WO1993000908A1 - Elam-1 receptor ligands as diagnostic compounds - Google Patents

Elam-1 receptor ligands as diagnostic compounds Download PDF

Info

Publication number
WO1993000908A1
WO1993000908A1 PCT/GB1992/001216 GB9201216W WO9300908A1 WO 1993000908 A1 WO1993000908 A1 WO 1993000908A1 GB 9201216 W GB9201216 W GB 9201216W WO 9300908 A1 WO9300908 A1 WO 9300908A1
Authority
WO
WIPO (PCT)
Prior art keywords
group
compound
gal
elam
glc
Prior art date
Application number
PCT/GB1992/001216
Other languages
French (fr)
Inventor
Alan William John Stuttle
Original Assignee
Antisoma Limited
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Antisoma Limited filed Critical Antisoma Limited
Publication of WO1993000908A1 publication Critical patent/WO1993000908A1/en

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/0491—Sugars, nucleosides, nucleotides, oligonucleotides, nucleic acids, e.g. DNA, RNA, nucleic acid aptamers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2123/00—Preparations for testing in vivo

Definitions

  • the present invention relates to diagnostic compounds, in other words compounds which can be used in the diagnosis and investigation of disease.
  • One aspect of the invention provides a compound comprising a first portion adapted to bind specifically the ELAM-1 receptor and a second portion capable of being detected in
  • the first portion comprises a saccharide structure of Formula (I) :
  • X 1 is GIcNac or Glc
  • R 1 is a hydrogen atom forming an OH group at the 3 position on the galactose group or is a NeuAc(2 ⁇ 3) group
  • R 2 is hydrogen forming an OH group at the 3 position on the X 1 group or is a Fuc(l ⁇ 3) group thereon.
  • the sugars may be a or ⁇ but ⁇ is preferred for all except NeuAc and Fuc, for which a is preferred.
  • the saccharide structure above can be an isolated di-, tri- or tetra-saccharide or it can form part of a larger molecule such as a polysaccharide, mucopolysaccharide or glycolipid.
  • the compound of the invention has a molecular weight of less than 50 kD, more preferably less than 30 kD, 20 kD, 10 kD or 5 kD.
  • the compound is an oligo- or polysaccharide or a glycolipid, and preferably less than 50% of the molecular weight (more preferably less than 20% or 5% and most preferably none) is accounted for by amino acid moieties.
  • the saccharide forms part of a molecule which comprises a saccharide structure of Formula (II) :
  • X 1 and R 1 are as defined above, n is 1 or 2, X 2 is (l ⁇ 3)Gal(l ⁇ 4)X 3 (R 3 ) , R 2 and the or each R 3 are independently a hydrogen atom forming an OH group on the 3 position of the X 3 ring or a Fuc(l ⁇ 3) group thereon, and the or each X 3 is in each case GIcNac or Glc.
  • X 1 is GlcNac/3.
  • compound (1) is a compound of Formula (I) in which R 1 is hydrogen, X 1 is GlcNac/3, R 2 is Fuc ⁇ (l ⁇ 3) , n is two, the first X 3 is GIcNac/? and the second is Glc ⁇ , the first R 3 is Fuc ⁇ (l ⁇ 3) and the second is hydrogen, and the core saccharide structure is joined (l ⁇ l) to a Cer group.
  • Compound (2) is a compound of Formula (II) in which R 1 is NeuAc ⁇ (2 ⁇ 3), X 1 is GlcNacj3, R 2 is Fuc ⁇ (l ⁇ 3), n is two, the first X 3 is GlcNacj3, the first R 3 is Fuc ⁇ (l ⁇ 3), the second X 3 is Glcj3, the second R 3 is hydrogen and the core saccharide group is joined (l ⁇ l) to a Cer group.
  • the Cer (ceramide) group renders the compound suitable for location in a cell or liposome membrane but, in the context of the present invention, we prefer to replace the ceramide group with a peptide group of 1-10 amino acids.
  • Such a peptide group is preferably lipophilic; for example, greater than 60% of it may be made up of methionine, leucine, isoleucine, valine, tryptophan, phenylalanine, alanine, glycine, cysteine, threonine, tyrosine, histidine or lysine residues.
  • the structure can be an isolated oligosaccharide of from four to ten sugar rings, as indicated by Formula (II) , or it can form part of a larger molecule, for example a polysaccharide, mucopolysaccharide or glycolipid.
  • the saccharide structure of Formula (I) or (II) preferably has up to 10 (for example 1, 5, or 8) amino acids at one end or both ends of it.
  • the invention embraces the use of neutral and ⁇ (2,3) sialylated, ⁇ (l,3) fucosylated lactosaminoglycans (eg NeuAc ⁇ 2 ⁇ 3-Ga l ⁇ 1 ⁇ 4 (Fuc ⁇ l-»3)GIcNac, abbreviated to sLex, and Gal3l ⁇ 4 (Fuc ⁇ l ⁇ 3)GIcNac, abbreviated to Lex) for the purpose of detecting and diagnosing inflammatory/infectious/- neoplastic lesions by virtue of localising on activated endothelial cells.
  • neutral and ⁇ (2,3) sialylated, ⁇ (l,3) fucosylated lactosaminoglycans eg NeuAc ⁇ 2 ⁇ 3-Ga l ⁇ 1 ⁇ 4 (Fuc ⁇ l-»3)GIcNac, abbreviated to sLex, and Gal3l ⁇ 4 (Fuc ⁇ l ⁇ 3)GIcNac, abbreviated to
  • the molecules may be labelled with ⁇ -emitting isotopes for detection by gamma camera, nmr enhancers for detection by magnetic resonance imaging or with positron-emitting isotopes for detection by positron emission tomography (PET) .
  • PET positron emission tomography
  • Glc is D-glucose
  • GIcNac is N-acetyl-D-glucosamine
  • Gal is D- galactose
  • Fuc is L-fucose
  • NeuAc is N-acetylneuraminic acid.
  • Cer is a ceramide group.
  • These carbohydrates are commercially available, or may be custom synthesised, and can be labelled by a variety of procedures, for example: esterification of hydroxyl bonds to form a structure capable of complexing directly with a radioisotope or nmr enhancer; reaction of the carbohydrate with amino diacetic acid (IDA) in organic solvent to form an N-linked glycoside derivative which would be capable of complexing with a radioisotope via the nitrogen and oxygen atoms of the IDA group; or coupling of the carbohydrate to amino acids which may be labelled directly (eg cysteine, tyrosine) or labelled via a bifunctional chelating agent (eg lysine) .
  • IDA amino diacetic acid
  • the second portion usually comprises a radioactive atom for scintigraphic studies, for example technetium 99m ( 99m Tc) , iodine-123 ( 123 I) or indium-Ill ( ⁇ In), or a label for nuclear magnetic resonance (nmr) imaging (also known as magnetic resonance imaging, mri) , such as gadolinium, manganese or iron, or a positron-emitting isotope such as iodine-124, fluorine-19, carbon-13 , nitrogen-15 or oxygen- 17.
  • the second portion is part of the amino acid extension at one or both ends of the saccharide referred to above.
