thereof; and n is 2 - 6 such that the value of n will not exceed the valence of moiety B.

[0043] Moiety B of the compound of the present invention may be selected from the group consisting of:

wherein linkage of the nitrogen of the oligosaccharide unit to B is illustrated by the squiggle bond.

[0044] The compound may be of the general formula (Id), a pharmaceutically acceptable salt thereof, an ester thereof, a solvate thereof or a hydrate thereof:

(Id)

wherein:

each R ¹ is independently selected from the group consisting of OH , a hexose monosaccharide, a 6-deoxy-hexose monosaccharide, a pentose monosaccharide, and a methyl pentose monosaccharide;

each R ² is independently selected from the group consisting of OH, NHAc, and NHC(0)R ³ ; each R ³ is independently selected from C1-C1 ₀ aliphatic optionally interrupted by O, S, or N , C3-C20 cycloaliphatic, C ₂ -C ₂ o heterocyclic aliphatic, C ₅ -C ₂ o aryl, C ₂ -C ₂ o heteroaryl, and combinations thereof;

each R ⁴ is independently selected from the group consisting of H, C1-C1 ₀ aliphatic, CH ₂ C(0)NHCH ₂ C(0)OH, CH ₂ C(0)NHCH ₂ C(0)0(C Cio aliphatic), and CH ₂ C(0)NH(C C10 aliphatic);

R ⁷ is absent or is selected from C1-C1 ₀ aliphatic optionally interrupted by O, S, or N, C ₃ -C ₂₀ cycloaliphatic, C ₂ -C ₂₀ heterocyclic aliphatic, C ₅ -C ₂₀ aryl, C ₂ -C ₂₀ heteroaryl, and combinations thereof,

provided that:

i) when each R ⁴ is H or CH ₂ C(0)NHCH ₂ C0 ₂ Me

[0045] Each R ¹ may be independently selected from the group consisting of OH, a hexose monosaccharide, a 6-deoxy-hexose monosaccharide, a pentose monosaccharide, and a methyl pentose monosaccharide; each R ² may be OH; each R ⁴ may be independently selected from the group consisting of H, CH ₂ C(O)NHCH ₂ C(O)O(Ci-Ci ₀ aliphatic), and CH ₂ C(0)NH(C Cio aliphatic); R ⁷ may be absent or may be selected from C1-C1 ₀ aliphatic optionally interrupted by O, S, or N, C3-C20 cycloaliphatic, C ₂ -C ₂ o heterocyclic aliphatic, C ₅ -C ₂₀ aryl, C ₂ -C ₂₀ heteroaryl, and combinations thereof.

[0046] R ⁷ of the compound of the present invention may be absent or selected from C1-C1 ₀ aliphatic, C1-C1 ₀ aliphatic ether, C1-C5 aliphatic thioether, C ₆ -Ci ₀ aryl, and combinations thereof.

[0047] R ¹ of the compound of the present invention may be selected from the group consisting of OH, L-fucose, L-rhamnose, and L-arabinose. R ¹ may be OH or L-fucose.

[0048] R ⁴ of the compound of the present invention may be CH ₂ C(O)NH(Ci-Ci ₀ aliphatic).

[0049] In one embodiment, when each R ¹ and R ² are OH, each R ⁴ is not H.

[0050] In yet a further aspect the present invention provides for a pharmaceutical composition comprising a compound according to the present invention and a pharmaceutically acceptable carrier.

[0051] As used herein, the term C _x -C _y aliphatic refers to linear, branched, saturated and unsaturated hydrocarbon chains comprising C _x -C _y carbon atoms (and includes C _x -C _y alkyl, C _x -C _y alkenyl and C _x -C _y alkynyl). The carbon atoms of the hydrocarbon chain may optionally be substituted one or more times with at least one of a cyano group, a nitro group, a halogen, a Ci- C10 ether, a C1-C10 thioether, a C1-C10 ester, C1-C10 ketone, C1-C10 ketimine, C1-C10 sulfone, d- C10 sulfoxide, a C1-C10 primary amide or a Ci-C ₂₀ secondary amide.

[0052] Similarly, references to C _x -C _y alkyl, C _x -C _y alkenyl and C _x -C _y alkynyl include linear and branched C _x -C _y alkyl, C _x -C _y alkenyl and C _x -C _y alkynyl optionally substituted one or more times with at least one of a cyano group, a nitro group, a halogen, a C1-C1 ₀ ether, a C1-C1 ₀ thioether, a C1-C10 ester, C1-C10 ketone, C1-C10 ketimine, C1-C10 sulfone, C1-C10 sulfoxide, a C1-C10 primary amide or a Ci-C ₂₀ secondary amide.

[0053] As used herein, the term "C _x -C _y cycloaliphatic" refers to unfused, fused, spirocyclic, polycyclic, saturated and unsaturated hydrocarbon rings comprising C _x -C _y carbon atoms (and includes C _x -C _y cycloalkyl, C _x -C _y cycloalkenyl and C _x -C _y cycloalkynyl). The carbon atoms of the hydrocarbon ring may optionally be substituted one or more times with at least one of a cyano group, a nitro group, a halogen, a C1-C10 ether, a C1-C10 thioether, a C1-C10 ester, C1-C10 ketone, C1-C10 ketimine, C1-C10 sulfone, C1-C10 sulfoxide, a C1-C10 primary amide or a C1-C20 secondary amide.

[0054] Similarly, references to C _x -C _y cycloalkyl, C _x -C _y cycloalkenyl and C _x -C _y cycloalkynyl embrace compounds in which the carbon atoms of the rings may optionally substituted one or more times with at least one of a cyano group, a nitro group, a halogen, a C ₁ -C ₁₀ ether, a C ₁ -C ₁₀ thioether, a C1-C10 ester, C1-C10 ketone, C1-C10 ketimine, C1-C10 sulfone, C1-C10 sulfoxide, a C C ₁₀ primary amide or a C ₁ -C ₂₀ secondary amide.

[0055] As used herein, the term aryl/aromatic refers to an aromatic carbocyclic structure which is monocyclic or polycyclic and is unfused or fused. The carbon atoms of the aromatic ring may optionally be substituted one or more times with at least one of a cyano group, a nitro group, a halogen, a C1-C10 ether, a C1-C10 thioether, a C1-C10 ester, C1-C10 ketone, C1-C10 ketimine, d- C10 sulfone, C1-C10 sulfoxide, a C1-C10 primary amide or a C1-C20 secondary amide.

[0056] As used herein, the term heterocycle refers to cyclic compounds having as ring members atoms of at least two different elements. The cyclic compounds may be monocyclic or polycyclic and unfused or fused.

