Variability of Heparins and Heterogeneity of Low Molecular Weight Heparins

ABSTRACT

Chemical and physical characteristics, building blocks, constitutive disaccharides, sulfation degree, and biological activities of heparins (UFHs) and of low molecular weight heparins (LMWHs) obtained by different depolymerization processes are examined comparatively in terms of structure characteristics, content of 1,6-anhydro rings, and other fingerprints. The heterogeneity of different LMWHs depends on different manufacturing processes and on particular specifications of pharmacopoeias. The reported examples prove that the variability among samples of LMWHs manufactured by the same process is quite limited. Most of the variability is derived from the parent UFH. In contrast, fingerprint groups and residues are specific to the depolymerization process and their extent can be roughly controlled through the process parameters.

Heparin - low molecular weight heparins - end residues - sulfation pattern - 1,6-anhydro rings - trisaccharide - fingerprints - disaccharide building blocks - heterogeneity of LMWHs

REFERENCES

• 1 Casu B. Structure and active domains of heparin. In: Garg HG, Linhardt RJ, Hales CA Chemistry and Biology of Heparin and Heparan Sulfate. Oxford, UK; Elsevier Ltd 2005: 1-28
• 2 Linhardt R J. Heparin: structure and activity.  J Med Chem. 2003;  46 2551-2564
• 3 Bianchini P, Liverani L, Mascellani G, Bruna P. Heterogeneity of unfractionated heparins studied in connection with species, source, and production processes.  Semin Thromb Hemost. 1997;  23 3-10
• 4 Bianchini P, Nader H B, Takahashi H K, Osima B, Straus A H, Dietrich C P. Fractionation and identification of heparin and other mucopolysaccharides by a new discontinuous electrophoretic method.  J Chromatogr. 1980;  196 455-462
• 5 Bianchini P, Osima B, Parma B et al.. Fractions and structural features of two heparin families with high antithromotic, antilipemic and anticoagulant activities.  Arzneim-Forsch Drug Res. 1985;  35 1215-1219
• 6 Mourier P, Viskov C. Method of determining specific groups forming heparins.  PCT FR2003/002782 WO2004/02787.
• 7 Bell O, Safir P O. Citizen Petition to FDA, September 14, 2006. http://Available at: www.fda.gov/OHRMS/Dockets/Dockets/03P0064/03P-0064.S6P003.TOC.HTM Accessed February 9, 2007
• 8 Bisio A, Guerrini M, Yates F A, Torri G, Casu B. NMR identification of structural environments for 2,3 or 2,3,6 tri-sulfated glucosamine residues in heparin with high and no affinity for antithrombin.  Glycoconj J. 1997;  14(suppl 1) S89
• 9 Mourier P AJ, Viskov C. Chromatographic analysis and sequencing approach of heparin oligosaccharides using cetyltrimethyl ammonium dynamically coated stationary phases.  Anal Biochem. 2004;  332 299-313
• 10 Mascellani G, Liverani L, Bianchini P. Analysis of heparin origin by HPLC quantitation of disaccharide components.  Farmaco. 1996;  51(4) 247-254
• 11 Casu B, Torri G. Structural characterization of low molecular weight heparins.  Semin Thromb Hemost. 1999;  25 17-25
• 12 Rey R N, Perlin A S. Base catalized conversion of the α-L-galacturonic acid, and related reactions.  Carbohydr Res. 1990;  200 437-447
• 13 Yamada S, Watanabe M, Sugahara K. Conversion of N-sulfated glucosamine to N-sulfated mannosamine in an unsaturated heparin disaccharide by non-enzymatic, base-catalyzed C-2 epimerization during enzymatic preparation.  Carbohydr Res. 1998;  309 261-268
• 14 Toida T E, Vlahov I R, Smith A E et al.. C-2 epimerization of N-acetylglucosamine in an oligosaccaride derived from heparan sulphate.  J Carbohydr Chem. 1996;  15 351-360
• 15 Bell O, Safir P O. Citizen petition to FDA Februrary 19, 2003.  http://Available at: www.fda.gov/OHRMS/Dockets/Dockets/03P0064/03P0064.HTM Accessed February 27, 2003; 
• 16 Mourier P, Viskov C. Method for determining specific group constituting heparins or low molecular weight heparin. US2005/0119477A1.  Chem Abst. 2005;  142 89363
• 17 Mascellani G, Guerrini M, Torri G, Liverani L, Spelta F, Bianchini P. Characterization of di- and monosulfated, unsaturated heparin disaccharides with terminal N-sulfated 1,6-anhydro-β-D-glucosamine or N-sulfated 1,6-anhydro-β-D-mannosamine residues.  Carbohydr Res. 2007;  342 835-842
• 18 Ragazzi M, Ferro D R, Provaroli A et al.. Conformation of the unsaturated uronic acid residues of glycosaminoglycal disaccharides.  J Carbohydr Chem. 1993;  12 523-525