  • the compounds of the invention may be prepared in a sterile, non-pyrogenic medium and injected into the bloodstream of a patient at a dose to be determined in the usual way by the physician or radiologist. After a sufficient period " for a good balance to have been reached between (i) specificity of binding to activated endothelium compared to non-specific distribution and (ii) total amount of compound on activated endothelium, the compound is imaged in a conventional way, according to the nature of the second portion of the compound.
  • Phenyl 3,4, 6-tri- ⁇ -acetyl-2-deoxy-2-phthalimido-l-thio-/3- D-glucopyranoside (1).
  • To a stirred solution of 1,3,4,6- tetra-0-acetyl-2-deoxy-2-phthalimido-)3-D-glucopyranose (l:20g) in CH 2 Cl 2 (250 ml) is added trimethyl(phenylthio) - silane (25 ml) and tri ethylsilyl triflate (21 ml) .
  • Stirring is continued for 72 h at room temperature.
  • After neutralisation with triethylamine the reaction mixture is diluted with CHC1 3 , washed with water, dried, and concentrated. The residue is applied to a column of silica gel and eluted with 1 30-40% gradient of EtOAc in hexane. Evaporation gives an amorphous solid which on crystallisation from MeOH furnishes the title
  • Phenyl 4,6-0-benzylidene-2-deoxy-2-phthalimido-l-thio-/3-D- glucopyranoside (2) A solution of l(llg) in 0.02M sodium methoxide (200 ml) is stirred for 3 h at room temperature. The base is neutralised with Amberlite IR-120 (H ⁇ ) cation- exchange resin, the resin suspension is filtered, and the filtrate concentrated to give a solid residue. To a stirred solution of this solid in N,N-dimethylformamide (75 ml) is then added 4-toluenesulfonic acid monohydrate (0.2 g) and ⁇ , ⁇ -dimethoxytoluene (15 ml) .
  • Phenyl 6- ⁇ -benzyl-2-deoxy-2-phthalimido-l-thio- ⁇ -D- glucopyranoside (3) To a cold (0°C, bath) , stirred mixture of 2 (7.5 g) , sodium cyanoborohydride (10.5 g) , and powdered 3A molecular sieves (10 g) in dry oxolane (75 ml) is added, dropwise, a saturated solution of HC1 in ether
  • the later fractions contain the pure unreacted compound 4 (0.8 g) .
  • a solution of compound 7 (0.65 g) in 0.02 M sodium methoxide in MeOH (20 ml) is stirred for 4 h at room temperature.
  • the base is neutralised with Amberlite IR-120 (H " ) cation-exchange resin, the resin suspension is filtered, and the filtrate concentrated.
  • a mixture of 7 (0.3 g) and 10% Pd-C (0.5 g) in glacial acetic acid (30 ml) is shaken under hydrogen at -345 kPa for 2 days at room temperature.
  • the suspension is filtered through a bed of Celite, the solids are thoroughly washed with glacial acetic acid, and the combined filtrate and washings are then concentrated under reduced pressure.
  • the crude product is applied to a column of silica gel and eluted with 5:4:1 CHCl 3 -MeOH-water.
  • the fractions corresponding to 8 are concentrated and lyophilised to give an a
  • the disaccharide /3-D-Gal- ⁇ (1-4) -D-Glc (lactose) is synthesised and modified by attachment of an amine group and subsequent condensation with a peptide containing one or more cysteine residues or a single cysteine residue.
  • Reaction of the amino lactoside with F-Moc protected cysteine results in the production of a wide variety of side chains and is therefore not the preferred method of synthesis. However, this may only present a problem in liquid phase reactions.
  • the peptide or amino acid residue may be coupled to the lactoside using conventional solid phase peptide synthesis techniques.
  • An alternative approach is to attach the carbohydrate to a solid phase, extend the carbohydrate by four carbon atoms, convert the NH 2 group to COOH and make the active ester with succinimide.
  • Example 2 the formation of j3-lactosides of the ⁇ - Galp(l-4) - ⁇ -Glc-OR type
  • Such a lactoside forms part of the ultimate pentasaccharide Gal/3 (1-4 ) -[L-Fuc ⁇ (1-3) ] -GlcNAc/3 (1-3) Gal/3 (1-4) -Glc/3-OR where R is a group capable of coordinating 99 ⁇ n Tc.
  • Group R specifically may be a cysteine-rich peptide, linked gl cosidically to the glucose unit through a spacer arm.
  • the choice of starting material for the carbohydrate was ⁇ - D-lactose, Galp/3(1-4)-Glcj ⁇ -OH(13) . This disaccharide was to be efficiently converted into 2-aminoethyl-lactoside (18) ( Figure 1) .
  • Lactose was first converted to ⁇ - acetobromolactose (14) by established procedure in 76% yield 10 . Displacement of the bromide with chloroethanol using lead carbonate as catalyst gave the required chloroethyl lactoside (15) in good yield”. The product was subsequently treated with sodium azide in dimethylformamide as solvent and heated to 90°C over two hours. The R f value (the movement made by a compound on a thin layer chromatography plate) was the same for the newly formed azidoethyl lactoside (16) as for the chloride.
  • the amine moiety could then be condensed with the carboxyl terminus of a cysteine-rich peptide or a suitably protected cysteine amino acid.
  • Our initial exploration was therefore to determine if any difficulties would be encountered in the synthesis of 2-aminoethyl lactoside (18) . Therefore, de-acetylation of azide (16) was achieved by stirring with sodium methoxide solution. Adjustment of pH to neutral [Amberlite resin- 120(H)] and evaporation to dryness provided the deprotected azide (17)". Reduction of the azido to amino functionality was achieved by brief hydrogenation using 5% Palladium on carbon as catalyst 10 .
  • the active ester of the cysteine amino acid was formed using DCC and N-hydroxysuccinimide.
  • the isolated active ester could then be coupled with the amine in dichloro ethane with the minimum of by-products formed.
  • the active ester (25) of the HO-Cys(tBu) -Fmoc was prepared by cooling of solution of the amino acid and N- hydroxysuccinimide (24) in dry tetrahydrofuran to 0°C. Addition of DCC at this temperature followed and the mixture was allowed to stand at 4°C for 18 hours. The white precipitate of N,N-dicyclohexylurea was filtered off and the filtrate evaporated under reduced pressure to give a white solid in 84% yield ( Figure 3) .
  • Example 2 As an alternative strategy to the final deprotection of the glycoa ino acid in Example 2 involves a lengthening of the linker arm and a reversal of the carboxy terminus of the amino acid for the amino terminus, and similarly for the lactoside involes reaction of the amino lactoside with succinic anhydride (27) ( Figure 5) .
  • the active ester of the carboxy lactoside can then be formed and coupled with the amino terminus of the amino acid or peptide. This also provides a way of adding the carbohydrate portion to the cysteine-rich peptide in the solid phase.

