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US 2013/0296540 A1Published application
Chemoenzymatic synthesis of structurally homogeneous ultra-low molecular weight heparins
AssigneeThe University of North Carolina at Chapel Hill
InventorsYongmei Xu, Jian Liu
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Abstract
Methods for preparing synthetic heparins are provided. Synthetic heparin compounds, including ultralow molecular weight heparin compounds are provided. Also provided are methods of chemoenzymatically synthesizing structurally homogeneous ultra-low molecular weight heparins. Heparin compounds provided herein can have anticoagulant activity.
Claims29
A method of synthesizing a heparin compound, comprising:
- providing a saccharide substrate;
- elongating the saccharide substrate to a saccharide of a desired or predetermined length;
- performing an epimerization reaction;
- and
- performing one or more sulfation reactions,
- whereby a heparin compound is synthesized.
3 more elements
A method of synthesizing a heparin compound, comprising:
- providing a disaccharide substrate;
- elongating the disaccharide substrate to a tetrasaccharide;
- elongating the tetrasaccharide to a hexasaccharide or heptasaccharide, wherein the hexasaccharide or heptasaccharide comprises a N-sulfotransferase substrate residue;
- converting the N-sulfotransferase substrate residue on the hexasaccharide or heptasaccharide to a N-sulfo glucosamine (GlcNS) residue;
- performing an epimerization reaction;
- and
- performing one or more sulfation reactions selected from the group consisting of a 2-O-sulfation reaction, a 6-O-sulfation reaction, a 3-O-sulfation reaction, and combinations thereof,
- whereby a heparin compound is synthesized.
5 more elements
A method of synthesizing a heparin compound, comprising:
- providing a disaccharide substrate;
- elongating the disaccharide substrate to a tetrasaccharide using enzymes N-acetyl glucosaminyl transferase and heparosan synthase-2, and substrates glucuronic acid (GlcUA) and N-trifluoroacetyl glucosamine (GlcNTFA);
- elongating the tetrasaccharide to a heptasaccharide using enzymes N-acetyl glucosaminyl transferase and heparosan synthase-2, and substrates glucuronic acid (GlcUA), N-trifluoroacetyl glucosamine (GlcNTFA), and N-acetylated glucosamine (GlcNAc);
- converting N-trifluoroacetyl glucosamine (GlcNTFA) residue(s) on the heptasaccharide to N-sulfo glucosamine (GlcNS) residues using N-sulfotransferase (NST), 3′-phosphoadenosine 5′-phosphosulfate (PAPS), triethylamine, CH3OH, and H2O;
- epimerizing the heptasaccharide using C5-epimerase (C5-epi);
- sulfating the heptasaccharide using 2-O-sulfotransferase (2-OST) and 3′-phosphoadenosine 5′-phosphosulfate (PAPS);
- sulfating the heptasaccharide using 6-O-sulfotransferase (6-OST) and 3′-phosphoadenosine 5′-phosphosulfate (PAPS);
- and
- sulfating the heptasaccharide using 3-O-sulfotransferase (3-OST) and 3′-phosphoadenosine 5′-phosphosulfate (PAPS);
- wherein a heparin compound is synthesized.
7 more elements
A heparin compound comprising the following structure:
A method of synthesizing a heparin compound, comprising:
- providing a disaccharide substrate;
- elongating the disaccharide substrate to a tetrasaccharide using enzymes N-acetyl glucosaminyl transferase and heparosan synthase-2, and substrates glucuronic acid (GlcUA) and N-trifluoroacetyl glucosamine (GlcNTFA);
- elongating the tetrasaccharide to a hexasaccharide using enzymes N-acetyl glucosaminyl transferase and heparosan synthase-2, and substrates glucuronic acid (GlcUA) and N-trifluoroacetyl glucosamine (GlcNTFA);
- converting N-trifluoroacetyl glucosamine (GlcNTFA) residue(s) on the hexasaccharide to N-sulfo glucosamine (GlcNS) residues using N-sulfotransferase (NST), 3′-phosphoadenosine 5′-phosphosulfate (PAPS), triethylamine, CH3OH, and H2O;
- elongating the hexasaccharide to a heptasaccharide using enzyme N-acetyl glucosaminyl transferase and substrate N-trifluoroacetyl glucosamine (GlcNTFA);
- epimerizing the heptasaccharide using C5-epimerase (C5-epi);
- sulfating the heptasaccharide using 2-O-sulfotransferase (2-OST) and 3′-phosphoadenosine 5′-phosphosulfate (PAPS);
- converting N-trifluoroacetyl glucosamine (GlcNTFA) residue(s) on the heptasaccharide to N-sulfo glucosamine (GlcNS) residues using N-sulfotransferase (NST), 3′-phosphoadenosine 5′-phosphosulfate (PAPS), triethylamine, CH3OH, and H2O;
- sulfating the heptasaccharide using 6-O-sulfotransferase (6-OST) and 3′-phosphoadenosine 5′-phosphosulfate (PAPS);
- and
- sulfating the heptasaccharide using 3-O-sulfotransferase (3-OST) and 3′-phosphoadenosine 5′-phosphosulfate (PAPS);
- wherein a heparin compound is synthesized.
9 more elements
A heparin compound comprising the following structure:
A heparin compound comprising the following structure:
- wherein R2 is OH, OSO3H, OCH3, or OCH2CH3;
- R3 is NHSO3H or NHCOCH3;
- R4 is OH, OSO3H, OCH3, or OCH2CH3;
- R5 is NHSO3H or NHCOCH3;
- and R6 is OH, OSO3H, OCH3, or OCH2CH3.
2 more elements
Dependencies are inferred from the claim text.