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Semin Thromb Hemost 2002; 28(4): 335-342
DOI: 10.1055/s-2002-34302
Copyright © 2002 by Thieme Medical Publishers, Inc., 333 Seventh Avenue, New York, NY 10001, USA. Tel.: +1(212) 584-4662

Chemical Derivatization as a Strategy to Study Structure-Activity Relationships of Glycosaminoglycans

Benito Casu, Annamaria Naggi, Giangiacomo Torri
  • G. Ronzoni Institute for Chemical and Biochemical Research, Milan, Italy
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Publication History

Publication Date:
23 September 2002 (online)

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ABSTRACT

Sulfated glycosaminoglycans (GAGs) are amenable to a number of chemical modifications that modulate their biological activity. N-sulfate groups can be exposed and N-acylated (usually N-acetylated), specific O-sulfate groups can be removed, and free hydroxyl groups (either preexisting in the original GAG or exposed by desulfation) can be sulfated. Heparin/heparan sulfate, chondroitin sulfate, and dermatan sulfate have been variously desulfated or sulfated to afford novel GAGs with protein binding and associated biological properties different from those of the original GAGs. Regiospecific sulfation of N-acetyl heparosan (E. coli K5 polysaccharide) afforded a number of derivatives, some endowed with antithrombotic activity and others with antimetastatic properties. Most of the activities could be correlated with typical sulfation patterns along each GAG backbone. Glycol splitting of nonsulfated glucuronic residues (including a critical residue in the pentasaccharide sequence of the active site for antithrombin) leads to substantial loss of anticoagulant activity of heparin. Partial removal of sulfate groups at position 2 of iduronic acid residues followed by glycol splitting of all nonsulfated uronic acid residues afforded nonanticoagulant, antiangiogenic heparins.

KEYWORDS

Glycosaminoglycans - sulfation/desulfation - glycol splitting - structure-activity relationships

