Novel Cyclic Peptides for Targeting EGFR and EGRvIII Mutation for Drug Delivery

Abstract

The epidermal growth factor–epidermal growth factor receptor (EGF-EGFR) pathway has become the main focus of selective chemotherapeutic intervention. As a result, two classes of EGFR inhibitors have been clinically approved, namely monoclonal antibodies and small molecule kinase inhibitors. Despite an initial good response rate to these drugs, most patients develop drug resistance. Therefore, new treatment approaches are needed. In this work, we aimed to find a new EGFR-specific, short cyclic peptide, which could be used for targeted drug delivery. Phage display peptide technology and biopanning were applied to three EGFR expressing cells, including cells expressing the EGFRvIII mutation. DNA from the internalized phage was extracted and the peptide inserts were sequenced using next-generation sequencing (NGS). Eleven peptides were selected for further investigation using binding, internalization, and competition assays, and the results were confirmed by confocal microscopy and peptide docking. Among these eleven peptides, seven showed specific and selective binding and internalization into EGFR positive (EGFR+ve) cells, with two of them—P6 and P9—also demonstrating high specificity for non-small cell lung cancer (NSCLC) and glioblastoma cells, respectively. These peptides were chemically conjugated to camptothecin (CPT). The conjugates were more cytotoxic to EGFR+ve cells than free CPT. Our results describe a novel cyclic peptide, which can be used for targeted drug delivery to cells overexpressing the EGFR and EGFRvIII mutation.

A war on many fronts: cross disciplinary approaches for novel cancer treatment strategies

“…Trade theory has conceptually been used to identify cellular trade-offs when performing different biological tasks ( Hausser and Alon, 2020 ; Boddy et al, 2018 ). On the other hand, competition assays have focused on studying the ability of different molecules to bind to a receptor and produce a downstream effect on cancer progression ( Yu et al, 2020 ; Cheng et al, 2022 ; Furman et al, 2022 ). Contrary to Lotka-Volterra interactions that assess population dynamics, economic competition assays focus on resource competition and cellular interactions based on the exchange of signals.…”

Material and Design Toolkit for Drug Delivery: State of the Art, Trends, and Challenges

“…The dual ligand modification improved the therapeutic Two peptides, i.e., P6 and P9, also displayed high specificity for nonsmall cell lung cancer and glioblastoma cells, respectively. 215 Although peptides have many advantages over other ligands, their application has been hindered by problems, such as poor stability and unexpected immunogenicity. Aside from this, peptides are sensitive to enzymatic environments and are readily degraded.…”

“…Of the 11 peptides, seven peptides displayed specific and selective binding and showed internalization into EGFR-positive (EGFR+) cells. Two peptides, i.e., P6 and P9, also displayed high specificity for nonsmall cell lung cancer and glioblastoma cells, respectively …”

Methods for Engineering Binders to Multi-Pass Membrane Proteins

“…After four rounds of subtractive panning, fourteen unique clones with preferential cell-binding abilities were identified. Using a similar strat-egy, Furman, et al successfully enriched cyclic peptides able to mediate binding and internalization into cells expressing the epidermal growth factor receptor [65].…”