[0057] As used herein, the term heteroaromatic/heteroaryl refers to an aromatic heterocyclic structure having as ring members atoms of at least two different elements. The heterocycle may be monocyclic or polycyclic and unfused or fused. The carbon atoms of the heteroaromatic ring may optionally be substituted one or more times with at least one of a cyano group, a nitro group, a halogen, a C1-C10 ether, a C1-C10 thioether, a C1-C10 ester, C1-C10 ketone, C1-C10 ketimine, C1-C10 sulfone, C1-C10 sulfoxide, a C1-C10 primary amide or a C1-C20 secondary amide.

[0058] The compounds of the present invention may be found or isolated in the form of prodrugs, esters, salts, hydrates or solvates - all of which are embraced by the present invention.

[0059] Where suitable, it will be appreciated that all optional and/or preferred features of one embodiment of the invention may be combined with optional and/or preferred features of another/other embodiment(s) of the invention.

Brief Description of the Drawings

[0060] Additional features and advantages of the present invention are described in, and will be apparent from, the detailed description of the invention and from the drawings in which:

[0061] Figure 1 illustrates the effect of the compounds of the present invention on the binding of C. jejuni to immobilized chicken large intestine mucin; and

[0062] Figure 2 illustrates the ability of compounds of the present invention to bind C. jejuni already bound to chicken large intestine mucin, thereby releasing bound C. jejuni from the chicken large intestine mucin. Detailed Description of the Drawings

[0063] It should be readily apparent to one of ordinary skill in the art that the examples disclosed herein below represent generalised examples only, and that other arrangements and methods capable of reproducing the invention are possible and are embraced by the present invention.

General synthetic routes to glycoclusters

[0064] Synthetic routes to bivalent glycoclusters containing lactose are commenced from lactosyl amine 1 (Scheme 1 ). The Ugi reaction of lactosyl amine 1 in the presence of formaldehyde, methyl isocyanoacetate, and isophthalic acid and subsequent Zemplen deacetylation resulted in the bivalent compound 2. When other dicarboxylic acid such as oxalic acid and 2,6-naphthalenedicarboxylic acid were used instead of terephthalic acid, compounds 3 and 4 were obtained, respectively (Scheme 2). When ie f-butyl isocyanide was reacted with lactosamine 1 , formaldehyde and the appropriate dicarboxylic acid in methanol, the bivalent compounds 5 and 6 were obtained as examples (Scheme 2).

Scheme 1

[0065] As a further illustration of the invention the synthesis of bivalent glycoclusters based on fucosyl lactose are shown (Scheme 3-6). Firstly the synthesis of trisaccharide 9 was achieved via glycosylation of acceptor 8 with fucosyl donor 7 (Wang, Z.; Zhou, L. Y.; El-Boubbou, K.; Ye, X. S.; Huang, X. F. J. Org. C em. 2007, 72, 6409-6420.)- For the synthesis of the acceptor 8 an approach originally described by K. L. Matta and co-workers was used (Abbas, S. A.; Barlow, J. J.; Matta, K. L. Carbohydr. Res. 1981 , 88, 51 -60). Two promoter systems NIS/TfOH and benzensulfinylpiperidine (BSP)/ Tf ₂ 0 have been investigated for the glycosylation to give the trisaccharide 9 in yields of 46% and 70%, respectively. In terms of reaction time and yield, the BSP/Tf ₂ 0 system was substantially better than the NIS/TfOH system for glycosidation. The assignment of configuration to the glycosidic linkage between the fucosyl residue and the lactose residue was assigned based on the size of the coupling constant (Jr,2" = 3.3 Hz) in the 1 H NMR spectrum and the signal for C-1 " occurring at 95.2 ppm in the 13C NMR spectrum. A small amount of β-fucosyl anomer was isolated with the C-1 " chemical shift occurring at 102.6 ppm in the ¹³ C NMR spectrum.

[0066] After debenzoylation of compound 9 with methanolic sodium methoxide, hydrolysis of the acetonide groups in 60% acetic acid at 60 °C provided the tri-O-benzyl trisaccharide 11 which was directly acetylated to afford compound 12. However the glycosidic bond between benzylated fucosyl building block and the acetylated lactosyl building block was very vulnerable to acidic conditions. Direct and indirect methods to introduce an azide at the anomeric position, including the use of TMSN ₃ & SnCI ₄ , 33% HBr in AcOH, and BiBr ₃ /TMSBr (Montero, J. L;

Winum, J. Y.; Leydet, A.; Kamal, M.; Pavia, A. A.; Roque, J. P. Carbohydr. Res. 1997, 297, 175- 180) all led to the cleavage of the fucosidic bond. To address this problem, the benzyl groups in compound 11 were removed by catalytic hydrogenolysis to give the free trisaccharide 14.

Subsequent acetylation followed by reaction of the product with SnCI ₄ and TMSN ₃ and then catalytic hydrogenation gave the amine 17 (Scheme 4) via 15 and 16.

Scheme 3

Scheme 4

[0067] Coupling of 17 with terephthaloyi chloride in the presence of DIPEA and subsequent Zemplen deacetylation gave 19 via 18 (Scheme 5). Since the glycosyl amine 10 also contained a small amount of the oanomer, a mixture of diamides were obtained from the coupling reaction. The bivalent compound 19 was thus obtained after HPLC-based separation of the deacetylated diamide mixture (Scheme 5).

Scheme 5

[0068] The Ugi reaction did not succeed for the peracetylated-fucosyllactosamine 17. It is suggested that the acetylated 2'-fucosyl residue reduces the nucleophilicity of the anomeric amine group, even though the fucose reside is somewhat remote from the amine. To address this problem, the benzyl groups on the fucosyl building block were preserved throughout the synthesis. Hence compound 11 was treated with a saturated solution of ammonium hydrogen carbonate as previously reported (Christiansenbrams, I.; Meldal, M.; Bock, K. J. Chem. Soc, Perkin Trans. 1 1993, 1461 -1471 ) to give a β-D-glycosylamine which was used in the subsequent steps. Thus for temporary amine protection, the fucosyl lactosamine 20 was treated with fluoren-9-lymethosylcarbonyl succinimide ester (Fmoc-OSu) in pyridine and acetylation of the hydroxyl groups was achieved by addition of acetic anhydride to give 21. The Fmoc was deprotected by morpholine in DMF from 21 to afford the fucosyllactosamine 22, which was subsequently reacted with formaldehyde, methyl isocyanoacetate, and terephthalic acid to give 23. Subsequent Zemplen deacetylation and hydrogenolysis resulted in bivalent fucosyllactoside 24 (Scheme 6).

Scheme 6

Experimental details [0069] /V,/V'-Di(/3-D-galactopyranosyl-(1→4)-/3-D-glucopyranose)- /V,/V'-di[(1 - methoxycarbon-yl)methylamino-2-oxoethyl]isophthalamide (2).