REFERENCES

  • 1 Kjellén L, Lindahl U. Proteoglycans: structure and interactions.  Ann Rev Biochem . 1991;  60 443-475
    CrossrefPubMedGoogle Scholar
  • 2 Lindahl U. Heparan sulfate: a polyanion with multiple messages.  Pure Appl Chem . 1997;  69 1897-1902
    PubMedGoogle Scholar
  • 3 Casu B, Lindahl U. Structure and biological interactions of heparin and heparan sulfate.  Adv Carbohydr Chem Biochem . 2001;  57 159-206
    PubMedGoogle Scholar
  • 4 Lane D A, Björk I, Lindahl U. Heparin and Related Polysaccharides.  New York: Plenum Press 1992
    Google Scholar
  • 5 Lindahl U, Lidholt K, Spillmann D, Kjellén L. More to ``heparin'' than anticoagulation.  Thromb Res . 1994;  75 1-32
    CrossrefPubMedGoogle Scholar
  • 6 Harenberg J, Casu B. Nonanticoagulant Actions of Glycosaminoglycans.  New York: Plenum Press; 1996
    Google Scholar
  • 7 Lever R, Page C P. Novel development opportunities for heparin.  Nature Rev Drug Discov . 2002;  2 140-148
    PubMedGoogle Scholar
  • 8 Casu B. Protein binding of sulfated glycosaminoglycans: searching for specificity. In: Harenberg J, Casu B, eds. Nonanticoagulant Actions of Glycosaminoglycans New York: Plenum Press 1996: 89-99
    Google Scholar
  • 9 Conrad H E. Heparin Binding Proteins.  New York: Academic Press; 1998
    Google Scholar
  • 10 Hileman R E, Fromm J R, Weiler J M, Linhardt R J. Glycosaminoglycan-protein interactions: definition of consensus sites in glycosaminoglycan binding proteins.  Bioessays . 1998;  20 156-167
    CrossrefPubMedGoogle Scholar
  • 11 Van Boeckel A A C, Petitou M. The unique antithrombin binding domain of heparin: a lead to new synthetic antithrombotics.  Angew Chem Int Ed Engl . 1993;  32 1671-1818
    PubMedGoogle Scholar
  • 12 Petitou M, Hérault J-P, Bernat A. Synthesis of thrombin-inhibiting heparin mimetics without side effects.  Nature . 1999;  398 417-422
    CrossrefPubMedGoogle Scholar
  • 13 Casu B, Torri G, Naggi A. Modulation of sulfation patterns of glycosaminoglycans. In: Fraser-Reid B, Tatsuta K, Thiem J, eds. Glycoscience Chemistry and Chemical Biology. Vol 3. Heidelberg: Springer Verlag; 2000: 1895-1904
    Google Scholar
  • 14 Torri G. New NMR spectroscopic approaches for structural analysis of glycosaminoglycans. In: Harenberg J, Casu B, eds. Nonanticoagulant Actions of Glycosaminoglycans New York: Plenum Press 1996: 15-25
    Google Scholar
  • 15 Casu B, Guerrini M, Naggi A. Characterization of sulfation patterns of beef and pig mucosal heparins by nuclear magnetic resonance spectroscopy.  Arzneim Forsch Drug Res . 1999;  46 472-477
    PubMedGoogle Scholar
  • 16 Guerrini M, Bisio A, Torri G. Combined quantitative 1H and 13C nuclear magnetic resonance spectroscopy for characterization of heparin preparations.  Semin Thromb Hemost . 2001;  27 473-482
    Article in Thieme ConnectPubMedGoogle Scholar
  • 17 Linhardt R J, Wang H, Ampofo S A. New methodologies in heparin structure analysis and the generation of LMW heparins. In: Lane DA, Björk I, Lindahl U, eds. Heparin and Related Polysaccharides New York: Plenum Press 1992: 37-47
    Google Scholar
  • 18 Thurnbull J H, Hopwood J T, Gallagher A. Strategy for rapid sequencing of heparan sulfate and heparin saccharides.  Proc Natl Acad Sci USA . 1999;  96 2698-2703
    CrossrefPubMedGoogle Scholar
  • 19 Vivès R R, Pye D A, Salvimirta M. Sequence analysis of heparan sulphate and heparin oligosaccharides.  Biochem J . 1999;  339 767-773
    CrossrefPubMedGoogle Scholar
  • 20 Venkataraman G, Shriver Z, Raman R, Sasisekharan R. Sequencing complex polysaccharides.  Science . 1999;  286 537-542
    CrossrefPubMedGoogle Scholar
  • 21 Inohue Y, Nagasawa K. Selective N-desulfation of heparin with dimethyl sulfoxide containing water or methanol.  Carbohydr Res . 1976;  46 87-95
    CrossrefPubMedGoogle Scholar
  • 22 Levvy G A, McAllan A. The N-acetylation and estimation of hexosamines.  Biochem J . 1959;  73 127-132
    PubMedGoogle Scholar
  • 23 Shaklee P N, Conrad H E. Hydrazinolysis of heparin and other glycosaminoglycans.  Biochem J . 1984;  217 187-197
    PubMedGoogle Scholar
  • 24 Loyd A G, Embery G, Fowler J. Preparation of [35S]sulfamate derivatives for studies on heparin degrading enzymes of mammalian origin.  Biochem Pharmacol . 1971;  20 637-664
    CrossrefPubMedGoogle Scholar
  • 25 Casu B. Structure and biological activity of heparin.  Adv Carbohydr Chem Biochem . 1985;  43 51-132
    PubMedGoogle Scholar
  • 26 Naggi A, Torri G, Casu B. ``Supersulfated'' heparin fragments, a new type of low-molecular weight heparin.  Biochem Pharmacol . 1986;  36 1895-1900
    PubMedGoogle Scholar
  • 27 Nagasawa K, Uchiyama H, Wajima N. Chemical sulfation of preparations of chondroitin 4- and 6-sulfate and dermatan sulfate. Preparation of chondroitin sulfate E-like materials from chondroitin 4-sulfate.  Carbohydr Res . 1986;  158 195-190
    PubMedGoogle Scholar
  • 28 Ogamo A, Metori A, Uchiyama H, Nagasawa K. Reactivity toward chemical sulfation of hydroxyl groups of heparin.  Carbohydr Res . 1989;  193 165-172
    CrossrefPubMedGoogle Scholar
  • 29 Naggi A, Torri G, Angiuli P. Sulfamino-galactosaminoglycans, a new class of semi-synthetic polysaccharides Preparation, characterization, and lipase-releasing properties. In: Biomedical and Biotechnological Advances in Industrial Polysaccharides New York: Gordon & Breach Science Publisher 1989: 101-108
    Google Scholar
  • 30 Naggi A. Simulation of glycosaminoglycan structure by chemical modifications of E coli polysaccharides K5 and K4. In: Harenberg J, Casu B, eds. Nonanticoagulant Actions of Glycosaminoglycans New York: Plenum Press 1996: 59-64
