Oliveira Stephan N M C G, Bezerra Francisco F, Piquet Adriana A, Sales Rodrigo A, Valle Gabrielly C T, Capillé Nina V, Tovar Ana M F, Mourão Paulo A S
Laboratório de Tecido Conjuntivo, Hospital Universitário Clementino Fraga Filho and Instituto de Bioquímica Médica Leopoldo de Meis, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-913, Brazil.
ACS Omega. 2024 May 10;9(21):23111-23120. doi: 10.1021/acsomega.4c02128. eCollection 2024 May 28.
Low-molecular-weight heparin represent a significant advancement in anticoagulant therapy with enoxaparin being a prominent example obtained exclusively through the fragmentation of porcine intestinal heparin. However, escalating demand and limited resources have raised concerns about enoxaparin supplementation. The current challenge involves exploring alternative heparin sources for large-scale enoxaparin production with bovine intestinal heparin emerging as a promising option. Our study demonstrates that enoxaparin derived from the available bovine heparin preparation differs significantly from the reference compound. Yet, the implementation of a straightforward purification step yields a preparation termed "high-anticoagulant bovine heparin". Fragmentation of this purified product through β-elimination produces enoxaparin akin to the standard from a porcine origin. To ensure physicochemical similarity, we employed various spectroscopic, enzymatic, and chromatographic tests to compare the new bovine-derived enoxaparin with the original porcine compound. Biological activity was confirmed through coagulation assays and assessments using an animal model of venous thrombosis. Our study affirms that the β-elimination reaction cleaves the bovine heparin chain without preferential breaks in regions with different sulfation patterns. Additionally, we scrutinized decasaccharides purified from enoxaparin preparations, providing a comprehensive demonstration of the similarity between products obtained from porcine and bovine heparin. In summary, our findings indicate that an enoxaparin equivalent to the original porcine-derived product can be derived from bovine heparin, given that the starting material undergoes a simple purification step.
低分子量肝素是抗凝治疗的一项重大进展，依诺肝素就是一个突出例子，它完全通过猪肠肝素的片段化获得。然而，需求不断增加和资源有限引发了对依诺肝素补充的担忧。当前的挑战是探索用于大规模生产依诺肝素的替代肝素来源，牛肠肝素成为一个有前景的选择。我们的研究表明，从现有的牛肝素制剂中获得的依诺肝素与参考化合物有显著差异。然而，实施一个简单的纯化步骤可得到一种称为“高抗凝牛肝素”的制剂。通过β-消除对该纯化产物进行片段化可产生与猪源标准品类似的依诺肝素。为确保物理化学相似性，我们采用了各种光谱、酶和色谱测试，将新的牛源依诺肝素与原始猪化合物进行比较。通过凝血试验和使用静脉血栓形成动物模型的评估来确认生物活性。我们的研究证实，β-消除反应可切割牛肝素链，且在具有不同硫酸化模式的区域不会出现优先断裂。此外，我们仔细研究了从依诺肝素制剂中纯化的十糖，全面证明了从猪和牛肝素获得的产物之间的相似性。总之，我们的研究结果表明，只要起始材料经过简单的纯化步骤，就可以从牛肝素中获得与原始猪源产品等效的依诺肝素。