Abstract

(Radio)labelled ligands which bind to the ELAM-1 receptor expressed on activated endothelium are used to aid the diagnosis or imaging of inflammation and neoplastic lesions. Such ligands include compounds of formula (I): R1-Gal(1←4)X1(R2) wherein X1 is GlcNac or Glc, R1 is a hydrogen atom forming an OH group at the 3 position on the galactose group or is a NeuAc(2←3) group and R2 is hydrogen forming an OH group at the 3 position on the X1 group or is a Fuc(1←3) group thereon.

Description

ELAM-1 RECEPTOR LIGANDS AS DIAGNOSTIC COMPOUNDS
The present invention relates to diagnostic compounds, in other words compounds which can be used in the diagnosis and investigation of disease.
Background and Prior Art
During disease states such as inflammation and infection it is widely documented that leucocytes will marginate out of the circulation through the vessel wall and localise within the affected area. It is now becoming apparent that cell adhesion to activated endothelium is a prerequisite for leucocyte margination and that this is partially mediated by the endothelial-leucocyte adhesion molecule ELAM-1. A recent paper by Lowe et al (1990 Cell 63, 475-484) suggests that the ELAM-1 receptor recognises carbohydrates characterised by neutral (Lex) and sialylated (sLex) Lewis x antigens expressed on the leucocyte membrane. Lowe et al
speculated that ligands for other members of the LEC- CAM/SELECTIN family of adhesion molecules, once they had been identified, might be useful in therapy.
Current nuclear medicine techniques for the diagnosis and/or localisation of inflammatory and/or infectious lesions rely on targeting the radioisotope to granulocytes. This is usually achieved by separation of granulocytes from other blood formed elements in vitro , followed by radiolabelling via a lipophilic chelate. Anti-granulocyte monoclonal antibodies have been used for imaging purposes with limited success. Recently, human immunoglobulin has been launched for imaging of inflammatory lesions, and is thought to act by targeting Fc receptors on the granulocyte membrane.
Phillips et al (1990 Science 250, 1130-1131) showed that ELAM-1 mediates cell adhesion by recognition of a carbohydrate ligand, namely sialyl-Lex, and speculated that blocking of the ELAM-1-mediated cell adhesion could be of therapeutic value in controlling inflammation and metastasis.
Summary of the Invention
One aspect of the invention provides a compound comprising a first portion adapted to bind specifically the ELAM-1 receptor and a second portion capable of being detected in
vivo by remote sensing techniques when the compound is bound to the said receptor.
Preferably the first portion comprises a saccharide structure of Formula (I) :
(I) RI-Gal(l→4)X1(R:i)
wherein X1 is GIcNac or Glc, R1 is a hydrogen atom forming an OH group at the 3 position on the galactose group or is a NeuAc(2→3) group and R2 is hydrogen forming an OH group at the 3 position on the X1 group or is a Fuc(l→3) group thereon.
The sugars (Fuc, Gal, NeuAc, GlcNAc and Glc) (throughout this specification) may be a or β but β is preferred for all except NeuAc and Fuc, for which a is preferred. The saccharide structure above can be an isolated di-, tri- or tetra-saccharide or it can form part of a larger molecule such as a polysaccharide, mucopolysaccharide or glycolipid. Preferably, the compound of the invention has a molecular weight of less than 50 kD, more preferably less than 30 kD, 20 kD, 10 kD or 5 kD. Preferably, the compound is an oligo- or polysaccharide or a glycolipid, and preferably less than 50% of the molecular weight (more preferably less than 20% or 5% and most preferably none) is accounted for by amino acid moieties.
Preferably the saccharide forms part of a molecule which comprises a saccharide structure of Formula (II) :
(II) R1-Gal(l→4)X1(R2)-(X2)n
wherein X1 and R1 are as defined above, n is 1 or 2, X2 is (l→3)Gal(l→4)X3(R3) , R2 and the or each R3 are independently a hydrogen atom forming an OH group on the 3 position of the X3 ring or a Fuc(l→3) group thereon, and the or each X3 is in each case GIcNac or Glc.
Preferably, X1 is GlcNac/3.
Particular examples of compounds including saccharide structures of Formula (I) or (II) include two from the Phillips et al paper referred to above:
(1) Galj3(1→4) [Fucα(1→3) ]GlcNac/3(1→3)Galj3(1→4) [Fucα(1→3) ] GlcNac-j3(1→3)Galβ(1→4)GlcjS(l→)Cer, and
(2) NeuAcα(2→3) Gal/3(1→4) [Fucα(1→3) ]GlcNaqβ(1→3)Gal/3(1→4) - [Fucα(l-3) ]GlcNac/3(1→3)Gal/3(1→4) Glcβ(l→)Cer
Thus, compound (1) is a compound of Formula (I) in which R1 is hydrogen, X1 is GlcNac/3, R2 is Fucα(l→3) , n is two, the first X3 is GIcNac/? and the second is Glcβ, the first R3 is Fucα(l→3) and the second is hydrogen, and the core saccharide structure is joined (l→l) to a Cer group. Compound (2) is a compound of Formula (II) in which R1 is NeuAcα(2→3), X1 is GlcNacj3, R2 is Fucα(l→3), n is two, the first X3 is GlcNacj3, the first R3 is Fucα(l→3), the second X3 is Glcj3, the second R3 is hydrogen and the core saccharide group is joined (l→l) to a Cer group. The Cer (ceramide) group renders the compound suitable for location in a cell or liposome membrane but, in the context of the present invention, we prefer to replace the ceramide group with a peptide group of 1-10 amino acids. Such a peptide group is preferably lipophilic; for example, greater than 60% of it may be made up of methionine, leucine, isoleucine, valine, tryptophan, phenylalanine, alanine, glycine, cysteine, threonine, tyrosine, histidine or lysine residues.
The structure can be an isolated oligosaccharide of from four to ten sugar rings, as indicated by Formula (II) , or it can form part of a larger molecule, for example a polysaccharide, mucopolysaccharide or glycolipid. Thus, the saccharide structure of Formula (I) or (II) preferably has up to 10 (for example 1, 5, or 8) amino acids at one end or both ends of it.
Thus, the invention embraces the use of neutral and α(2,3) sialylated, α(l,3) fucosylated lactosaminoglycans (eg NeuAcα2→3-Ga lβ1→4 (Fucαl-»3)GIcNac, abbreviated to sLex, and Gal3l→4 (Fucαl→3)GIcNac, abbreviated to Lex) for the purpose of detecting and diagnosing inflammatory/infectious/- neoplastic lesions by virtue of localising on activated endothelial cells. For this purpose the molecules may be labelled with γ-emitting isotopes for detection by gamma camera, nmr enhancers for detection by magnetic resonance imaging or with positron-emitting isotopes for detection by positron emission tomography (PET) .
The sugar abbreviations used herein are conventional: Glc is D-glucose, GIcNac is N-acetyl-D-glucosamine, Gal is D- galactose, Fuc is L-fucose and NeuAc is N-acetylneuraminic acid. Cer is a ceramide group.