[0070] Isophthalic acid (20 mg, 0.12 mmol), lactosyl amine 1 [Leyden, R.; Velasco-Torrijos, T.; Andre, S.; Gouin, S.; Gabius, H. J.; Murphy, P. V. J. Org. C em. 2009, 74, 9010-9026 and Aravind, S.; Park, W. K. C; Brochu, S.; Roy, R. Tetrahedron Lett. 1994, 35, 7739-7742] (152 mg, 0.24 mmol), and formaldehyde (23 μΙ_ of a 37% soln, 0.29 mmol) were suspended in MeOH (5 mL) and the mixture was stirred at rt for 1 h. Methyl isocyanoacetate (23 μΙ_, 0.24 mmol) was then added and the mixture was stirred at rt overnight. The reaction was then heated to 45 °C for 12 h and then solvent was removed under reduced pressure. Chromatography of the residue (CH ₂ CI ₂ -CH ₃ OH, gradient elution, 70:1 to 60:1 to 50:1 ) gave the protected intermediate as a white amorphous solid (129 mg, 65%). Zemplen deacetylation of this intermediate and purification by BioGel P-2 gel column (eluent: H ₂ 0) and C18 reverse column (gradient elution, H ₂ 0 to H ₂ O-CH ₃ OH, 95:5) gave 2 as an interconverting mixture of EE and EZ isomers (3.75:1 ); [a] ²⁰ _D +27.3 (c 0.6, D ₂ 0); ¹ H-NMR (500 MHz, D ₂ 0) data for EE isomer δ 7.72 (s, 1 H), 7.71 (s,

2H), 7.68 - 7.63 (m, 1 H), 4.76 (d, J = 8.8 Hz, 2H, H-1 ), 4.36 (d, J = 7.6 Hz, 2H, H-1 '), 4.31 (d, J = 1 1 .9 Hz, 4H), 4.07 (d, J = 3.5 Hz, 4H), 3.90 (d, J = 12.7 Hz, 2H), 3.85 (d, J = 3.3 Hz, 2H), 3.79 - 3.56 (m, 20H), 3.50-3.42 (m, 6H); selected ¹ H NMR data for EZ isomer δ 5.70 (d, J = 9.5 Hz, 2H), 4.42 (d, J = 7.2 Hz, 2H); ¹³ C NMR (125 MHz, D ₂ 0) δ 174.0 (C), 171 .8 (C), 134.0 (C), 130.0, 129.7, 102.8, 87.2, 77.5, 76.7, 75.3, 74.1 , 72.4, 70.8, 69.7, 68.5 (each CH), 61 .0 (CH ₂ ), 59.9 (CH ₂ ), 52.8( CH ₃ ) , 44.8 (CH ₂ ), 41 .3 (CH ₂ ); HRMS-ESI: calcd for C42H ₆₂ N ₄ 028Na: 1093.3448; Found: 1093.3466.

[0071] yV,yV'-Di(j8-D-galactopyranosyl-(1→4)-j8-D-glucopyranose)- yV,yV'-di[(1 - methoxycarbon-yl) methylamino-2-oxoethyl]oxalamide (3).

[0072] Oxalic acid (7 mg, 0.08 mmol), lactosyl amine 1 (100 mg, 0.16 mmol), and formaldehyde (16 μΙ_ of a 37% soln, 0.19 mmol) were suspended in MeOH (5 mL) and the mixture was stirred at rt for 1 h. Methyl isocyanoacetate (15 μί, 0.16 mmol) was then added and the mixture was stirred at rt overnight. The reaction was then heated to 45 °C for 12 h and then solvent was removed under reduced pressure. Chromatography of the residue (CH ₂ CI ₂ -CH ₃ OH, gradient elution, 80:1 to 60:1 to 50:1 ) gave the protected intermediate as a white amorphous solid (84 mg, 67%). Zemplen deacetylation of this intermediate and purification by BioGel P-2 gel column (eluent: H ₂ 0) and C18 reverse column (elution, H ₂ 0) gave 3 as an interconverting mixture of EE and EZ isomers (3.1 :1 ); [a] ²⁰ _D +30.8 (c 0.5, D ₂ 0); ¹ H-NMR (500 MHz, D ₂ 0) data for EE isomer δ

4.83 (d, J = 8.8 Hz, 2H, H-1 ), 4.33 (d, J = 7.8 Hz, 2H, H-1 '), 4.20 (ABq, J = 16.7 Hz, 4H), 3.99- 3.90 (m, 4H), 3.86 - 3.75 (m, 4H), 3.74 - 3.48 (m, 24H), 3.41 (dd, J = 7.8, 10.0 Hz, 2H, H-2'); selected ¹ H NMR data for EZ isomer δ 5.51 (d, J = 9.4 Hz, 2H); ¹³ C NMR (125 MHz, D ₂ 0) δ 171 .8 (C), 170.5 (C), 165.1 (C), 102.8, 86.4, 77.3, 76.8, 75.3, 74.1 , 72.5, 70.9, 69.4, 68.5 (each CH), 61 .0 (CH ₂ ), 59.8 (CH ₂ ), 52.8 ( CH ₃ ), 43.7 (CH ₂ ), 41 .2 (CH ₂ ); HRMS-ESI: calcd for

C ₃ 6H ₅₈ N ₄ 028Na: 1017.3135; Found: 1017.3108.

[0073] 2,6-[/V,/V'-Di(/3-D-galactopyranosyl-(1→4)-/3-D-glucopyran ose)-/V,/V'-di(1 - methoxycarbo- nyl)methylamino-2-oxoethyl]Naphthalenedicarboxylamide(4).

[0074] 2,6-Naphthalenedicarbox-ylic acid (18 mg, 0.08 mmol), lactosyl amine 1 (100 mg, 0.16 mmol), and formaldehyde (16 μΙ_ of a 37% soln, 0.19 mmol) were suspended in MeOH (5 mL) and the mixture was stirred at rt for 1 h. Methyl isocyanoacetate (15 μΙ_, 0.24 mmol) was then added and the mixture was stirred at rt overnight. The reaction was then heated to 45 °C for 12 h and then solvent was removed under reduced pressure. Chromatography of the residue (CH ₂ CI ₂ -CH ₃ OH, gradient elution, 70:1 to 60:1 to 50:1 ) gave the protected intermediate as a white amorphous solid (89 mg, 66%). Zemplen deacetylation of this intermediate and purification by BioGel P-2 gel column (eluent: H ₂ 0) and C18 reverse column (gradient elution, H ₂ 0 to H ₂ O-CH ₃ OH, 95:5) gave 4 as an interconverting mixture of EE and EZ isomers (4:1 ); [a] ²⁰ _D +36 (c 0.2, D ₂ 0); ¹ H-NMR (500 MHz, D ₂ 0) data for EE isomer δ 8.09 (s, 2H), 8.03 (d, J =

