T6496 targeting EGFR mediated by T790M or C797S mutant: machine learning, virtual screening and bioactivity evaluation study

Area of Science:

• Medicinal Chemistry
• Oncology
• Computational Drug Discovery

Background:

• Acquired resistance to epidermal growth factor receptor (EGFR) inhibitors is a significant challenge in non-small cell lung cancer (NSCLC) treatment.
• Emergence of resistance mutations like T790M and C797S necessitates the development of novel therapeutic agents.
• Targeting specific EGFR mutations is crucial for overcoming acquired drug resistance.

Purpose of the Study:

• To discover novel small molecules capable of inhibiting drug-resistant EGFR variants (EGFR^T790M/L858R and EGFR^{T790M/C797S/L858R}).
• To identify potential anticancer agents that can overcome EGFR-mediated drug resistance in lung cancer.
• To evaluate the preclinical efficacy and mechanism of action of newly identified compounds.

Main Methods:

• Utilized ROC-guided machine learning and virtual screening of over 220,000 compounds to identify potential drug candidates.
• Employed cluster analysis and binding model analysis to select promising compounds.
• Conducted kinase assays, cellular assays (H1975 cell lines), apoptosis assays (AO and JC-1 staining), and ROS experiments to evaluate compound activity and mechanism.

Main Results:

• Identified six potential compounds with higher sensitivity to EGFR than c-Met.
• Compound T6496 demonstrated significant inhibition of EGFR^T790M/L858R (IC₅₀ = 3.30 μM) and EGFR^{T790M/C797S/L858R} (IC₅₀ = 8.72 μM).
• T6496 exhibited potent anticancer activity against H1975 cells (IC₅₀ = 2.7 μM) and induced apoptosis in a time- and concentration-dependent manner, with favorable toxicity profiles.

Conclusions:

• Compound T6496 is a promising novel agent that can overcome acquired resistance to EGFR inhibitors caused by T790M or C797S mutations.
• T6496 demonstrates potent antitumor effects and induces apoptosis, suggesting its potential as a therapeutic candidate for resistant lung cancer.
• Further structural optimization of T6496 is warranted for clinical development.