PF-4136309

PF-4136309 is potent in inhibiting CCR2 mediated signaling events such as intracellular calcium mobilization and ERK (extracellular signal-regulated kinase) phosphorylation. ^{<a class="sref" href="#s_ref">[1]</a>}

500,000 HPBMCs in serum free DMEM media are incubated with or without PF-4136309 and warmed to 37 °C. 400 μL of warmed 10 nM MCP-1 is added to the bottom chamber in all wells except the negative control. An 8 micron membrane filter is placed on top and the chamber lid is closed. Cells are then added to the holes in the chamber lid which are associated with the chamber wells below the filter membrane. The whole chamber is incubated at 37 °C, 5% CO2 for 30 min. The cells are then aspirated off, the chamber lid opened, and the filter gently removed. The filter is air dried and stained with Wright Geimsa stain. Filters are counted by microscopy.  Antagonist potency is determined by comparing the number of cells that migrate to the bottom chamber in wells which contained antagonist, to the number of cells which migrate to the bottom chamber in MCP-1 control wells.

PF-4136309 exhibits >100-fold selectivity over other homologous chemokine receptors, a free fraction of 24% in human serum and 15% in mouse serum, and an oral bioavailability of 47% in mice, suitable as a tool compound for target validation in rodent models.^{<a class="sref" href="#s_ref">[2]</a>}