Heavy Heparin: A Stable Isotope‐Enriched, Chemoenzymatically‐Synthesized, Poly‐Component Drug

Abstract

Heparin is ah ighly 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-Nacetylation, chemical N-sulfation, enzymatic epimerization, and enzymatic sulfation with recombinant heparin biosynthetic enzymes afforded perdeuteroheparin comparable to pharmaceutical heparin. Aseries of applications for heavy heparin and its heavy biosynthetic intermediates are demonstrated, including generation of stable isotope labeled disaccharide standards, development of an on-radioactive NMR assayf or glucuronosyl-C5-epimerase,a nd background-free quantification of in vivo half-life following administration to rabbits.W e anticipate that this approach can be extended to produce other isotope-enriched glycosaminoglycans.

Synthetic heparan sulfate standards and machine learning facilitate the development of solid-state nanopore analysis

“…We prepared a carefully designed group of chemoenzymatically synthesized HS GAG chains and have synthesized these GAGs following a similar strategy as described in our previous publication (34). Biosynthetically enriched heparosan precursor was obtained from microbial culture.…”

“…NSH to NS6S2S Sample Preparation Major Pathway. We have synthesized GAGs following similar strategy as our previous publication (34). Biosynthetically enriched heparosan precursor is obtained from microbial culture.…”

Heparan Sulfate Glycomimetics via Iterative Assembly of “Clickable” Disaccharides

“…To understand the structure-activity relationships of the sulfation patterns in HS, Seeberger, [6,7,8] Linhardt, [9,10,11] Liu, [12,13,14] Boons, [15,16] Weigel, [17,18] DeAngelis, [19,20] Hung, [21,22,23] Huang, [24,25,26,27] and others [28,29,30,31,32,33,34] have developed chemical and chemoenzymatic synthetic approaches to welldefined HS oligosaccharides that have significantly advanced our understanding of HS biology. Nevertheless, the chemical synthesis of HS oligosaccharides often requires exquisite synthetic route design and skillful carbohydrate chemistry manipulations, resulting in high production cost and scalability challenges.…”

Recent advances in biotechnology for heparin and heparan sulfate analysis

“…Recently, biotechnological versions of UFH, LMWH and ULMWH have reached the stage of commercial development [16,17,18]. These products rely on chemoenzymatic synthesis making their preparation more cost competitive than products like fondaparinux, which relies on chemical synthesis.…”