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Semin Thromb Hemost 1997; 23(2): 99-107
DOI: 10.1055/s-2007-996077
Copyright © 1997 by Thieme Medical Publishers, Inc.

Synthesis and Biological Effects of N-Alkylamine-Labeled Low-Molecular-Mass Dermatan Sulfate

Reinhard Malsch* , Marco Guerrini† , Carlo Berti† , Annamaria Naggi† , Giangiacomo Torri† , Benito Casu† , Job Harenberg*
  • From the *1st Department of Medicine, Faculty of Clinical Medicine Mannheim, University of Heidelberg, Mannheim, Germany, and
  • †Institute di Chimica e Biochimica “G. Ronzoni,” Milan, Italy.
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Publication History

Publication Date:
08 February 2008 (online)

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Abstract

Dermatan sulfate (DS) is a component of connective tissue and catalyzes the heparin cofactor II-mediated inhibition of thrombin. Low-molecular-mass dermatan sulfates (LMMDS) are produced to prolong the antithrombotic activity of this substance. Cleavage of DS by nitrous acid leads to an LMMDS with a terminal 2,5-anhydrotalose (At) group at the reducing end which can react with primary amines. Tyramine (Tyr) was bound to the terminal At of LMMDS using reductive amination.

LMMDS-tyr is produced using DS. LMMDS desacetglated were produced using totally deaminated DS. These compounds were employed as a model for the characterization of DS using NMR spectroscopy. The purity of the compounds was checked using capillary electrophoresis. The structure of the products was proven by 1H- and 13C-NMR spectroscopy.

LMMDS-Tyr was radiolabeled with 125I for use in a radioimmunoassay. The anti-Xa activity and antithrombin activity of the tyramine-labeled DS are very low. The clotting assays Heptest, aPTT, thrombin time, and ecarin time indicate a highly anticoagulant-active substance. The heparin cofactor II-mediated inhibition of thrombin is similar to the parent compound.

LMMDS were labeled “endpoint-attached.” They are a new tool to understand the actions of DS in biologic systems.

Keywords:

Dermatan sulfate - low-molecular-mass dermatan sulfate - dermatan sulfate-tyramine - radiolabeled dermatan sulfate - NMR spectroscopy