8.5 Hz, 2H), 7.59 (d, J = 8.5 Hz, 2H), 4.74 (d, J = 8.9 Hz, 2H, H-1 ), 4.35 - 4.23 (m, 6H), 4.02 (s, 4H), 3.87 (d, J = 1 1 .9 Hz, 2H), 3.76 (d, J = 3.0 Hz, 2H), 3.72 - 3.45 (m, 20H), 3.39 - 3.23 (m, 6H); selected ¹ H N MR data for EZ isomer δ 5.66 (d, J = 9.0 Hz, 2H); ¹³ C NMR (125 MHz, D ₂ 0) δ 175.0 (C), 171 .93(C), 171 .87 (C), 132.9 (C), 132.4 (C), 129.7, 127.0, 124.7, 102.7, 87.4, 77.5, 76.9, 75.3, 74.1 , 72.4, 70.8, 69.7, 68.4 (each CH), 60.9 (CH ₂ ), 60.0 (CH ₂ ), 52.8 ( CH ₃ ),

44.7(CH ₂ ), 41 .3 (CH ₂ ); HRMS-ESI: calcd for C ₄₆ H ₆₄ N ₄ 0 ₂₈ Na: 1 143.3605; Found: 1 143.3601 .

[0075] yV,yV'-Di(j8-D-galactopyranosyl-(1→4)-j8-D-glucopyranose)- yV,yV'-di[(tert- butylcarbamoyl) methyljterephthalamide (5).

[0076] Terephthalic acid (13.6 mg, 0.08 mmol), lactosyl amine 1 (100 mg, 0.16 mmol), and formaldehyde (16 μΙ_ of a 37% soln, 0.19 mmol) were suspended in MeOH (5 mL) and the mixture was stirred at rt for 1 h. tert-Butyl isocyanide (19 μΙ_, 0.16 mmol) was then added and the mixture was stirred at rt overnight. The reaction was then heated to 45 °C for 10 h and then solvent was removed under reduced pressure. Chromatography of the residue (CH ₂ CI ₂ -CH ₃ OH, gradient elution, 70:1 to 60:1 to 50:1 ) gave the protected intermediate as a white amorphous solid (98 mg, 77%). Zemplen deacetylation of this intermediate and purification by BioGel P-2 gel column (eluent: H ₂ 0) and C18 reverse column (gradient elution, H ₂ 0 to H ₂ 0-CH ₃ OH, 95:5) gave 5 as an interconverting mixture of EE and EZ isomers (4:1 ); [a] ²⁰ _D +28.0 (c 0.1 , D ₂ 0); ¹ H-

NMR (500 MHz, D ₂ 0) data for EE isomer δ 7.56 (s, 4H), 4.69 (d, J = 8.8 Hz, 2H, H-1 ), 4.29 (d, J = 7.8 Hz, 2H, H-1 '), 4.06 (ABq, J = 16.3 Hz, 4H), 3.85 (d, J = 12.5 Hz, 2H), 3.78 (d, J = 3.3 Hz, 2H), 3.70-3.49 (m, 14H), 3.44-3.36 (m, 6H), 1.23 (s, 18H); selected ¹ H N MR data for EZ isomer δ 7.50 (d, J = 7.8 Hz, 2H), 7.48 (d, J = 7.8 Hz, 2H), 5.62 (d, J = 9.3 Hz, 2H), 4.35 (d, J = 7.8 Hz, 2H); ¹³ C NMR (125 MHz, D ₂ 0) δ 174.1 (C), 169.9 (C), 135.9 (C), 127.5, 102.8, 87.2, 77.5, 76.7, 75.3, 74.0, 72.4, 70.8, 69.7, 68.5 (each CH), 61 .0 (CH ₂ ), 59.9 (CH ₂ ), 51 .5 (C), 45.5 (CH ₂ ), 27.6 (CH ₃ ); HRMS-ESI: calcd for C ₄ 4H ₇ oN ₄ 024Na: 1061 .4278; Found: 1061 .4293

[0077] yV,yV'-Di(j8-D-galactopyranosyl-(1→4)-j8-D-glucopyranose)- yV,yV'-di[(tert- butylcarbamoyl) methyljisophthalamide (6).

[0078] Isophthalic acid (13.6 mg, 0.08 mmol), lactosyl amine 1 (100 mg, 0.16 mmol), and formaldehyde (16 μΙ_ of a 37% soln, 0.19 mmol) were suspended in MeOH (5 mL) and the mixture was stirred at rt for 1 h. tert-Butyl isocyanide (19 μΙ_, 0.16 mmol) was then added and the mixture was stirred at rt overnight. The reaction was then heated to 45 °C for 12 h and then solvent was removed under reduced pressure. Chromatography of the residue (CH ₂ CI ₂ -CH ₃ OH, gradient elution, 80:1 to 60:1 to 50:1 ) gave the protected intermediate as a white amorphous solid (101 mg, 79%). Zemplen deacetylation of this intermediate and purification by BioGel P-2 gel column (eluent: H ₂ 0) and C18 reverse column (gradient elution, H ₂ 0 to H ₂ O-CH ₃ OH, 95:5) gave 6 as an interconverting mixture of EE and EZ isomers (4:1 ); [a] ²⁰ _D +22.3 (c 0.27, D ₂ 0); ¹ H-

NMR (500 MHz, D ₂ 0) data for EE isomer δ 7.62 - 7.53 (m, 4H), 4.63 (d, J = 8.9 Hz, 2H, H-1 ), 4.25 (d, J = 7.8 Hz, 2H, H-1 '), 4.02 (ABq, J = 16.4 Hz, 4H), 3.82 - 3.78 (m, 2H), 3.75 (d, J = 3.2 Hz, 2H), 3.68 - 3.45 (m, 14H), 3.42 - 3.26 (m, 6H), 1 .20 (s, 18H); selected ¹ H N MR data for EZ isomer δ 5.57 (d, J = 9.3 Hz, 2H), 4.32 (d, J = 7.3 Hz, 2H), 1 .00 (s, 18H); ¹³ C NMR (125 MHz, D ₂ 0) δ 174.1 (C), 169.9 (C), 134.1 (C), 130.0, 129.5, 102.8, 87.3, 77.5, 76.7, 75.3, 74.1 , 72.4, 70.8, 69.7, 68.5 (each CH), 61 .0 (CH ₂ ), 59.9 (CH ₂ ), 51 .5 (C), 45.7 (CH ₂ ), 27.6 (CH ₃ ); HRMS- ESI: calcd for C ₄₄ H ₇₀ N ₄ O ₂₄ Na: 1061 .4278; Found: 1061 .4257.

