Heavy Heparin: A Stable Isotope-Enriched, Chemoenzymatically-Synthesized, Poly-Component Drug
Heavy Heparin: A Stable Isotope-Enriched, Chemoenzymatically-Synthesized, Poly-Component Drug
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DOI：
10.1002/anie.201900768
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发表时间：
2019-04-23
影响因子：
16.6
通讯作者：
Linhardt, Robert J.
中科院分区：
文献类型：
--
作者：
Cress, Brady F.;Bhaskar, Ujjwal;Linhardt, Robert J.
肝素是一种高度硫酸化的复合多糖，是广泛使用的抗凝药物。在这项工作中，我们从氘化培养基中微生物培养物中获得的生物合成富集的肝素前体中化学酶法合成了全氘化肝素。使用重组肝素生物合成酶进行化学脱 N-乙酰化、化学 N-硫酸化、酶促差向异构化和酶促硫酸化，得到与药用肝素相当的全氘代肝素。展示了重质肝素及其重质生物合成中间体的一系列应用，包括生成稳定同位素标记的二糖标准品、开发葡萄糖醛酸基-C5-差向异构酶的非放射性核磁共振测定，以及对兔子给药后体内半衰期的无背景定量。我们预计这种方法可以扩展到生产其他富含同位素的糖胺聚糖。
Heparin is a highly sulfated, complex polysaccharide and widely used anticoagulant pharmaceutical. In this work, we chemoenzymatically synthesized perdeuteroheparin from biosynthetically enriched heparosan precursor obtained from microbial culture in deuterated medium. Chemical de-N-acetylation, chemical N-sulfation, enzymatic epimerization, and enzymatic sulfation with recombinant heparin biosynthetic enzymes afforded perdeuteroheparin comparable to pharmaceutical heparin. A series of applications for heavy heparin and its heavy biosynthetic intermediates are demonstrated, including generation of stable isotope labeled disaccharide standards, development of a non-radioactive NMR assay for glucuronosyl-C5-epimerase, and background-free quantification of invivo half-life following administration to rabbits. We anticipate that this approach can be extended to produce other isotope-enriched glycosaminoglycans.