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Synthetic heparin pentasaccharide depolymerization by heparinase I: molecular and biological implications.
1Department of Pathology, Loyola University Medical Center, Maywood, IL 60153, USA.
Summary
Synthetic pentasaccharide SR90107/ORG31540, used for venous thromboembolism, is effectively neutralized by heparinase I. This enzyme breaks down the pentasaccharide, suggesting its use in managing overdoses. A methylated analog showed resistance to heparinase I.
Area of Science:
- Biochemistry
- Pharmacology
Background:
- Synthetic pentasaccharides like SR90107/ORG31540 are developed for venous thromboembolism prophylaxis.
- These compounds target anti-factor Xa (AXa) activity with minimal anti-factor IIa (AIIa) effects.
Purpose of the Study:
- To investigate the effect of heparinase I on the AXa activity of synthetic pentasaccharide SR90107/ORG31540.
- To determine if heparinase I can neutralize the pentasaccharide's anticoagulant effects and identify breakdown products.
Main Methods:
- Incubation of SR90107/ORG31540 with varying concentrations of heparinase I.
- Analysis of AXa activity to assess neutralization.
- High-performance liquid chromatography (HPLC) and high-performance size exclusion chromatography (HPSEC) to identify breakdown products.
Main Results:
- Heparinase I demonstrated a significant neutralizing effect on SR90107/ORG31540's AXa activity.
- HPLC and HPSEC revealed concentration-dependent cleavage of the pentasaccharide into trisaccharide and disaccharide fragments.
- The disaccharide fragment showed a double bond, indicating susceptibility to enzymatic digestion.
Conclusions:
- Synthetic pentasaccharide SR90107/ORG31540 is digestible by heparinase I.
- Heparinase I can serve as a neutralizing agent for pentasaccharide overdose.
- A methylated analog (SanOrg34006) is resistant to heparinase I, suggesting potential for long-acting formulations.