[0079] 0-(2,3,4-Tri-0-benzyl)-a-L-fucopyranosyl)-(1→2)-0-(6-0-ben zoyl-3,4-0- isopropylidene-j8-D-galactopyranosyl)-(1→4)-2,3:5,6-di-0-i sopropylidene-D-glucose dimethyl acetal (9).

[0080] Fucosyl donor 7 (1 .26 g, 2.34 mmol), disaccharide building block 8 (1 .30 g, 2.12 mmol), BSP (252 mg, 1.17 mmol) and 4 A MS were stirred in dry CH ₂ CI ₂ (25 mL) at room temperature for 1 h under N ₂ . The reaction mixture was cooled to -78°C, followed by the addition of Tf ₂ 0 (219 μΙ_, 1 .27 mmol) and TTBP (598 mg, 2.34 mmol). Then the temperature was increased gradually from -78 °C to 0 °C within 2h and the mixture was stirred at 0 °C for an additional 4h. The reaction was quenched with triethylamine (3 mL) and diluted with CH ₂ CI ₂ . The reaction mixture was filtered and washed sequentially with sat. aq. Na ₂ S ₂ 0 ₃ , sat. aq. NaHC0 ₃ , H ₂ 0, and brine, dried (Na ₂ S0 ), filtered, and concentrated. The crude mixture was purified by flash column chromatography (PE-EtOAc, gradient elution, 7:1 to 6:1 ) to give the title compound 9 as a white foam (1 .54 g, 66%), R _f 0.5 (PE-EtOAc, 2.5:1 ) and β-fucosyl anomer (0.10 g), α/β = 15:1 . ¹ H- NMR (500 MHz, CDCI ₃ ) δ 8.05 (d, J = 7.5 Hz, 2H), 7.57 (t, J = 7.5 Hz, 1 H), 7.45 (t, J = 7.5 Hz, 2H), 7.40 - 7.24 (m, 15H), 5.60 (d, J = 3.3 Hz, H-1 "), 4.98 (d, J = 1 1 .6 Hz, 1 H), 4.87 (d, J = 1 1 .9 Hz, 1 H), 4.75 - 4.73 (m, 3H), 4.70 ( d, J = 8.0 Hz, 1 H, H-1 '), 4.65 (d, J = 1 1 .6 Hz, 1 H), 4.56 - 4.48 (m, 3H), 4.34 (d, J = 5.9 Hz, 1 H, H-1 ), 4.25 (dd, J = 1 1 .0, 5.8 Hz, 1 H), 4.21 (t, J = 6.0 Hz, 1H), 4.16 (dd, J= 5.6, 2.0 Hz, 1H), 4.10-4.00 (m, 7H), 3.90 (dd, J= 8.3, 6.7 Hz, 1H), 3.75 (dd, J= 7.8, 6.7 Hz, 1H), 3.66 (s, 1H), 3.36 (s, 3H), 3.33 (s, 3H), 1.49 (s, 3H), 1.43 (s, 3H), 1.37 (s, 6H), 1.33 (s,3H), 1.27 (s, 3H), 1.11 (d, J = 6.5Hz, 3H); ¹³ C NMR (125 MHz, CDCI ₃ )5166.3 (C), 139.3 (C), 139.1 (C), 138.9 (C), 133.1, 129.7, 128.4, 128.4, 128.3, 128.1, 128.1, 127.6, 127.4, 127.4, 127.3, 127.3 (each CH), 110.3 (C), 110.0 (C), 108.6 (C), 105.1 (CH, C-1), 101.3 (CH, C- 1'), 95.2 (C-1"), 80.2, 79.2, 78.1, 77.7, 77.5, 76.5, 75.4, 75.0 (each CH), 74.8 (CH ₂ ), 74.1 (CH),

73.8 (CH), 73.2 (CH ₂ ), 72.7 (CH ₂ ), 70.9 (CH), 66.5 (CH), 65.2 (CH ₂ ), 63.8 (CH ₂ ), 56.0 (CH ₃ ),

52.9 (CH ₃ ), 29.7 (CH ₃ ), 27.8 (CH ₃ ), 27.2 (CH ₃ ), 26.9 (CH ₃ ), 26.5 (CH ₃ ), 25.2 (CH ₃ ), 16.9 (CH ₃ ); LRMS (ESI) 1051.5 [M+Na] ⁺ ; HRMS-ESI: calcd for C ₅₇ H ₇₂ 0i ₇ Na: 1051.4667; Found: 1051.4664.

[0081 ] 0-(2,3,4-Tri-0-acetyl-a-L-fucopyranosyl)-(1→2)-0-(3,4,6-tr i-0-acetyl-/3-D- galactopyran osyl)-(1→4)-1,2,3,6-tetra-0-acetyl-a/j8-D-glucopyranose (15).

[0082] A Solution of compound 10 (2.12 g, 2.33 mmol) in 30 mL aq 60% acetic acid was heated for 6h at 60°C. The reaction mixture was then diluted with toluene and concentrated. Then to a solution of the residue in THF/H ₂ 0/AcOH (4:2:1, 14 mL), 10% Pd/C (50 mg) was added. The suspension was stirred under an atmosphere of hydrogen for 2 days at ambient temperature. When the reaction was completed, the mixture was followed to filtered over Celite and concentrated. The residue was co-concentrated with toluene (3 * 30 mL) to remove the acetic acid and water. Then the deprotected trisaccharide was dissolved in pyridine/acetic anhydride (2:1 , 30 mL) which was stirred overnight under an atmosphere of nitrogen at ambient temperature. After which the solvent was removed and the residue was participated by CH ₂ CI ₂ (100 mL) and water (25 mL). The organic phase was washed by water (25 mL 2), dried by Na ₂ S0 ₄ and concentrated. The crude residue was purified by flash chromatography (PE-EtOAc, gradient elution, 4:1 to 3:1) to give the title compound 15 as a white foam (1.67 mg, 79%), R _f 0.35 (PE-EtOAc, 1.5:1), which were mixture of anomers (β/α=1:1); ¹ H NMR (500 MHz, CDCI ₃ ) data for the β anomer: δ 5.69 (d, J = 8.3 Hz, 1 H, H-1) , 5.37 (d, J= 3.5 Hz, 1H, H-1"), 5.31- 5.27 (m, 2H), 5.21 (t, J= 9.5 Hz, 1H), 5.18-5.12 (m, 1H), 5.10 (t, J= 8.3 Hz, 1H), 5.00-4.95 (m, 2H), 4.50-4.38 (m, 3H), 4.27 (dd, J= 12.2, 5.6 Hz, 1H), 4.18-4.06 (m, 2H), 3.91 -3.80 (m, 4H), 2.18-1.97 (10s, 30H), 1.22 (d, J= 6.5 Hz, 3H); ¹ H NMR data for the a anomer: δ 6.30 (d, J= 3.7 Hz, 1H, H-1), 5.41 (t, J= 9.9 Hz, 1H), 5.37 (d, J= 3.5 Hz, 1H, H-1"), 5.31-5.27 (m, 2H), 5.18-5.12 (m, 1H), 5.05 (dd, J= 10.4, 3.7 Hz, 1H), 1H, 5.00 -4.95 (m, 2H), 4.50-4.38 (m, 3H), 4.27 (dd, J= 12.2, 5.6 Hz, 1H), 4.18-4.06 (m, 2H), 3.91 -3.80 (m, 4H), 2.18-1.97 (10s, 30H), .22 (d, J = 6.5 Hz, 3H); mixture ¹³ C NMR (125 MHz, CDCI ₃ ) data for both a and β anomer: δ 171.1, 170.67, 170.65, 170.56, 170.51, 170.33, 170.27, 170.1, 169.98, 169.96, 169.8, 169.7, 169.6, 169.3, 168.8, 168.7 (each C), 100.2, 99.9, 95.7, 73.9, 73.8, 73.7, 73.4, 73.3, 71.7, 71.11, 71.09, 70.9, 70.8, 70.7, 70.3, 69.2, 69.1 , 68.1 , 68.0, 67.5, 67.3, 67.0, 65.0, 64.9 (each CH), 62.1, 61.9, 61.0, 60.8 (each CH ₂ ), 21.1, 21.0, 20.9, 20.8, 20.7, 20.65, 20.6, 20.5, 15.6, 15.4 (each CH ₃ ); Selected ¹³ C NMR data for the a anomer: δ 89.0 (CH, C-1 ); Selected ¹³ C NMR data for the β anomer: δ 91 .5 (CH, C-1 ); LRMS (ESI) 926.3 [M+Na] ⁺ ; HRMS-ESI: calcd for

