PF-8380 is an effective and orally available autotaxin inhibitor (IC50: 2.8 nM, in isolated enzyme assay; 101 nM, in the human whole blood). It modulates lysophosphatidic acid (LPA) levels in vivo/vitro by directly inhibiting autotaxin; reduces LPA levels both in plasma and at the site of inflammation.

ATX:2.8 nM

Pre-treatment with PF-8380 before radiotherapy can inhibit radiation-induced angiogenesis in tumor endothelial cells and delay the growth and development of intracranial glioma tumors. In a rat air pouch inflammation model, PF8380 (30 mg/kg, p.o.) reduced inflammatory pain sensitivity and decreased LPA levels in plasma and inflamed tissues by more than 95% within 3 hours.

PF-8380 inhibits autotoxin in rat substrates (IC50: 1.16 nM) and demonstrates inhibition in human whole blood with an IC50 of 101 nM after 2 hours of treatment [1]. Furthermore, PF-8380 suppresses autotoxin in GBM cells, subsequently reducing their invasion and migration capabilities while enhancing radiation sensitization.

FS-3 substrate is solubilized in assay buffer at 500 μM and frozen at -20°C in single-use aliquots for up to 4 weeks. Recombinant autotaxin is diluted in Tris-buffered saline (140 mM NaCl, 5 mM KCl, 1 mM CaCl2, 1 mM MgCl2, 50 mM Tris, pH 8.0) and incubated with compound in DMSO or DMSO alone (final 1% DMSO) for 15 min at 37°C, and the reaction is started with the addition of FS-3 at a final concentration of 1 μM. The reaction is allowed to proceed at 37°C for 30 min and monitored at 520 nm until the uninhibited control compared with a no-enzyme control gave a Z′≥0.5. IC50s are determined in triplicate by using a four-parameter fit[1].

GL261 or U87-MG cells are plated in triplicate onto 6 cm plates and allowed to grow to 70% confluence. The semi-confluent cell layer is scratched with a sterile 200 μL pipette tip to create a scratch devoid of cells and plates are washed once with PBS to remove non-adherent cells and debris. For radiosensitization drug studies, cells are treated with 1 μM PF-8380 or DMSO for 45 min prior to irradiation with 4 Gy, and then incubated at 37°C in 5% CO2. Control plates are monitored for cell migration (20–24 h). Cells are fixed with 70% ethanol and stained with 1% methylene blue. To quantify migration, cells in three randomly selected high power fields (HPFs) in the scratched area are counted and normalized for surrounding cell density.(Only for Reference)

DMSO: < 1 mg/mL (insoluble or slightly soluble)