Cell experiment:

Mononuclear cells are incubated for 30 min with BW-A 78U at the concentration of 10 nM to 10 μM. The cells are then stimulated with lipopolysaccharide (10 μg/mL) overnight at 37℃ in an atmosphere of 5% CO2 at 100% humidity. Cell-free supernatants are collected, centrifuged (2000 g), and stored frozen at -20℃ before TNF-α determination. TNF-α concentrations in cell culture supernatants are determined by specific ELISA using a commercial kit. Sensitivity of the assay is 1 pg/mL. The absorbance at 450 nm is assessed with an ELISA reader[1].

BW-A 78U is a PDE4 inhibitor with an IC50 of 3 μM.

BW-A 78U is a PDE4 inhibitor with an IC50 of 3 μM. BW-A 78U fails to significantly inhibit arachidonate release. BW-A 78U is ineffective to inhibit the lipopolysaccharide (LPS)-induced TNF-α release[1].

[1]. Boichot E, et al. Anti-inflammatory activities of a new series of selective phosphodiesterase 4 inhibitors derivedfrom 9-benzyladenine. J Pharmacol Exp Ther. 2000 Feb;292(2):647-53.