C ₃ 8H ₅₂ N ₃ 0 ₂₅ Na: 931 .2695; Found: 931 .2686.

[0083] 0-(2,3,4-Tri-0-acetyl-a-L-fucopyranosyl)-(1→2)-0-(3,4,6-tr i-0-acetyl-/3-D- galactopyran osyl)-(1→4)-1 ,2,3,6-tetra-0-acetyl-a/j8-D-glucopyranosyl azide (16).

[0084] Compound 15 (1 .35 g, 1 .49 mmol) was dissolved in CH ₂ CI ₂ (30 ml_, anhydrous) under an atmosphere of N ₂ . To this solution was added TMSN ₃ (0.54 ml_, 4.46 mmol) followed by the dropwise addition of SnCI ₄ (88 μΙ_, 0.74 mmol). After 20h, the reaction was deemed complete by TLC analysis and the solution was diluted with CH ₂ CI ₂ , quenched by the addition of saturated NaHC0 ₃ solution (10 mL) and left stirring for a further 30 min. The biphasic solution was then filtered through Celite and the organic layer extracted, washed with saturated NaHC0 ₃ solution (20 mL x 2), H ₂ 0 (20 mL x 2), dried (Na ₂ S0 ₄ ) and concentrated in vacuo. The crude residue was purified by flash chromatography (PE- EtOAc=3:1 ) to give the title compound 16 as a white foam (1 .26 g, 95%), R _f 0.57 (PE-EtOAc, 1 :1 ); ¹ H N MR (500 MHz, CDCI ₃ ) data for the β anomer: 5 5.38 (d, J = 3.0 Hz, 1 H, H-1 "), 5.30 (dd, J = 12.5, 3.0 Hz, 2H), 5.18 - 5.09 (m, 2H), 4.98 (m, 2H), 4.92 (t, J = 9.5 Hz, 1 H), 4.64 (d, J = 9.0 Hz, 1 H, H-1 ), 4.54 (d, J = 12.0 Hz, 1 H), 4.1 1 -4.37 (m, 1 H), 4.10 (d, J = 7.5 Hz, 1 H, H-1 ')4.29 (dd, J = 12.0, 6.0 Hz, 1 H), 4.15 (dd, J = 1 1 .1 , 6.5 Hz, 1 H), 4.08 (dd, J = 1 1 .1 , 7.0 Hz, 1 H), 3.90 - 3.80 (m, 3H), 3.78 - 3.71 (m, 1 H), 2.16 (d, J = 1 .4 Hz, 6H), 2.12 (s, 3H), 2.1 1 - 2.03 (m, 9H), 2.02 - 1 .95 (m, 9H), 1.21 (d, J = 6.5 Hz, 3H);

Selected ¹ H NMR data for the a anomer: δ 5.35 (d, J = 3.0 Hz, 1 H), 5.21 (t, J = 9.2 Hz, 1 H), 5.10 (d, J = 8.0 Hz, 1 H), 4.86 (t, J = 9.1 Hz, 1 H), 4.63 (d, J = 8.7 Hz, 1 H), 3.83 - 3.66 (m, 1 H). ¹³ C NMR (125 MHz, CDCI) for the β anomer: δ 170.7 (C), 170.6 (C), 170.6 (C), 170.39 (C), 170.1 (C), 170.0 (C), 169.7 (C), 169.3 (C), 100.2 (CH), 95.5 (CH), 88.0 (CH), 75.2 (CH), 73.9 (CH), 73.4 (CH), 71 .8 (CH), 71 .3 (CH), 71 .0 (CH), 70.8 (CH), 70.5 (CH), 68.0 (CH), 67.4 (CH), 67.0 (CH), 64.9 (CH), 62.0 (CH ₂ ), 60.8 (CH ₂ ), 20.8 (CH ₃ ), 20.7 (CH ₃ ), 20.67 (CH ₃ ), 20.66 (CH ₃ ), 20.64 (CH ₃ ), 20.63 (CH ₃ ), 20.61 (CH ₃ ), 20.59 (CH ₃ ), 15.6 (CH ₃ ); Selected ¹³ C NMR data for the a anomer: δ 101 .1 (CH), 87.7 (CH), 75.8 (CH), 74.8 (CH), 72.5 (CH), 71.0 (CH), 70.9 (CH), 70.8 (CH), 69.1 (CH), 66.6, 61 .7 (CH ₂ ), 60.4 (CH ₂ ), 14.2 (CH ₃ ); LRMS (ESI) 909.3(M+NH ₄ ⁺ ), 914.3 [M+Na] ⁺ ; HRMS-ESI: calcd for C ₃₆ H ₄₉ N ₃ 0 ₂₃ Na: 914.2655; Found: 914.2621 .

[0085] 0-(2,3,4-Tri-0-acetyl-a-L-fucopyranosyl)-(1→2)-0-(3,4,6-tr i-0-acetyl-/3-D- galactopyran osyl)-(1→4)-1 ,2,3,6-tetra-0-acetyl-a/j8-D-glucopyranosyl amine (17).

