EGFR-IN-173 is an orally active, broad-spectrum mutant EGFR tyrosine kinase inhibitor that targets EGFR19del, L858R/T790M, and the triple mutation C797S, with an IC50 of 1.19 nM, effectively inhibiting the EGFR19del/T790M/C797S mutant. It demonstrates over 100-fold selectivity for mutated EGFR compared to the wild type [wild type IC50= 19.362 μM]. EGFR-IN-173 significantly suppresses cell migration, induces apoptosis in non-small cell lung cancer (NSCLC) cells, inhibits EGFR phosphorylation, and blocks downstream pathways (MAPK/ERK, AKT, STAT3). It shows antitumor activity in NSCLC and Ba/F3 xenograft models and is applicable in NSCLC research.

EGFR-IN-173 (compound D10) exhibits strong antiproliferative effects against various EGFR mutants, with an IC50 of 0.69 nM in HCC827 cells (EGFR 19del), 0.242 μM in H1975 cells (EGFR L858R/T790M), 0.192 μM in Ba/F3-EGFR 19del/T790M/C797S cells, and 1.303 μM in Ba/F3-EGFR L858R/T790M/C797S cells, demonstrating over 100-fold selectivity for mutant EGFR (wild-type IC50 = 19.362 μM). It occupies the active site of EGFR, forming crucial hydrogen bonds with Lys728 and Ser797. At concentrations of 10-100 nM over 14 days, EGFR-IN-173 dose-dependently inhibits colony formation of non-small cell lung cancer HCC827 cells, impeding their long-term proliferation. Additionally, EGFR-IN-173 (10-100 nM, 0-48 hours) effectively inhibits the migration of non-small cell lung cancer H1975 cells in a concentration-dependent manner. It induces G0/G1 phase arrest in HCC827 cells at concentrations of 20-200 nM over 48 hours, in a dose-dependent fashion. Furthermore, EGFR-IN-173 (1-2000 nM, 48 hours) triggers both early and late apoptosis at varying concentrations by inducing hallmark morphological changes such as chromatin condensation, nuclear fragmentation, and apoptotic body formation. At 0.01-1 μM over 48 hours, it dose-dependently suppresses EGFR phosphorylation and inhibits downstream pathways (MAPK/ERK, AKT, STAT3), thereby initiating apoptosis and degradation of signaling proteins like ERK1/2.

EGFR-IN-173, administered orally at doses of 25 and 50 mg/kg once daily for 12 days, demonstrates significant tumor growth inhibition in the HCC827 xenograft mouse model. Additionally, EGFR-IN-173 at a dose of 50 mg/kg, given orally once daily for 17 days, shows some inhibitory effect on the triple mutation EGFR 19del/T790M/C797S in the Ba/F3-EGFR 19del/T790M/C797S xenograft mouse model.