In vitro activity: AZD7986 (formerly known as INS 1007) is a novel, second generation, highly potent, reversible, and selective DPP1 (Dipeptidyl peptidase 1) inhibitor pIC50s of 6.85, 7.6, 7.7, 7.8, and 7.8 in human, mouse, rat, dog and rabbit, respectively. It has the potential for COPD (chronic obstructive pulmonary disease) treatment with predicted human PK properties suitable for once daily human dosing.

Kinase Assay: Activation of neutrophil serine proteases (NSPs) is assessed in vitro using primary bone marrow derived CD34+ neutrophil progenitor cells. Cells are cultured in media supplemented with rhSCF and rhIL-3 for 7 days, and then for a further 7 days in the presence of G-CSF and different concentrations of AZD7986 (38 pM to 10 μM). After harvesting and lysis with 10% Triton X-100 buffer, cell lysates are kept at -20°C until NSP activity analysis.

Cell Assay: Cellular potency is studied using the DPP1-expressing monocytic U937 cell line. Briefly, cells grown in RPMI are plated on 384-well polypropylene v-bottom plates at a density of 5×105 cells/mL per well. Added to this is 10 μL of AZD7986 at 37°C for 60 min, followed by 350 μM Gly-Phe-AFC. The well fluorescence is read using a multilabel plate reader. Data are analyzed to calculate pIC50 values.