EGFR-IN-176 is an orally active and ATP-competitive inhibitor specifically targeting mutated forms of EGFR, particularly the C797S-mediated EGFR triple mutant. It effectively inhibits the activation of the AKT signaling pathway and induces apoptosis in Ba/F3 and PC-9 cells, which express EGFR19del/T790M/C797S or EGFRL858R/T790M/C797S mutations, respectively. EGFR-IN-176 selectively blocks EGFR signaling in cell lines with the EGFR triple mutation, without affecting wild-type EGFR expressed in A431 cells. Additionally, EGFR-IN-176 effectively inhibits ALK enzyme activity with an IC50 of less than 0.5 nM and is useful in the study of non-small cell lung cancer.

EGFR-IN-176 (Compound 31r, 0.0045 nM-10,000 nM, 48 hours) demonstrates highly selective inhibitory activity on the growth of cells with EGFR triple mutations in vitro, including Ba/F3 EGFR 19del/T790M/C797S (IC50 = 0.001 nM) and Ba/F3 EGFR L858R/T790M/C797S (IC50 = 0.04 nM), while showing lower cytotoxicity towards wild-type EGFR cells A431 (IC50 > 10,000 nM). At 500 nM over 7 days, EGFR-IN-176 effectively suppresses proliferation of PC-9 EGFR 19del/T790M/C797S and PC-9 EGFR L858R/T790M/C797S, and ultimately diminishes the self-renewal and colony formation abilities of cancer cells. Additionally, at 250 nM for 0-24 hours, it efficiently inhibits the migration capacity of the same PC-9 EGFR mutants. Furthermore, treatment with 500 nM for 48 hours induces apoptosis in PC-9 EGFR 19del/T790M/C797S and PC-9 EGFR L858R/T790M/C797S cells.

EGFR-IN-176 (Compound 31r), administered orally at dosages of 40-80 mg/kg daily over 21 days, significantly inhibits tumor growth in a dose-dependent manner, leading to observable tumor regression in PC-9 xenograft mice.