    Google Scholar
  • 31 Casu B, Grazioli G, Razi N. Heparin-like compounds prepared by chemical modification from E K5. Carbohydr Res .  1994;  263 271-284
    CrossrefPubMedGoogle Scholar
  • 32 Razi N, Feyzi E, Björk I. Structural and functional properties of heparin analogues obtained by chemical sulfation of Escherichia coli capsular polysaccharide K5.  Biochem J . 1995;  309 465-472
    PubMedGoogle Scholar
  • 33 Casu B, Grazioli G, Hannesson H H. Biologically active heparan sulfate-like species by combined chemical and enzymic modification of Escherichia coli polysaccharide K5.  Carbohydr Lett . 1994;  1 107-114
    PubMedGoogle Scholar
  • 34 Naggi A, Torri G, Casu B. Towards a biotechnological heparin through a combined chemical and enzymatic modification of the Escherichia coli K5 polysaccharide.  Semin Thromb Hemost . 2001;  27 437-441
    Article in Thieme ConnectPubMedGoogle Scholar
  • 35 Naggi A, De Cristofano A, Bisio A., et al. Generation of anti-factor Xa active, 3-O-sulfated glucosamine-rich sequences by controlled desulfation of oversulfated heparins.  Carbohydr Res . 2001;  336 283-290
    CrossrefPubMedGoogle Scholar
  • 36 Liu Z, Perlin A S. Regioselectivity in sulfation of some chemically-modified heparins, and observations on their cationic-binding characteristics.  Carbohydr Res . 1992;  236 121-133
    CrossrefPubMedGoogle Scholar
  • 37 Kovenski J, Cirelli A F. Chemical modification of glycosaminoglycans. Selective 2-sulfation of D-glucuronic acid in heparan sulfates.  Carbohydr Res . 1997;  303 119-122
    CrossrefPubMedGoogle Scholar
  • 38 Jaseja M, Rej R N, Sauriol F, Perlin A S. Novel regio- and stereoselective modifications of heparin in alkaline solution. Nuclear magnetic resonance spectroscopic evidence.  Can J Chem . 1989;  67 1449-1456
    PubMedGoogle Scholar
  • 39 Rej R N, Perlin A S. Base-catalyzed conversion of the α-L-iduronic acid 2-sulfate unit of heparin into a unit of α-L-galacturonic acid, and related reactions.  Carbohydr Res . 1992;  200 437-447
    PubMedGoogle Scholar
  • 40 Piani S, Casu B, Marchi E. Alkali-induced optical rotation changes in heparins and heparan sulfates and their relation to iduronic acid containing sequences.  J Carbohydr Chem . 1993;  12 507-521
    PubMedGoogle Scholar
  • 41 Holme K R, Liang W, Yang Z. A detailed evaluation of the structural and biological effects of alkaline O-desulfation reactions of heparin. In: Harenberg J, Casu B, eds. Nonanticoagulant Actions of Glycosaminoglycans New York: Plenum Press 1996: 139-162
    Google Scholar
  • 42 Santini F, Bisio A., Guerrini M. Modification under basic conditions of the minor sequences of heparin containing 2,3 or 2,3,6 sulfated D-glucosamine residues.  Carbohydr Res . 1997;  302 103-108
    CrossrefPubMedGoogle Scholar
  • 43 Casu B, Petitou M, Provasoli M, SinaĀ P. Conformational flexibility: a new concept for explaining binding and biological activity of iduronic acid-containing glycosaminoglycans.  Trends Biochem Sci . 1998;  13 221-225
    PubMedGoogle Scholar
  • 44 Casu B. Structure, shape and functions of glycosaminoglycans. In: Harenberg J, Heene DL, Stehle G, Schettler G, eds. New Trends in Haemostasis Heidelberg: Springer Verlag 1990: 2-11
    Google Scholar
  • 45 Casu B, Guerrini M, Naggi A. Short heparin sequences spaced by glycol-split uronate residues are antagonists of fibroblast growth factor-2 and angiogenesis inhibitors (in press).  Biochemistry. 2002; 
    PubMedGoogle Scholar
  • 46 Pisano C, Cervoni M L, Chiarucci I. Antiangiogenic and antitumoral activity of novel heparin derivatives devoid of anticoagulant effects. National Cancer Institute-European Organization for Research and Treatment of Cancer-American Association for Cancer Research Symposium (NCI-EORTC- AACR), 2002 (Abst)
    Google Scholar
  • 47 Bisio A, Casu B, Menozzi C. Physico-chemical characterization and interaction with heparin cofactor-2 of ``disulfated'' galactosaminoglycans. Proceedings 3rd Italian Carbohydrate Symposium, Grado, 1991;86 (Abst)
    Google Scholar
  • 48 Maruyama T, Toida T, Imanari T, Yu G, Linhardt R J. Conformational changes and anticoagulant activity of chondroitin sulfate following its O-sulfonation.  Carbohydr Res . 1998;  306 35-43
    CrossrefPubMedGoogle Scholar
  • 49 Gigli M, Ghiselli G, Torri G. A comparative study of low-density lipoprotein interaction with glycosaminoglycans.  Biochim Biophys Acta . 1993;  1167 211-217
    PubMedGoogle Scholar
  • 50 Kostoulas G, Horler D, Naggi A. Electrostatic interactions between human leukocyte elastase and sulfated glycosaminoglycans: physiological implications.  Biol Chem . 1997;  378 1481-1489
    PubMedGoogle Scholar
  • 51 Pye D A, Vivès R R, Hyde P, Gallagher J T. Regulation of FGF-1 mitogenic activity by heparan sulfate oligosaccharides is dependent on specific structural features: differential requirements for the modulation of FGF-1 and FGF-2.  Glycobiology . 2000;  10 1183-1192
    CrossrefPubMedGoogle Scholar
  • 52 Lundin L, Larsson H, Kreuger J. Selectively desulfated heparin inhibits fibroblast growth factor-induced mitogenicity and angiogenesis.  J Biol Chem . 2000;  275 24653-24600
    CrossrefPubMedGoogle Scholar
  • 53 Naggi A, Perez M, Torri G. Modulation of heparanase-inhibiting activity of heparin through selective desulfation, graded N-acetylation, and glycol splitting. Abstract 21st International Carbohydr Symposium, Cairns, Australia 2002
    Google Scholar