title: Oligosaccharide mimics containing galactose and fucose specifically label tumour cell surfaces and inhibit cell adhesion to fibronectin.
abstract: With the aim of establishing a versatile and easy synthesis of branched saccharides for biological applications, we used molecular-dynamics simulations to model Lewis(y) to two classes of di- or triantennary saccharide mimetics. One set of mimetics was based on 1,3,5-tris(hydroxymethyl)cyclohexane (TMC) as the core, the other on furan, and both were derivatised with galactose and/or fucose. The TMC-based saccharides were biotinylated, while the furan disaccharides were treated with maleimide-activated biotin in a Diels-Alder fashion to yield oxazatricyclodecanes (OTDs). These were then assayed as cell-surface labels in human colon (SW480 and CaCo-2), liver (PLC), Glia (U333 CG 343) and ovary (SKOV-3) tumour cell lines. Discrete staining patterns were observed in all cells, usually at one or two poles of the cells, particularly with the asymmetric 3-beta-L-fucopyranosyloxymethyl-4-beta-D-galactopyranosyloxymethyl-OTD. Normal SV40-transformed fibroblasts (SV80) showed no staining. Adhesion of the highly metastatic mouse melanoma line B16 F10 to fibronectin was inhibited by 80 % by the TMC-digalactoside and by 30 % by 3,4-bis-(beta-D-galactopyranosyloxymethyl)furan. None of the saccharide mimetics inhibited the adhesion of the less metastatic B16 F1 line. Migration of B16 F10 cells through Matrigel was greatly inhibited by the TMC-digalactoside and weakly inhibited by the TMC-trigalactoside. The saccharide mimetics that had shown the best structural agreements with the terminal saccharides of Lewis(y) in the molecular dynamics simulation were also the most biologically potent compounds; this underlines the predictive nature of molecular dynamics simulations. The use of the non-saccharide cores enabled us to adapt spacer lengths and terminal saccharides to optimise the structures to bind more avidly to cell-surface lectins.
authors.affiliation: Molecular Toxicology, Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 280, 69120 Heidelberg, Germany.
authors.forename: Evelyn Y-L
authors.initials: EY
authors.lastname: Kim
authors.forename: Claas
authors.initials: C
authors.lastname: Gronewold
authors.forename: Amitava
authors.initials: A
authors.lastname: Chatterjee
authors.forename: Claus-Wilhelm
authors.initials: CW
authors.lastname: von der Lieth
authors.forename: Christian
authors.initials: C
authors.lastname: Kliem
authors.forename: Birgit
authors.initials: B
authors.lastname: Schmauser
authors.forename: Manfred
authors.initials: M
authors.lastname: Wiessler
authors.forename: Eva
authors.initials: E
authors.lastname: Frei
mesh_terms: D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism*; D002465:Cell Movement; D003094:Collagen / Q000378:metabolism; D003198:Computer Simulation; D004338:Drug Combinations; D005353:Fibronectins / Q000378:metabolism*; D005643:Fucose / Q000737:chemistry* / Q000378:metabolism; D005690:Galactose / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D007797:Laminin / Q000378:metabolism; D051379:Mice; D018716:Molecular Mimicry; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009844:Oligosaccharides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism*; D011509:Proteoglycans / Q000378:metabolism
chemical_list: D004338:Drug Combinations; D005353:Fibronectins; D007797:Laminin; D009844:Oligosaccharides; D011509:Proteoglycans; C059485:matrigel; D005643:Fucose; D003094:Collagen; D005690:Galactose