DGKα/ζ-IN-2 is a potent, orally active, and highly selective inhibitor of DGKα/ζ, exhibiting IC50 values of 23 nM for DGKα and 1.2 nM for DGKζ. It shows excellent selectivity over other DGK isoforms such as DGKβ and DGKγ. In the OT-1 mouse model, DGKα/ζ-IN-2 demonstrates significant and dose-dependent immune activation under antigen-presenting conditions. This compound can be utilized in research related to anti-tumor immunity.

DGKα/ζ-IN-2 (compound 17) exhibits remarkable selectivity across multiple kinases (HPK1, EPHA2, FLT3, KDR, KIT, PDGFRβ, and TRKA) with IC₅₀ values > 10 μM. This compound effectively promotes IL-2 production in Jurkat cells (EC₅₀ = 64 nM) and human whole blood (EC₅₀ = 447 nM), showcasing significant functional activity. DGKα/ζ-IN-2 also demonstrates balanced in vitro ADME properties, including low human microsomal clearance rate (0.6 L/h/kg), high Caco-2 permeability (1.2 × 10⁻⁶ cm/s), and excellent solubility in both fasted state simulated intestinal fluid (> 1000 μg/mL) and simulated gastric fluid (659 μg/mL).

DGKα/ζ-IN-2, administered orally in single doses of 5, 25, and 50 mg/kg, induces robust, dose-dependent activation of CD8+ T cells in the presence of antigen presentation in an OT-I mouse model.