A drug discovery platform to identify compounds that inhibit EGFR triple mutants

Area of Science:

• Molecular Biology
• Pharmacology
• Oncology

Background:

• Receptor tyrosine kinases (RTKs) are critical in disease pathogenesis.
• Traditional drug discovery relies on in vitro kinase assays, which are limited in identifying compounds targeting resistant mutations.
• Drug resistance necessitates novel therapeutic strategies and screening methods for mutated RTKs.

Purpose of the Study:

• To introduce MaMTH-DS (mammalian membrane two-hybrid drug screening), a live-cell platform for high-throughput screening.
• To identify small molecules that target functional protein-protein interactions of RTKs, particularly mutant forms.
• To discover novel inhibitors for drug-resistant oncogenic mutations, such as in epidermal growth factor receptor (EGFR).

Main Methods:

• Development and application of the MaMTH-DS live-cell platform.
• Screening of small molecules targeting functional protein-protein interactions of RTKs.
• Application to an oncogenic, TKI-resistant EGFR mutant.

Main Results:

• Identification of four mutant-specific compounds targeting the oncogenic EGFR mutant.
• Two identified compounds were undetectable by conventional in vitro kinase assays.
• One compound exhibited a novel mechanism of action against mutant EGFR, distinct from classical tyrosine kinase inhibitors (TKIs).

Conclusions:

• MaMTH-DS is a powerful live-cell platform for discovering novel RTK inhibitors.
• This method complements traditional drug screening by identifying compounds targeting functional interactions and novel mechanisms.
• MaMTH-DS facilitates the discovery of therapeutics for drug-resistant mutations in RTKs.