[0086] Compound 16 (1 .2 g, 1.35 mmol) was dissolved in EtOAc to which was added 10% Pd- C (0. 10 g). The reaction was left to stir overnight under a positive pressure of H ₂ . Then the reaction was diluted with EtOAc and filtered through Celite. Removal of the solvent gave 17 as a white foam (1 .16 g, quantitative yield, mixture of anomers, β:α = 10:1 ) which was used without further purification; R _f 0.24, (EtOAc-CH ₂ CI ₂ , 9:1 ); ¹ H-NMR (500 MHz, CDCI ₃ ) data for the β anomer: 5 5.38 (d, J = 3.9 Hz, 1 H), 5.29 (dd, J = 10.3, 2.8 Hz, 2H), 5.16-5.13 (m, 2H), 5.01 - 4.94 (m, 2H), 4.78 (t, J = 9.4 Hz, 1 H), 4.48 (dd, J = 1 1 .9, 1 .8 Hz, 1 H), 4.45 - 4.38 (m, 2H), 4.24 (dd, J = 12.0, 6.1 Hz, 1 H), 4.17-4.1 1 (m, 3H), 4.1 1 - 4.04 (m, 1 H), 3.88 - 3.82 (m, 2H), 3.78 (t, J = 9.5 Hz, 1 H), 3.66 - 3.59 (m, 1 H), 2.16 (s, 3H), 2.15 (s, 3H), 2.12 (s, 3H), 2.08 (s, 3H), 2.07 (s, 3H), 2.06 (s, 3H), 2.00 (s, 3H), 1.98 (, 3H), 1.97 (s, 3H), 1.22 (d, J = 6.5 H, 3H). Selected ¹ H NMR data for the a anomer: δ 5.35 (d, J = 3.0 Hz, 1 H), 5.23 (t, J = 9.4 Hz, 1 H), 4.73 (t, J = 9.4 Hz, 1 H), 4.28 (dd, J = 12.0, 5.5 Hz, 1 H); ¹³ C NMR (125 MHz, CDCI) for the β anomer: δ 170.7, 170.7, 170.6, 170.34, 170.30, 170.1 , 170.03, 170.01 , 169.67 (each C), 100.2, 95.5, 85.0, 74.7, 74.0, 73.5, 72.4, 72.0, 71 .4, 71 .1 , 70.7, 68.1 , 67.5, 67.1 , 64.8 (each CH), 62.7 (CH ₂ ), 60.9 (CH ₂ ), 20.9, 20.9, 20.8, 20.67, 20.66, 20.65, 20.63, 20.60, 20.59, 15.53 (each CH ₃ ); LRMS (ESI) 866.2 [M+H] ⁺ ; HRMS-ESI: calcd for CseHssNiOzs: 866.2921 ; Found: 866.2930.

[0087] W,yV'-Di[0-2,3,4-tri-0-acetyl-a-L-fucopyranosyl-(1→2)-0-(3 ,4,6-tri-0-acetyl-j8-D- galactopyranosyl)-(1→4)-1 ,2,3,6-tetra-0-acetyl-j8-D-glucopyranosyl)]terephthalamide (18).

[0088] The lactosyl amine 17 (1 12 mg, 0.13 mmol) and DIPEA (0.16 ml_, 0.19 mmol) in dry THF (10 mL) were added dropwise at r.t. into freshly recrystallized terephthaloyl chloride (26 mg, 0.065 mmol) in dry THF. The reaction was stirred overnight and then the solvent was removed. Chromatography of the residue (EtOAc-PE, gradient elution, 1 :1 to 2:1 ) gave a white amorphous solid (105 mg, 87% including trace anomer); R _f 0.75 (PE-EtOAc, 1 :4); [α] ²⁰ _β -77.2 (c

1 .1 , CHCI ₃ ); ¹ H-NMR (500 MHz, CDCI ₃ ) δ 7.82 (s, 4H), 7.03 (d, J = 9.0 Hz, 2H), 5.43 - 5.34 (m,

6H), 5.34 (d, J = 2.6 Hz, 2H), 5.30 (d, J = 3.6 Hz, 2H), 5.19 (dd, J = 1 1 .0, 3.2 Hz, 2H), 5.03 - 4.95 (m, 6H), 4.55 - 4.37 (m, 6H), 4.36 - 4.25 (m, 2H), 4.18 (dd, J = 1 1 .1 , 6.6 Hz, 2H), 4.08 (dd, J = 1 1 .1 , 6.8 Hz, 2H), 3.93 - 3.78 (m, 8H), 2.17 (s, 6H), 2.14 (s, 6H), 2.13 (s, 6H), 2.1 1 (s, 6H), 2.10 (s, 6H), 2.05 (s, 6H), 2.00 (s, 6H), 1 .98 (s, 6H), 1 .98 (s, 6H), 1 .25 (d, J = 6.7 Hz, 6H); ¹³ C NMR (125 MHz, CDCI ₃ ) δ 171 .7, 170.7, 170.6, 170.5, 170.3, 170.2, 170.0, 169.7, 169.6, 165.9, 136.2 (each C), 127.7, 99.9, 95.7, 79.0, 74.9, 74.1 , 73.4, 71 .7, 71 .4, 71 .1 , 71 .0, 70.8, 68.1 , 67.4, 67.0, 65.0 (each CH), 62.3 (CH ₂ ), 61 .0 (CH ₂ ), 20.9, 20.8, 20.8, 20.7, 20.7, 20.7, 20.6, 20.6, 15.6 (each CH ₃ ); LRMS (ESI) 1883.4 [M+ Na] ⁺ ; HRMS-ESI: calcd for CsoH^NzCUs Na: 1883.5656; Found: 1883.5609.

[0089] /V,/V'-Di(a-L-fucopyranosyl-(1→2)-j8-D-galactopyranosyl-(1 →4)-j8-D-glucopyranosyl) terephthalamide (19).

