Heparin synthesis

Zsebo KM, Metcalfe DD. Geissler EN. Galli SJ Induction of mast cell proliferation, maturation, and heparin synthesis by the rat c-kit ligand, stem dS cell factor. Proc Natl Acad Sci USA 1991 88 6382. 

An Approach for Heparin Synthesis Based on 3-Buten-2-yl Glycosides. 625... 

Heparin inhibits the formation of fibrin clots, inhibits the conversion of fibrinogen to fibrin, and inactivates several of the factors necessary for the clotting of blood. Heparin cannot be taken orally because it is inactivated by gastric acid in the stomach therefore, it must be given by injection. Heparin has no effect on clots that have already formed and aids only in preventing the formation of new blood clots (thrombi). The LMWHs act to inhibit clotting reactions by binding to antithrombin HI, which inhibits the synthesis of factor Xa and the formation of thrombin. 

Grainger DW, Kim SW, and Feijen J. Poly(dimethyl siloxane)-poly(ethylene oxide)-heparin block copolymers. I Synthesis and characterization. J Biomed Mater Res, 1988, 22, 231-242. 

Figure 1. Synthesis of F-D and predicted reaction with heparin.

In other studies, mast cells have been shown to be abundant in the marrow of osteoporotic patients, and heparin, which is contained within the secretory granules of connective tissue (peritoneal)-type mast cells, has been shown to enhance bone resorption and to inhibit bone-cell replication and collagen synthesis in vitro [130]. Moreover, heparin is known to bind growth factors such as fibroblast growth factor and may therefore be important in limiting their availability [134], Taken together, these various studies suggest a possible involvement of mast cells in the homeostasis of bone, but much more work is needed before any definitive conclusions can be drawn. 

Probably, one of the most valuable advances in this field has dealt with the first chemoenzymatic synthesis of the stable isotope-enriched heparin from a uniformly double labelled 13C, 15N /V-acetylheparosan from E. coli K5. Heteronuclear, multidimensional nuclear magnetic resonance spectroscopy was employed to analyze the chemical composition and solution conformation of N-acety 1 hcparosan, the precursors, and heparin. Isotopic enrichment was found to provide well-resolved 13C spectra with the high sensitivity required for conformational studies of these biomolecules. Stable isotope-labelled heparin was indistinguishable from heparin derived from animal tissues and might be employed as a novel tool for studying the interaction of heparin with different receptors.30... 

The anticoagulant fondaparinux, a synthetic analogue of the terminal fragment of heparin, is synthesized using multiple protection/deprotection steps that result in a route of up to 50 steps. There is, as yet, no enzymatic system that approaches the capability to make such a molecule." As this modified pentasaccharide is a natural product, it should, in theory, be accessible through a series of biotransformations, but we currently lack the biocatalytic tools to achieve more than a few steps and would stiU need to use some protection steps to avoid multiple products. Enzymatic synthesis in vivo depends largely on the levels and selectivities of glycosylating enzymes to achieve multistep reactions, a situation that has been mimicked in vitro for simpler systems." ... 

This process starts with the synthesis of novel chemical compounds. Substances with complex structures may be obtained from various sources, e.g., plants (cardiac glycosides), animal tissues (heparin), microbial cultures (penicillin G), or human cells (urokinase), or by means of gene technology (human insu-Un). As more insight is gained into structure-activity relationships, the search for new agents becomes more clearly focused. 

Histamine is synthesized from the amino acid histidine by an action of the enzyme histidine decarboxylase (Fig. 38.1). Following synthesis, histamine is either rapidly inactivated or stored in the secretory granules of mast cells and basophils as an inactive complex with proteases and heparin sulfate or chondroitin sulfate. 

The sulfonated calix[8]arene 27 (Structures 8), which can be viewed as a simplified y-CD with the carbohydrate backbone replaced by an aryl moiety, mimicked heparin in the stimulation of heparan sulfate synthesis from cultured endothelial cells [62]. 

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