EGFR/HER2/DHFR-IN-1 is a highly selective and potent anticancer compound targeting MCF-7 breast cancer cells, acting as a multi-inhibitor for EGFR/HER2 kinase and DHFR enzymes, with IC50 values of 0.153 μM, 0.108 μM, and 0.291 μM, respectively. It induces cell cycle arrest at the G1/S phase and apoptosis in cells [1].

EGFR/HER2-IN-7 (compound 39) exhibits broad-spectrum cytotoxic efficacy with an IC50 of 1.83 μM against MCF-7 breast cancer cell lines, demonstrating strong anti-cancer properties [1]. It induces apoptosis and G1/S phase cell cycle arrest in various cancer cell lines, including HepG2, MCF-7, HCT-116, PC-3, and HeLa, at 0-1 mM over 72 hours. The compound inhibits EGFR/HER2 kinase and DHFR, leading to S phase and G1 phase arrest, respectively. EGFR/HER2-IN-7 significantly reduces cancer cell growth with IC50 values of 3.48 μM (HepG2), 1.83 μM (MCF-7), 6.08 μM (HCT-116), 12.74 μM (PC-3), and 4.78 μM (HeLa), highlighting its potential as a versatile oncology therapeutic agent [1].

Caspase-3, a lysosomal enzyme pivotal for apoptosis, serves as a crucial biomarker for identifying apoptotic cells. Compound EGFR/HER2-IN-7, administered at a dosage of 10 mg/kg via intraperitoneal injection to Swiss albino female mice bearing breast cancer without lactating mammary glands, demonstrated significant anti-breast cancer efficacy over a duration of 20 days. The treatment not only reduced tumor volume by 76.5% and body weight by 17.4% but also notably increased Caspase-3 immunoexpression, with a Caspase-3 score of 1.33, underscoring its potential as a therapeutic agent against breast cancer.