[0090] Compound 18 (40 mg, 21 .5 μηηοΙ, including trace anomer) was dissolved in methanol (5 mL) to which a catalytic amount of NaOMe (0.1 mL of a 0.2 M solution in MeOH) was added and the resulting solution was stirred for 1 h at room temperature. Amberlite IR-120 (plus) was added to neutralize pH = 7, after which the resin was removed by filtration and washed with water. The solvent removed on a rotary evaporator to give 6 as a yellow solid (23 mg, 97%). The compound was further purified by Semipreparative HPLC (isocratic elution with water-CH ₃ CN, 97:3, flow rate 10 mL/min, t _R = 22 min) and after lyophilisation was a white solid; [a] ²⁰ _D -41 .5 (c

0.325, D ₂ 0); ¹ H-NMR (500 MHz, D ₂ 0) δ 7.82 (s, 4H), 5.20 (d, J = 2.5 Hz, 2H, H-1 "), 5.09 (d, J = 9.2 Hz, 2H, H-1 ), 4.44 (d, J = 7.6 Hz, 2H, H-1 '), 4.14 (dd, J = 13.1 , 6.4 Hz, 2H), 3.85 (d, J = 1 1 .5 Hz, 2H), 3.77-3.49 (m, 28H), 1 .15 (d, J = 6.5 Hz, 6H); ¹³ C NMR (125 MHz, D ₂ 0) δ 171 .0 (C), 136.4 (C), 127.9, 100.2, 99.3, 79.8, 77.0, 76.2, 75.2, 75.1 , 73.5, 71 .6, 71 .5, 69.6, 69.1 , 68.1 , 66.9 (each CH), 61 .1 (CH ₂ ), 59.9 (CH ₂ ), 15.2 (CH ₃ ); HRMS-ESI : calcd for C ₄ 4H ₆₈ N ₂ 0 ₃ o Na:

1 127.3755; Found: 1 127.3728.

[0091 ] 0-(2,3,4-Tri-0-benzyl-a-L-fucopyranosyl)-(1→2)-0-(3,4,6-tr i-0-acetyl-/3-D- galactopyran osyl)-(1→4)-1 ,2,3,6-tetra-0-acetyl-j8-D-glucopyranosyl amine (22).

[0092] Compound 1 1 (450 mg, 0.59 mmol) was dissolved in CH ₃ OH/H ₂ 0 (30 mL, 2: 1 ) and treated with an excess of ammonium hydrogencarbonate for 7 days at 30 °C. The solution was concentrated to half its original volume and diluted with water. This procedure was repeated twice and then water was removed. The resulting residue was co-concentrated with toluene (25 mL x 3). Then the residue was suspended in pyridine (25 mL), Fmoc-OSu (21 1 mg, 0.65 mmol) was added and the mixture was stirred overnight at ambient temperature. Acetic anhydride (12 mL) was added and the mixture was stirred for another 12h. After concentration and co- evaporation with toluene, the residue was purified by chromatography (EtOAc- PE, gradient elution, 5: 1 to 2: 1 ) to give colourless oil 21 . Then the obtained oil 21 was dissolved in DMF (5 mL) and morpholine (5 mL) was added. After 25 min the solution was diluted with toluene (10 mL) and concentrated. The residue was purified by chromatography (CH ₂ CI ₂ -CH ₃ OH, gradient elution, 80: 1 to 70: 1 ) to give colourless oil 22 (237 mg, 40% for three steps), R _f 0.33 (CH ₂ CI ₂ - CH ₃ OH, 40: 1 ). ¹ H NMR (500 MHz, CDCI ₃ ) δ 7.34-7.26 (m, 15H), 5.32 (d, J = 2.5 Hz, 1 H, H-4'), 5.30 (s, 1 H), 5.22 (d, J = 3.4 Hz, 1 H, H-1 "), 5.13 (t, J = 9.6 Hz, 1 H, H-3), 5.05 (dd, J = 10.0, 3.4 Hz, 1 H, H-3'), 4.97 (d, J = 1 1 .6 Hz, 1 H), 4.75 (t, J = 9.5 Hz, 1 H, H-2), 4.72 - 4.61 (m, 5H), 4.49 (d, J = 1 1 .5 Hz, 1 H), 4.38 (d, J = 7.6 Hz, 1 H, H-1 '), 4.17-4.02 (m, 6H), 3.89 - 3.74 (m, 4H), 3.69 (s, 1 H), 3.57 - 3.46 (m, 1 H), 2.09-2.07 (4s, 12H), 2.04 (s, 3H), 1 .79 (s, 3H), 1 .19 (d, J = 6.4 Hz, 3H); ¹³ C NMR (125 MHz, CDCI ₃ ): 170.4, 170.35, 170.31 , 170.0, 169.9, 169.8, 138.7, 138.6, 138.4 (each C), 128.4, 128.3, 128.2, 127.7, 127.6, 127.54, 127.50, 127.3, 100.9, 97.9, 84.9, 79.4, 77.7, 76.3 (each CH), 74.8 (CH ₂ ), 74.0 (CH), 73.8 (CH), 73.4 (CH ₂ ), 73.1 (CH), 73.0 (CH), 72.9 (CH ₂ ), 72.6 (CH), 72.1 (CH), 70.5(CH), 67.22 (CH), 67.20 (CH), 62.5 (CH ₂ ), 61 .0 (CH ₂ ), 20.91 , 20.88, 20.8, 20.7, 20.6, 16.5 (CH ₃ ). HRMS-ESI : calcd for C ₅ i H ₆₄ NO ₂₀ Na: 1032.3841 ; Found: 1032.3837.

Biological Testing

[0093] The effect of some synthetic compounds on the binding of C. jejuni to purified chicken large intestine (LI) mucin

[0094] 5 μg aliquots of purified chicken large intestine mucin were immobilised onto methanol activated Polyvinylidene Fluoride (PVDF) membrane. Each test compound was suspended in dH ₂ 0 to a final concentration of 1 mg/ml. C. jejuni was resuspended in either Phosphate- buffered Saline (PBS), 10μg ml chicken large intestine mucin as a positive control or one of the test compounds of interest to a final concentration of 4x10 ⁵ cfu/ml and then overlayed onto the membrane. C.jejuni binding was detected using polyclonal antibodies (see Figure 1 ).

[0095] Compounds wgn-2 and compound 19 inhibited the ability of C. jejuni to bind to immobilised chicken large intestine mucin at a concentration of 1 mg/ml (see Figure 1 ). It is envisaged that compounds 2, 3, 4, 5, 6, and 24 will exhibit improved inhibition of the binding between C. jejuni and mucin.

Effect of compounds of the present invention on immobilised chicken mucin C. jejuni bound thereto

[0096] 5 μg aliquots of purified chicken large intestine mucin were immobilised onto a methanol activated Polyvinylidene Fluoride (PVDF) membrane. C. jejuni was resuspended in Phosphate- buffered Saline (PBS) to a final concentration of 4x10 ⁵ cfu/ml and then overlayed onto the membrane for 1 h. The membrane was washed twice with PBS and then one of PBS, 1 mg/ml of wgn-2, or 1 mg/ml of compound 19 was added to each slot and the blot was incubated for a further hour. Bound C. jejuni was detected using polyclonal antibodies (see Figure 2).

[0097] Compounds wgn-2 and 19 resulted in a reduction in C. jejuni bound to immobilised chicken large intestine mucin (see Figure 2).

[0098] The words "comprises/comprising" and the words "having/including" when used herein with reference to the present invention are used to specify the presence of stated features, integers, steps or components but do not preclude the presence or addition of one or more other features, integers, steps, components or groups thereof.

[0099] It is appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable subcombination.