These carbohydrates are commercially available, or may be custom synthesised, and can be labelled by a variety of procedures, for example: esterification of hydroxyl bonds to form a structure capable of complexing directly with a radioisotope or nmr enhancer; reaction of the carbohydrate with amino diacetic acid (IDA) in organic solvent to form an N-linked glycoside derivative which would be capable of complexing with a radioisotope via the nitrogen and oxygen atoms of the IDA group; or coupling of the carbohydrate to amino acids which may be labelled directly (eg cysteine, tyrosine) or labelled via a bifunctional chelating agent (eg lysine) .
The second portion usually comprises a radioactive atom for scintigraphic studies, for example technetium 99m (99mTc) , iodine-123 (123I) or indium-Ill (ιπIn), or a label for nuclear magnetic resonance (nmr) imaging (also known as magnetic resonance imaging, mri) , such as gadolinium, manganese or iron, or a positron-emitting isotope such as iodine-124, fluorine-19, carbon-13 , nitrogen-15 or oxygen- 17. Conveniently, the second portion is part of the amino acid extension at one or both ends of the saccharide referred to above.
The compounds of the invention may be prepared in a sterile, non-pyrogenic medium and injected into the bloodstream of a patient at a dose to be determined in the usual way by the physician or radiologist. After a sufficient period "for a good balance to have been reached between (i) specificity of binding to activated endothelium compared to non-specific distribution and (ii) total amount of compound on activated endothelium, the compound is imaged in a conventional way, according to the nature of the second portion of the compound.
The invention will now be illustrated by reference to the following non-limiting example.
Example 1
General Methods. Solutions in organic solvents are generally dried with anhydr. Na2S04. Dichloroethane, dichloromethane, and N,N-dimethylformamide are dried over 4A molecular sieves.
Phenyl 3,4, 6-tri-θ-acetyl-2-deoxy-2-phthalimido-l-thio-/3- D-glucopyranoside (1). To a stirred solution of 1,3,4,6- tetra-0-acetyl-2-deoxy-2-phthalimido-)3-D-glucopyranose (l:20g) in CH2Cl2 (250 ml) is added trimethyl(phenylthio) - silane (25 ml) and tri ethylsilyl triflate (21 ml) . Stirring is continued for 72 h at room temperature. After neutralisation with triethylamine the reaction mixture is diluted with CHC13, washed with water, dried, and concentrated. The residue is applied to a column of silica gel and eluted with 1 30-40% gradient of EtOAc in hexane. Evaporation gives an amorphous solid which on crystallisation from MeOH furnishes the title compound.
Phenyl 4,6-0-benzylidene-2-deoxy-2-phthalimido-l-thio-/3-D- glucopyranoside (2). A solution of l(llg) in 0.02M sodium methoxide (200 ml) is stirred for 3 h at room temperature. The base is neutralised with Amberlite IR-120 (H~) cation- exchange resin, the resin suspension is filtered, and the filtrate concentrated to give a solid residue. To a stirred solution of this solid in N,N-dimethylformamide (75 ml) is then added 4-toluenesulfonic acid monohydrate (0.2 g) and α,α-dimethoxytoluene (15 ml) . The stirring is continued for 16 h at room temperature. The acid is neutralised with a little triethylamine and the solution concentrated under reduced pressue. The residue is purified on a column of silica gel using a 25-30% gradient of EtOAc in hexane to furnish 2.
Phenyl 6-θ-benzyl-2-deoxy-2-phthalimido-l-thio-β-D- glucopyranoside (3) . To a cold (0°C, bath) , stirred mixture of 2 (7.5 g) , sodium cyanoborohydride (10.5 g) , and powdered 3A molecular sieves (10 g) in dry oxolane (75 ml) is added, dropwise, a saturated solution of HC1 in ether
(45 ml), and stirring is continued for 20 min. Tic (4:1 CHC13 acetone) reveals the disappearance of 2 and the presence of a major product migrating more slowly than 2. A trace of a still slower-migrating contaminant (presumably from the complete cleavage of the acetal group of 2) is also evident. The mixture is diluted with CHC13, and the solids are filtered through Celite and washed with CHC13. The combined filtrate and washings are washed with cold water, cold sat. NaHC03 and water, dried, and concentrated under reduced pressure. The residue is purified on a column of silica gel eluted with 1:1 hexane-EtOAc. The fractions corresponding to the proudct 3 are concentrated to give an amorphous solid.
Phenyl O-(2,3, ,6-tetra-0-acetyl-/3-D-galactopyranosyl)- (l→4)-6-0-benzyl-2-deoxy-2-phthalimido-l-thio-/3-D- glucopyranoside (5). A solution of 3 (4.2 g, 8.5 mmol) and 2, 3 ,4 , 6-tetra-O-acetyl-β-D-galactopyranosyl fluoride (5.3 g, 15.1 mmol) in 5:1 CH2Cl2-toluene (84 ml) is stirred for 0.5 h at -15°C with 4A molecular sieves (8 g) , protected from light and moisture under an argon atmosphere. A mixture of stannous chloride (1.75 g, 9.3 mmol) and silver trifluoromethanesulfonate (2.45 g, 9.6 mmol) is then added, and the reaction mixture is allowed to gradually warm to room temperature, with stirring continued for an additional 2 h. Examination by tic (1:1 hexane-EtOAc) shows the presence of one major spot migrating faster than the starting material, along with minor products. The mixture is filtered through Celite and the solids are thoroughly washed with CHC13. The combined filtrate and washings are washed with sat. aq. NAHC03, filtered through Celite to remove precipitated inorganic material, washed with water, dried, and concentrated under reduced pressure. The residue is applied to a column of silica gel and eluted with a 30-40% gradient of EtOAc in hexane. On concentration the fractions corresponding to 4 give an amorphous solid.
Phenyl 0-(2,3,4,6-tetra-0-acetyl-ϊ-D-galactopyranosyl)- (1→4) -θ-{ (2,3,4-tri-θ-benzyl-α-L-fucopyranosyl)-(l-*3) }-6-θ- benzyl-2-deoxy-2-phthalimido-l-thio-/3-D-glucopyranoside (5). A solution of compound 4 (3.0 g, 3.6 mmol) and methyl 2,3,4-tri-O-benzyl-l-thio-β-L-fucopyranoside (2.1 g, 4.5 mmol) in 5:1 dichloroethane-N,N-dimethylformamide (120 ml) is stirred for 0.5 h with 4A molecular sives (12 g) under protection from light and moisture. Tetrabutylammonium bromide (2.25 g, 7.0 mmol and CuBr2 (1.63 g, 7.0 mmol) is then added and the mixture is stirred for 2 days at room temperature. Further amounts of thiofucopyranoside (1.05 g) , tetrabutylammonium bromide (1.25 g) and CuBr2 (0.8 g) is added, and the stirring is continued for a total of 4 days; tic (1:1 hexane-EtOAc) then shows the presence of one major spot migrating faster than the starting material. The mixture is filtered through Celite, the solids are thoroughly washed with CHC13 and the combined filtrate and washings are washed with aq. NaHC03 and water, dried, and concentrated to a small volume. The concentrate is applied to a column of silica gel and eluted with a 30-40% gradient of EtOAc in hexane. The earlier fractions contain the faster-migrating compound 5. On concentration, these fractions afford a solid, which is crystallised from ether.
