Heparin in Acid and Alkaline Environments—A Study of the Correlations between Hydrodynamic Properties and Desulphation

This work evaluated the hydrodynamic properties of heparin hydrolysed at temperatures ranging from 40 °C to 80 °C in buffered acid and alkaline environments. The correlation between hydrodynamic parameters led to the conclusion that polymer conformational changes appeared to be minimal until chain depolymerisation, initiated at pH 1 and 80 °C. However, the synergy of conformational changes, even if minimal, and sulphate loss observed at pH 1, pH 3 and pH 12 (various temperatures) resulted in a loss of the antifactor Xa activity. Therefore, the ‘contribution’ of conformational changes should be added to the generally recognized effect of desulphation towards the activity of heparin. This is of significance as the processing of medical heparin is complex, and requires adjustment of several physical and chemical factors, including pH and temperature.

[1]  Alan M. Smith,et al.  Monitoring the stability of heparin: NMR evidence for the rearrangement of sulfated iduronate in phosphate buffer. , 2023, Carbohydrate polymers.

[2]  James A. London,et al.  Chemical Modification of Glycosaminoglycan Polysaccharides , 2021, Molecules.

[3]  Alan M. Smith,et al.  Hydrolytic Degradation of Heparin in Acidic Environments: Nuclear Magnetic Resonance Reveals Details of Selective Desulfation. , 2021, ACS Applied Materials and Interfaces.

[4]  E. Yates,et al.  Introduction to the Molecules Special Edition Entitled ‘Heparan Sulfate and Heparin: Challenges and Controversies’: Some Outstanding Questions in Heparan Sulfate and Heparin Research , 2019, Molecules.

[5]  R. Beynon,et al.  The heparin-binding proteome in normal pancreas and murine experimental acute pancreatitis , 2018, bioRxiv.

[6]  M. Parrilli,et al.  A review of chemical methods for the selective sulfation and desulfation of polysaccharides. , 2017, Carbohydrate polymers.

[7]  E. Kellenbach,et al.  From Farm to Pharma: An Overview of Industrial Heparin Manufacturing Methods , 2017, Molecules.

[8]  R. Linhardt,et al.  Heparin's solution structure determined by small‐angle neutron scattering , 2016, Biopolymers.

[9]  R. Woods,et al.  Uncovering the Relationship between Sulphation Patterns and Conformation of Iduronic Acid in Heparan Sulphate , 2016, Scientific Reports.

[10]  R. Linhardt,et al.  Heparin and anticoagulation. , 2016, Frontiers in bioscience.

[11]  C. Page,et al.  Pharmacology of Heparin and Related Drugs , 2016, Pharmacological Reviews.

[12]  M. Hricovíni.  Solution Structure of Heparin Pentasaccharide: NMR and DFT Analysis. , 2015, The journal of physical chemistry. B.

[13]  R. Linhardt,et al.  Heparin and related polysaccharides: synthesis using recombinant enzymes and metabolic engineering , 2015, Applied Microbiology and Biotechnology.

[14]  G. Torri,et al.  Re-visiting the structure of heparin. , 2015, Carbohydrate research.

[15]  Stephen E. Harding,et al.  On hydrodynamic methods for the analysis of the sizes and shapes of polysaccharides in dilute solution: A short review , 2014 .

[16]  Z. Shriver,et al.  USP compendial methods for analysis of heparin: chromatographic determination of molecular weight distributions for heparin sodium , 2014, Analytical and Bioanalytical Chemistry.

[17]  V. Pomin.  Solution NMR conformation of glycosaminoglycans. , 2014, Progress in biophysics and molecular biology.

[18]  V. Pomin.  NMR chemical shifts in structural biology of glycosaminoglycans. , 2014, Analytical chemistry.

[19]  S. Perkins,et al.  The Solution Structure of Heparan Sulfate Differs from That of Heparin , 2013, The Journal of Biological Chemistry.

[20]  M. Ralet,et al.  The effect of neutral sugar distribution on the dilute solution conformation of sugar beet pectin , 2012 .

[21]  E. Yates,et al.  A highly efficient tree structure for the biosynthesis of heparan sulfate accounts for the commonly observed disaccharides and suggests a mechanism for domain synthesis. , 2012, Molecular bioSystems.

[22]  S. Perkins,et al.  Molecular architecture of heparin and heparan sulfate: Recent developments in solution structural studies , 2011 .

[23]  S. Harding,et al.  The kinetics of chitosan depolymerisation at different temperatures , 2009 .

[24]  N. Nagai,et al.  6-O-Sulfation of Heparan Sulfate Differentially Regulates Various Fibroblast Growth Factor-dependent Signalings in Culture* , 2008, Journal of Biological Chemistry.

[25]  Elaine Gray,et al.  Heparin and low-molecular-weight heparin , 2008, Thrombosis and Haemostasis.

[26]  J. Turnbull,et al.  The Activities of Heparan Sulfate and its Analogue Heparin are Dictated by Biosynthesis, Sequence, and Conformation , 2008, Connective tissue research.

