Development of Quinazoline-Tetrahydrofuranyl-Pyridinyl Hybrids as Dual EGFR and HER۲ Inhibitors to Combat Lapatinib Resistance in Breast Cancer

Lapatinib resistance in HER۲-positive breast cancer poses a major therapeutic challenge. In this study, we developed novel quinazoline-tetrahydrofuranyl-pyridinyl hybrids as dual EGFR and HER۲ inhibitors to address this resistance. Among the synthesized compounds, compounds ۱۳e, ۱۳i, and ۱۳j were selected for detailed in vitro evaluation. ۱۳i exhibited superior dual inhibitory activity, with IC۵۰ values of ۲۷.۴ ± ۱.۸ nM for EGFR and ۱۸.۲ ± ۱.۹ nM for HER۲, outperforming lapatinib (IC۵۰:۲۹.۷ ± ۱.۳ nM for EGFR, ۱۹.۳ ± ۱.۵ nM for HER۲). In cytotoxicity assays, ۱۳i demonstrated potent antiproliferative effects, with IC۵۰ values of ۱.۹۵ ± ۰.۱۷ µM in BT-۴۷۴, ۲.۵۵ ± ۰.۲۳ µM in SKBR۳, and ۵.۹۲ ± ۰.۲۶ µM in lapatinib-resistant LAP-SKBR۳ cells, compared to lapatinib’s ۲۱.۶ ± ۱.۵۴ µM in LAP-SKBR۳. Apoptosis assays revealed that ۱۳i induced late apoptosis in ۵۱.۳ ± ۴.۴% of LAP-SKBR۳ cells, which was significantly higher than lapatinib’s ۱۵.۲ ± ۱.۴%. DNA fragmentation assays confirmed extensive genomic cleavage in resistant cells treated with ۱۳i, in contrast to the marginal effects of lapatinib. Additionally, ۱۳i at ۲xIC₅₀ reduced migration and invasion in LAP-SKBR۳ cells to levels significantly lower than lapatinib at IC۵۰, with a dose-dependent decrease in migrated and invaded cells. Western blot analysis showed that ۱۳i markedly reduced p-EGFR, p-HER۲, and p-ERK۱/۲ levels in LAP-SKBR۳ cells compared with lapatinib, confirming robust pathway inhibition. These findings highlight ۱۳i’s potent anti-tumor activity via dual kinase inhibition, apoptosis induction, and metastasis suppression, making it a promising candidate for further preclinical studies on lapatinib-resistant HER۲-positive breast cancer.

کلیدواژه ها:

breast cancer ، Lapatinib ، Drug resistance ، EGFR ، HER۲ ، Quinazoline hybrids