The later fractions contain the pure unreacted compound 4 (0.8 g) .
Methyl O-(2,3,4,6-tetra-θ-acetyl-/3-D-galactopyranosyl)- (l→4)-0-{ (2,3,4-tri-0-benzyl-x-l-fucopyranosyl)-(l→3) }-0- (6-0-benzyl-2-deoxy-2-phthalimido-?-D-glucopyranosyl) - (l→3)-2,4,6-tri-θ-benzyl-/3-D-galactopyranoside (6). A solution of compound 5 (0.6 g, 0.5 mmol), methyl 2,4,6-tri- O-benzyl-β-D-galactopyranoside16 (0.49 g, 0.86 mmol), and N- iodosuccinimide (0.4 g, 1.8 mmol) in CH2C12 (40 ml) is stirred with 4A molecular sieves (6.0 g) for 0.5 h at -50° to -60° under an argon atmosphere. Then, a dilute solution of trifluoromethanesulfonic acid (0.024 ml in 20 ml of CH2C12) is added dropwise. After 10 min at -50° to -60° tic in 1:1 hexane-EtOAc shows the disappearance of the phenylthio donor 5 and the formation of one major product migrating just below this donor. The acid is then neutralised with a few drops of triethylamine. The mixture is filtered through Celite, the solids are thoroughly washed with CHC13, and the combined filtrate and washings are washed successively with water, sat. NaHC03 solution, and 10% Na2S203, dried, and concentrated in vacuo . The residue is purified on a column of silica gel with a 30-40% gradient of EtOAc in hexane to provide 6. Methyl-O-β-D-galactopyranosyl) -(1-3) }-θ-(2-acetamido-6-o- benzyl-2-deoxy-/3-D-glucopyranosyl) - (1-3) -2,4, 6-tri-o- benzyl-β-D-galactopyranoside (7) . A solution of compound 7 (0.65 g) in 0.02 M sodium methoxide in MeOH (20 ml) is stirred for 4 h at room temperature. The base is neutralised with Amberlite IR-120 (H") cation-exchange resin, the resin suspension is filtered, and the filtrate concentrated. The solid so obtained is heated under reflux for 16 h in a mixture of EtOH (70 ml) and hydrazine hydrate (5.0 ml). The liquids are then evaporated to give a residue, which is dissolved in pyridine (40 ml) and acetic anhydride (20 ml) and stirred overnight at room temperature. Solvent and reagent are removed under reduced pressure, then the residue is applied to a column of silica gel and eluted with 19:1 CHC13 acetone. The material is subjected to O-deacetylation without further purification.
To accomplish this it is suspended in 0.02 M sodium methoxide (20 ml) and stirred overnight at room temperature. The base is neutralised by IR-120 (H") cation- exchange resin. The resin is filtered off and thoroughly washed with MeOH, and the filtrate and washings are combined and concentrated. The residue is purified on a silica gel column using 49:1 MeOH-CHCl3 as the eluent to give 7.
Methyl 0-j3-D-galactopyranosyl-(1—4) -0-{-α-L-fucopyranosyl- (1—3) }-0-(2-acetamido-2-deoxy-_-D-glucopyranosyl) -(1—3) -β- D-galactopyranoside (8). A mixture of 7 (0.3 g) and 10% Pd-C (0.5 g) in glacial acetic acid (30 ml) is shaken under hydrogen at -345 kPa for 2 days at room temperature. The suspension is filtered through a bed of Celite, the solids are thoroughly washed with glacial acetic acid, and the combined filtrate and washings are then concentrated under reduced pressure. The crude product is applied to a column of silica gel and eluted with 5:4:1 CHCl3-MeOH-water. The fractions corresponding to 8 are concentrated and lyophilised to give an amorphous solid.
Methyl O-(2,3,4,6-tetra-0-acetyl-/3-D-galactopyranosyl) -
(1-4) -θ-{ (2,3, 4-tri-O-benzyl-α-L-fucopyranosyl) -(1—3) }-θ-
( 6-o-benzyl-2-deoxy-2-phthalimido-J-D-glucopyranosyl) -
(1—6) -o-{ (2,3,4, 6-tetra-O-acetyl-0-D-galactopyranosyl) - (1—3) }-2-acetamido-2-deoxy-α-0-galactopyranoside (10).
Compound 5 (0.6 g 0.48 mmol) is reacted with methyl O-
(2,3,4, 6-tetra-0-acetyl-3-D-galactopyranosyl) - (1-3) -2- acetamido-2-deoxy-α-D-galactopyranoside9 (0.34 g 0.6 mmol) in CH2C12 (40 ml) in the presence of N-iodosuccinimide (0.28 g 1.23 mmol, trifluoromethanesulphonic acid (0.034 ml in 20 ml CHCl-,) , and 4A molecular sieves (6.0 g) in a manner analogous to that described for the preparation of 6.
After processing as for 6 the crude product is applied to a column of silica gel and eluted with a 20-30% gradient of acetone in CHC13. Evaporation of the fractions corresponding to the product yielded 9. Methyl O-(2,3,4, 6-tetra-O-acetyl-jS-D-galactopyranosyl)- (1-4) -θ-{ (2,3,4-tri-o-benzyl-α-L-fucopyranosyl)-(l-3) }-o- (2-acetamido-6-O-benzy1-2-deoxy-0-D-glucopyranosy1)-(1—6)- 0-{(2,3,4, 6-tetra-0-acetyl-/?-D-galactopyranosyl) -(1-3) }-2- acetamido-4-0-acetyl-2-deoxy-α-D-galactopyranoside (10) . Compound 9 (0.55 g) is treated with 0.02 M sodium methoxide in MeOH and stirred for 4 h at room temperature. After processing as described for the initial O-deacetylation of 6 the product is treated with hydrazine hydrate - EtOH (see 6—7) and then stirred with 2:1 pyridine-acetic anhydride (60 ml) overnight at room temperature. The residue from this treatment is purified on a column of silica gel by elution with a 30-40% gradient of acetone in CHC13. Upon concentration the fractions corresponding to the product gave compound 10 as a solid.
Methyl θ-/J-D-galactopyranosy1-(1-4)-O-{α-L-f copyranosy1-
(1—3) }-o-(2-acetamido-2-deoxy-/3-D-glucopyranosyl)-(1—6) -O-
{β-D-galactopyranosyl- (1—3) }-2-acetamido-2-deoxy-α-D- galactopyranoside (11). Compound 10 (0.36 g) is stirred in 0.05 M ethanolic sodium methoxide (50 ml) for 16 h at room temperature. The solution is deionised with Amberlite IR- 120 (H~) cation-exchange resin, filtered, and concentrated under reduced pressure. The residue is dissolved in glacial acetic acid (30 ml) and shaken with 10% Pd-C (0.6 g) under hydrogen at -345 kPa exactly as described for the preparation of compound 8. After purification over a silica gel column with 4:5:1 CHCl,-MeOH-water as the eluent 12 (0.15 g 70%) is obtained as an amorphous solid.
The disaccharide /3-D-Gal-α(1-4) -D-Glc (lactose) is synthesised and modified by attachment of an amine group and subsequent condensation with a peptide containing one or more cysteine residues or a single cysteine residue. Reaction of the amino lactoside with F-Moc protected cysteine (as used in solid phase peptide synthesis) results in the production of a wide variety of side chains and is therefore not the preferred method of synthesis. However, this may only present a problem in liquid phase reactions. The peptide or amino acid residue may be coupled to the lactoside using conventional solid phase peptide synthesis techniques. An alternative approach is to attach the carbohydrate to a solid phase, extend the carbohydrate by four carbon atoms, convert the NH2 group to COOH and make the active ester with succinimide.
Following formation of the derivatised lactoside, this is linked to the trisaccharide portion obtained as above via a 1-3 linkage.
Example 2: the formation of j3-lactosides of the β- Galp(l-4) -β-Glc-OR type
