Title : From in silico design to xenograft validation: Synthesis and evaluation of an Anti-EGFR antibody-derived peptide-drug conjugate for NSCLC
Abstract:
Background: Non-small cell lung cancer (NSCLC) accounts for approximately 85% of lung cancer cases and remains a leading cause of cancer related mortality worldwide. Epidermal growth factor receptor (EGFR) is a clinically validated therapeutic target in NSCLC. Current EGFR-targeted therapies include tyrosine kinase inhibitors (TKIs), and several FDA approved monoclonal antibodies. While these therapies have improved patient outcomes, challenges including resistance, systemic toxicity, limited tumor selectivity continue to drive the development of alternative targeted drug delivery approaches. Peptide-drug conjugates (PDCs) have emerged as a promising class of targeted therapeutics. Recently, computer aided drug design has become an attractive tool for rational discovery of peptides as it is cost effective, rapid, and enhance accuracy.
Objective: This study aimed to design a linear peptide carrier from an antibody-EGFR binding interface using in-silico modeling, to develop a novel EGFR-targeting PDC, and evaluate its therapeutic potential against NSCLC.
Methods: A lead peptide sequence was derived from the antibody Fab region based on the antibody-EGFR binding interface using molecular dynamics (MD) and MM/GBSA (ΔG) calculations. The sequence was optimized through in-silico mutational screening to enhance binding affinity against EGFR using MD and MM/GBSA (Schrödinger). Three peptides were selected for solid-phase peptide synthesis, in-vitro assays. Antitumor efficacy of PDC was assessed in an H1975 NSCLC xenograft model after once-weekly dosing for five weeks.
Results: Among three peptides, PE3 showed enhanced EGFR-mediated cellular internalization and target engagement. PE3 was conjugated with DA-1, a proprietary auristatin-based microtubule inhibitor, developed in our laboratory, generated a metabolically stable PDC with potent antitumor activity, inducing complete tumor regression in the H1975 xenograft model without off-target toxicity.
Conclusion: This work demonstrates the potential of antibody-derived peptides as carrier for selective drug delivery and supports further preclinical investigation of PE3-DA1 as a targeted therapeutic strategy for EGFR-positive NSCLC and other EGFR-overexpressing malignancies.
