title: Synthesis of tailor-made glycoconjugate mimetics of heparan sulfate that bind IFN-gamma in the nanomolar range.
abstract: We have recently described the preparation of three building blocks for the combinatorial synthesis of heparan sulfate (HS) fragments. Herein we show that one of these building blocks (disaccharide 4) allows the preparation, in high yields and with total alpha stereoselectivity, of tetra-, hexa- and octasaccharides from the heparin (HP) regular region, by using 2+2, 2+4 and 4+4 glycosylation strategies, respectively. These oligosaccharides were processed into sulfated derivatives bearing an allyl moiety in the anomeric position. The UV-promoted conjugation of these compounds with alpha,omega-bis(thio)poly(ethylene glycol) spacers of three different lengths allowed us to prepare nine benzylated glycoconjugates. After final deprotection, the glycoconjugates 1 a-c, 2 a-c and 3 a-c were obtained and their ability to inhibit the interaction between IFN-gamma and HP was tested by using surface plasmon resonance detection. Compound 3 b, containing two HP octasaccharides linked by a 50-A linker was able to inhibit the IFN-gamma/HP interaction with an IC(50) value of approximately 35 nM. In addition, the nine glycoconjugates were perfect tools in the study to ascertain the topology of the IFN-gamma binding site on HS. Compounds 1 a-c, 2 a-c and 3 a-c, by mimicking the alternating sulfated and nonsulfated regions found in HS, thus comprise the first example of a library of synthetic HS mimetics giving access to the "second level of molecular diversity" found in HS.
authors.affiliation: Laboratoire de Chimie Organique Multifonctionnelle, UMR CNRS-UPS 8614 Glycochimie Moléculaire, Bat. 420, Université Paris Sud, 91405 Orsay Cedex, France.
authors.forename: André
authors.initials: A
authors.lastname: Lubineau
authors.forename: Hugues
authors.initials: H
authors.lastname: Lortat-Jacob
authors.forename: Ollivier
authors.initials: O
authors.lastname: Gavard
authors.forename: Stéphane
authors.initials: S
authors.lastname: Sarrazin
authors.forename: David
authors.initials: D
authors.lastname: Bonnaffé
mesh_terms: D000595:Amino Acid Sequence; D032701:Biomimetics; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000737:chemistry; D006497:Heparan Sulfate / Q000138:chemical synthesis* / Q000737:chemistry*; D007202:Indicators and Reagents; D007371:Interferon-gamma / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D013056:Spectrophotometry, Ultraviolet
chemical_list: D006023:Glycoproteins; D006497:Heparan Sulfate; D007202:Indicators and Reagents; D007371:Interferon-gamma