PB1113 - Exploring the Synergistic Physical-Chemical Factors Behind Anticoagulant Properties in Heparins from Various Animal Sources

Background: Heparin, a vital anticoagulant in medical practice for almost a century, has been traditionally sourced from various animal tissues. Notably, the transition from bovine lung (HBL) to porcine intestine (HPI) marked a significant advancement in production efficiency and anticoagulant efficacy. The present study delves into the physical-chemical characteristics of heparins, including the emerging source from ovine intestine (HOI), to unravel the complexities influencing their anticoagulant activities.

Aims: This research aims to comprehensively understand the correlation between the physical-chemical attributes and anticoagulant effects of HBL, HPI, and HOI. By exploring molecular size, composition, and their collective impact, the study seeks to elucidate the nuanced factors contributing to their distinct anticoagulant properties.

Methods: To assess the structure, we employed analytical size-exclusion chromatography, NMR spectrometry analyses, disaccharide analyses with SAX-HPLC chromatography, and oligosaccharides analyses with size-exclusion chromatography. For activity evaluation, fluorimetric analyses of heparin-antithrombin interactions, activated partial thromboplastin time assay (APTT), in vitro anti-FXa and anti-FIIa activities, and in vivo antithrombotic activity were conducted.

Results: Analysis reveals variations in molecular size and composition among HBL, HPI, and HOI. While HPI and HOI exhibit similar and heightened anticoagulant potency compared to HBL, the study underscores the pivotal role of α-GlcN,3,6-trisulfated units in enhancing AT binding, particularly in HOI. Structural analyses through SEC and NMR techniques corroborate the overall similarity with minor yet discernible differences among the heparins.

Conclusion(s): It found that the anticoagulant effect is influenced by a synergistic blend of physical-chemical factors, particularly emphasizing molecular size and the presence of α-GlcN,3,6-trisulfated units. The practical implications suggest considering these heparins differently for medical use, proposing analytical methods for differentiation. The study also highlights the potential of the new pharmaceutical heparins as starting materials for low molecular weight heparins, calling for detailed investigations, especially for ovine origin.