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

EGFR/HER2-IN-7 (compound 39) shows remarkable broad spectrum cytotoxic potency, with an IC 50 value of 1.83 μM against MCF-7 breast cancer cell lines [1]. EGFR/HER2-IN-7 (1.83 μM) induces cell apoptosis by arreating cell cycle at G1/S [1]. Cell Cycle Analysis [1] Cell Line: HepG2 hepatocellular carcinoma, MCF-7 breast cancer, HCT-116 colorectal carcinoma, PC-3 prostate and Hea cervical epithelioid carcinoma Concentration: 0-1 mM Incubation Time: 72 hours Result: Inhibited EGFR/HER2 kinase and DHFR, while DHFR inhibition caused cell cycle arrest at the S phase while EGFR/HER2 kinase inhibition caused arrest at the G1 phase. Apoptosis Analysis [1] Cell Line: HepG2 hepatocellular carcinoma, MCF-7 breast cancer, HCT-116 colorectal carcinoma, PC-3 prostate and Hea cervical epithelioid carcinoma Concentration: 0-1 mM Incubation Time: 72 hours Result: Inhibited cancer cells growth and induced apopsosis with IC 50 s of 3.48 μM (HepG2), 1.83 μM (MCF-7), 6.08 μM (HCT-116), 12.74 μM (PC3), 4.78 μM (Hela), respectively.

Caspase-3 is a lysosomal enzyme involved in apoptosis, and is used as a biomarker for detection of apoptotic cells [1]. EGFR/HER2-IN-7 (compound 39) (10 mg/kg; i.p.; once daily; 20 d) shows anti-breast cancer activity in vivo and increases caspase-3 immunoexpression in brease cancer mice [1]. Animal Model: Breast cancer with non-lactating mammary glands animal model in Swiss albino female mice (8-weeks-old) [1] Dosage: 10 mg/kg Administration: Intraperitoneal injection; once daily; 20 days Result: Inhibited tumor volume with reduction rate of 76.5%. Reduced body weight with loss rate of 17.4%. Showed the Caspase-3 score of 1.33.