Such a lactoside forms part of the ultimate pentasaccharide Gal/3 (1-4 ) -[L-Fucα(1-3) ] -GlcNAc/3 (1-3) Gal/3 (1-4) -Glc/3-OR where R is a group capable of coordinating 99ιnTc. Group R specifically may be a cysteine-rich peptide, linked gl cosidically to the glucose unit through a spacer arm. The choice of starting material for the carbohydrate was β- D-lactose, Galp/3(1-4)-Glcjβ-OH(13) . This disaccharide was to be efficiently converted into 2-aminoethyl-lactoside (18) (Figure 1) . Lactose was first converted to α- acetobromolactose (14) by established procedure in 76% yield10. Displacement of the bromide with chloroethanol using lead carbonate as catalyst gave the required chloroethyl lactoside (15) in good yield". The product was subsequently treated with sodium azide in dimethylformamide as solvent and heated to 90°C over two hours. The Rf value (the movement made by a compound on a thin layer chromatography plate) was the same for the newly formed azidoethyl lactoside (16) as for the chloride.
The displacement was therefore monitored by 360 MHz 3H.n.m.r infra-red spectrometry which indicated complete conversion again with good crude yield. Our initial plan was to displace the anomeric hydroxyl group (the hydroxyl at carbon 1, see Figure 1) with chloroethyl functionality which could be converted into the required 2-aminoethyl linker arm on the disaccharide (13) .
The amine moiety could then be condensed with the carboxyl terminus of a cysteine-rich peptide or a suitably protected cysteine amino acid. Our initial exploration was therefore to determine if any difficulties would be encountered in the synthesis of 2-aminoethyl lactoside (18) . Therefore, de-acetylation of azide (16) was achieved by stirring with sodium methoxide solution. Adjustment of pH to neutral [Amberlite resin- 120(H)] and evaporation to dryness provided the deprotected azide (17)". Reduction of the azido to amino functionality was achieved by brief hydrogenation using 5% Palladium on carbon as catalyst10.
The classic coupling reagent for solid phase peptide synthesis has been dicyclohexylcarbodiimide (DCC) (9) in spite of well known side reactions12,13. Recent developments have led to reports of new activating agents for peptide bond formation in the solid phase such as 2-(lH-benzo- triazol-l-yl)-l,l,3, 3-tetramethyluronium hexafluoro- phosphate (HBTU) (20) and 2-(lH-benzotriazol-1-yl) -1, 1, 3, 3- tetramethyluronium tetrafluoroborate (TBTU) (21) . These reagents are reported as having high reactivity without undesired side reactions with harmless by-products13.
It was decided that the new activating agents would be ideal for the condensation of the cysteine rich peptide at the carboxyl terminus with the amino terminus of our 2- aminoethyl-lactoside. However, the use of the unprotected lactoside was considered to be problematic in that the activating agents, HBTU, TBTU and DCC are known to effect esterification as well as amidation14. Therefore it was decided to reduce the fully acylated azide to give acylated amine. The hydroxyl would then be protected leaving the amino group to condense with the cysteine rich peptide. Our next step was to effect the reduction of the acylated azide (16) to amine (22) . Again, the best conditions for this reduction were found to be methanol/Pd on carbon at 414 KPa (Figure 2) . This method proved superior to the use of SnCl or NaBH4. The 2-amino acylated lactoside was filtered through Celite and the solvent reduced in vacuo.
Addition of cold ether gave the amine in good yield as an off-white solid.
The active ester of the cysteine amino acid was formed using DCC and N-hydroxysuccinimide. The isolated active ester could then be coupled with the amine in dichloro ethane with the minimum of by-products formed.
The active ester (25) of the HO-Cys(tBu) -Fmoc was prepared by cooling of solution of the amino acid and N- hydroxysuccinimide (24) in dry tetrahydrofuran to 0°C. Addition of DCC at this temperature followed and the mixture was allowed to stand at 4°C for 18 hours. The white precipitate of N,N-dicyclohexylurea was filtered off and the filtrate evaporated under reduced pressure to give a white solid in 84% yield (Figure 3) .
With the active ester in hand, we were ready to attempt the coupling with 2-aminoethyl-hepta-0-acetyl-/3-D-lactoside
(12) . The active ester was dissolved in dichloromethane and cooled in an ice/ ater bath. To the stirred, cooled solution was added amine lactoside and the solution stirred with cooling for 2 hours. The solution was then allowed to reach room temperature and was stirred for a further twelve hours. TLC (thin layer chromatography) showed the presence of a new intense product which was both ultra-violet- and H2S04-visualised with an Rf of 0.36. The new spot was isolated by flash column chromatography in 50% yield. This yield has not been optimised at present. The compound was characterised by 360 MHz Ηnmr and i3Cnmr. The sequence of reactions to the fully protected glyco-amino acid is simple to perform, is high yielding and can be performed on a relatively large scale (Figure 4) .
Deprotection of the glyco-amino acid using methods mentioned earlier in the report gives the desired ligand compound for use as a 99mTc coordinator.
Example 3
As an alternative strategy to the final deprotection of the glycoa ino acid in Example 2 involves a lengthening of the linker arm and a reversal of the carboxy terminus of the amino acid for the amino terminus, and similarly for the lactoside involes reaction of the amino lactoside with succinic anhydride (27) (Figure 5) .
The active ester of the carboxy lactoside can then be formed and coupled with the amino terminus of the amino acid or peptide. This also provides a way of adding the carbohydrate portion to the cysteine-rich peptide in the solid phase.
REFERENCES
1. A. M.S. Marr et al (1968) Biochem. J. no, 789-79.
2. K.o. Lloyd et al (1966) Biochemistry 5, 1489-1501.
3. H. Yang and S. Hakomori (1971) J. Biol. Chem. 246, 1192-1200.
4. B.L. Slomiany ei al (1975) Eur. J. Biochem. 56, 353-
358.
5. s. Sato et al (1987) Carbohydr. Res. 167, 197-210.
6. H. Lonn (1985) Carbohydr. Res. 139, 115-121.
7. M. Nilsson and T. Norberg (1988) Carbohydr. Res.
183, 71-82.
8. B. Classon et al (1989) J. Carbohydr. Chem. 8, 543-
551.
9. C.F. Piskorz and K.L. Matta Unpublished results.
10. R.S. Bhatt (1975) PhD Thesis, Queen Elizabeth College, University of London.
11. L. Hough et al (1991) Reel. Trav. Chim. Pays-Bas 110, 450. 12. J.C. Shehan and G.P. Hess (1955) J. Amer. Chem.
Soc. 11, 1067.
13. R. Knorr et al (1984) Synthesis 572.
14. J. March Advanced Organic Chemistry, John Wiley &
Sons, New York, 3rd Ed. 15. A.K.G. Povey (1990) PhD Thesis, King's College London, University of London. 16. K. Kohata et al (1984) Carbohydr. Res . 132, 127-135.

