Structure and anticoagulant activity of fucosylated glycosaminoglycan degraded by deaminative cleavage

Carbohydr Polym. 2013 Nov 6;98(2):1514-23. doi: 10.1016/j.carbpol.2013.07.063. Epub 2013 Aug 6.

Abstract

Fucosylated glycosaminoglycans (FGs) are complex glycosaminoglycans that exhibit potent anticoagulant activity. To study the relationship between molecular size and biological activity, oligosaccharides with (2,5)-anhydro-D-talose units at new reducing ends were prepared by hydrazine deacetylation and nitrous acid depolymerization. The product chemical structures were analyzed by one- and two-dimensional NMR methods. Additionally, anticoagulant activities were evaluated by clotting assay and chromogenic substrate cleavage. The results demonstrated that under mild deacetylation and deaminative cleavage conditions, both products were relatively homogeneous and sulfated fucose branch types and sulfate substituents remained stable. These depolymerized FGs with different molecular sizes had potent intrinsic anticoagulant activities, which were similar to those that were obtained by free-radical depolymerization with similar molecular weights. Decreasing molecular weight may weaken activity but not significantly affect factor Xase and heparin cofactor II (HCII)-mediated thrombin inhibition.

Keywords: 2,4-O-disulfated fucose; 3-O-sulfated fucose; 4,6-O-disulfated N-acetylgalactosamine; 4-O-sulfated fucose; APTT; Anticoagulant activity; DD; DFG; Deacetylation; Deaminative cleavage; EC(50); FG; Fuc2S4S; Fuc3S; Fuc4S; GAG; GalN; GalNAc4S6S; GlcUA; Glycosaminoglycan; HCII; LMWH; Sea cucumber; UFH; activated partial thromboplastin time; anTal; anhydro-d-talose; deacetylation degree; depolymerized fucosylated glycosaminoglycan; fucosylated glycosaminoglycan; galactosamine; glucuronic acid; glycosaminoglycan; half maximal effective concentration; heparin cofactor II; low-molecular-weight heparin; unfractionated heparin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anticoagulants / chemistry*
  • Anticoagulants / isolation & purification
  • Anticoagulants / pharmacology
  • Blood Coagulation / drug effects*
  • Cysteine Endopeptidases / chemistry
  • Enzyme Assays
  • Fucose / chemistry*
  • Glycosaminoglycans / chemistry*
  • Glycosaminoglycans / isolation & purification
  • Glycosaminoglycans / pharmacology
  • Heparin Cofactor II / chemistry
  • Humans
  • Hydrazines / chemistry
  • Lactones / chemistry
  • Magnetic Resonance Spectroscopy
  • Molecular Weight
  • Neoplasm Proteins / chemistry
  • Nitrous Acid / chemistry
  • Sea Cucumbers / chemistry*
  • Structure-Activity Relationship
  • Thrombin / antagonists & inhibitors
  • Thrombin / chemistry

Substances

  • Anticoagulants
  • Glycosaminoglycans
  • Hydrazines
  • L-talo-gamma-lactone
  • Lactones
  • Neoplasm Proteins
  • hydrazine
  • Fucose
  • Heparin Cofactor II
  • Thrombin
  • Cysteine Endopeptidases
  • cancer procoagulant
  • Nitrous Acid