[27]  J. García de la Torre,et al.  Equivalent radii and ratios of radii from solution properties as indicators of macromolecular conformation, shape, and flexibility. , 2007, Biomacromolecules.

[28]  S. Cui.  Food Carbohydrates : Chemistry, Physical Properties, and Applications , 2005 .

[29]  S. Sun.  Physical Chemistry of Macromolecules , 2004 .

[30]  S. Finet,et al.  Conformation of heparin studied with macromolecular hydrodynamic methods and X-ray scattering , 2003, European Biophysics Journal.

[31]  R. Linhardt.  2003 Claude S. Hudson Award address in carbohydrate chemistry. Heparin: structure and activity. , 2003, Journal of medicinal chemistry.

[32]  S. Harding,et al.  A hydrodynamic study of the depolymerisation of a high methoxy pectin at elevated temperatures , 2002 .

[33]  D. Rabenstein.  Heparin and heparan sulfate: structure and function. , 2002, Natural product reports.

[34]  R. Linhardt,et al.  Heparin-protein interactions. , 2002, Angewandte Chemie.

[35]  E. Racine.  Differentiation of the Low‐Molecular‐Weight Heparins , 2001, Pharmacotherapy.

[36]  Michael B. Smith.  Carbohydrates , 2001, Public Health Nutrition.

[37]  M. Forster,et al.  Conformation and dynamics of heparin and heparan sulfate. , 2000, Glycobiology.

[38]  M. Salmivirta,et al.  Selectively Desulfated Heparin Inhibits Fibroblast Growth Factor-induced Mitogenicity and Angiogenesis* , 2000, The Journal of Biological Chemistry.

[39]  P. Schuck.  Sedimentation analysis of noninteracting and self-associating solutes using numerical solutions to the Lamm equation. , 1998, Biophysical journal.

[40]  M. Tombs,et al.  An introduction to polysaccharide biotechnology , 1997 .

[41]  S. Harding,et al.  The intrinsic viscosity of biological macromolecules. Progress in measurement, interpretation and application to structure in dilute solution. , 1997, Progress in biophysics and molecular biology.

[42]  R. Linhardt.  Chemical and Enzymic Methods for the Depolymerization and Modification of Heparin , 1995 .

[43]  S. Sun.  Physical Chemistry of Macromolecules: Basic Principles and Issues , 1994 .

[44]  M. Forster,et al.  N.m.r. and molecular-modelling studies of the solution conformation of heparin. , 1993, The Biochemical journal.

[45]  R. Rej,et al.  Base-catalyzed conversion of the a-l-iduronic acid 2-sulfate unit of heparin into a unit of a-l-galacturonic acid, and related reactions , 1990 .

[46]  G. Torri,et al.  Evidence for Conformational Equilibrium of the Sulfated L-Iduronate Residue in Heparin and in Synthetic Heparin Mono- and Oligosaccharides: NMR and Force-Field Studies. , 1987 .

[47]  Benito Casu,et al.  Evidence for conformational equilibrium of the sulfated L-iduronate residue in heparin and in synthetic heparin mono- and oligo-saccharides: NMR and force-field studies , 1986 .

[48]  M. Bienkowski,et al.  Structural characterization of the oligosaccharides formed by depolymerization of heparin with nitrous acid. , 1985, The Journal of biological chemistry.

[49]  S. Stivala,et al.  Macromolecular properties of heparin in dilute solution: 2. Dimensional parameters as a function of pH, ionic strength and desulphation , 1974 .

[50]  S. Stivala,et al.  Physicochemical studies of fractionated bovine heparin. 3. Some physical parameters in relation to biological activity. , 1967, Archives of biochemistry and biophysics.

[51]  J. J. Carr,et al.  Investigations on the chemistry of heparin. I. Desulfation and acetylation. , 1960, Archives of biochemistry and biophysics.

[52]  H. Kowarzyk.  Structure and Function. , 1910, Nature.

[53]  Barbara Mulloy,et al.  Heparin - A Century of Progress , 2012, Handbook of Experimental Pharmacology.

[54]  R. Linhardt,et al.  Engineering of routes to heparin and related polysaccharides , 2011, Applied Microbiology and Biotechnology.

[55]  S. Harding.  Analysis of polysaccharides by ultracentrifugation. Size, conformation and interactions in solution , 2005 .

[56]  C. Mohan.  Buffers A guide for the preparation and use of buffers in biological systems , 2003 .

[57]  R. Linhardt,et al.  Accelerated stability studies of heparin. , 1996, Journal of pharmaceutical sciences.

[58]  D. D. Perrin,et al.  Preparation of Buffer Solutions , 1974 .

[59]  H. Inoue.  Antithrombotic Therapy , 2010 .

[60]  E. Jorpes,et al.  On the relationship between the sulphur content and the anticoagulant activity of heparin preparations. , 1939, The Biochemical journal.