Claims

1. A compound comprising a first portion adapted to bind specifically the ELAM-1 receptor and a second portion capable of being detected in vivo by remote sensing techniques when the compound is bound to the said receptor.
2. A compound according to Claim 1 wherein the first portion comprises a saccharide structure of Formula
(I) :
(I) R'-Gal(l→4)X'(R2)
wherein X1 is GIcNac or Glc, R1 is a hydrogen atom forming an OH group at the 3 position on the galactose group or is a NeuAc(2→3) group and R2 is hydrogen forming an OH group at the 3 position on the X1 group or is a Fuc(1→3) group thereon.
3. A compound according to Claim 2 wherein the first portion comprises a saccharide structure of Formula (II):
(II) R'-Gal(^4)X'(R2)-(X2)n wherein X1 and R1 are as defined above, n is 1 or 2, X2 is (l→3)Gal(l→4)X3(R3) , R2 and the or each R3 are independently a hydrogen atom forming an OH group on the X3 ring or a Fuc(l→3) group thereon, and the or each X3 is in each case GIcNac or Glc.
4. A compound according to any one of the preceding claims wherein the second portion comprises a radioactive atom, an nmr-enhancing atom or a positron-emitting atom.
5. A method of identifying activated endothelium in a patient comprising administering to the patient a compound according to any one of the preceding claims and detecting localised concentrations of the said second portion of the compound.
PCT/GB1992/001216 1991-07-06 1992-07-06 Elam-1 receptor ligands as diagnostic compounds WO1993000908A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB9114657.1 1991-07-06
GB919114657A GB9114657D0 (en) 1991-07-06 1991-07-06 Diagnostic compounds

Publications (1)

Publication Number Publication Date
WO1993000908A1 true WO1993000908A1 (en) 1993-01-21

Family

ID=10697961

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB1992/001216 WO1993000908A1 (en) 1991-07-06 1992-07-06 Elam-1 receptor ligands as diagnostic compounds

Country Status (2)

Country Link
GB (1) GB9114657D0 (en)
WO (1) WO1993000908A1 (en)

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997007809A1 (en) * 1995-08-23 1997-03-06 The Regents Of The University Of California Sulfated disaccharide inhibitors of selectins methods for synthesis and therapeutic use
US5658880A (en) * 1993-06-16 1997-08-19 Glycomed Incorporated Sialic acid/fucose based medicaments
US5660992A (en) * 1993-06-16 1997-08-26 Glycomed Incorporated Sialic acid/fucose based assay reagents and assay methods
US5679321A (en) * 1993-06-17 1997-10-21 Glycomed Incorporated Sialic acid/fucose based medicaments
US5789385A (en) * 1993-06-16 1998-08-04 Glycomed Incorporated Sialyl Lewisx mimetics containing phenyl backbones
US5837689A (en) * 1993-06-16 1998-11-17 Glycomed Incorporated Sialyl lewis-x mimetics containing naphthyl backbones
EP0919563A2 (en) * 1997-10-31 1999-06-02 Health Research, Inc. Synthetic Core 2-Like Branched Structures as Ligands for Selectins
US9902690B2 (en) 2013-12-27 2018-02-27 Novus International, Inc. Ethoxylated surfactants
US10584306B2 (en) 2017-08-11 2020-03-10 Board Of Regents Of The University Of Oklahoma Surfactant microemulsions

