Journal: Chembiochem : a European journal of chemical biology

Article Title: Gucosamine-6-sulfamate Analogs of Heparan Sulfate as Inhibitors of Endosulfatases

doi: 10.1002/cbic.201000401

Figure Lengend Snippet: (A) Biochemical evaluation of inhibitors against Sulf-1 and Sulf-2 using 4-MUS assay. Bars: percent inhibition in the presence of 5 mM inhibitor relative to uninhibited controls (*3a and 3b show full inhibition at 5mM, and bars are labelled with half maximal biochemical activity, or IC50, values). (B) Selectivity of sulfamate inhibitors. GlcN-6-sulfamates 3a and 3b are more selective for the Sulfs than PARS and are more potent than general inhibitors 1 and 2 (Bars: percent activity in the presence of 500µM inhibitor relative to uninhibited control). (C) Anti-inflammatory activity of Inhibitors. Production of IL-6 by IL-1β stimulation is a proinflammatory response in arthritis. Data show this response is inhibited by up to 50% by sulfamate-based sulfatase inhibitors (Bars: percent the IL-6 response is attenuated in the presence of 200 µM inhibitor relative to uninhibited control in IL-1β stimulated fibroblast-like synoviocytes).

Article Snippet: Cell-free culture supernatants or human recombinant IL-6 standard (25 µL) was added to the plates and incubated for 2 hours followed by the addition of 1 µg biotinylated goat antihuman IL-6 detection antibody (R & D Systems) for 1 hour.

Techniques: Inhibition, Activity Assay