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991019502A1 (en) * 1990-06-15 1991-12-26 Cytel Corporation Intercellular adhesion mediators
WO1992002527A1 (en) * 1990-07-30 1992-02-20 Glycomed, Incorporated New carbohydrate-based anti-inflammatory agents
WO1992008489A1 (en) * 1990-11-08 1992-05-29 Biogen, Inc. Treatment for inflammatory bowel disease

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991019502A1 (en) * 1990-06-15 1991-12-26 Cytel Corporation Intercellular adhesion mediators
WO1991019501A1 (en) * 1990-06-15 1991-12-26 Cytel Corporation Intercellular adhesion mediators
WO1992002527A1 (en) * 1990-07-30 1992-02-20 Glycomed, Incorporated New carbohydrate-based anti-inflammatory agents
WO1992008489A1 (en) * 1990-11-08 1992-05-29 Biogen, Inc. Treatment for inflammatory bowel disease

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
Cell, Vol. 63, November 1990 John B. Lowe et al.: "ELAM-1-Dependent Cell Adhesion to Vascular Endothelium Determined by a Transfected Human Fucosyltransferase cDNA ", *
SCIENCE, Vol. 250, 1990 Gerd Walz et al.: "Recognition by ELAM-1 of the Sialyl-Le Determinant on Myeloid and Tumor Cells ", *
SCIENCE, Vol. 250, 1990 M. Laurie et al.: "ELAM-1 Mediates Cell Adhesion by Recognition of a Carbohydrate Ligand, Sialyl-Le ", *

Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5789385A (en) * 1993-06-16 1998-08-04 Glycomed Incorporated Sialyl Lewisx mimetics containing phenyl backbones
US5658880A (en) * 1993-06-16 1997-08-19 Glycomed Incorporated Sialic acid/fucose based medicaments
US5660992A (en) * 1993-06-16 1997-08-26 Glycomed Incorporated Sialic acid/fucose based assay reagents and assay methods
US5837689A (en) * 1993-06-16 1998-11-17 Glycomed Incorporated Sialyl lewis-x mimetics containing naphthyl backbones
US5750508A (en) * 1993-06-16 1998-05-12 Glycomed Incorporated Sialic acid/fucose based medicaments
US5679321A (en) * 1993-06-17 1997-10-21 Glycomed Incorporated Sialic acid/fucose based medicaments
US5783693A (en) * 1993-11-19 1998-07-21 The Regents Of The University Of California Methods for synthesizing sulfated disaccharide inhibitors of selectins
WO1997007809A1 (en) * 1995-08-23 1997-03-06 The Regents Of The University Of California Sulfated disaccharide inhibitors of selectins methods for synthesis and therapeutic use
US5977080A (en) * 1995-08-23 1999-11-02 The Regents Of The University Of California Sulfated disaccharide inhibitors of selectins, methods for synthesis and therapeutic use
EP0919563A2 (en) * 1997-10-31 1999-06-02 Health Research, Inc. Synthetic Core 2-Like Branched Structures as Ligands for Selectins
EP0919563A3 (en) * 1997-10-31 2001-08-29 Health Research, Inc. Synthetic Core 2-Like Branched Structures as Ligands for Selectins
US9902690B2 (en) 2013-12-27 2018-02-27 Novus International, Inc. Ethoxylated surfactants
US10584306B2 (en) 2017-08-11 2020-03-10 Board Of Regents Of The University Of Oklahoma Surfactant microemulsions

Also Published As

Publication number Publication date
GB9114657D0 (en) 1991-08-21

Similar Documents

Publication Publication Date Title
Ramphal et al. Structure-activity relationships of sialyl Lewis x-containing oligosaccharides. 1. Effect of modifications of the fucose moiety
Vankar et al. Chemistry of glycosphingolipids—carbohydrate molecules of biological significance
US5326752A (en) Substituted lactose and lactosamine derivatives as cell adhesion inhibitors
JPH10279589A (en) Sialic acid glycoside, antigen, immunoadsorbemt and their preparation
CA2197336A1 (en) Sialic acid/fucose based medicaments
WO1996005209A9 (en) Sialic acid/fucose based medicaments
Kretzschmar et al. Large scale synthesis of linker-modified sialyl LewisX, LewisX and N-acetyllactosamine
JPH05507487A (en) Oligosaccharide-protein complex and method for producing the same
Bélot et al. Unexpected stereochemical outcome of activated 4, 6-O-benzylidene derivatives of the 2-deoxy-2-trichloroacetamido-D-galacto series in glycosylation reactions during the synthesis of a chondroitin 6-sulfate trisaccharide methyl glycoside
WO1993000908A1 (en) Elam-1 receptor ligands as diagnostic compounds
Cao et al. Chemical preparation of sialyl Lewis x using an enzymatically synthesized sialoside building block
US6462183B1 (en) Protected aminosugars
Saksena et al. Immunogens related to the synthetic tetrasaccharide side chain of the Bacillus anthracis exosporium
Cipolla et al. Stereoselective synthesis of α-C-glycosides of N-acetylgalactosamine
Csíki et al. Synthesis of glycosaminoglycan oligosaccharides. Part 4: Synthesis of aza-l-iduronic acid-containing analogs of heparan sulfate oligosaccharides as heparanase inhibitors
US5344870A (en) Sialic acid glycosides, antigens, immunoadsorbents, and methods for their preparation
Herzner et al. Spacer-separated sialyl LewisX cyclopeptide conjugates as potential E-selectin ligands
WO1994000477A1 (en) Substituted lactose derivatives as cell adhesion inhibitors
WO2023216732A1 (en) Chemical synthesis method for helicobacter pylori core lipopolysaccharide oligosaccharide antigen carbohydrate chain
Lio et al. Chemoenzymatic synthesis of spacer-linked oligosaccharides for the preparation of neoglycoproteins
Zhao et al. Stereoselective synthesis of a branched α-decaglucan
Bi et al. Synthesis of a trisaccharide repeating unit of the O-antigen from Burkholderia cenocepacia and its dimer
Kunetskiy et al. Synthesis of blood group A and B (type 2) tetrasaccharides. A strategy with fucosylation at the last stage
JPH075622B2 (en) Method for producing synthetic N-linked glycoconjugate
Ogawa et al. Synthesis of Ether‐and Imino‐Linked Octyl N‐Acetyl‐5a′‐carba‐β‐lactosaminides and‐isolactosaminides: Acceptor Substrates for α‐(1→ 3/4)‐Fucosyltransferase, and Enzymatic Synthesis of 5a′‐Carbatrisaccharides

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): JP US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IT LU MC NL SE

122 Ep: pct application non-entry in european phase